Cristina Carranza-Rodríguez

Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

BackgroundCoxiella burnetii is a highly clonal microorganism which is difficult to culture, requiring BSL3 conditions for its propagation. This leads to a scarce availability of isolates worldwide. On the other hand, published methods of characterization have delineated up to 8 different genomic groups and 36 genotypes. However, all these methodologies, with the exception of one that exhibited limited discriminatory power (3 genotypes), rely on performing between 10 and 20 PCR amplifications or sequencing long fragments of DNA, which make their direct application to clinical samples impracticable and leads to a scarce accessibility of data on the circulation of C. burnetii genotypes.ResultsTo assess the variability of this organism in Spain, we have developed a novel method that consists of a multiplex (8 targets) PCR and hybridization with specific probes that reproduce the previous classification of this organism into 8 genomic groups, and up to 16 genotypes. It allows for a direct characterization from clinical and environmental samples in a single run, which will help in the study of the different genotypes circulating in wild and domestic cycles as well as from sporadic human cases and outbreaks. The method has been validated with reference isolates. A high variability of C. burnetii has been found in Spain among 90 samples tested, detecting 10 different genotypes, being those adaA negative associated with acute Q fever cases presenting as fever of intermediate duration with liver involvement and with chronic cases. Genotypes infecting humans are also found in sheep, goats, rats, wild boar and ticks, and the only genotype found in cattle has never been found among our clinical samples.ConclusionsThis newly developed methodology has permitted to demonstrate that C. burnetii is highly variable in Spain. With the data presented here, cattle seem not to participate in the transmission of C. burnetii to humans in the samples studied, while sheep, goats, wild boar, rats and ticks share genotypes with the human population.

Mumps Orchitis in the Post-vaccine Era (1967-2009): A Single-center Series of 67 Patients and Review of Clinical Outcome and Trends

Since the introduction of the mumps vaccine, the age of appearance of mumps infection has shifted from children to adolescents and young adults, groups with a higher incidence of disease complications and sequelae. During the years 2000-2001, the Gran Canaria Island was part of an epidemic of mumps. In that period, our institution attended 67 cases of serologically confirmed acute mumps orchitis, the most serious complication of mumps infection in young postpubertal males. We conducted a descriptive and prospective study of this cohort and extensively reviewed the literature from 1967 (the year the first mumps vaccine was introduced) to 2009. Fifty-six patients were admitted because of general impairment and were treated with alpha-interferon. Sixty-six patients presented parotitis previous to orchitis (interval from parotitis to orchitis, 4.9 d). Orchitis was unilateral in 89.5% and bilateral in 10.4% of cases. More than 98% of patients had orchitis-associated fever. Nine patients had clinical and biochemical data showing acute mumps meningitis, and 11 had subclinical pancreatitis. The mean duration of symptoms was 4.6 days (range, 1-9). During the acute phase, more than 41% of the evaluated testes had a volume >25 mL. Acute hormonal disturbances were highly prevalent. These included decreased levels of testosterone and inhibin B with low or normal levels of gonadotropins in 35% of subjects, and, to our knowledge not previously reported, an atypical hormonal pattern consisting of low levels of free testosterone and inhibin B, along with increased measures of luteinizing hormone but low or normal follicle-stimulating hormone levels (11% of cases). During the follow-up period (mean, 331 d) a high incidence of sperm disturbance was found. Abbreviations: CRP = C-reactive protein, CSF = cerebrospinal fluid, ESR = erythrocyte sedimentation rate, FSH = follicle-stimulating hormone, LH = luteinizing hormone, MMR = measles, mumps, and rubella triple vaccine, NSAID = nonsteroidal antiinflammatory drugs, WHO = World Health Organization.

Helicobacter pylori and cagA seroprevalence in sub-Saharan inmigrants recently arrived to Gran Canaria (Spain)

Non-ulcerous dyspepsia is common among sub-Saharan people migrating into Spain. Given the high prevalence of H. pylori (HP) infection in their countries of origin, we studied the prevalence of infection in this population, and specifically the prevalence of infection by the more virulent, cagA-positive strains (CAP). 140 sub-Saharan immigrants recently arrived to Gran Canaria (Canary Islands, Spain) were studied. 80.7% were male, with a mean age of 24.2 y. 90.7% tested seropositive for HP and 72.2% of them carried antibodies against the ‘pathogenicity island’ cagA. We did not find any relationship between the presence of these antibodies and the clinical variables studied. We can conclude that HP infection is virtually universal in this population, with a high percentage of infection by CAP strains.

