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PubMed2006

Porcine endogenous retrovirus integration sites in the human genome: features in common with those of murine leukemia virus.

Porcine endogenous retroviruses (PERV) are a major concern when porcine tissues and organs are used for xenotransplantation. PERV has been shown to infect human cells in vitro, highlighting a potential zoonotic risk. No pathology is associated with PERV in its natural host, but the pathogenic potential might differ in the case of cross-species transmission and can only be inferred from knowledge of related gammaretroviruses. We therefore investigated the integration features of the PERV DNA in the human genome in vitro in order to further characterize the risk associated with PERV transmission. In this study, we characterized 189 PERV integration site sequences from human HEK-293 cells. Data showed that PERV integration was strongly enhanced at transcriptional start sites and CpG islands and that the frequencies of integration events increased with the expression levels of the genes, except for the genes with the highest levels of expression, which were disfavored for integration. Finally, we extracted genomic sequences directly flanking the integration sites and found an original 8-base statistical palindromic consensus sequence [TG(int)GTACCAGC]. All these results show similarities between PERV and murine leukemia virus integration site selection, suggesting that gammaretroviruses have a common pattern of integration and that the mechanisms of target site selection within a retrovirus genus might be similar.

3' Flanking Region↗
PubMed2005

Swabbing of waiting room magazines reveals only low levels of bacterial contamination.

Previous studies have shown that toys in waiting rooms of general practice surgeries can be contaminated with potentially pathogenic bacteria. The question was raised as to whether magazines might also be sources of contamination. Swabbing of the front page of 15 magazines from 11 general practice surgeries, followed by analysis for total and specific bacteria, revealed low levels of contamination. Among targeted groups of pathogens only two colonies of Staphylococcus aureus were detected. Magazines do not seem to be potentially important vectors of bacterial transfer in the setting examined.

Bacteria↗
PubMedDate not supplied

Lack of pathogenicity of five serotype 2 infectious bursal disease viruses in chickens.

The pathogenic potential of five strains of serotype 2 infectious bursal disease virus (IBDV) for specific-pathogen-free chickens was examined. There were no gross or microscopic lesions in the inoculated chickens. Bursa-to-body-weight ratios of IBDV-infected chickens were not significantly different from those of uninfected controls. Virus-neutralizing antibodies to IBDV of serotype 2, but not serotype 1, were detected in infected chickens. This study indicated that the serotype 2 viruses examined were infectious but not pathogenic in chickens.

Animals↗
PubMed1999

Swinepox virus as a vaccine vector for swine pathogens.

Several small and large viruses (e.g., adenovirus, poxvirus, and herpesviruses) have been investigated as vaccine vectors. Each viral system has its advantages and disadvantages. One major advantage for viral vector vaccines is their ability to elicit a protective cell-mediated immunity as well as a humoral response to the antigen delivered by the vector. One major problem to using recombinant viruses as vaccines is the pathogenic potential of the parent virus. Therefore, it is important that along with the optimal expression of the foreign genes and ability to provide protection, the pathogenicity of the vector virus must be reduced during genetic manipulation without affecting its multiplication. The requirements to develop a viral vector, for example, swinepox virus, are a cell culture system that will support the growth of the virus, a suitable nonessential region(s) in the virus genome for insertion of foreign DNA so that virus replication is not affected, a foreign gene(s) that encodes for an immunogenic protein of a swine pathogen, strong transcriptional regulatory elements (promoters) necessary for optimal expression of the foreign genes, a procedure for delivering the foreign gene(s) into the nonessential locus, and a convenient method of distinguishing the recombinant viruses from the parent wild-type virus. Using this methodology, recombinant swinepox virus vaccines expressing pseudorabies virus antigens have been developed and shown to provide protection against challenge. These studies and evidence of local infection of the oral tract by swinepox virus indicate its potential as a recombinant vector for providing immunity against various swine pathogens including those that infect the respiratory and gastrointestinal tracts.

Animals↗
PubMed1996

Ashkenazi Jews, sulfur gases, and ulcerative colitis.

