D. Di Cave

Acanthamoeba T4 and T15 genotypes associated with keratitis infections in Italy

Thus far there is little data available concerning Acanthamoeba associated amoebic keratitis (AK) from Italy. In order to understand the incidence of Acanthamoeba in patients with ocular infections and to characterize the isolates at the molecular level, ocular specimens and contact lenses or lens case solutions from 140 patients were analysed by culture and by an 18S rRNA (Rns) gene-based PCR method. Nineteen (13.6%) patients showed Acanthamoeba culture positive samples. Eleven out of the 14 genetically characterized isolates were assigned to the T4 genotype. Three isolates, two of them from patients with keratitis responding to specific anti-Acanthamoeba therapy, were identified as belonging to the T15 genotype. This finding represents the first association between the T15 genotype and human amoebic keratitis. PCR amplification of the 18S ribosomal DNA proved to be a sensitive method, potentially able to detect Acanthamoeba without the need of long culture incubation, and thus considerably useful for clinical applications.

Molecular analysis based on mtLSU-rRNA and DHPS sequences of Pneumocystis jirovecii from immunocompromised and immunocompetent patients in Italy.

Pneumocystis jirovecii is an opportunistic fungus predominantly reported in immunocompromised individuals, who develop severe interstitial pneumonia (PcP). However, it is known that asymptomatic or mild pulmonary infections, defined as colonization, are widely observed in the general adult population. So far, genetic and epidemiological data of P. jirovecii infections in Italy are rather scarce and limited to defined geographical regions, mainly regarding isolates from HIV-infected patients. The aim of this study was to evaluate the polymorphisms at the mtLSU-rRNA and the DHPS loci by the screening and genotyping of a cohort of patients from two major hospitals in Rome (Italy). The study included 263 patients divided into two groups, all enrolled consecutively from January 2006 to December 2010: (i) 38 immunocompromised subjects including 25 HIV-infected; (ii) 225 immunocompetent patients. Sixty-seven out of 263 patients (25.5%) were found positive after PCR amplification of the mtLSU-rRNA gene. Overall, genotyping at mtLSU-rRNA locus revealed that the genotype 2 was the most frequent. Sequences of the DHPS gene were obtained from 21 patients, 9 from immunocompromised patients (6 from HIV infected individuals), 12 from immunocompetent ones. Considering the most common DHPS mutations usually detected at amino acid positions 55 and 57 and potentially related to drug resistance, all samples analyzed showed the wild-type signatures. These are the first data in Italy on prevalence and genotypes of P. jirovecii regarding colonized immunocompetent adults. Further multicenter analyses on P. jirovecii infection will be necessary to better define the specific epidemiology of the disease in the Italian populations.

Genetic characterization of Giardia duodenalis by sequence analysis in humans and animals in Pemba Island, Tanzania

Giardia duodenalis represents one of the most widespread human enteric parasites: about 200million people in Asia, Africa and Latin America are infected. Giardia exerts a deep impact on public health because of high prevalence and possible effects on growth and cognitive functions in infected children. The major aim of this study was to detect and genetically characterize G. duodenalis in both human and animal fecal samples collected in Pemba Island, in the archipelago of Zanzibar (Tanzania), in order to deepen the knowledge of genotypes of Giardia in this area. Between October 2009 and October 2010, we collected 45 human fecal samples from children from 2 primary schools and 60 animal fecal samples: 19 from zebus (Bos primigenius indicus) and 41 from goats (Capra hircus). Detection and genetic identification were performed by multilocus analysis of ssu-rDNA and gdh genes. In humans we found a higher prevalence of assemblage B (sub-assemblage BIV), in goats of assemblage E and in zebus of assemblage A. Our study represents an important contribution to the epidemiological knowledge of G. duodenalis in this area of Tanzania.

Pharmacological treatments in asthma-affected horses: A pair-wise and network meta-analysis

