[Comparative biochemical studies on the pathogenic and non-pathogenic amebae of the genus Naegleria].
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Th paper reviews the dinoflagellate general invading marine and freshwater fishes, presents their diagnostic features, points out the difference from tunicate-invading genus Oodinium Chatton, 1912 and comments on their pathogenicity. The genus Crepidoodinium Lom and Lawler g.n. is proposed for Oodinium cyprinodontum Lawler, 1967 from marine cyprinodontids, and Piscinoodinium g.n. is proposed to accommodate freshwater Oodinium pillularis Schäperclaus, 1954 and O. limneticum Jacobs, 1964. The most important diagnostic character is the structure of the attachment organelle which is depicted along with some other morphological features.
The analysis of a wide range of experimental models indicates that none of them could be regarded as an adequate model of human candidiasis. Hence, at present, the reproduction of separate components of candidiasis is necessary under various effects on the body.
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This critical review deals with our present knowledge on the classification, taxonomy and pathogenicity of the different species of the genus Haemophilus occurring in humans and animals, including the five biotypes of H. influenzae and the 3 biotypes of H. para-influenzae. The first, second, third and fourth stage tests for differentiation of the species are summarized in several tables.
Enterococci are members of the healthy human intestinal flora, but are also leading causes of highly antibiotic-resistant, hospital-acquired infection. We examined the genomes of a strain of Enterococcus faecalis that caused an infectious outbreak in a hospital ward in the mid-1980s (ref. 2), and a strain that was identified as the first vancomycin-resistant isolate in the United States, and found that virulence determinants were clustered on a large pathogenicity island, a genetic element previously unknown in this genus. The pathogenicity island, which varies only subtly between strains, is approximately 150 kilobases in size, has a lower G + C content than the rest of the genome, and is flanked by terminal repeats. Here we show that subtle variations within the structure of the pathogenicity island enable strains harbouring the element to modulate virulence, and that these variations occur at high frequency. Moreover, the enterococcal pathogenicity island, in addition to coding for most known auxiliary traits that enhance virulence of the organism, includes a number of additional, previously unstudied genes that are rare in non-infection-derived isolates, identifying a class of new targets associated with disease which are not essential for the commensal behaviour of the organism.
The natural occurrence and host range of species of the insect pathogenic fungal genus Strongwellsea [Zygomycota: Entomophthorales] were studied by extensive sampling and examination of adult Diptera [Cyclorrhapha]. The host range for Strongwellsea spp. was significantly enlarged. Three families were documented as new hosts: Muscidae (three species), Calliphoridae (one species), and Sarcophagidae (one species). Further, within the family Anthomyiidae six new host species were recorded and three new host species were documented in the Fanniidae. Strongwellsea castrans was identified as the pathogen in the Anthomyiids, while records from Fanniidae belonged to S. magna. The records of S. magna were the first outside the type locality (California). Primary conidia morphology indicated that muscid and calliphorid species were infected by three undescribed species of Strongwellsea. For the sarcophagid fly, no conidia were encountered, so the Strongwellsea species could not be identified. The tested sampling methods had each different advantages. Sweep netting and diagnosis in situ gave the best opportunity to sample a high number of infected dipterans per time unit spend, while sweep netting followed by incubation in the laboratory was the only method for the documentation of resting spores. The prevalence of S. castrans in the cabbage root fly Delia radicum was obtained by two methods: Samples collected by sweep net and incubated and water trap samples. Water trap captures gave higher prevalences of conidial infections than sweep-net captures. Measured prevalences of Strongwellsea spp. infections are therefore highly dependent on sampling method. The occurrence of resting spores of S. castrans in D. radicum was almost exclusively restricted to females and varied during the season. In samples from 1988 through 1993, no infected females in June contained resting spores, while 43.0% of the S. castrans-infected females from samples in August contained resting spores. During September and October, a decreasing proportion of S. castrans-infected D. radicum contained resting spores. The results document that species from the genus Strongwellsea are common fungal pathogens of adult flies from different families, occasionally with high prevalences. It also appears that the two described species of Strongwellsea, S. castrans and S. magna, have a range of dipterous host species that may always belong to a single family, Anthomyiidae and Fanniidae, respectively. Our data shows also that the host family Muscidae may be exploited by two new species of Strongwellsea. Copyright 1999 Academic Press.
