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Evidence that orthotopic transposition following rat heterotopic small bowel transplantation corrects overgrowth of potentially pathogenic bacteria.

An overgrowth of pathogenic organisms occurs following rat heterotopic small bowel transplantation. This study assessed whether the bacterial microflora return to normal following subsequent orthotopic transposition of the graft. After 14 days the heterotopic graft was placed into continuity following resection of 15 cm of the host midintestinal loop. Quantitative and qualitative analyses of the intraluminal bacteria were performed studying the resected host intestine, the heterotopic graft at 14 days, and the graft 14 days after transposition. A group of normal rats were used as controls. An overgrowth of Staphylococcus epidermidis evident in the heterotopic graft at 14 days returned to a more normal bacterial profile following orthotopic transposition. These findings suggest that early interposition of a small bowel graft into an orthotopic position may prevent an alteration in the small bowel ecology toward potentially pathogenic organisms capable of translocation.

Abdominal Muscles↗

Detection of Campylobacter jejuni and other potential pathogens in travellers' diarrhoea in Bangladesh.

To determine the distribution of pathogens associated with travellers' diarrhoea in Bangladesh, we have studied 269 expatriates who presented with diarrhoea to our clinic, over a one-year period. Patients were interviewed about their history of diarrhoea and a stool specimen examined for parasites and cultured for Campylobacter jejuni, Shigella sp., Salmonella sp., Vibrio and Aeromonas hydrophila and Plesiomonas shigelloides. C. jejuni and Shigella sp. were each isolated from 45 patients (17%) and P. shigelloides, A. hydrophila and V. cholerae non 01 from 7, 2 and 4 patients respectively. Patients with C. jejuni developed diarrhoea earlier after their arrival in Bangladesh than patients with Shigella. Fever and dysentery were significantly more common in patients with C. jejuni and Shigella. Infections with C. jejuni were less frequently associated with dysentery than infections with Shigella sp. Both C. jejuni and Shigella are commonly associated with travellers' diarrhoea in Bangladesh, and P. Shigelloides, A. hydrophila and V. cholerae non 01 may have an infrequent but potentially pathogenic role.

Adolescent↗

Mycobacterium palustre sp. nov., a potentially pathogenic, slowly growing mycobacterium isolated from clinical and veterinary specimens and from Finnish stream waters.

Taxonomic studies were performed on a phenotypically homogeneous group of 13 mycobacteria isolated from clinical, veterinary and stream-water samples. The methods applied included chromatographic analyses of bacterial lipids, biochemical tests and sequencing of the 16S rDNA and the internal transcribed spacer 1 (ITS1) region. Positive results in urease, Tween 80 hydrolysis and pyrazinamidase tests and a negative result in a semi-quantitative catalase test, combined with the ability to grow at 42 degrees C, distinguished this group among the yellow-pigmented, slowly growing mycobacteria. Unique fatty acid and mycolic acid profiles in chromatographic analyses and the results of gene sequencing indicated that the novel isolates represent a previously undescribed species, for which the name Mycobacterium palustre sp. nov. is proposed. The fatty acid profile obtained by GLC was characterized by the presence of several methyl-branched fatty acid markers. The most prominent markers were 2-methyleicosanoic, tetracosanoic and hexacosanoic acids. According to 16S rDNA sequencing, M. palustre is phylogenetically closest to Mycobacterium kubicae, a recently described species. M. palustre gives a false-positive result in a hybridization test with the AccuProbe Mycobacterium avium complex. One of the strains was isolated from a lymph-node biopsy from a child with cervical lymphadenitis. Thus, M. palustre should be listed among potential inducers of paediatric lymphadenitis. The veterinary isolates originated from the lymph nodes of slaughter pigs. The majority of the strains were recovered from natural waters, which highlights the role of the environment as a source of potentially pathogenic mycobacteria. The type strain of M. palustre is strain E846T (= DSM 44572T = ATCC BAA-377T).

Animals↗

Vertical distribution of potentially pathogenic free-living amoebae in freshwater lakes.