Fecal pathogens have been suspected to cause ulcerative colitis, yet none have been identified. Meanwhile, the 400 species comprising normal colonic flora have received little attention as potential pathogens. Sulfate-reducing bacteria (SRB), a frequent colonic commensal, have been identified in greater numbers in people with ulcerative colitis. The bacteria produce hydrogen sulfide, a toxic compound with the potential to cause colonic injury and possibly ulcerative colitis. If these bacteria are pathogenic, high-risk populations may harbor greater numbers of these organisms in their colons. We compared a group with a high incidence of ulcerative colitis, Ashkenazi Jews, to a control population to assess carriage rates for SRB. Breath samples were collected to indirectly determine colonic conditions. No difference was found between the two groups studied.

Adolescent↗
PubMed1994

Methionine uptake and cytopathogenicity of viable but nonculturable Shigella dysenteriae type 1.

A pathogenic strain of Shigella dysenteriae type 1 was selected for study to elucidate the physiology and potential pathogenicity of organisms in the viable but nonculturable (VBNC) state in the environment. Studies in our laboratory have shown that S. dysenteriae type 1 survives in laboratory microcosms in the VBNC state for long periods of time, i.e., more than 6 months. VBNC cells of S. dysenteriae type 1 were found to retain cytopathogenicity for cultured HeLa cells. To determine whether VBNC S. dysenteriae type 1 expressed protein after loss of culturability, 35S-labelled methionine was added to suspensions of VBNC cells. Total cellular proteins were extracted and examined by autoradiography. Results indicate that VBNC S. dysenteriae type 1 is capable of both active uptake of methionine and incorporation of methionine into protein. Amino acid uptake and protein synthesis substantiate the viability of cells of S. dysenteriae type 1 in the VBNC state, i.e., although the cells are unable to be cultured on laboratory media by standard bacteriological methods, the cells remain metabolically active. Furthermore, VBNC cells of S. dysenteriae type 1 may pose a potential public health hazard that has not yet been recognized.

Biological Transport, Active↗
PubMed1997

Convection warmers--not just hot air.

We sought to determine whether the forced air convection warmers (nine Bair Huggers, Augustine Medical, and one Warm Touch, Mallinkrodt Medical) used in our operating theatres could be a source of microbial pathogens. Agar plates were placed directly in the air stream of the warmers. Four of these grew potentially pathogenic organisms. When the warmers were set to blow through perforated blankets, no growth occurred. Three of the warmers were swabbed and sites of colonisation were found in their hoses. After fixing a microbial filter to the end of the hose, organisms were no longer detectable. We conclude that these warming devices are a potential source of nosocomial infection. They should only be used in conjunction with perforated blankets, should have their microbial filters changed regularly and their hoses sterilised. The inclusion of a microbial filter into the nozzle of the hose could be incorporated into the design of the warmer.

Air Microbiology↗
PubMed1984

Chronic inflammatory polyneuropathy. Reduction of nerve conduction velocities in monkeys by systemic passive transfer of immunoglobulin G.

In chronic (relapsing) inflammatory polyneuropathy (CRIP), successful treatment with plasma exchange has led to the concept of pathogenic humoral factors. In 6 patients with CRIP, 5 of whom improved after plasma exchange, the potential pathogenic role of circulating immunoglobulin (Ig) fractions was tested by applying the systemic passive transfer model to marmoset monkeys. After continuous treatment with intramuscular injections for 2-8 weeks, monkeys injected with the crude immunoglobulin fractions or with purified IgG from 5 of the 6 patients showed a significant and partially reversible reduction of the motor nerve conduction velocity (mean 34%, P less than 0.001) when compared with pre-treatment values. In control animals the reduction was 4%. Morphological examination revealed only minor ultrastructural changes of the myelin sheath. Immunocytochemistry revealed that human IgG was able to cross the blood-nerve barrier. It is concluded that the circulating IgG-fraction of patients with CRIP contains a factor that may contribute to the disordered nerve function after crossing the blood-nerve barrier. It may be the removal of this particular factor which is responsible for the rapid recovery of nerve conduction in patients after plasma exchange.

Animals↗
PubMedDate not supplied

Causative agents of acute diarrhoea in the first 3 years of life: hospital-based study.

