Human infection by a "fish tapeworm", Diphyllobothrium latum, in a non-endemic country.

We document a case of locally acquired "broad- or fish-tapeworm" infection caused by Diphyllobothrium latum in a 27-year-old Spanish man, confirmed by molecular analysis (COI gene). The patient had naturally expelled a worm of 110 cm in length, but the physical examination did not yield any remarkable findings, and the patient did not suffer from any particular symptoms. Laboratory test results were normal except for a remarkable increase in the red blood cell count, and an evident decrease in the mean corpuscular volume and mean corpuscular hemoglobin. Vitamin B12 and folic acid values were in the normal range without signs of anemia. It was suggested that these anomalies in erythrocyte formation might not be related to the parasite, and analysis of the patient's anamnestic data revealed that the infection could only have been caused by the ingestion of imported fish, although no light could be shed on the specific source of infection. From a public health viewpoint, this human case of fish-borne zoonosis is exemplary, suggesting that not only is control of fish and fish product quality essential, but also increased awareness of the general population with regard to changes in culinary habits.

Comparison of two methods skipping cell lysis and protein extraction for identification of bacteria from blood cultures by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry.

OBJECTIVE: Matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS) is widely used for fast identification of bacteria from blood cultures (BC). We compared the performance of two procedures, one including a pre-enrichment step in brain heart infusion and the other a direct method using vacutainer separator gel tubes (DI), for identification of bacteria from blood cultures by MALDI-TOF MS. METHODS: We first prepared a training set of 20 simulated bacteremia specimens, including 10 Gram-negative and 10 Gram-positive species. A total of 145 non-consecutive BCs flagged as positive (68 Gram-negative rods, and 77 Gram-positive cocci) were prospectively analyzed (validation set). RESULTS: A total of 82% and 49% of isolates were correctly identified to the species level by the respective methods. CONCLUSIONS: The pre-enrichment method outperformed the DI method for identification of virtually all bacterial species included in the panels.

[Systemic cryptococcosis and pneumocystosis in an HIV-positive patient]. / Criptococosis sistémica y neumocistosis en un paciente VIH-positivo.

Cryptococcosis is one of the most common opportunistic infections in AIDS patients. Neurological symptoms are the most frequent clinical presentation of this fungal infection, and pulmonary involvement is clinically much less evident. We report the case of a patient who was treated in the emergency room for acute respiratory failure but who did not survive. Microbiologic and histopathologic studies demonstrated simultaneous pulmonary infections with Cryptococcus neoformans and Pneumocystis carinii. We discuss this clinical presentation of cryptococcosis with no neurological manifestations, and the importance of concurrent infections by several opportunistic organisms that may go unrecognized.

[Corneal ulcer caused by Nocardia asteroides after penetrating keratoplasty]. / Ulcera corneal por Nocardia asteroides tras queratoplastia penetrante.

We report the case of a 43-years-old patient, to whom a corneal transplantation was made because he presented a Salzmann nodular degeneration in his left eye. The patient was observed every week and his development during the following months was good. Nine weeks later he was attended at the emergency room of the hospital, with an intensive secretion and partial loss of vision in the operated eye. It was detected a peripheral ulcer of a diffused borders with a loss of epithelium and anterior stroma in the superior temporal part of the cornea. Five days later, the microbiological cultures confirmed the presence of Nocardia asteroides. In spite of the initial good evolution of the ulcer treated topically with a 20% sulfacetamide and trimetoprim-sulfadiacine p.o., the graft ended unsuccessfully.

[Enzymatic activities of Campylobacter jejuni, C. coli, and C. lari]. / Actividades enzimáticas de Campylobacter jejuni, C. coli y C. lari.

