Francisco Javier Castillo

Respiratory cryptosporidiosis: Case series and review of the literature

SummaryFive case of intestinal cryptosporidiosis with pulmonary involvement in patients with AIDS are reported. The diagnosis was based on the recognition of acid-fast oocysts in sputum or aspirated bronchial material and stool specimens. Coughing and excess secretions were present in all cases. Four patients had other associated pulmonary pathogens: twoMycobacterium tuberculosis, oneMycobacterium fortuitum and one Cytomegalovirus +Pneumocystis carinii; all of them had a previous (three cases) or simultaneous (one case) diagnosis of intestinal cryptosporidiosis, presenting with diarrhoea and vomiting. In the fifth patientCryptosporidium was the only pulmonary pathogen found in a bronchial aspirate, and the onset of diarrhoea was 1 month after respiratory detection. Fifty-seven cases of respiratory cryptosporidiosis have been reported since 1980. In 17 of them, no other pathogen was found. Diarrhoea was present in 77% of the patients, cough in 77%, dyspnea in 58%, expectoration in 54%, fever in 45%, thoracic pain in 33%.ZusammenfassungWir berichten über fünf Fälle von intestinaler Kryptosporidiose mit pulmonaler Beteiligung bei AIDS Patienten. Die Diagnose wurde durch den Nachweis von säurefesten Oozysten im Sputum oder aspiriertem Bronchialsekret und im Stuhl gestellt. Bei allen Patienten bestand Husten und Auswurf. Bei vier Patienten fand sich gleichzeitig eine Assoziation mit anderen Infektionen, in zwei Fällen durchMycobacterium tuberculosis, in einem Fall durchMycobacterium fortuitum und in einem Fall durch Cytomegalovirus plusPneumocystis carinii. Bei allen Patienten war eine vorbestehende (drei Fälle) oder gleichzeitige intestinale Kryptosporidiose vorhanden, die mit Durchfall und Erbrechen einherging. Bei dem fünften Patienten war im Bronchialsekret ausschließlichCryptosporidium nachzuweisen, die Diarrhoe setzte erst einen Monat nach Entdeckung des Erregers im Respirationstrakt ein. Seit 1980 wurden 57 Fälle von respiratorischer Kryptosporidiose publiziert. In 17 Fällen fand sich kein anderer Erreger. In je 77% der Fälle waren Diarrhoe und Husten, in 58% Atemnot, in 54% Auswurf, 45% Fieber und in 33% Schmerzen aufgetreten.

Evaluation of an immunochromatographic dip strip test for simultaneous detection of Cryptosporidium spp , Giardia duodenalis , and Entamoeba histolytica antigens in human faecal samples

Immunochromatographic (IC) tests may play an important role in the future diagnosis of parasitic diseases because of their speed and simplicity of use. A recently developed test to detect Cryptosporidium spp, Giardia duodenalis and Entamoeba histolytica was evaluated. Microscopy and PCR were the “gold standard” reference techniques and the results of this IC test were compared with those obtained with ELISA and IC single test for the three parasites. One hundred sixty stool samples were assayed. Using microscopy, 22 samples were diagnosed as positive for Cryptosporidium spp., 31 for Giardia duodenalis, 41 for Entamoeba histolytica/dispar, and 68 had a negative diagnosis for the three parasites. Results of IC tests show sensitivities of 70–72% for Cryptosporidium, 90–97% for Giardia and 62.5% for Entamoeba histolytica. Specificities were of 93.6–94.9%, >99% and 96.1%, respectively. In all diagnoses, agreement with microscopy and PCR was over 90%, except in the triple test and microscopy in E. histolytica detection that was 76.3%, due to the inability of microscopy to differentiate E. histolytica from nonpathogenic species such as E. dispar or E. moshkovskii. The triple stool immunoassays provide adequate sensitivities and specificities for use in outbreak situations, for screening proposals and for massive assays in endemic areas where a large number of samples must be analysed or as complementary test for individual diagnosis.

Evaluation of an Immunochromatographic Dip-Strip Test for the Detection of Cryptosporidium Oocysts in Stool Specimens

Abstract.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 1,000 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.

Detection and characterization of a ST6 clone of vanB2-Enterococcus faecalis from three different hospitals in Spain.

Thirteen vancomycin-resistant and teicoplanin-susceptible Enterococcus faecalis isolates were recovered from unrelated patients in three Spanish hospitals from November 2009 to December 2010. All isolates carried the vanB2 gene, showed indistinguishable or closely-related PFGE patterns and were ascribed to the sequence type ST6 (included into the high-risk clonal-complex CC2). They showed a multiresistance phenotype (erythromycin, tetracycline, ciprofloxacin and high-level-resistance to streptomycin, gentamicin and kanamycin) and harboured the aac(6’)-aph(2”), ant(6)-Ia, and tet(M)+/−tet(L) genes. All isolates produced gelatinase and harboured the gelE gene, but not the esp or hyl genes. The inclusion of the vanB2 gene into the Tn5382 transposon was demonstrated in one isolate. Clonal dissemination of vanB2-containing the E. faecalis strain is demonstrated.