During the 2 years of the study, 402 patients and an equal number of age- and sex-matched controls were investigated for the presence of diarrhoeal pathogens. Pathogenic organisms were recovered from 277 (68.9%) patients and 97 (24.1%) controls. In the patient group, possible bacterial pathogens were found in 210 (52.2%) cases. Enteropathogenic Escherichia coli (EPEC) was the most frequently found potential pathogen, being recovered in 132 cases (32.8%) with serotypes 026, 086, 0111 and 0124 being the most frequently identified. Other bacterial pathogens identified were enterotoxigenic E. coli (ETEC) 57 (14.2%), Shigella 13 (3.2%) and Salmonella eight (2%). Rotavirus was identified in 33 (8.2%) cases. Mixed bacterial and viral infections were also seen in 26 (6.5%) cases. In the control group, enteric pathogens were recovered from 97 (24.1%) specimens. The most common bacterial pathogen found in this group was again EPEC (40, 9.9%), with serogroups 018, 044, 0111 and 0126 being the most frequent. Giardia lamblia and Entamoeba histolytica were found in 31 (7.7%) and 10 (2.5%) controls, respectively. Rotavirus was found in 16 (4%) controls. The results of both centres where the study was performed (Karachi and Rawalpindi) were compared.

Bacterial Infections↗
PubMed2003

Microbial identification in the management of periodontal diseases. A systematic review.

BACKGROUND: Our understanding of the complexity of the oral microbiota continues to improve as new technologies, such as the analysis of 16S rRNA bacterial genes, are utilized. Despite the difficult of cataloguing all microorganisms and determining their pathogenic potential, some species, mostly members of the resident oral microbiota, have been identified as likely periodontal pathogens. However, for microbial diagnosis to be of value, it needs to affect disease diagnosis and/or treatment planning as well as result in superior treatment outcomes and/or provide an economic benefit to the patient. RATIONALE: The purposes of this systematic review were to determine if microbial identification influences periodontal patient management and whether treatment outcomes are better compared to patients whose treatment plans are developed without this information. FOCUSED QUESTION: In patients with periodontal diseases, does microbial identification influence patient management compared to treatment prescribed without this information? SEARCH PROTOCOL: The MEDLINE database was searched for clinical studies in English from 1991 through 2002 by 2 investigators. Hand searches were performed on the Journal of Clinical Periodontology, Journal of Periodontology, Journal of Periodontal Research, Oral Microbiology and Immunology, and Periodontology 2000. In addition, directors of diagnostic laboratories were contacted about unpublished data. SELECTION CRITERIA INCLUSION CRITERIA: Articles in which bacterial identification influenced patient treatment were preferred as were those reporting longitudinal data demonstrating a direct relationship between the presence or absence of certain bacteria and subsequent alterations in clinical variables. Because of the limited number of studies, all articles, including case reports, were considered. EXCLUSION CRITERIA: Review articles without original data were excluded, although references were examined for possible inclusion. Articles reporting data showing associations between certain microorganisms and disease or health that did not affect treatment were excluded. Clinical trials testing antibacterial agents for their ability to enhance mechanical debridement were not included since bacterial identification had little effect on drug selection or experimental group assignment. Articles dealing with implants rather than natural teeth were omitted. DATA ANALYSIS AND COLLECTION: The heterogeneity of the published data precludes any meaningful pooling of data or meta-analysis. The pertinent literature, including relevant variables of plaque, gingivitis, and bleeding on probing scores; probing depth; clinical attachment level; number of lost teeth; and microbial changes; and patient-centered outcomes including decrease in morbidity, reduced need for surgery, and duration and cost of treatment are summarized. MAIN RESULTS: 1. There was a lack of articles with a high evidence rating; most pertinent articles were either case reports or case series without controls. 2. Because reports were heterogeneous regarding study design, patient selection, and data collection, meta-analysis was not feasible and results are summarized in tabular format. 3. This report is based on a total of 24 studies, representing a total patient population of approximately 835. 4. Thirteen studies reported on microbiological identification as an aid in treatment planning. 5. Eleven studies reported a differential clinical response depending on the detection or lack of detection of specific organisms. REVIEWERS' CONCLUSIONS: 1. The published material suggests that microbiological monitoring may be useful in management of selected patients who do not respond to standard therapy. 2. Some practitioners consider microbial identification a valuable adjunct to managing patients with certain forms of periodontitis, although there is a lack of strong evidence to this effect. 3. Additional research is needed to address this issue.