Susceptibility to nalidixic acid has been considered for a long time as an important test in the identification of the different species of the genus Campylobacter. However, due to the increasing development of resistance new laboratory tests are needed to carry out an accurate identification to the species level in combination with other typing schemes. We have evaluated the enzymatic activity of 180 Campylobacter spp. strains isolated from clinical specimens performing the Api ZYM system (bioMérieux) in order to assay whether different enzymatic patterns could help to characterize these microorganisms. Thirteen of the 19 enzymatic activities detected by the system (lipase-C14, valine arylamidase, cystine arylamidase, trypsin, chymiotrypsin, alpha-galactosidase, beta-galactosidase, beta-glucuronidase, alpha-glucosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase, alpha-manosidase and alpha-fucosidase) were negative for all the strains tested, whereas 3 enzymes (alkaline phosphatase, acid phosphatase and naphtol-A-S-BI-phosphohydrolase) were detected in 96.6% of the strains. Esterase-C1, esterase lipase-C8 and leucin arylamidase showed a variable reactivity depending on isolates. Enzymatic activity patterns clearly differentiate all the C. lari strains from other Campylobacter species. No significant differences were detected among the enzymatic activities of C. jejuni and C. coli strains. Our study suggests that the Api ZYM system is easy to perform and a valuable method to be applied in the characterization of the campylobacteria as a complement to other biotyping and serotyping schemes.

[The liver and AIDS]. / Hígado y SIDA.

Hepatic disorders in AIDS are very common, although the injuries observed are usually non-specific. This is the reason why the real usefulness of hepatic biopsy in this patients is being currently discussed. In this work, such aspect and the hepatic injuries observed in patients with AIDS are discussed. Current indications for hepatic biopsy are summarized, as well as its later manipulation in order to obtain maximum profitability of it.

Global study of viral diarrhea in hospitalized children in Spain: results of structural surveillance of viral gastroenteritis net work (VIGESS-net) 2006-2008.

BACKGROUND AND OBJECTIVES: Detection and characterization of gastroenteritis cases (viruses) was conducted during eleven years through the surveillance VIGESS-net, which was created in an effort to conduct a structured surveillance of rotavirus genotypes co-circulating in Spain. STUDY DESIGN AND RESULTS: This phase includes the study of 2048 fecal samples from children <5 years old, hospitalized in fifteen different hospitals throughout Spain from March 2006 to March 2008. Of them, 821 (40.1%) samples were rotavirus positive. Rotavirus was identified as the only etiological agent in 694 (33.9%) cases and in 127 (6.2%) was found as coinfection with other enteric viruses, mainly with noroviruses. Predominant G genotypes detected were G1 (49.8%) followed by G9 (32.9%), G3 (2.6%), G8 (1.0%), G4 (0.4%), G6 (0.2%) and G12 (0.2%). The G2 was encountered in 3.2% of cases. Rotavirus mixed G-types infections occurred in 3.9% of cases. The main G/P combinations were G1P[8] (51.9%) and G9P[8] (35.6%), which predominates alternatively in the first and second period of the study. More rare combinations occur in less than 7.4% of cases. CONCLUSION: The diversity of rotavirus circulating strains suggests to maintain a surveillance system through different regions of the country.

Viral proteins VP2, VP6, and NSP2 are strongly precipitated by serum and fecal antibodies from children with rotavirus symptomatic infection.

Rotavirus-specific IgA has been correlated with immune protection against rotavirus reinfection and symptomatic disease. Systemic and mucosal antibody responses were determined by an enzyme-linked immunosorbent assay in 11 infants with severe rotavirus gastroenteritis. Geometric mean titers of antirotavirus serum IgG and IgA antibodies were significantly higher during the convalescence of the disease (P < 0.001 vs. acute-phase titers). Rotavirus-specific fecal sIgA antibodies increased 4 times during the convalescence in 9 (81.8%) children (P < 0.001). The serum IgG and IgA antibody and fecal sIgA antibody responses to individual rotavirus polypeptides were characterized by radioimmunoprecipitation assay (RIPA) using Staphylococcus aureus protein A and the lectin jacalin to precipitate IgG- and IgA-immune complexes, respectively. The main IgG response was directed toward the structural viral proteins VP2, VP4, and VP6 and toward the nonstructural protein NSP2. Serum IgA reactivity was detected by RIPA in all serum samples, with major responses to VP2, VP6, and NSP2. Interestingly, fecal sIgA in convalescent samples reacted strongly toward NSP2 and VP6. These data reinforce the antigenic importance of rotaviral proteins other than VP4 and VP7, such as VP2, VP6, and NSP2, as main targets in the immune response to rotavirus.

