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1.
Rev Esp Quimioter ; 34(5): 496-499, 2021 Oct.
Article in Spanish | MEDLINE | ID: mdl-34286949

ABSTRACT

OBJECTIVE: Mycoplasma genitalium is associated with persistent/recurrent sexually transmitted infections. The aim of this work was to estimate the prevalence and azithromycin resistance of M. genitalium in general population that was attended at Primary Care of Santiago de Compostela Health Care Area. METHODS: The study was carried out in 2019 in general population of Santiago de Compostela Health Care Area. Real-time multiplex PCR was used for screening of sexually transmitted infections associated pathogens and detection of mutations in the 23S rRNA gene. RESULTS: A total of 502 women and 532 men were studied. The prevalence of M. genitalium was 2,4% in men and 2,9% in women. Overall azithromycin resistance was 20% all of them detected in men. The mutations found were A2059G, A2058G and A2058T. CONCLUSIONS: Although the proportion of M. genitalium infection is low, the high percentage of azithromycin resistance detected supports the relevance of these data in order to the right management of the patients with sexually transmitted diseases and, so as, to avoid the emergence of resistance in other pathogens of the urogenital tract.


Subject(s)
Mycoplasma Infections , Mycoplasma genitalium , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Azithromycin/pharmacology , Azithromycin/therapeutic use , Delivery of Health Care , Drug Resistance, Bacterial/genetics , Female , Humans , Macrolides , Male , Mycoplasma Infections/drug therapy , Mycoplasma Infections/epidemiology , Mycoplasma genitalium/genetics , Prevalence , Primary Health Care
6.
Toxicon ; 60(5): 864-73, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22683532

ABSTRACT

Commission Regulation (EC) N° 2074/2005 recognises the biological method as the reference method for Paralytic Shellfish Poisoning (PSP) toxins detection in molluscs. It was amended by Commission Regulation (EC) N° 1664/2006 that accepted the so-called Lawrence method as an alternative to the reference method. The goal of this study was to compare AOAC Official Methods of Analysis 959.08 (Biological method) and 2005.06 (Prechromatographic Oxidation and Liquid Chromatography with fluorescence detection) in samples with different toxin profiles. The influence of extraction solvent in the total samples toxicity was also evaluated. A total of 40 samples including mussels, clams, scallops, razor-clams, cockles, oysters and barnacles were analysed by both official methods. Samples were selected with Alexandrium and Gymnodinium toxic profiles, from different origin and including several presentations: fresh, frozen, canned and boiled. Acetic and hydrochloric acid extractions were performed in all samples and the extracts were simultaneously analysed by both methods. Most samples were naturally contaminated and two samples were spiked. Comparison of both official methods, mouse bioassay (MBA) with HCl extraction and Liquid Chromatography with fluorescence detection (HPLC-FLD) with acetic acid extraction, led to an 85% of consistent results regarding compliance with legal limit, including samples below and above it. The linear correlation coefficient was r² = 0.69 and the paired t test (two tails, α = 0.05) indicated that there were not significant differences among both sets of data. Nevertheless, toxicity differences were found in several samples. In 15 out of 18 shellfish with a Gymnodinium toxic profile, higher toxicity levels were obtained by MBA. This fact was more evident in 7 samples, partially related to the lack of standards and the impossibility of analysing dc-NEO, C1, 2 and GTX6 at the beginning of the study. However, other factors concerning the extraction and SPE clean-ups steps may also contribute. By contrast, 9 samples presented a much higher total toxicity by HPLC-FLD than by MBA. These higher results obtained by HPLC-FLD could not only be due to the use of the highest toxicity equivalency factor (TEF) for isomers oxidated into products that coelute when total toxicity of these samples were calculated. Further analyses of results obtained by HPLC-FLD and by MBA with both extracts were done separately.