Bacteria↗
PubMed2005

[Health risks associated with the migration of Atlantic salmon (Salmo salar L.): an epidemiological surveillance programme in Northern Spain].

The authors present the results of an epidemiological surveillance programme involving Atlantic salmon (Salmo salar) populations in north-eastern Spain. The study investigates the risk factors critical in disease transmission, which include the presence of potentially pathogenic micro-organisms, the innate cell-mediated immune response of the salmon, and the potential interactions between the two. Also taken into account are the biological and migratory cycles of these salmon populations. The results showed that Aeromonas salmonicida and Saprolegnia parasitica were the micro-organisms that played the most important role in the transmission of the disease by virtue of their pathological characteristics, their occurrence in large numbers, and their capacity to transmit disease. With regard to the cell-mediated immune response, the authors observed a wide variability between individuals in some cell groups, as well as certain seasonal and age differences. These may be factors that predispose the animals to infection or to becoming asymptomatic carriers, with the attendant risk of transmitting infection in the course of migration. This could be especially relevant in the case of salmon infected with A. salmonicida and S. parasitica, since it has been observed that such salmon have a decreased number of leucocytes compared to non-infected salmon.

Aeromonas salmonicida↗
PubMed2006

Genotyping of pathogenic Acanthamoebae isolated from clinical samples in Greece--report of a clinical isolate presenting T5 genotype.

Amoebae belonging to the genus Acanthamoeba are potentially pathogenic to humans, causing mainly amoebic keratitis. Pathogenic ability of the 15 known Acanthamoeba genotypes is under investigation. We report that four out of five cases of amoebic keratitis studied in Greece, present T4 sequence type, while the remaining one presents T5 sequence type (Acanthamoeba lenticulata), which is the second most frequent genotype found among environmental samples. Thus, it is confirmed, for the first time to our knowledge, that A. lenticulata can cause keratitis. However the reason that it is under represented in clinical samples compared to environmental ones is unknown.

Acanthamoeba↗
PubMed2001

Molecular determinants of virulence, cell tropism, and pathogenic phenotype of infectious bursal disease virus.

Infectious bursal disease viruses (IBDVs), belonging to the family Birnaviridae, exhibit a wide range of immunosuppressive potential, pathogenicity, and virulence for chickens. The genomic segment A encodes all the structural (VP2, VP4, and VP3) and nonstructural proteins, whereas segment B encodes the viral RNA-dependent RNA polymerase (VP1). To identify the molecular determinants for the virulence, pathogenic phenotype, and cell tropism of IBDV, we prepared full-length cDNA clones of a virulent strain, Irwin Moulthrop (IM), and constructed several chimeric cDNA clones of segments A and B between the attenuated vaccine strain (D78) and the virulent IM or GLS variant strain. Using the cRNA-based reverse-genetics system developed for IBDV, we generated five chimeric viruses after transfection by electroporation procedures in Vero or chicken embryo fibroblast (CEF) cells, one of which was recovered after propagation in embryonated eggs. To evaluate the characteristics of the recovered viruses in vivo, we inoculated 3-week-old chickens with D78, IM, GLS, or chimeric viruses and analyzed their bursae for pathological lesions 3 days postinfection. Viruses in which VP4, VP4-VP3, and VP1 coding sequences of the virulent strain IM were substituted for the corresponding region in the vaccine strain failed to induce hemorrhagic lesions in the bursa. In contrast, viruses in which the VP2 coding region of the vaccine strain was replaced with the variant GLS or virulent IM strain caused rapid bursal atrophy or hemorrhagic lesions in the bursa, as seen with the variant or classical virulent strain, respectively. These results show that the virulence and pathogenic-phenotype markers of IBDV reside in VP2. Moreover, one of the chimeric viruses containing VP2 sequences of the virulent strain could not be recovered in Vero or CEF cells but was recovered in embryonated eggs, suggesting that VP2 contains the determinants for cell tropism. Similarly, one of the chimeric viruses containing the VP1 segment of the virulent strain could not be recovered in Vero cells but was recovered in CEF cells, suggesting that VP1 contains the determinants for cell-specific replication in Vero cells. By comparing the deduced amino acid sequences of the D78 and IM strains and their reactivities with monoclonal antibody 21, which binds specifically to virulent IBDV, the putative amino acids involved in virulence and cell tropism were identified. Our results indicate that residues Gln at position 253 (Gln253), Asp279, and Ala284 of VP2 are involved in the virulence, cell tropism, and pathogenic phenotype of virulent IBDV.