[Virus-specific serum and fecal antibodies response in children with acute rotavirus gastroenteritis]. / Respuesta de anticuerpos séricos y coproanticuerpos específicos en niños con gastroenteritis aguda por rotavirus.

BACKGROUND: The analysis of the immune response to rotavirus infection and the characterization of the viral antigens recognized by specific antibodies are of great concern in evaluating the protection against rotavirus. MATERIAL AND METHODS: The levels of rotavirus-specific fecal (sIgA) and serum antibodies (IgM, IgG and IgA) were evaluated by ELISA in 25 children with acute gastroenteritis for rotavirus, in 11 of them during the acute and convalescent phases. The specificity of serum antibodies to viral polypeptides was characterized by immunoblotting. RESULTS: Serum IgM antibodies with a geometric mean titer (GMT) of 1/3,973 were the predominant antibodies detected during the acute phase. In comparison, IgG and IgA serum antibodies and sIgA coproantibodies levels were higher in the convalescent phase (GMT = 1/5,799, 1/257 and 1/137 respectively). Significant differences were observed for all the isotypes of immunoglobulins evaluated during the infection and in the convalescence (p < 0.01). Rotavirus-specific serum antibodies recognized mainly the structural VP6, VP7 and VP3/VP4 proteins. Other polypeptides also detected were VP1, VP5 and the non-structural NS34 protein. CONCLUSIONS: Rotavirus infection produce an intense humoral immune response both in serum and in the gut. Specific antibodies react against structural proteins of the internal (VP6) and external (VP7) capsids of rotavirus.

First detection of group C rotavirus in children with acute diarrhea in Spain.

Group C rotavirus causes sporadic cases and outbreaks of acute diarrhea in children and adults in many countries, but has never been detected among children in Spain. In a recently conducted surveillance study to screen fecal specimens for bacteria and viruses from a cohort of 822 young children who were treated for acute diarrhea in Madrid, no pathogens were detected in fecal specimens from 238 (29%) children. In this study, we examined 147 of those specimens for group C rotavirus by EIA and PCR and found 22 (15%) were positive. Our findings demonstrate that group C rotavirus is an important cause of childhood diarrhea in Spain.

Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.

Our aim was to evaluate the reverse transcription and polymerase chain reaction (RT/PCR) technique for the detection of rotavirus shedding by infected children as a routine diagnostic procedure, in comparison to the enzyme-linked immunosorbent assay (ELISA), electron microscopy (EM) and polyacrylamide gel electrophoresis (PAGE) of rotavirus double-stranded RNA. Two-hundred and twenty stool specimens were collected from infants and young children with diarrhoea, and 10-20% faecal suspensions were made. Several methods of rotavirus dsRNA extraction were assayed. Electrophoretic analysis of viral RNA was carried out on 10% polyacrylamide gels followed by silver staining. RT/PCR was performed using oligonucleotide primers specific for both 3' and 5' ends of the rotavirus gene encoding VP7 which are highly conserved among group A rotaviruses. Following RNA extraction with phenol-chloroform and ethanol precipitation, RT/PCR could detect rotaviral RNA in only 11 of 25 samples known to contain rotaviruses by conventional methods. The purification of RNA extracts by CF11 cellulose and the application of the RNAID method were equally effective in extracting RNA and/or removing inhibitory substances from the faecal samples. RT/PCR led to the detection of 66 positive samples from 220 specimens tested (30%), whilst 64 specimens were positive by ELISA (29%), 59 (26.8%) by PAGE and 56 (25.4%) by EM. In our study, RT/PCR was 100 times more sensitive than the ELISA test in detecting rotaviruses serially diluted in a faecal suspension. Although RT/PCR is theoretically much more sensitive than ELISA, PAGE and EM for detection of rotaviruses, great care must be taken to remove inhibitory substances from the enzymatic reactions. We do not consider that RT/PCR should replace immunoassays with high sensitivity and specificity for rotavirus testing in faecal samples, although this technique has other applications, like the search for rotavirus in different clinical specimens (sera, cerebrospinal fluid, respiratory secretions, etc.) and in environmental samples, as well as the typing of viral strains using serotype-specific primers.