The vertical distribution of thermotolerant (37 degrees C and 45 degrees C) free-living amoebae (FLA) in warm monomictic lakes was determined in relation to the onset of thermal stratification and associated physical and chemical changes. The position of abiotic or biotic particulate layers in the water column was located by using a submersible horizontal beam transmissometer that measures attenuance, or the absorption and scattering of light by particulates in the water column. During mixis, the vertical distribution of amoebae was sporadic with significant numbers of FLA only occurring in clay layers caused by runoff after heavy rains. With the onset of thermal stratification in the lakes, phytoplankton layers began to form. Few amoebae were isolated from layers containing flagellated phytoplankton; however, significant (P less than 0.005) numbers of FLA were isolated from two particulate layers dominated by the filamentous blue-green algae Aphanizomenon and Lyngbya, respectively. By late June, a persistent detrital or decomposition layer formed in the lower metalimnion, as well as a hypolimnetic iron layer where the Fe2+ state was predominant. In this midsummer period, 13 Naegleria fowleri were isolated, with three from the detrital layer and seven from the iron layer. The presence of attenuation zones was found to be the best indicator of the vertical distribution of FLA in the water column, and such layers represent an important, previously undescribed habitat for potentially pathogenic FLA.

Amoeba↗

Molecular identification of potential pathogens in water and air of a hospital therapy pool.

Indoor warm-water therapy pool workers in a Midwestern regional hospital were diagnosed with non-tuberculosis pulmonary hypersensitive pneumonitis and Mycobacterium avium infections. In response, we conducted a multiseason survey of microorganisms present in this therapy pool water, in biofilms associated with the pool containment walls, and in air immediately above the pool. The survey used culture, microscopy, and culture-independent molecular phylogenetic analyses. Although outfitted with a state-of-the-art UV-peroxide disinfection system, the numbers of bacteria in the therapy pool water were relatively high compared with the potable water used to fill the pool. Regardless of the source, direct microscopic counts of microbes were routinely approximately 1,000 times greater than conventional plate counts. Analysis of clone libraries of small subunit rRNA genes from environmental DNA provided phylogenetic diversity estimates of the microorganisms collected in and above the pool. A survey of >1,300 rRNA genes yielded a total of 628 unique sequences, the most common of which was nearly identical to that of M. avium strains. The high proportion of clones with different Mycobacterium spp. rRNA genes suggested that such organisms comprised a significant fraction of microbes in the pool water (to >30%) and preferentially partition into aerosols (to >80%) relative to other waterborne bacteria present. The results of the study strongly validate aerosol partitioning as a mechanism for disease transfer in these environments. The results also show that culture protocols currently used by public health facilities and agencies are seriously inadequate for the detection and enumeration of potential pathogens.

Air Microbiology↗

Characterization and pathogenic potential of Listeria monocytogenes isolates from the smoked fish industry.

This study was designed to evaluate the hypothesis that some of the Listeria monocytogenes subtypes associated with foods, specifically smoked fish, may have an attenuated ability to cause human disease. We tested this hypothesis by using two different approaches: (i) comparison of molecular subtypes found among 117 isolates from smoked fish, raw materials, fish in process, and processing environments with subtypes found among a collection of 275 human clinical isolates and (ii) the evaluation of the cytopathogenicity of industrial isolates. Ribotyping and PCR-restriction fragment length polymorphism typing of the hlyA and actA genes differentiated 23 subtypes among the industrial isolates and allowed classification of the isolates into three genetic lineages. A significantly higher proportion of human isolates (69.1%) than industrial isolates (36.8%) were classified as lineage I, which contains human sporadic isolates and all epidemic isolates. All other industrial isolates (63.2%) were classified as lineage II, which contains only human sporadic isolates. Lineage I ribotypes DUP-1038B and DUP-1042B represented a significantly higher proportion of the human isolates than industrial isolates (5.1%). Lineage II ribotypes DUP-1039C, DUP-1042C, and DUP-1045, shown previously to persist in the smoked fish processing environment, represented nearly 50% of the industrial isolates, compared to 7.6% of the human isolates. Representatives of each subtype were evaluated with a tissue culture plaque assay. Lineage I isolates formed plaques that were significantly larger than those formed by lineage II isolates. Isolates from the smoked fish industry representing three ribotypes formed no plaques or small plaques, indicating that they had an impaired ability to infect mammalian cells. While L. monocytogenes clonal groups linked to human listeriosis cases and outbreaks were isolated, our data also suggest that at least some L. monocytogenes subtypes present in ready-to-eat foods may have limited human-pathogenic potential.

