Drift of tetM determinant in urogenital microbiocenosis containing mycoplasmas during treatment with a tetracycline antibiotic

We studied the correlation between genetic transfer of tetM determinant in Tn916 conjugative transposon by urogenital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) and changes in the bacterial repertoire during treatment with a tetracycline antibiotic. Basic conditions favoring the nonspecific transfer of tetM determinant into mollicute cells are determined and the allele polymorphism of tetM determinant in clinical strains of M. hominis and U. urealyticum is evaluated. The structure of tetM gene in clinical mycoplasma and ureaplasma strains is characterized by a peculiar mosaic pattern and differs from all previously described alleles of this gene. The results suggest that tetracycline resistance in mollicutes is determined by mechanisms alternative to genetic transfer of tetM determinant.     

Detection of Mycoplasma hominis antigen in clinical specimens by using a four-layer modification of enzyme immunoassay (EIA)

A 4-layer modification of enzyme immunoassay (EIA) was developed for the detection of Mycoplasma hominis antigen in clinical specimens. Microtiter plates were sensitized with rabbit anti-mycoplasma immunoglobulin, guinea pig anti-mycoplasma immunoglobulin was used as the secondary antibody, and horseradish peroxidase-conjugated anti-guinea pig immunoglobulin was used as the indicator antibody. The specificity of the assay was confirmed by using guinea pig immunoglobulins from preimmunization sera. The sensitivity of the assay is down to 10 ng/ml of antigen protein. Marked cross-reactivity was demonstrated for different strains within the species M. hominis, whereas the other genital mycoplasma species tested showed no reactivity in the assay. A comparison was made of EIA and conventional culture of vaginal specimens from 24 women. All 6 specimens positive by culture were also positive for M. hominis antigen by EIA. Antigen detection by EIA is a sensitive, rapid and simple method for the detection of M. hominis in clinical specimens.     

Immunohistochemical labelling of cytokines in lung lesions of pigs naturally infected with Mycoplasma hyopneumoniae

Mycoplasma hyopneumoniae (Mh) is the primary agent of porcine enzootic pneumonia (PEN), a chronic respiratory disease endemic to pig farms, and characterized histologically by infiltration of mononuclear cells in airways and prominent hyperplasia of the bronchus-associated lymphoid tissue (BALT). To gain further insight into the pathogenesis of PEN, cytokine expression in the lung, with particular attention to the BALT, was examined immunohistochemically in pigs naturally infected with Mh. An increase (P < 0.05) in proinflammatory and immunoregulatory cytokines (especially interleukin [IL]-2, IL-4 and tumour necrosis factor [TNF]-alpha, and to a lesser extent IL-1 [alpha and beta] and IL-6) was detected in the BALT, which showed intense lymphoid hyperplasia. IL-1beta and TNF-alpha were also detected in the bronchoalveolar exudate of infected pigs, and IL-6 and IL-8 were demonstrated in mononuclear cells of the alveolar septa. The results showed that in Mh infection, macrophage and lymphocyte activation results in the expression of a number of cytokines capable of inducing lung lesions and lymphoreticular hyperplasia of the BALT.     

Relative contribution of vertical,within-farm and between-farm transmission of Mycoplasma synoviae in layer pullet flocks

ABSTRACT

In this study, the relative contribution of vertical transmission, within-farm transmission and between-farm transmission of Mycoplasma synoviae in layer pullet flocks was quantified using logistic regression analysis. Data from 311 Dutch pullet flocks, of which 172 (55%) were positive for M. synoviae, were included in the study. Also the M. synoviae status of the parent stock of these flocks was included. The M. synoviae status was determined with the M. synoviae rapid plate agglutination test. Data analysis showed that vertical transmission was the most important transmission route for M. synoviae in layers as is demonstrated by an odds ratio of 5.8 (P?=?0.000). A positive association with M. synoviae infections was found for layer pullet flocks on a multi-house farm where at least one other flock was M. synoviae-positive compared to single-house farms (odds ratio 3.1, P?=?0.022), while a negative association was found when no other M. synoviae-positive flocks were present (odds ratio?=?0.2, P?=?0.003). No association was found between M. synoviae status of pullet flocks and poultry farm density. Odds ratios were 0.54 (P?=?0.288) and 0.34 (P?=?0.073), respectively, for medium and highest poultry farm density compared to lowest poultry farm density. This is the first time that the relative contribution of horizontal and vertical transmission of M. synoviae has been quantified. These results can be extrapolated to M. synoviae control in general, and emphasize the importance of M. synoviae control in parent stock and practical channelling.     

建立不同民族永生细胞株质量控制方法的探讨

目的以建立普米族、独龙族和怒族永生细胞为基础,探讨EB病毒转化细胞、细胞培养、冻存、复苏以及支原体检测等建立永生细胞株的技术要点及质量检测方法.方法采用EB病毒转化技术建立3个民族永生细胞株;培养法和PCR法对细胞株进行支原体污染检测;染色体G显带及核型分析细胞株的遗传稳定性.结果成功建立了3个民族B淋巴细胞永生细胞株,转化率分别为98%、86%和76%.对已建株保存的3个民族的永生细胞进行复苏培养,复苏成活率为100%.支原体污染检测均为阴性.染色体计数和G带分析显示,细胞株经早期传代培养后,仍然保持二倍体特征,未发现染色体结构畸变.结论本研究中传代培养、细胞冻存、细胞复苏和支原体污染防范等一整套技术过程是满足建库要求的.同时为大规模永生细胞库的建立和进行相应的质量监测提供了科学的依据.另外,本研究还对一些影响转化的因素和可能的机理进行了探讨.     