With the increase in the number of immunocompromised hosts, the number of fungal pathogens has increased markedly. Identification and classification, especially of yeast species and strains, is often difficult when based solely on phenotypic characteristics. Since it became clear that different fungal pathogens require specific treatment strategies, there is a need for simple, rapid and reliable methods to identify fungal isolates. Polymerase chain reaction (PCR) fingerprinting was successfully applied here to identify yeast isolates. Microsatellite [(GTG)5; (GACA)4] and minisatellite [(5'GAGGGTGGCGGTTCT 3'), derived from the core-sequence of the phage M13] specific primers were used as single primers in the PCR to amplify hypervariable interrepeat DNA sequences from over 200 European, American and Australian clinical isolates within the genus Candida. Each species, represented by its type strain, could be identified by a specific multilocus pattern, allowing for the assignment of all the isolates to the appropriate species. Intra-species variation in the multilocus profiles was about 20% compared to inter-species variation, which was up to 80%. Anamorph-teleomorph pairs could be identified by highly homologous PCR fingerprint patterns. PCR fingerprinting was more discriminatory when compared with routinely used biochemical tests (Vitek YBC and API ID 32C). PCR fingerprinting has proven to be a powerful tool for the identification of medically important yeasts. It is rapid, sensitive, reliable, highly reproducible, stable in vitro and in vivo, and applicable to large scale experiments. Potential applications include: yeast taxonomy, epidemiology, environmental surveys, and improvement of the diagnosis of mycotic diseases.
Comparative analysis of 70 16S rDNA sequences representing 20 Rhizobium species (including pathogenic Agrobacterium spp.) was conducted using Maximum Likelihood to establish relationships of species using multiple sequences. There is no significant internal division of the Rhizobium clade to suggest that it represents more than one genus. Plant pathogenic (Agrobacterium) species are distributed within the genus. The analysis supported the synonymy of some species (Rhizobium gallicum and Rhizobium mongolense) and the need for comparative investigations of the tumorigenic and nodulating properties of Rhizobium tropici and Rhizobium rhizogenes. Misidentification of some sequences may conceal one or more putative novel species. Some sequences appear to be misidentified because of faulty sequencing or incomplete or inadequate analysis.
This study describes fluorogenic 5' nuclease PCR assays suitable for rapid, sensitive, high-throughput detection of the human-pathogenic microsporidial species Encephalitozoon hellem, E. cuniculi and E.intestinalis. The assays utilize species-specific primer sets and a genus-specific dual fluorescent-labeled probe that anneals to a region within the Encephalitozoon 16S rRNA gene. The assay design theoretically permits the probe to be used either with one set of primers for species-level determination or with a combination of all three primer sets for a genus-level screening of samples. The linear range of all three species-specific calibration curves that were developed using serial ten-fold dilutions of genomic DNA isolated from hemacytometer counted spores was determined to be between 10(4) and 10(-1) spores per PCR sample. The coefficients of variation were < or =5.2% over the entire 5-log span of each calibration curve. When DNA isolated from flow cytometric enumerated spores from each of the three Encephalitozoon species was used to evaluate the quantitative capability of the species' respective calibration curves, the results from 34 out of 36 (94%) samples were within 2 standard deviations. The species-specificity of each assay was confirmed using DNA isolated from 10(4) spores from each of the other two Encephalitozoon species as well as DNA extracted from numerous other protozoa, algae and bacteria.
An investigation of the parenteral pathogenic potential of aerobic spore-forming bacteria (genus Bacillus) in mice revealed that in the case of i.p. inoculation (0.5 ml suspension, 10(6)-10(7) viable cells per ml) only B. cereus was pathogenic, killing appr. one half of the inoculated animals (293 out of 577). Other bacilli (B. subtilis, B. megaterium, B. pumilus etc.) administered intraperitoneally did not produce any lethal effect. If inoculated intracerebrally, B. cereus killed appr. 90% of the mice (138 out of 155), while for other bacilli the rate was appr. 50% only (260 out of 529). It is suggested that most of the aerobic spore-forming organisms cultured from cases of hospital infections are B. cereus which, however, does not completely exclude the presence of other species.