Animals↗

Role of Mycoplasma penetrans endonuclease P40 as a potential pathogenic determinant.

Recently, we reported the purification to homogeneity and characterization of Ca(2+)- and Mg(2+)-dependent endonuclease P40 produced by Mycoplasma penetrans (M. Bendjennat, A. Blanchard, M. Loutfi, L. Montagnier, and E. Bahraoui, J. Bacteriol. 179; 2210-2220, 1997), a mycoplasma which was isolated for the first time from the urine of human immunodeficiency virus-infected patients. To evaluate how this nuclease could interact with host cells, we tested its effect on CEM and Molt-4 lymphocytic cell lines and on peripheral blood mononuclear cells. We observed that 10(-7) to 10(-9) M P40 is able to mediate a cytotoxic effect. We found that 100% of cells were killed after 24 h of incubation with 10(-7) M P40 while only 40% cytotoxicity was obtained after 72 h of incubation with 10(-9) M P40. Phase-contrast microscopy observations of P40-treated cells revealed morphological changes, including pronounced blebbing of the plasma membrane and cytoplasmic shrinkage characteristic of programmed cell death, which is in agreement with the internucleosomal fragmentation of P40-treated cell DNA as shown by agarose gel electrophoresis. We showed that (125)I-radiolabeled or fluorescein isothiocyanate-labeled P40 was able to bind specifically in a dose-dependent manner to the cell membrane of CEM cells, which suggested that the cytotoxicity of P40 endonuclease was mediated by its interaction with the cell surface receptor(s). The concentration of unlabeled P40 required to inhibit by 50% the formation of (125)I-P40-CEM complexes was about 3 x 10(-9) M, indicating a high-affinity interaction. Both P40 interaction and cytotoxicity are Ca(2+) dependent. Our results suggest that the cytotoxicity of M. penetrans observed in vitro is mediated at least partially by secreted P40, which, after interaction with host cells, can induce an apoptosis-like death. These results strongly suggest a major role of mycoplasmal nucleases as potential pathogenic determinants.

Calcium↗

Quantitation of pathogenic potential of Staphylococcus aureus.

Numerical estimates of the pathogenicity of Staphylococcus aureus strains were made for phage-typed strains from a relative incidence of significant to nonsignificant isolates from hospital patients. For a specific phage-patterned strain, the number of isolates from significant (wounds, abscesses, blood, etc.) sites was divided by the number of isolates from nonsignificant (respiratory tract, body surfaces, etc.) sites. This value, multiplied by 100, was the index of infection potential (IIP). IIP values for the S. aureus strains studied ranged from a low of 8 to a high of 50. The average IIP for all phage-patterned strains that occurred 50 or more times was 20. There was an inverse relationship between length of the phage pattern (number of the 26 typing phages that lysed the strain) and pathogenicity. Those strains with shorter phage patterns had higher IIP values and were more pathogenic. Strains lysed by one phage had an average IIP of 27, whereas those lysed by 18 phages had an average IIP of 14.

Bacteriophage Typing↗

Quantifying the potential pathogens transmission of the blowflies (Diptera: Calliphoridae).

To quantify the potential capability of transporting and passing infective pathogens of some blowflies (Diptera: Calliphoridae), Mihályi's danger-index was calculated for seven species. The original equation was modified to include synanthropic information to discriminate between asynanthropic, hemisynanthropic, and eusynanthropic status. Three groups were recognized, of which Phaenicia cluvia and Musca domestica proved the flies with lowest index value (D = 2.93 and 3.00 respectively); Cochliomyia macellaria, Chrysomya albiceps and Sarconesia chlorogaster presented a significantly higher index value (p<0.10; D = 4.28, 4.44 and 5.66 respectively) and C. megacephala, C. vicina and P. sericata appear to represent the heaviest potential sanitary risk with the highest index value (p<0.10; D = 15.54, 16.88 and 12.49 respectively).