Evaluation of the WIDER I system for antimicrobial susceptibility testing of clinical isolates of Haemophilus influenzae and Streptococcus pneumoniae

The aim of this study was to evaluate the WIDER I system for susceptibility testing of Haemophilus influenzae and Streptococcus pneumoniae . MICs of 12 antimicrobials against 42 H. influenzae and 58 S. pneumoniae strains were determined using 1W MIC panels and compared with those obtained by microdilution. Overall essential agreements were >99%. Very major errors were not detected. Major errors occurred with ampicillin (1.7% H. influenzae ). Minor errors were 2.3% (amoxicillin–clavulanate, cefuroxime, chloramphenicol), 7.1% (ampicillin) and 16.7% (clarithromycin) for H. influenzae , and 1.7% (chloramphenicol, erythromycin, meropenem), 3.4% (amoxicillin–clavulanate, cefuroxime, tetracycline) and 8.6% (levofloxacin) for S. pneumoniae . The WIDER I system is a reliable method for susceptibility testing of H. influenzae and S. pneumoniae .     

Does infection with Chlamydia pneumoniae and/or Helicobacter pylori increase the expression of endothelial cell adhesion molecules in humans?

Objective   To investigate if Chlamydia pneumoniae and/or Helicobacter pylori seropositivity is associated with elevated levels of soluble endothelial cell adhesion molecules (sCAMs) as markers of atherosclerotic activity.
Methods   Immunoglobulin A (IgA) and IgG antibodies to the two bacteria, soluble intercellular cell adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1) and E-selectin were measured in coronary heart disease (CHD) patients ( n  = 193) and age- and sex-matched controls ( n  = 193). Two different serological methods were used for the detection of Chlamydia antibodies: Labsystems microimmunofluorescence to detect species-specific C. pneumoniae antibodies and Medac's recombinant enzyme-linked immunosorbent assay to detect genus-specific lipopolysaccharide antibodies.
Results   The concentrations of sICAM-1 and E-selectin were higher in CHD patients with positive vs. negative Chlamydia lipopolysaccharide IgA ( P  = 0.044 for both). H. pylori antibodies alone did not predict raised levels of sCAMs, but in CHD patients sICAM-1 was increased with IgA seropositivity to both bacteria compared to double seronegativity ( P  = 0.034). Concentrations of sVCAM-1 were elevated in CHD patients with double IgA seropositivity compared to those with Chlamydia lipopolysaccharide IgA seropositivity alone ( P  = 0.018).
Conclusion   Our results may indicate that C. pneumoniae contributes to increased inflammation in CHD, and that this contribution is even more pronounced when present in combination with H. pylori IgA antibodies.     

Comparison of nasal swab culture, quantitative culture of nasal mucosal tissue and PCR in detecting Streptococcus pneumoniae carriage in rats

It is not known how well nasopharyngeal swab culture represents pneumococcal carriage status. We tested this by comparing swab culture to quantitative culture and quantitative PCR of mucosal tissue in a rat model of pneumococcal carriage. Quantitative culture and quantitative PCR identified significantly more carriers compared to swab culture (differences 15% and 33%, 95% CI 1-28% and 16-47%, p=0.04 and 0.001, respectively). The sensitivity and specificity of swab culture was 75/92% and 63/100% compared to quantitative tissue culture and quantitative PCR, respectively. The quantitative estimates of culture and PCR were very similar (Pearson correlation coefficient 0.79, p<0.001). In conclusion, even a well-controlled swab sampling markedly underestimates pneumococcal carriage rate, and simultaneous use of quantitative culture and PCR increases the number of positive samples by about one third.     

Immunologic reactivity of (C57BL/6 × A/Sn)F1 mice in mixed Mycoplasma-virus infection

Mixed infection of hybrid mice, highly resistant to Rauscher virus, with this virus andMycoplasma arthritidis was accompanied by progressive inhibition of populations of splenic rosetteforming (REC) and plaque-forming (PFC) cells and led to induction of malignant erythroblastosis, cytologically identical with Rauscher's leukemia. During mixed infection of the hybrid mice withAcholeplasma laidlawii and Rauscher virus the immune response was almost completely suppressed on the 21st day and considerable splenomegaly was observed, but by the 62nd day of infection the RFC and PFC populations and also the weight of the spleens had regained the control level. The possible role of mycoplasmas in the induction and development of Rauscher's leukemia is discussed.N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician O. V. Baroyan.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 9, pp. 327–329, September, 1979.     

Changes in Streptococcus pneumoniae serotypes causing invasive disease with non-universal vaccination coverage of the seven-valent conjugate vaccine

The pneumococcal seven-valent conjugate vaccine (PCV7) has been administered in Portugal since late 2001 through the private sector. To evaluate the impact of PCV7 use, the serotypes and antimicrobial susceptibility of pneumococci causing invasive disease in Portugal during 2003–2005 were determined and compared with available data for the period 1999–2002. Changes in serotype distribution compatible with the introduction of PCV7 were shown for children ≤5 years of age from 2003 onwards and for adults from 2004 onwards. PCV7 use with coverage of 43% of children with four doses in the 2004 birth cohort, although substantially below universal coverage, seems to have contributed to greatly reducing the proportion of invasive infections due to vaccine serotypes  4, 6B, 14 and 23F. Similarly, significant indirect effects on the serotype distribution of pneumococci causing infections in adults were noted, with reductions in the proportion of invasive infections caused by serotypes  4, 5 and 14. These changes were accompanied by an increase in the proportion of two non-vaccine serotypes: 19A isolates in all age groups and 7F isolates in adults. Whereas serotypes  6B, 14 and 19A were associated with multidrug resistance, isolates expressing serotypes  4 and 7F were fully susceptible for the most part. There were no changes in the proportion of resistant isolates within each serotype and, in spite of the changes in serotype prevalence, there was not an overall reduction in the proportion of infections caused by resistant pneumococci.