Seroprevalence of Rickettsia typhi and Rickettsia conorii infections in the Canary Islands (Spain)

OBJECTIVE The aim of this work was to evaluate the prevalence of past infection due to Rickettsia typhi and Rickettsia conorii in the Canary Islands (Spain). METHODS A representative sample of the population of the seven islands, formed of 662 people aged between 5 and 75 years (368 females, 294 males), was analyzed. Epidemiological data were obtained by direct survey. The detection of serum IgG antibodies against both microorganisms was based on an indirect immunofluorescence test, considered positive if the titers were ≥ 1/80. RESULTS Of the analyzed population 3.9% had IgG antibodies against R. typhi and 4.4% against R. conorii. Out of these positive samples, only three were positive for both species. The seroprevalence was similar in both sexes. Positive results were found in all age groups, but a higher rate was noticed in those aged 46 years and older (p<0.05). R. typhi was found to be more prevalent in rural areas of all islands, as well as in farmers. CONCLUSIONS Our results confirm the presence of antibodies against the causative agents of murine typhus and Mediterranean spotted fever in the Canary Islands. Indirect data suggest that the detection of antibodies to R. conorii might be due to a cross-reaction between these species.

The Rapid-Heat LAMPellet Method: A Potential Diagnostic Method for Human Urogenital Schistosomiasis

Background Urogenital schistosomiasis due to Schistosoma haematobium is a serious underestimated public health problem affecting 112 million people - particularly in sub-Saharan Africa. Microscopic examination of urine samples to detect parasite eggs still remains as definitive diagnosis. This work was focussed on developing a novel loop-mediated isothermal amplification (LAMP) assay for detection of S. haematobium DNA in human urine samples as a high-throughput, simple, accurate and affordable diagnostic tool to use in diagnosis of urogenital schistosomiasis. Methodology/Principal Findings A LAMP assay targeting a species specific sequence of S. haematobium ribosomal intergenic spacer was designed. The effectiveness of our LAMP was assessed in a number of patients´ urine samples with microscopy confirmed S. haematobium infection. For potentially large-scale application in field conditions, different DNA extraction methods, including a commercial kit, a modified NaOH extraction method and a rapid heating method were tested using small volumes of urine fractions (whole urine, supernatants and pellets). The heating of pellets from clinical samples was the most efficient method to obtain good-quality DNA detectable by LAMP. The detection limit of our LAMP was 1 fg/µL of S. haematobium DNA in urine samples. When testing all patients´ urine samples included in our study, diagnostic parameters for sensitivity and specificity were calculated for LAMP assay, 100% sensitivity (95% CI: 81.32%-100%) and 86.67% specificity (95% CI: 75.40%-94.05%), and also for microscopy detection of eggs in urine samples, 69.23% sensitivity (95% CI: 48.21% -85.63%) and 100% specificity (95% CI: 93.08%-100%). Conclusions/Significance We have developed and evaluated, for the first time, a LAMP assay for detection of S. haematobium DNA in heated pellets from patients´ urine samples using no complicated requirement procedure for DNA extraction. The procedure has been named the Rapid-Heat LAMPellet method and has the potential to be developed further as a field diagnostic tool for use in urogenital schistosomiasis-endemic areas.

Red blood cell disorders in recently arrived African immigrants to Gran Canaria, Spain

BACKGROUND In the last decade immigration to Europe has increased, with Africa being the source of a large number of immigrants. In addition to infections, this group has other less known health problems, such as erythrocyte abnormalities. METHODS The objectives of this study were: the systematic evaluation of red cell abnormalities in 200 newly arrived asymptomatic African immigrants on the Canaries; the systematic evaluation of haemoglobinopathies and their characterization in this population; and the relationship of red blood cell disorders and parasitic infections. RESULTS Of the studied immigrants 53 (26.5%) had red cell disorders according to their CBC parameters (Hb and/or MCV). In 48 people (24.0%) one or more etiologic diagnoses were made. Specifically, in order of frequency, a total of 26 structural haemoglobinopathies, 14 α-thalassemias, 2 β-thalassemias and 14 iron deficiencies were diagnosed. There was a statistically significant association between the presence of anemia, microcytosis, structural haemoglobinopathies or α thalassemia and sub-Saharan origin. However, no statistically significant association between the abovementioned parameters and eosinophilia or helminthic infection was observed. CONCLUSIONS These results suggest that, even in the presence of normal Hb and MCV values, including haemoglobinopathies in the initial screening of newly arrived sub-Saharan immigrants would be very useful.