Amino Acid Sequence↗
PubMed1989

[Efficacy of a PVP-iodine compound on selected pathogens of the oral cavity in vitro].

The study was designed to test the efficacy of a PVP-iodine antiseptic agent on potentially pathogenic oral bacteria. A 0.5% concentration of the test compound was sufficient to inhibit the growth of 7 out of 8 tested strains. Only bacteroides gingivalis continued to grow during exposure to a 0.5 and 1% concentration of the test compound. Quantitative suspension experiments demonstrated that the onset of action occurred within five minutes. Quantitative suspension experiments with protein loading resulted in a lower efficacy of the tested agent. Results indicate that the tested compound may be recommended as an oral disinfectant.

Bacteria↗
PubMed2005

Microarray-based detection of genetic heterogeneity, antimicrobial resistance, and the viable but nonculturable state in human pathogenic Vibrio spp.

The morbidity and mortality associated with Vibrio-mediated waterborne diseases necessitates the development of sensitive detection technologies that are able to elucidate the identity, potential pathogenicity, susceptibility, and viability of contaminating bacteria in a timely manner. For this purpose, we have designed a single multiplex PCR assay to simultaneously amplify 95 diagnostic regions (encompassing species/serogroup-specific, antimicrobial resistance, and known toxin markers) and combined it with a long oligonucleotide microarray to create a platform capable of rapidly detecting and discriminating the major human pathogenic species from the genus Vibrio: V. cholerae, V. parahaemolyticus, V. vulnificus, and V. mimicus. We were able to validate this strategy by testing 100 geographically and temporally distributed isolates and observed an excellent concordance between species- and serotype-level microarray-based identification and traditional typing methods. In addition to accurate identification, the microarray simultaneously provided evidence of antibiotic resistance genes and mobile genetic elements, such as sulfamethoxazole-trimethoprim constins and class I integrons, and common toxin (ctxAB, rtxA, hap, hlyA, tl, tdh, trh, vvhA, vlly, and vmhA) and pathogenicity (tcpA, type III secretion system) genes that are associated with pathogenic Vibrio. The versatility of this method was further underscored by its ability to detect the expression of known toxin and virulence genes from potentially harmful viable but nonculturable organisms. The results suggest that this molecular identification method provides rapid and definitive information that would be of value in epidemiological, environmental, and health risk assessment surveillance.

Anti-Infective Agents↗
PubMed2006

Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk.

Fifty-six Escherichia coli strains, serogrouped as EPEC, isolated from three different brands of pasteurised milk commercialised in Rio de Janeiro, Brazil, were tested for enteropathogenicity markers. Most of the strains (71.4%) were adherent to HEp-2 cells. The adherent strains were distributed among 7 EPEC serogroups (O26, O55, O111, O114, O125, O127, O128, O158). Although almost half of these strains (33.9%) presented unrecognisable adherence phenotypes, classical adherence patterns (localised-like, aggregative and diffuse adherence) described for E. coli and epidemiologically associated with diarrheagenic strains were observed. None of the strains showed typical localised adherence, usually associated with EPEC strains, but 4 of them displayed a localised-like adherence (LAL) phenotype, characterised by fewer and less compact microcolonies but that is still associated with diarrheagenic strains as well as strains of non-human origin. Indeed, 3 of these 4 strains were able to elicit the attaching-effacing lesion (FAS-positive), the central feature of EPEC pathogenesis, and hybridised with bfpA and eae DNA probes. The other LAL-positive strain hybridised with the bfpA probe but gave negative results for the eae probe and FAS assays. Interestingly, all LAL-positive strains produced amplicons of 200 bp in the PCR for bfpA, instead of the expected 326 bp fragment. PCR reactions for stx1 and stx2, two shiga-toxin-encoding genes, gave negative results. Typing of LEE-associated genes by PCR showed the profile eae (beta), tir (beta), espA (alpha) and espB (alpha) for one of the LAL-positive strain. The most prevalent adherence phenotype was the aggregative pattern which is observed in strains epidemiologically associated with persistent diarrhea. Additionally, one strain promoted complete detachment of the Hep-2 cell monolayer after 3 h of infection which might be related to the production of citotoxins, a feature that has been increasingly observed in clinical strains. The possession of EPEC-related O and H antigens is no longer deemed an essential characteristic of true pathogenic EPEC strains, emphasising the importance of routinely screen for virulence markers in E. coli strains isolated from foods. Our results are in accordance with data from the literature that demonstrate that environmental strains display atypical features but yet are capable of eliciting the classical A/E lesion and thus must be considered as potentially pathogenic. Further, our results demonstrate the potential of pasteurised milk as a vehicle for transmission of diarrheagenic E. coli in Brazil.

