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Biomedical subjects

A Clavel

Publications and source records attributed to A Clavel.

At least 19 recordsLinked to original sources

A specific primer pair for the diagnosis and identification of Acanthamoeba astronyxis by random amplified polymorphic DNA-polymerase chain reaction.

Random amplified polymorphic DNA (RAPD) is a useful tool for species identification. The obtained band patterns can be used for specific primer pair design that is useful for species identification. In this study, a distinctive 485-bp band in Acanthamoeba astronyxis band patterns was found, using the OPC20 primer (ACTTCGCCAC). The band specificity was confirmed by hybridization, using it as a probe, against all OPC20 amplifications from different Acanthamoeba species. Once the fragment was sequenced, we used it to design a specific primer pair that was useful for the identification of different isolates as A. astronyxis species.

Acanthamoeba↗

Evaluation of an immunochromatographic dip-strip test for the detection of Cryptosporidium oocysts in stool specimens.

The study presented here examined the efficacy of a commercially available qualitative immunochromatographic assay for detecting Cryptosporidium oocysts in stool samples. A total of 75 samples were tested, including 50 positive for Cryptosporidium spp. by acid-fast stain, 20 positive for other parasites ( Blastocystis hominis, Endolimax nana, Entamoeba coli, Giardia lamblia, Ascaris lumbricoides, Strongyloides stercoralis and Trichuris trichiura), and five negative samples. The observed sensitivity was 98%, while specificity was 100%; the detection threshold was near 1000 oocysts/ml. Correctly diagnosed positive samples included Cryptosporidium parvum genotypes 1 and 2, whereas the single false-negative sample corresponded to a Cryptosporidium meleagridis infection.

Animals↗

[Antimicrobial resistance of clinical strains of Salmonella enterica isolated in Zaragoza].

In order to identify any changes in the incidence of Salmonella enterica serotypes and their resistance to a variety of antimicrobial agents, we conducted a retrospective study of all the strains isolated from stool samples at Hospital Clínico Universitario Lozano Blesa in Zaragoza from 1997 to 2000. We observed an increase in the number of isolates of Salmonella and Campylobacter and a decrease in other enteropathogens. Enteritidis was the most frequently isolated serotype (55.2%), showing an increasing tendency (from 44.1% in 1997 to 60.6% in 2000). Hadar, glostrup and virchow showed the highest rate of resistance to nalidixic acid. Enteritidis also showed an important increase in resistance to nalidixic acid (from 17.6% in 1997 to 41.4% in 2000). Typhimurium showed the highest resistance levels to ampicillin, chloramphenicol and cotrimoxazole. No resistance to fluoroquinolones or to cefotaxime was detected, with the exception of 0.5% of the S. enteritidis strains, which showed resistance to fluoroquinolones.

Drug Resistance, Bacterial↗

The eaeA gene is not found in Hafnia alvei from patients with diarrhea in Aragón, Spain.

A total of 102 Hafnia alvei clinical strains isolated from different patients with diarrhea has been tested, using polymerase chain reaction and dot-blot hybridization, for the enteropathogenic Escherichia coli attaching and effacing A (eaeA) gene to establish their role as a causative agent of diarrhea in our environment. None of them was positive for the eaeA gene. We cannot consider the eaeA gene as the virulence-associated factor implicated in the H. alvei strains isolated from diarrheal feces in our region.

Adhesins, Bacterial↗

Western blot applied to the diagnosis and post-treatment monitoring of human hydatidosis.

The serologic diagnosis of hydatidosis (caused by Echinococcus granulosus) can be made by different techniques, although the lack of standardization of the antigens affects the sensitivity, specificity and concordance among the different tests. We have applied the Western-Blot (WB) technique, associated with a purified antigen from sheep hydatid fluid, at 60 samples of serum from 14 patients suffering echinococcosis in different bodily locations, monitored for 3 years. The WB test enabled the detection of antibodies in the pre-surgical samples for proteins of 12-14, 16, 20, 24-26, 34, 39 and 42 kDa in molecular weight in 15-96% of the patients. The combination involving 2 of the 3 proteins of 20, 39 and 42 kDa has made it possible to diagnose 100% of the cases. The antibodies specific to proteins 39 and 42 kDa disappeared in less than one year in the patients cured after surgery, while in patients with persistent or recurrent parasitism the bands present before surgery persisted or other new ones appeared. The WB with purified antigens proved to be highly useful in the diagnosis and post-surgical monitoring of hydatidosis patients. The antigen used is proposed as a standard antigen for the diagnosis and follow-up of pre- and postsurgical hydatidosis.