Algorithms↗

Incidence of Chlamydia trachomatis and other potential pathogens in neonatal conjunctivitis.

OBJECTIVE: Ocular infection in neonatology is a permanent and important health problem. To improve primary attention, prevention, and control, the study of the potential bacterial etiology of all consecutive cases of conjunctivitis was incorporated as a regular procedure in primary care from July 1995 to December 1998. MATERIALS AND METHODS: Prof. A. Posadas Hospital (Great Buenos Aires) has an average of 4294 births per year. This report analyzes the results obtained in 332 infants (age range, 0-30 d) with conjunctivitis. Clinical conjunctivitis was diagnosed in inpatients and outpatients by the same specialized staff. Isolation and characterization of bacteria were done by conventional microbiologic methods, including specific search for Neisseria gonorrhoeae and Chlamydia trachomatis. Chlamydia trachomatis was studied by antigen immunodetection and polymerase chain reaction, and genotyped by restriction fragment length polymorphism. RESULTS: Conjunctivitis had an incidence (cases per 1000 live births) of 39.6 in 1995, 25.3 in 1996, 15.4 in 1997, and 15.2 in 1998. Microbial growth was detected in 167 (50.3%) of 332 cases. Ocular C. trachomatis infection was detected in 26 cases (7.83%). Five of seven isolates in tissue cultures belonged to type E and two to type G. Bacteria from respiratory ecology were the main isolates: Haemophilus influenzae (16.9%), Streptococcus pneumoniae (12.3%), and Staphylococcus aureus (8.7%). Haemophilus influenzae isolates were not serotyped and 17.2% of them were b-lactamase producers. In 15 cases both H. influenzae and S. pneumoniae were isolated together. Of S. pneumoniae, 4.9% were oxacillin resistant. CONCLUSIONS: There has been a decline in the total number of cases of neonatal conjunctivitis, but the disease is still an important health problem. Chlamydia trachomatis also shows a decreasing profile with an incidence of (cases per 1000 live births) 4.39 in 1995, 1.85 in 1996, 1.01 in 1997, and 0.78 in 1998, and a tendency to show more incidence in spring-summer and significant accumulation of cases in babies between 7 and 9 days of age. Haemophilus influenzae alone (12.3%) or associated with S. pneumoniae (4.5%) appears as a prevalent potential bacterial pathogen. A significant accumulation of H. influenzae and S. pneumoniae cases occurs in winter. In 47.6% of cases, there was no bacterial growth. No significant seasonal differences in percentage of negative cultures or among the three-day age groups were detected. Neisseria gonorrhoeae was not found associated with ophthalmia neonatorum in this series.

Age Factors↗

Human culture and monkey behavior: Assessing the contexts of potential pathogen transmission between macaques and humans.

An in-depth understanding of the contexts and patterns of human-macaque interactions can play an important role in assessing and managing the potential for pathogen transmission. The Padangtegal Monkey Forest in Bali, Indonesia, and the Upper Rock Nature Reserve in Gibraltar are two sites that have been relatively well studied in regard to human-macaque interactions. This article presents an overview of interaction patterns between humans and macaques at these sites along with a description of the cultural, demographic, and contextual differences between local and nonlocal humans at the sites. Differences in these two sites' interaction patterns included bite rates, the role of food in aggressive interactions, and the context in which the interactions took place. Similarities included overrepresentation by adult male macaques in interactions and a substantial impact by local cultural and demographic factors. These similarities and differences are interpreted as resulting from differences in macaque species and behaviors, and human demography, culture, and behavioral patterns.

Adolescent↗

Effect of growth of Bacteroides fragilis at different redox levels on potential pathogenicity in a HeLa cell system: demonstration by confocal laser scanning microscopy.