Subject(s)
Biological Assay/methods , Chromatography, High Pressure Liquid/methods , Saxitoxin/analysis , Saxitoxin/toxicity , Shellfish/analysis , Shellfish/standards , Animals , European Union , Mice , Molecular Structure , Saxitoxin/chemistry , Species Specificity
12.
Toxicon ; 51(5): 765-73, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18243269

ABSTRACT

The contamination of different types of shellfish by okadaic acid (OA)-group toxin esters is an important problem that presents serious risk for human health. During previous investigations carried out in our laboratory by liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS), the occurrence of a high percentage of esters in relation to the total OA equivalents has been observed in several shellfish species. The determination of these kinds of toxins using LC/MS or other chemical methods requires a hydrolysis step in order to convert the sterified compounds into the parent toxins, OA, dinophysistoxins-1 (DTX-1) and dinophysistoxins-2 (DTX-2). Most of the hydrolysis procedures are based on an alkaline hydrolysis reaction. However, despite hydrolysis being a critical step within the analysis, it has not been studied in depth up to now. The present paper reports the results obtained after evaluating the hydrolysis process of an esterified form of OA by using a standard of 7-O-acyl ester with palmitoyl as the fatty acid (palOA). Investigations were focused on checking the effectiveness of the hydrolysis for palOA using methanol as solvent standard and matrices matched standards. From the results obtained, no matrix influence on the hydrolysis process was observed and the quantity of palOA converted into OA was always above 80%. The analyses of different Spanish shellfish samples showed percentages of palOA in relation to the total OA esters ranging from 27% to 90%, depending on the shellfish specie.


Subject(s)
Food Analysis/methods , Mollusca/chemistry , Mollusk Venoms/chemistry , Okadaic Acid/analogs & derivatives , Okadaic Acid/chemistry , Shellfish , Animals , Chromatography, Liquid , Esters , Hydrolysis , Molecular Structure , Spain , Tandem Mass Spectrometry
13.
Toxicon ; 49(8): 1129-34, 2007 Jun 15.
Article in English | MEDLINE | ID: mdl-17368701

ABSTRACT

By the end of 2005, a toxic episode of phytoplankton origin in bivalve shellfish led to the closing down of several shellfish production areas in Galicia (northwestern region of Spain). During this time, different kinds of shellfish were collected and analysed by LC-MS/MS to search for the following lipophilic toxins: okadaic acid (OA), dinophysistoxins (DTXs), pectenotoxins (PTXs), azaspiracids (AZAs) and spirolides. Samples were analysed before alkaline hydrolysis in order to investigate the presence of free OA and DTXs, AZAs, PTXs and spirolides, and after alkaline hydrolysis to detect OA and DTXs esters. All of the samples were found to be contaminated with OA and/or DTX-2, as well as esterified forms of these diarrhetic shellfish poison (DSP) toxins, at levels around and above European regulatory limit (160 microg of okadaic acid equivalents/kg). The analyses of mussels and razor clam also revealed the presence of 13-desmethyl spirolide C (SPX-1) at levels below 31 microg/kg. Likewise, in many of the samples different levels of pectenotoxin-2 secoacid (PTX-2sa) were detected. DSP toxin esters represent practically the 100% of the total OA equivalents for scallops, clams, razor clams and cockles.


Subject(s)
Bivalvia/microbiology , Dinoflagellida/chemistry , Environmental Monitoring/statistics & numerical data , Marine Toxins/analysis , Phytoplankton/chemistry , Shellfish/microbiology , Animals , Chromatography, Liquid , Okadaic Acid/analysis , Spain , Species Specificity , Spiro Compounds/analysis , Tandem Mass Spectrometry
14.
J Chromatogr A ; 1140(1-2): 78-87, 2007 Jan 26.
Article in English | MEDLINE | ID: mdl-17140589