Adult↗

Astrovirus infection among children with gastroenteritis in the city of Zaragoza, Spain.

The incidence of astrovirus infection in children less than 10 years of age with gastroenteritis in the city of Zaragoza, Spain, was analysed during a 12-month period. A total of 718 stool samples obtained from 534 children were examined. In 401 samples no routinely searched for pathogenic organism was detected; these specimens were then tested for the presence of astrovirus antigens. Astrovirus was detected in the samples of 15 (5.5%) patients, a detection rate similar to that recognised for Yersinia enterocolitica and Cryptosporidium spp. All children with confirmed astrovirus infection had diarrhoea (median duration, 9 days), 11 experienced loss of appetite, 5 vomiting, and 4 fever. The incidence of astrovirus infection reported here indicates that Spanish children suffering from gastroenteritis should be routinely screened for the presence of astrovirus.

Astroviridae Infections↗

Emerging and reemerging pathogens.

From 1973 to 1995, 29 new and reemerging pathogenic microbes were recognized. However, in discussions about emerging infectious diseases, the focus is often on the clinical effects of the host-parasite relationship, rather than the examination of the biology of the pathogen. Many of what we refer to as emerging diseases are characterized better as 'diseases of human progress'. Thus, the aerosolization of water has played an important role in the emergence of Legionella pneumophila infections. New diseases are superimposed on endemic diseases such as diarrhoeal diseases, malaria and tuberculosis. In addition, many pathogens are becoming increasingly resistant to standard antimicrobial drugs, making treatment difficult and in some cases impossible. We summarize our experience on emerging parasitic diseases (primary amoebic meningoencephalitis, respiratory cryptosporidiosis, and diplogonoporiasis), and selected problems of bacterial resistance (MDR tuberculosis caused by Mycobacterium bovis and macrolide-resistance mechanisms of Streptococcus pneumoniae and S. pyogenes).

AIDS-Related Opportunistic Infections↗

Visualization of hydatid elements: comparison of several techniques.

Some techniques available at our laboratory were tested for their ability to aid in the morphological diagnosis of hydatid elements (Echinococcus granulosus ["Taenia echinococcus"]) isolated from cysts in humans and sheep. Unstained, methanol-fixed hooklets were fluorescent, most starkly so under violet light (excitation filter wavelength, 405 nm; long-pass filter wavelength, 495 nm). Auramine-rhodamine and Gram procedures failed to stain hooklets. Ziehl-Neelsen stain yielded indifferent results when organisms were viewed under transmitted light but resulted in a surprisingly intense red fluorescence when organisms were viewed under green light (excitation, 546 nm; long pass, 590 nm). Wheatley trichrome stain gave better and more uniform results than fuchsin. Ryan trichrome blue stain was the best under transmitted light; hooklets stained uniformly and intensely and were easily distinguishable from the background. Very satisfactory results were also obtained with a much simpler procedure (modified Baxby technique: no fixation, steaming hot 1% safranin for 2 min, and malachite green for 30 s). Therefore, Ryan and modified Baxby stains are recommended for the examination of E. granulosus under transmitted light. For fluorescence microscopy, Ziehl-Neelsen stain under green excitation light, or violet light with no staining, is also very useful. Epifluorescence microscopy is especially convenient for examining samples concentrated by filtration, as it renders the filter pores inconspicuous.

Animals↗

[Value of transesophageal programmed atrial stimulation in the evaluation of unexplained cerebrovascular accidents].