Elicitins form a family of structurally related proteins that induce the hypersensitive response (HR) in plants, particularly Nicotiana spp. The elicitin family is composed of several classes. Most species of the plant-pathogenic oomycete genus Phytophthora produce the well-characterized 10-kDa canonical elicitins (class I), such as INF1 of the potato and tomato pathogen Phytophthora infestans. Two genes, inf2A and inf2B, encoding a distinct class (class III) of elicitin-like proteins, also occur in P. infestans. Unlike secreted class I elicitins, class III elicitins are thought to be cell-surface-anchored polypeptides. Molecular characterization of the inf2 genes indicated that they are widespread in Phytophthora spp. and occur as a small gene family. In addition, Southern blot and Northern blot hybridizations using gene-specific probes showed that inf2A and inf2B genes and transcripts can be detected in 17 different P. infestans isolates. Functional secreted expression in plant cells of the elicitin domain of the infl and inf2 genes was conducted using a binary Potato virus X (PVX) vector (agroinfection) and Agrobacterium tumefaciens transient transformation assays (agroinfiltration), and resulted in HR-like necrotic symptoms and induction of defense response genes in tobacco. However, comparative analyses of elicitor activity of INF1, INF2A, and INF2B revealed significant differences in intensity, specificity, and consistency of HR induction. Whereas INF1 induced the HR in Nicotiana benthamiana, INF2A induced weak symptoms and INF2B induced no symptoms on this plant. Nonetheless, similar to INF1, HR induction by INF2A in N. benthamiana required the ubiquitin ligase-associated protein SGT1. Overall, these results suggest that variation in the resistance of Nicotiana spp. to P. infestans is shadowed by variation in the response to INF elicitins. The ability of tobacco, but not N. benthamiana, to respond to INF2B could explain differences in resistance to P. infestans observed for these two species.
The recent increase of the number of antimicrobials and isolation of antibiotic resistant strains from humans and environmental objects is indicative of the necessity of further investigation of antibiotic susceptibility of the representatives of the genus Vibrio pathogenic for man to provide rational therapy of the diseases due to them. Susceptibility of 160 strains of pathogenic vibrios of 9 species to 11 antibiotics and chemotherapeutic drugs was assayed by the method of serial dilutions in agar media. The isolates were shown to be highly susceptible to chloramphenicol, doxycycline, cefotaxime, nalidixic acid and ciprofloxacin which made it possible to consider them as the drugs of choice in the treatment of the diseases caused by the microorganisms. A tendency to form polyantibiotic resistant strains within every species of tested pathogenic vibrios was observed. It conditioned the prospects of further profound study of the phenomenon with the analysis of the genetic determination of antibiotic resistance markers in pathogenic vibrios.
Heme uptake is a common means of iron and porphyrin acquisition by many pathogenic bacteria. The genus Haemophilus includes several important pathogenic bacterial species that characteristically require hemin-, protoporphyrin-, or heme-substituted proteins as essential growth factors under aerobic conditions. However, the mechanism of heme transport is not understood for Haemophilus. We have cloned a DNA fragment from H. influenzae that allows an Escherichia coli hemA mutant to employ exogenous hemin or protoporphyrin IX as sole sources of porphyrin. DNA sequencing of the cloned DNA fragment suggested that a previously characterized gene (hel) encoding an antigenic, outer membrane lipoprotein e(P4) was responsible for the complementation activity. Construction of hel insertion mutations in strain H. influenzae Rd demonstrated that hel is essential for growth under aerobic conditions but not under anaerobic conditions. The aerobic growth defect of hel mutants could be reversed by providing exogenous hemin in the presence of outer membrane. The analysis of hybrids between e(P4) and beta-lactamase demonstrated that a domain of e(P4) near its NH2' terminus was required for its function in hemin use. Within this domain is a short amino acid sequence that displays similarity to H. influenzae hemin binding protein HbpA, hemin-binding motifs present in eukaryotic transcription activator heme-activated protein, and the heme containing proteins hemoglobin (alpha-chain) and cytochrome C3, suggesting that this region may be involved in hemin binding and/or transport.
Members of the genus Campylobacter are well recognised as enteric pathogens but have rarely been implicated as human placental pathogens. A case of septic abortion due to C jejuni is reported. This occurred in a previously healthy woman with a diarrhoeal illness. The presence of chorioamnionitis raises the possibility of ascending infection rather than septicaemic spread as the pathogenesis. Members of the genus Campylobacter have special culture requirements and their incidence as placental pathogens may therefore be underestimated.