During trauma, the intestinal anaerobe, Bacteroides fragilis, may enter into a pathogenic state. The process coincides with changing environmental conditions particularly the redox level in situ. To gain insight into this phenomenon B. fragilis was grown at different redox levels, and the invasive potential was examined using an in vitro model consisting of HeLa cell monolayers. The clinical strain AIP 5-86 was taken from a small collection of B. fragilis strains able to penetrate into tissue cell monolayers when selected by an acridine orange-crystal violet fluorescent staining technique. Following preliminary investigation by confocal laser scanning microscopy (CLSM), this particular strain was regarded as representative for examining the invasive potential. After growth in a defined medium under oxidizing, reducing or intermediate Eh7 conditions, the washed mid-log phase bacteria were allowed to interact with HeLa cell monolayers for 45 min at 37 degrees C. The results were extensively monitored by CLSM to follow the reactions in a stereoscopic dimension. In addition, the bacteria were examined by transmission and scanning electron microscopy before interaction to distinguish characteristics in surface configuration. The growth of the bacteria at particular redox levels seemed to influence their potential for pathogenicity. After growth at relatively high Eh, the bacteria easily penetrated into the HeLa cells, but not at low Eh, as determined by the laser scanning studies. Examination of the bacteria alone by transmission and scanning electron microscopy revealed small vesicles and a tendency to aggregate after growth at the low redox level while there were rather few vesicles and an implied dispersion at the high redox level. This leaves it open whether the invasiveness was based on the alterations found during growth of the bacteria. Different redox levels as well as the respective changes of the bacterial surface may help to discern the commensal from the pathogenic state of B. fragilis.

Bacteroides fragilis↗

Colonization with potentially pathogenic respiratory tract bacteria. A household study.

A group of 235 persons (180 adults and 55 children 0-15 years old) recorded symptoms of upper respiratory tract infection daily during two three-month periods (autumn 1986 and spring 1987). Samples for culture were taken from the nasopharynx and throat once during each period. Fifteen per cent of asymptomatic subjects harboured respiratory pathogens in the nasopharynx, as did 28% of those subjects with minor respiratory tract infections and 46% of those with more severe respiratory tract infections. Of children up to seven years of age, 58% were colonized with potential respiratory pathogens, which is important to keep in mind when evaluating culture reports from young children. Adults living with young children were colonized significantly more often than other adults. Branhamella catarrhalis was the most common pathogen.

Adolescent↗

Pseudomonas fluorescens as a potential pathogen: adherence to nerve cells.

In order to determine the infectious potential of the psychrotrophic bacterium Pseudomonas fluorescens, a species closely related to the opportunistic pathogen P. aeruginosa, we investigated the binding activity of this bacterium on primary cultures of rat neonate cortical neurons and glial cells, adrenal paraneurons and NG108-15 neuroblastoma cells. Incubated at concentrations of 10(6) and 10(8) CFU/mL, P. fluorescens MF37 exhibited a high binding activity on neurons in the same range as that of P. aeruginosa PAO1. A significant, but lower, adherence of P. fluorescens was also detected on glial cells and adrenal paraneurons. In contrast, when P. fluorescens MF37 or P. aeruginosa PAO1 were incubated with neuroblastoma cells, no binding was observed. In neurons, the association of P. fluorescens with the plasma membrane occurred both on neurites and cell body. Leakage of the cytoplasmic content was frequently noted. Studies performed using the fluorescent probe Hoechst 33258 revealed that in 10% of neurons, P. fluorescens induced the appearance of densely stained clusters of DNA that was typical of an early step of apoptosis. In glial cells exposed to P. fluorescens, marked changes in the morphology of the nucleus, including fragmentation into lobular structures and aggregation of DNA, were also reminiscent of the existence of a possible apoptotic mechanism. Taken together, these results reveal that P. fluorescens can bind to nerve cells and affect their physiology and, in agreement with recent clinical observations, suggest that P. fluorescens could behave as a pathogen.

Animals↗

Use of a genetic marker for wild-type potentially pathogenic infectious bursal disease viruses.