Certain embolic cerebrovascular accidents can be explained by the development of paroxysmal atrial fibrillation. When noninvasive complementary investigations are negative, programmed atrial stimulation can be proposed to detect increased atrial vulnerability. The objective of this study was to evaluate the reliability of this method performed via a transoesophageal approach in 59 subjects presenting with an embolic cerebrovascular accident and who were in sinus rhythm at the time of the accident. Seven of these patients had a history of paroxysmal atrial fibrillation (AF) or atrial tachycardia (AT) (group I). Three of these seven patients also presented AV nodal reentrant junctional tachycardia. The other 52 patients had no history of arrhythmia and their Holter recording did not reveal any episodes of sustained atrial tachycardia (group II). Transoesophageal programmed atrial stimulation used up to 2 extrastimuli under baseline conditions and during Isuprel infusion. The following results were obtained: sustained atrial tachycardia (> 1 min) was induced in all patients of group 1, 3 of them also presented inducible junctional tachycardias. 14 patients of group II (27%) presented inducible supraventricular tachycardia: atrial tachycardia in 7 cases. Patients in group II with inducible AT presented either heart disease (n = 3) or minor abnormalities on the Holter recording (runs of atrial premature complexes or sinus pauses (n = 3). Two of these patients subsequently developed sustained atrial fibrillation during follow-up. In 25 patients with normal Holter recording and no heart disease, programmed atrial stimulation induced junctional tachycardia in 4 cases. In conclusion, transoesophageal electrophysiological investigation is a useful way to identify various forms of supraventricular tachycardia able to explain an embolic cerebrovascular accident. The considerable incidence of inducible AV nodal reentrant junctional tachycardia must be emphasized, while the incidence of atrial fibrillation is much lower than during intracardiac investigations.

Adult↗

Use of a selective medium and a membrane filter method for isolation of Campylobacter species from Spanish paediatric patients.

A study was conducted to assess the value of a combination of two culture methods for isolation of Campylobacter spp. from Spanish children. Seven hundred twenty-nine diarrhoeal stool specimens from 599 patients were examined for Campylobacter spp. by culturing them on charcoal cefoperazone deoxycholate agar and on blood agar with a membrane filter. One hundred sixteen Campylobacter strains were isolated from a total of 108 specimens; 75 (64.6%) were Campylobacter jejuni, 32 (27.5%) were Campylobacter coli, 8 (6.8%) were non-typeable, and one (0.9%) was Campylobacter upsaliensis. Campylobacters were isolated from 99 positive samples using charcoal cefoperazone deoxycholate agar alone. The filtration technique alone yielded only 86 positive samples. Seven specimens yielded different Campylobacter spp. with different media. The only catalase-negative strain was recovered using the filter method. The combination of the selective medium with the filter method increased the isolation rate of Campylobacter strains by 14.1%. Isolation rates of campylobacters using the filter method were similar to those reported in European studies, in which a similar frequency of Campylobacter upsaliensis was observed. The addition of a filter method for routine laboratory isolation of campylobacters should be considered in selected age groups (in children < 10 years of age) or in areas where catalase-negative or weakly-positive Campylobacter strains may be of epidemiological significance.

Adolescent↗

Fuchsin fluorescence and autofluorescence in Cryptosporidium, Isospora and Cyclospora oocysts.

Cryptosporidium parvum and Isospora belli oocysts stained with carbol-fuchsin, as in a modified Ziehl Neelsen technique, fluoresce bright red under green light (546nm). Cryptosporidium oocysts tend to fluoresce more brightly the less intensely stained they appear under transmitted light; this is not the case with Isospora. Fuchsin-stained Cyclospora cayetanensis oocysts fluoresce rather dimly, but those not taking the dye retain their typical autofluorescence. Cryptosporidium and Isospora oocysts are also autofluorescent, appearing violet under u.v. light (365 nm), and green under violet (405 nm) and blue-violet light (436 nm). Their autofluorescence does not survive the staining procedure.

Animals↗

Visceral leishmaniasis with cutaneous lesions in a patient infected with human immunodeficiency virus.

We report a case of visceral leishmaniasis (VL) with cutaneous lesions in a patient infected with human immunodeficiency virus (HIV). The cutaneous lesions consisted of erythematous papules on the legs. Biopsy of one lesion showed abundant Leishmania amastigotes within epithelial cells of an eccrine sweat gland in the dermis. Leishmania organisms were also found in a blood smear. Rapid and complete clearance of the cutaneous lesions was achieved after antimony therapy. Cutaneous lesions in VL are being reported increasingly frequently in patients with HIV infection and their significance remains in discussion.

AIDS-Related Opportunistic Infections↗

[Quinolone resistance in Salmonella enterica].