ABSTRACT

An HPLC-FLD method, involving pre-chromatographic oxidation of the PSP toxins with hydrogen peroxide and periodate, has been AOAC validated through a collaborative trial and adopted as AOAC Official Method. This method could be a candidate for replacing the mouse bioassay (MBA) for the Official Control of PSP toxins at European level, once accepted by the legislation. An interlaboratory exercise has been organized by the CRLMB to evaluate its "fitness for purpose" for the Official Control of PSP toxins in the EU laboratories. Eighteen EU laboratories took part in the study and had to analyze six bivalve mollusc samples with several PSP toxic profiles. The performance of the participant laboratories in the application of this method was compared with that obtained at the collaborative trial. Information on problems/drawbacks encountered by participants in the application of this method was also sought. The HPLC validated method is only applicable for Official PSP Control for certain samples. This depends on sample PSP toxic profile. Results obtained for samples where only GTX2,3 and STX were present were satisfactory and in agreement with MBA results. Results obtained for a sample with a toxic profile dominated by GTX6 and suspected to contain also C1,2 and C3,4 were not satisfactory. GTX5 and dc-STX could be quantified, although the results achieved (total toxicity) were lower than those obtained by MBA. It can be also useful as a screening method, complementary to MBA, helping in the reduction of the animals used. However, the lack of several PSP standards, the fact that the method is not validated for all the PSP toxins, and several drawbacks found in its application are a handicap to fully implement it for Official PSP Control as a viable replacement for bioassay.


Subject(s)
Chromatography, High Pressure Liquid/methods , Marine Toxins/analysis , Neurotoxins/analysis , Animals , Bivalvia/chemistry , Laboratories/standards , Marine Toxins/poisoning , Paralysis/chemically induced , Quality Control , Shellfish/analysis
15.
Toxicon ; 48(8): 1068-74, 2006 Dec 15.
Article in English | MEDLINE | ID: mdl-17046040

ABSTRACT

During the months of November and December 2005, a harmful algal bloom in the northwestern region of Spain caused the accumulation of different algal toxins in several types of shellfish. Many of the shellfish were contaminated at levels above European regulatory limit, presenting serious risks for human health. The analysis of mussels sampled in the affected areas to search for lipophilic toxins, using liquid chromatography tandem mass spectrometry (LC-MS/MS), showed the presence of free okadaic acid (OA) and dinophysistoxin-2 (DTX-2) as well as esters of these toxins. The results also revealed the presence of minor amounts of 13-desmethyl spirolide C (SPX-1) in the analysed samples, although this toxin has never been reported before in Spain. The combination of different MS modes of operation, just as enhanced MS (EMS) and MS(3) experiments, allowed to confirm the first occurrence of spirolides in Spanish shellfish.


Subject(s)
Bivalvia/chemistry , Marine Toxins/isolation & purification , Shellfish , Spiro Compounds/isolation & purification , Animals , Chromatography, Liquid/methods , Marine Toxins/analysis , Mass Spectrometry/methods , Spain , Spiro Compounds/analysis , Spiro Compounds/chemistry
16.
Food Addit Contam ; 20(3): 276-80, 2003 Mar.
Article in English | MEDLINE | ID: mdl-12623653

ABSTRACT

Raw cow's milk collected from dairy farms in the province of Leon, Spain, was examined for aflatoxin M(1) (AFM(1)). The samples were analysed with a commercial competitive enzyme-linked immunosorbent assay (ELISA) kit and high-performance liquid chromatography (HPLC). The concentrations of AFM(1) in the milk extracts were initially estimated by ELISA, with recovery rates of 74.6-109% for artificially contaminated milk at levels of 10-80 ng x l(1). Samples found to contain more than 10 ng x l(1) were further quantified with HPLC. The mean recovery for this method was 89.3%. The quantification limit was 10 ng x l(1) for both ELISA and HPLC. Although AFM(1) was confirmed in only 3.3% of the samples, the concentrations in all these cases were lower than the maximum limit applicable to these products pursuant to European Union legislation. Both methods were validated with reference material certified by the Community Bureau of Reference.


Subject(s)
Aflatoxin M1/analysis , Food Contamination/analysis , Milk/chemistry , Animals , Chromatography, High Pressure Liquid , Enzyme-Linked Immunosorbent Assay , Food Analysis/methods , Spain
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