An amino acid mutation at residue 284 (Ala to Thr) in the VP2 protein of infectious bursal disease viruses (IBDVs) has been correlated with the ability to replicate in cell culture. In this study, we designed a molecular test for this mutation. The reverse transcriptase/polymerase chain reaction (RT/PCR) was used to amplify a 743-bp region of the VP2 gene that contained the codon for amino acid 284. The restriction endonuclease NgoMIV was selected for this study because the first three nucleotides of its six-base recognition sequence are the codon responsible for the amino acid alanine at residue 284. The RT/PCR products from 10 known pathogenic and 16 vaccine strains of IBDV were examined for the presence or absence of the NgoMIV site. We also examined 189 field strains of IBDV for the NgoMIV site. All 10 known pathogenic IBDV strains contained the NgoMIV site, indicating they contained alanine at residue 284. None of the vaccine strains had the NgoMIV site, suggesting they had threonine or another amino acid at residue 284. The results suggest that the presence of this NgoMIV site can be used as a marker for the identification of wild-type (nonvaccine) IBDV strains. The RT/PCR products from 152 (80.4%) of the field strains had the NgoMIV site and thus have the potential to be wild-type pathogenic viruses. The RT/PCR products from 37 (19.6%) of the field strains were not cleaved by NgoMIV and thus are potentially attenuated vaccine strains. Molecular diagnostic assays have been used to place IBDV strains into genetically related groups. The identification of this genetic marker now makes it possible to identify viruses that are wild-type strains that have the potential to be pathogenic viruses.

Amino Acid Sequence↗

Providencia stuartii, a hospital pathogen: potential factors for its emergence and transmission.

The emergence of Providencia stuartii as a hospital pathogen in a burn unit was demonstrated by routine infection surveillance. The organism was initially recognized in a burn wound and subsequently in urine or sputum. Compared to controls, those patients harboring P. stuartii were similar in age and percentage of body surface burned and were more likely to have been in one of the two burn unit rooms, (p less than 0.02). Infection with P. stuartii was independent of duration in the Intensive Care Unit or Burn Unit, and of number of visits to hydrotherapy or operating rooms (OR). Once patients were colonized with P. stuartii they had greater morbidity than non-colonized patients as evidenced by longer stays in the unit and increased visits to the OR for debridement. P. stuartii was isolated from air samples (5/14) more commonly than from the hands of personnel. In vitro tests suggested that extensive use of parenteral gentamicin and replacement of the antibacterial topical cream sulfamylon by silver sulfadiazine favored the emergence of P. stuartii over Pseudomonas aeruginosa as the predominant colonizing organism.

Adult↗

Monozygotic twins discordant for epilepsy differ in the levels of potentially pathogenic autoantibodies and cytokines.

Can autoantibodies (Ab's) and cytokines play a role in epilepsy?Monozygotic twins discordant for epilepsy (most probably Rasmussen's encephalitis (RE)), compared to 49 neurologically intact controls, were both found to contain in their serum (at the time of epilepsy diagnosis) significantly elevated levels of specific Ab's against peptide B (amino acids 372-395) of the ionotropic glutamate receptor of AMPA subtype 3 (i.e. GluR3B peptide). Interestingly, both twins also had clinically elevated levels of Ab's to double-stranded (ds) DNA, glutamic acid decarboxylase, nuclear antigens, beta2-glycoprotein I and cardiolipin, as in "classical" autoimmune diseases. Both twins also had significantly elevated levels of IFNgamma, TNFalpha, IL-4 and IL-10 in the serum, compared to the controls. Comparing the twins revealed that the epileptic twin had significantly higher levels of five of the above anti-self Ab's, but significantly lower levels of all four cytokines compared to her healthy sister. Importantly, the epileptic twin, alike three other RE patients tested herein, contained elevated levels of Ab's to GluR3B and dsDNA also in cerebrospinal fluid (CSF) (unavailable of the healthy twin). Our results suggest that the various autoimmune Ab's studied herein, all of which are known already to have a potential to be pathogenic in the nervous system and/or peripheral organs, may play a role in some types of epilepsy. The titer of such Ab's and of key cytokines may be crucial for either facilitating or arresting the development of epilepsy. Our findings also show that anti-GluR3B Ab's in serum are not necessarily detrimental (their presence in the CSF may be more dangerous), and that they are not a mere side effect of already existing epilepsy, as they were found herein in serum of a healthy individual. These findings and suggestions may be of clinical importance and call for further studies.

Autoantibodies↗