Nalidixic acid resistance is a marker of quinolone sensibility in Salmonella spp. In our laboratory we observed an increase of nalidixic acid resistant strains from 0.44% in 1990 to 13.7% in 1996. The purpose of this work was to study the evolution of nalidixic acid resistance in Salmonella spp., as well as the association with certain serotypes and the cross-resistance to others agents. A total of 1,782 isolates of Salmonella enterica were studied from fecal and blood sources over a 7-year period; 45 strains (2.52%) were resistant to nalidixic acid (MIC > 16 microgram/ml). Two isolates required 2 microgram/ml of ciprofloxacin for inhibition, while the remaining 43 strains required 1 microgram/ml. The most frequent serotypes in nalidixic acid resistant strains were the following: enteritidis (33.3%), hadar (31.11%) and virchow (15.5%). In 1996, enteritidis was the predominant serotype (13/31), and hadar serotype was found to have the greatest nalidixic acid resistance at 80% of strains. No differences of sensibility to other agents were detected between the nalidixic acid resistant group and nalidixic acid sensitive strains. This type of resistance appears regularly in uncommon serotypes such as hadar. The increasing incidence of resistant isolates in enteritidis in 1996 is of even greater concern.

Anti-Infective Agents↗

Short report: one case of nasal human myiasis caused by third stage instar larvae of Oestrus ovis.

This is a report of a case of human nasal myiasis caused by third instar larvae of the sheep nasal bot fly, Oestrus ovis. Female flies rarely deposit first instar larvae in the eye, nostrils, and external auditory canal of humans, where they usually survive only a few days without further development. One human infestation by third instar larvae of O. ovis has been recorded in an patient infected with human immunodeficiency virus (HIV) in the United Kingdom. Our report describes a new case of nasal myiasis caused by the third instar larvae of the sheep bot fly in a patient without HIV or signs of any other disease.

Adult↗

Diplogonoporiasis presumably introduced into Spain: first confirmed case of human infection acquired outside the Far East.

A 58-year-old man who was very fond of eating raw fish and had not left Zaragoza Province in Spain in the last 20 years excreted a short chain of gravid proglottids. Treatment with 50 mg/kg of paromomycin sulfate was divided into three doses, all given within the same day, followed by administration of a laxative at night. On days 1 and 2 post-treatment, several chains of degenerated proglottids were evacuated. Only eggs were expelled on days 3 and 4. A long complete strobila including the scolex, and immature, mature, and gravid segments was spontaneously discharged on day 25 after treatment. It was identified as Diplogonoporus balaenopterae, and was the first confirmed case of diplogonoporiasis outside the Far East (all previous cases were reported from Japan, except for one recent case reported from Korea). The origin of this case, outside of its normal geographic location, may be associated with the importation of fish into Spain. The viability of the infective larval stage is evidence of its resistance to export/import conditions. Treatment with paromomycin sulfate did not result in the whole worm being discharged but several short strobilar chains showed drug-induced degeneration.

Animals↗

Comparison of an acid-fast stain and a monoclonal antibody-based immunofluorescence reagent for the detection of Cryptosporidium oocysts in faecal specimens from cattle and pigs.

A commercially available direct immunofluorescence (IF) assay with monoclonal antibodies (Monofluo Kit Cryptosporidium, Diagnostics Pasteur, France) was compared with the modified Ziehl-Neelsen (MZN) acid-fast technique for the detection of Cryptosporidium oocysts in faecal samples from cattle and pigs. Stool specimens individually collected from 108 bovines and 90 pigs were examined in a blind test. The results of the two procedures corresponded (both positive or negative) in 102 (94.4%) cattle samples and 80 (88.9%) pig faecal samples. However, the remaining six (5.5%) cattle specimens and 10 (11.1%) pig stool samples, all of them harboring few oocysts (0-1 oocysts per 20 x field), were negative by MZN and positive by IF. False-negative results of the acid-fast stain occurred in suckling (17.2% of discrepant results) and weaned calves (2.9%) as well as weaned piglets (43.7%) and fattening pigs (10%). Stool specimens from the remaining age groups were negative by both techniques. The MacNemar's chi-square test showed that differences between both methods were statistically significant (P < 0.05). Compared with immunofluorescence procedure, the sensitivity of MZN technique in samples from cattle and pigs was 79.3% and 67.7% and the negative predictive value was 92.9% and 85.5% respectively. The specificity and positive predictive values of the acid-fast stain were 100% in both animal species. It is concluded that the monoclonal antibody-based immunofluorescence reagent evaluated is more efficient that the MZN technique, especially for detecting a low number of Cryptosporidium oocysts, in faecal specimens from both cattle and pigs.

Aging↗