Skin and soft tissue infections (patera foot) in immigrants, Spain.

An unusual skin and soft tissue infection of the lower limbs has been observed in immigrants from sub-Saharan Africa who cross the Atlantic Ocean crowded on small fishing boats (pateras). Response to conventional treatment is usually poor. Extreme extrinsic factors (including new pathogens) may contribute to the etiology of the infection and its pathogenesis.

Lesiones periungueales en un viajero procedente de Sudamérica

mas frecuente de enfermedades importadas, cuyo diagnostico y tratamiento es poco conocido. Dentro de los problemas frecuentes en viajeros destacan tres formas de presentacion, con incidencia variable segun las series: la fiebre, la diarrea y las lesiones cutaneas1. Dentro de las lesiones cutaneas, la tunguiasis supone hasta el 6% en algunas series2. La tunguiasis es una ectoparasitosis causada habitualmente por la penetracion de la hembra Tunga penetrans Caso clinico

Surveillance of strongyloidiasis in Spanish in-patients (1998–2014)

Background Strongyloides stercoralis is a parasite that causes strongyloidiasis, a neglected tropical disease. S. stercoralis is a soil-transmitted helminth that is widely distributed in tropical and subtropical regions of the world. Strongyloidiasis can occur without any symptoms or as a chronic infection characterized by mild, unspecific symptoms such as pruritus, abdominal pain or discomfort; respiratory impairment also may manifest as a potentially fatal hyperinfection or disseminated infection. Most studies on strongyloidiasis in Spain have been related to chronic forms in immigrants or travellers from endemic zones and have mainly analysed out-patient populations. Studies of the impact of strongyloidiasis cases admitted to hospitals in Spain are lacking. Therefore, the aim of this study was to analyse the impact of strongyloidiasis in hospital care in Spain. Methodology We designed a retrospective descriptive study using the Minimum Basic Data Set (MBDS, CMBD in Spanish) for inpatients with ICD-9: 127.2 (strongyloidiasis) diagnoses admitted to hospitals in the Spanish National Health System between 1998 and 2014. Principal findings A total of 507 hospitalizations with diagnosis of strongyloidiasis were recorded, 324 cases (63.9%) were males. The mean (±SD) age was 42.1±20.1 years. The impact of strongyloidiasis on the total population of Spain was 0.06 cases per 105 person-years, and the infection burden increased progressively over time (from 0.01 cases per 105 person-years in 1999 to 0.10 cases per 105 person-years in 2014). 40 cases (7.9%) died. The total cost was approximately €8,681,062.3, and the mean cost per patient was €17,122.4±97,968.8. Conclusions Our data suggest that strongyloidiasis is frequent in Spain and is increasing in incidence. Therefore, it would be desirable to improve the oversight and surveillance of this condition. Due to the fact that strongyloidiasis can be fatal, we believe that there is a need to establish risk categories for inclusion in national guidelines/protocols for screening individuals at risk of developing strongyloidiasis.

Immune Reconstitution Inflammatory Syndrome in HIV-Infected Immigrants.

Immune reconstitution inflammatory syndrome (IRIS) includes a group of potentially serious inflammatory processes that may be present in HIV-infected patients after initiating highly active antiretroviral therapy (HAART). Paradoxical IRIS is a worsening of symptoms, after an overwhelming response to a previously diagnosed opportunistic infection (OI); unmasking IRIS reveals a previously occult OI. The main objective of the study was to describe the epidemiological, clinical, and outcome data of HIV-infected immigrants, stratified according to high- or low-income countries of origin, who developed IRIS and to compare them with native-born Spanish patients. This retrospective study reviewed all patients with HIV infection admitted to the Unit of Infectious Diseases and Tropical Medicine between 1998 and 2014. IRIS was identified in 25/138 (18%) immigrant patients and 24/473 (5%) native-born Spanish patients infected with HIV. Most cases, 19/25 (76%), were of unmasking IRIS. The time elapsed between initiation of HAART and development of IRIS was significantly longer in patients with unmasking versus paradoxical IRIS. OIs, in particular due to mycobacteria, were the most frequently involved processes. Twenty percent of patients died. The comparison of immigrant and native-born patients found significant differences for both IRIS type (higher incidence of paradoxical forms among immigrants) and for the absence of malignancies in native-born patients. No significant differences were found when the data of immigrants from low- and high-income countries were compared.