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 ofCampylobacter spp. from Spanish children. Seven hundred twenty-nine diarrhoea] stool specimens from 599 patients were examined forCampylobacter spp. by culturing them on charcoal cefoperazone deoxycholate agar and on blood agar with a membrane filter. One hundred sixteenCampylobacter strains were isolated from a total of 108 specimens; 75 (64.6%) wereCampylobacter jejuni, 32 (27.5%) wereCampylobacter coli, 8 (6.8%) were non-typeable, and one (0.9%) wasCampylobacter 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 differentCampylobacter 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 ofCampylobacter 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 ofCampylobacter 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-positiveCampylobacter strains may be of epidemiological significance.

Prevalencia de Giardia duodenalis genotipo B en humanos en Zaragoza y León, España

OBJECTIVES To study Giardia duodenalis assemblages circulating in the health areas of two hospitals in Zaragoza and León (Spain). METHODS A total of 211 stool samples with Giardia were genotyped by PCR of the tpi gene. RESULTS Assemblage B was the most prevalent, both in Zaragoza (84,7%) and León (95,1%). The remaining isolates were identified as AII+B. CONCLUSIONS We detected the spread of G. duodenalis assemblage B in Zaragoza and in León, with an increase in its prevalence in Zaragoza compared to previous studies.

Astrovirus Infection Among Children with Gastroenteritis in the City of Zaragoza, Spain

Abstract 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.

Comparison of Local Features from Two Spanish Hospitals Reveals Common and Specific Traits at Multiple Levels of the Molecular Epidemiology of Metallo-β-Lactamase-Producing Pseudomonas spp.

ABSTRACT Twenty-seven well-characterized metallo-β-lactamase (MBL)-producing Pseudomonas strains from two distantly located hospitals were analyzed. The results revealed specific features defining the multilevel epidemiology of strains from each hospital in terms of species, clonality, predominance of high-risk clones, composition/diversity of integrons, and linkages of Tn402-related structures. Therefore, despite the global trends driving the epidemiology of MBL-producing Pseudomonas spp., the presence of local features has to be considered in order to understand this threat and implement proper control strategies.

Genotypic and phenotypic characterization of methicillin-resistant Staphylococcus aureus (MRSA) clones with high-level mupirocin resistance.

A high proportion of methicillin-resistant Staphylococcus aureus isolates recovered in one year period showed high-level mupirocin-resistance (HLMUPR-MRSA) in our environment (27.2%). HLMUPR-MRSA isolates were mainly collected from skin and soft tissue samples, and diabetes was the main related comorbidity condition. These isolates were more frequently found in vascular surgery. HLMUPR-MRSA was more resistant to aminoglycosides than mupirocin-susceptible MRSA, linked to the presence of bifunctional and/or nucleotidyltransferase enzymes with/without macrolide resistance associated with the msr(A) gene. Most of HLMUPR-MRSA isolates belonged to ST125/t067. Nine IS257-ileS2 amplification patterns (p3 was the most frequent) were observed in HLMUPR-MRSA isolates, suggesting the presence of several mupirocin-resistance-carrying plasmids in our environment and promoting the emergence of mupirocin resistance. The presence of the same IS257-ileS2 amplification pattern p3 in 65% of HLMUPR-MRSA, all of them ST125/t067, suggests a clonal spread in our hospital and community environment which could explain the high prevalence of HLMUPR-MRSA during the study period. An outbreak situation or an increase in mupirocin consumption was not observed.

Evaluation of four phenotypic methods to detect plasmid-mediated AmpC β-lactamases in clinical isolates

Four phenotypic methods (three dimensional test, AmpC test, cloxacillin synergy test and cefotetan/cefotetan-cloxacillin E-test) to detect plasmid-mediated AmpC β-lactamases (pAmpC) were compared in 125 clinical Enterobacteriaceae isolates with AmpC profile: 74 E. coli (blaCMY-2: 70; blaDHA-1: 4), five K. pneumoniae (blaCMY-2: 2; blaDHA-1: 3), six P. mirabilis (blaCMY-2: 6) and 40 negative isolates for pAmpC β-lactamases. All evaluated methods showed a good sensitivity (>95%) but low values of specificity (<60%) in E. coli, explained by an increase of AmpC expression caused by chromosomal ampC promoter/attenuator mutations (−42, −18, −1, +58, predominantly). The cefotetan/cefotetan-cloxacillin or cloxacillin synergy test may be advocated as phenotypic screening test, and the AmpC test as confirmatory test for detection of pAmpC in isolates that lack or minimally express chromosomally encoded AmpC β-lactamases. In the case of E. coli, the phenotypic evaluated tests were not able to differentiate between chromosomal ampC overexpression or acquisition of plasmid-encoded ampC genes.