BACKGROUNDnEquine asthma is a disease characterised by reversible airflow obstruction, bronchial hyper-responsiveness and airway inflammation following exposure of susceptible horses to specific airborne agents. Although clinical remission can be achieved in a low-airborne dust environment, repeated exacerbations may lead to irreversible airway remodelling. The available data on the pharmacotherapy of equine asthma result from several small studies, and no head-to-head clinical trials have been conducted among the available medications.nnnOBJECTIVESnTo assess the impact of the pharmacological interventions in equine asthma and compare the effect of different classes of drugs on lung function.nnnSTUDY DESIGNnPair-wise and network meta-analysis.nnnMETHODSnLiterature searches for clinical trials on the pharmacotherapy of equine asthma were performed. The risk of publication bias was assessed by funnel plots and Eggers test. Changes in maximum transpulmonary or pleural pressure, pulmonary resistance and dynamic lung compliance vs. control were analysed via random-effects models and Bayesian networks.nnnRESULTSnThe results obtained from 319 equine asthma-affected horses were extracted from 32 studies. Bronchodilators, corticosteroids and chromones improved maximum transpulmonary or pleural pressure (range: -8.0 to -21.4 cmH2 O; P<0.001). Bronchodilators, corticosteroids and furosemide reduced pulmonary resistance (range: -1.2 to -1.9 cmH2 O/L/s; P<0.001), and weakly increased dynamic lung compliance. Inhaled β2 -adrenoreceptor (β2 -AR) agonists and inhaled corticosteroids had the highest probability of being the best therapies. Long-term treatments were more effective than short-term treatments.nnnMAIN LIMITATIONSnWeak publication bias was detected.nnnCONCLUSIONSnThis study demonstrates that long-term treatments with inhaled corticosteroids and long-acting β2 -AR agonists may represent the first choice for treating equine asthma. Further high quality clinical trials are needed to clarify whether inhaled bronchodilators should be preferred to inhaled corticosteroids or vice versa, and to investigate the potential superiority of combination therapy in equine asthma.

Social determinants associated with Giardia duodenalis infection in southern Côte d’Ivoire

The purpose of this investigation was to analyze the association between different social determinants of health at the local scale and Giardia duodenalis infection in diverse settlements in southern Côte d’Ivoire. Stool samples from 306 individuals aged 1–16xa0years were collected from six rural villages and a small town. Five variables were categorized to classify the increasing risk levels of acquiring intestinal parasites. Giardia prevalences (%) and odds ratios (ORs) were evaluated. Correlation and regression analyses were conducted to determine the correlation coefficients and to propose predictive models based on social determinants to forecast the risk of giardiasis. The overall observed prevalence of Giardia was 21.6xa0%. When the analysis was conducted at the local level, the percentage of infected people varied from a minimum of 12.7 up to 36.4xa0%. A significant association (pu2009<u20090.001) was found between the selected social determinants and G. duodenalis prevalence in the different localities. Correlation and regression analyses allowed us to describe two predictive models to estimate the OR of Giardia transmission. This study helps to clarify the possible impact of different social determinants of health on the risk of giardiasis at the local scale. Both predictive models could be suitable in order to assess the likelihood of the transmission of intestinal parasitic infection in developing countries.

Identification and phylogenetic position of Naegleria spp. from geothermal springs in Italy

Naegleria spp. are free-living amoebae belonging to the family Vahlkampfiidae, in the class Heterolobosea. Among the recognized species, Naegleria fowleri causes primary amoebic meningoencephalitis (PAM), while two other species, Naegleria australiensis and Naegleria italica, have been reported as pathogenic in experimental animals. Due to the thermotolerance properties of some species, geothermal water sources including hot springs represent suitable habitats for their proliferation. The main aim of this study was a year-round sampling in two geothermal springs in Central Italy, to investigate the presence of Naegleria spp. using PCR/DNA sequencing based methods. The affinities between the sequences generated here and others reported in the literature were explored by using POY, which implements the concept of dynamic homology. Naegleria australiensis, Naegleria italica, and Naegleria lovaniensis, plus an unassigned Naegleria spp. were detected. Indels in the rDNA ITS1 and ITS2 turned out to be critical to distinguish the three species and confirmed their phylogenetic relationships. This is the first molecular report on the Naegleria spp. occurrence in geothermal waters in Central Italy, coupled with a fine genetic characterization.

Blastocystis in Côte d'Ivoire: molecular identification and epidemiological data.

Blastocystis is an enteric protozoan infecting humans and animals in both developed and developing countries at all latitudes. Despite this, data on Blastocystis infection are not available for several geographical areas, including many African countries. In this study, a survey was conducted on Blastocystis among humans and domestic animals in rural and urban localities in Côte d’Ivoire, in order to investigate the prevalence, the subtype distribution, and the zoonotic potential in association with sociodemographic factors, seasonality, symptoms, and co-infections. A total of 110 fecal samples were collected from patients living in four localities. Molecular and phylogenetic analyses were conducted for Blastocystis detection and subtyping. Positive samples from symptomatic patients were tested by Luminex xTAG® Gastrointestinal Pathogen Panel (GPP) to evidence the presence of other common intestinal pathogens. Overall, a prevalence of 58.2% was observed in humans and subtypes ST1(50.0%), ST2 (22.0%) and ST3 (28.1%) were identified. The prevalence values varied significantly among the sites but not in relation to the subtype. The seasonal rains significantly increase the infection rate in all localities. No significant differences in the ST distribution between asymptomatic and symptomatic subjects were observed. As regard the zoonotic transmission, an additional sampling was conducted in another village where fecal samples were simultaneously collected from humans and animals. Blastocystis STs 1–3 and ST7 were identified in eight humans and four chickens, respectively. This study provides the first exhaustive data on the prevalence and molecular epidemiology of Blastocystis in Côte d’Ivoire.