Animals↗
PubMed1995

An idiotype D23-bearing polyspecific, murine anti-DNA monoclonal antibody forms glomerular immune deposits. Pathogenic role of natural autoantibodies?

Identification of the immunochemical and structural properties of pathogenic anti-DNA antibodies is a major goal for understanding their origins and the mechanisms whereby they induce tissue lesions. Herein, we report on the production of an IgG2a,k anti-DNA monoclonal antibody (4B1), derived from a 12-month-old (NZB x NZW)F1 lupus mouse, able to form glomerular immune deposits. mAb 4B1 is a polyspecific antibody able to bind to ssDNA, actin, tubulin, cardiolipin and to laminin as shown by solid phase ELISAs. Indirect immunofluorescence labeling of HEp-2 cells gave a cytoplasmic staining pattern similar to that obtained with anti-cytoskeleton antibodies. Western blot analysis demonstrated that mAb 4B1 bore idiotype D23, previously shown to be characteristic of natural antibodies derived from normal mice. After injecting the 4B1-secreting hybridoma intraperitoneally into normal (NZW x BALB/c)F1 mice, glomerular immune deposits were observed along the capillary wall. These deposits contained mainly IgM, IgG2a and mAb 4B1, as demonstrated by direct immunofluorescence using a biotinylated-rat anti-4B1 idiotype mAb and kidney eluate analysis. Nucleotide sequence analysis of the VH and VL genes showed that mAb 4B1 is encoded by VH Q52, DSP2.9 and JH2 genes with minimal mutations and by VK8 very similar to the canonic D23 light chain, and JK1 germline genes. No arginine residues were observed in the VH CDR and both chains lacked N-segment addition. Thus, no structural characteristics deduced from the primary structure of mAb 4B1 could explain its pathogenic potential. However, the immunochemical and structural properties suggest that autoantibodies closely related to natural autoantibodies may be pathogenic.

Amino Acid Sequence↗
PubMed2005

Clinical evaluation of postpartum vaginal mucus reflects uterine bacterial infection and the immune response in cattle.

Bacteria contaminate the uterus of most dairy cattle after parturition and endometritis causes infertility. An endometritis score can be ascribed based on the vaginal mucus character and odour but it is not clear if the clinical score reflects the number of uterine bacteria or the inflammatory response. The present study tested the hypothesis that clinical evaluation of endometritis reflects the number of bacteria present in the uterus, and the acute phase protein response. Swabs (n = 328) were collected from the uterine lumen of dairy cattle, 21 and 28 days postpartum, vaginal mucus was scored for character and odour, and blood samples collected for acute phase protein measurement. Bacteria were identified following aerobic and anaerobic culture, and the bacterial growth density was scored semi-quantitatively. When bacteria were categorised by their expected pathogenic potential in the uterus, purulent or fetid odour vaginal mucus was associated with the growth density of pathogenic bacteria but not opportunist contaminants. When bacteria were analysed independently, Arcanobacterium pyogenes, Proteus and Fusobacterium necrophorum growth densities were associated with mucopurulent or purulent vaginal mucus. The bacterial growth densities for A. pyogenes, Escherichia coli, non-hemolytic Streptococci, and Mannheimia haemolytica were associated with a fetid mucus odour. Peripheral plasma concentrations of alpha(1)-acid glycoprotein were higher if there was a fetid compared with a normal vaginal mucus odour (1.50 +/- 0.09 mg/mL versus 1.05 +/- 0.02 mg/mL, P < 0.001), but did not differ significantly between vaginal mucus character scores. The evaluation of the character and odour of vaginal mucus reflects the number of bacteria in the uterus, and the acute phase protein response.

Actinomycetaceae↗