Molecular characterization of Acanthamoeba isolated in water treatment plants and comparison with clinical isolates.

A total of 116 samples (44 clinical specimens and 72 environmental samples) have been analyzed for the presence of Acanthamoeba. The environmental samples (ESs) were collected from four drinking water treatment plants (DWTP, n=32), seven wastewater treatment plants (n=28), and six locations of influence (n=12) on four river basins from the central area of Spain (winter-spring 2008). Water samples were concentrated by using the IDEXX Filta-Max(®) system. Acanthamoeba was identified in 65 of the 72 ESs by culture isolation (90.3%) and 63 by real-time PCR (87.5%), resulting in all sampling points (100%) positive for Acanthamoeba when considering both techniques and all the time period analyzed. Nine of the 44 clinical specimens were positive for Acanthamoeba. Seventeen Acanthamoeba strains (eight from four DWTP and nine from clinical samples) were also established in axenic-PYG medium. Twenty-four of the ESs and the 17 Acanthamoeba sp. strains were genotyped as T4/1, T4/8, and T4/9. The eight strains isolated from the DWTP samples were inoculated in nude mouse to ascertain their potential pathogenicity in this model. Animals that were inoculated died or showed central nervous system symptoms 9 days post-inoculation. Examination of immunofluorescence-stained brain and lung tissue sections showed multiple organisms invading both tissues, and re-isolation of throphozoites was successful in these tissues of all infected animals. For the first time, potentially pathogenic Acanthamoeba T4 has been detected in 100% of different types of water samples including tap water and sewage effluents in the central area of Spain suggesting a potential health threat for humans especially for the contact lens wearers.

Comparativa de tres técnicas de ElA indirecto para cuantificación de anticuerpos IgG a Helicobacter pylori / Comparision of three enzymo immunoassay for cuantification of immunoglobulin G against Helicobacter pylori

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[Keratitis caused by Acanthamoeba spp.: presentation of 10 cases]. / Queratitis por Acanthamoeba spp.: presentación de diez casos.

OBJECTIVE: To describe the clinical and microbiological characteristics of ten patients with Acanthamoeba spp. keratitis. METHODS: A retrospective review was made of the clinical charts of ten patients with both clinical keratitis and positive culture for Acanthamoeba spp. from 1992 (date of first diagnosed case in our hospital) on. Corneal tissue was cultured in Page media and also the contact lenses and maintenance or cleaning fluids used by each patient. RESULTS: The earliest clinical symptom observed in 60% of cases was foreign-body sensation. Before the parasitologic studies all the patients received different treatments for 4 days minimum to a maximum of 3 months. On ophthalmological examination the findings were: radial perineuritis in 4 cases, anular infiltrate in two, keratitis punctata in two and pseudodendritic ulcer in two more. Treatment employed was the following: propamidine and neomycin in 8 patients (four of them also received ketoconazole); biguanides in two. After a mean observation time of 7 months (minimum 4, maximum 10) the clinical status was favorable in 70% of cases. In three patients with delayed diagnosis of 2 to 3 months a keratoplasty was needed. COMMENTS: Acanthamoeba spp. keratitis is a process of difficult diagnosis and treatment. For early detection of this disease a narrow collaboration with the microbiologist is mandatory because the prognosis in this patients depends on the precocity of treatment.

Tularemia adquirida por picadura de garrapata en Castilla-León

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[Cat-scratch disease: series of 14 cases. The diagnostic usefulness of serology]. / Enfermedad por arañazo de gato: serie de 14 casos. Utilidad diagnóstica de la serología.

BACKGROUND: Disease by cat scratch (CSD) is a syndrome characterized by regional adenopathies following cat scratch or bite. Despite knowing the etiologic agent of the same, there is still controversy in the diagnosis of the syndrome in daily clinical practice. MATERIALS AND METHODS: The cases of CSD diagnosed from 1990 to 1997 with positive serology for Bartonella henselae performed by indirect immunofluorescence were reviewed. RESULTS: In 13 out of the 14 patients studied direct contact with cats was reported. Lymph node involvement corresponded to the following regions: axillary (57%), inguinal (28%), epitroclear (21%), cervical (14%), and periauricular (7%). In 35% of the cases, several territories were involved. In 3 patients the adenopathies were fluctuating. Eleven anatomopathologic studies were performed with 6 diagnoses of granulomatous adenitis, 3 reactive adenitis and 2 purulent adenitis. The serology for Bartonella henselae was positive in all the patients with a range of 1/64-1/256. The evolution was favorable in all the patients. DISCUSSION: Interpretation of the results of serology should be cautions and always related to the clinical and epidemiological picture of the patient. This technique should be considered as another criteria in the diagnosis of cat scratch disease to the detriment of the cutaneous test.

[Tularemia: report of 16 cases in the Castilla-Leon community]. / Tularemia: descripción de 16 casos procedentes de la Comunidad de Castilla-León.

OBJECTIVE: To report the clinico-epidemiological characteristics of 16 patients with the diagnosis of tularemia. METHOD: Retrospective review of clinical records of patients admitted to the hospital or examined at health centers in Vizcaya, with clinical course and epidemiology consistent with tularemia, from January to March 1998. CASE DEFINITION: Patient with suggestive clinical course and epidemiology (exposure to hares coming from the epizootic area) and positive serology (antibodies to Francisella tularensis > 1/160 in convalescent phase serum). RESULTS: Sixteen patients (8 males, 8 females) with a mean age of 53 years. The incubation period ranged from 1 and 8 days (mean: 5). Nine patients had the ulceroganglionar form, two the pharyngeal form, one the oculoganglionar form and one the typhoidal form. In three patients only cutaneous lesion or lesions were observed. The antibiotic treatment administered included streptomycin for five patients, tobramycin for 2 patients, and ciprofloxacin, azithromycin and amoxicillin (plus doxycycline) for other three patients. Three patients received initially antitermic drugs (with poor response) and later two of them, doxycycline. The administered antibiotic in the remaining three patients was unknown. The clinical course was satisfactory in all of them and so far no relapses have been detected. CONCLUSIONS: The ulceroglandular form, as it appears in literature, was the most common form in this series of patients with tularemia. Neither severe diseases nor complications were observed. Although streptomycin is considered the drug of choice, other antibiotics are likely equally effective, at least for the non complicated forms of the disease.

[Leptospirosis. Review of 11 cases]. / Leptospirosis: Revisión de 11 casos.

Eleven cases of leptospirosis diagnosed from 1988 to 1994 were retrospectively reviewed. The mean age of the patients was 52 years. Epidemiologic factors were found in 10 patients. Hepatorenal involvement was observed in 7 cases (64%), cardiac involvement in 3 (27%), bleeding episodes in 5 (45%) and central nervous system involvement in one case (9%). The Leptospira serogroups identified were: Icterohaemorrhage in 6 cases, Pomona in 1, Sejroe in and could not be determined in 3. One patient died because of multiorgan failure. The epidemiologic, clinical, analytical and therapeutic aspects are discussed.