Molecular epidemiology and genetic diversity of Blastocystis infection in humans in Italy.

In order to describe the molecular epidemiology of Blastocystis infection in Italy, 189 isolates, which had been collected during the years 2012-2014 from mildly symptomatic patients, or those affected by inflammatory bowel disease (IBD), irritable bowel syndrome (IBS) or chronic diarrhoea, or otherwise immunosuppressed, were subtyped by sequence analysis of the SSU rRNA gene (536 bp). Six subtypes (STs) were detected: ST1 (15·3%), ST2 (13·8%), ST3 (46·0%), ST4 (21·7%), ST6 (3·2%) and ST8 (0·5%). They clustered in distinct clades, as inferred from Bayesian inference phylogenetic and median joining network analyses. A high genetic differentiation was found at the inter-subtype level; it ranged from Jukes-Cantor (JC) distance = 0·02 (between ST1 and ST4) to JC = 0·11 (between ST6 and ST2). At the intra-ST level, a high genetic homogeneity was registered in ST4, whereas higher genetic variation was found in isolates corresponding to ST1 and ST2. Accordingly, high values of haplotype and nucleotide diversity were observed in ST1, ST2 and ST3. No association was found between patient gender and ST, whereas ST3 and ST1 were significantly more prevalent in patients aged 15-50 years. A significant occurrence of Blastocystis ST4 in patients suffering from IBS, IBD or chronic diarrhoea was observed; in addition, a slight significant association between ST1 and ST3 and IBS patients was found. Multiple correspondence analysis showed some significant contribution of different variables (subtypes, haplotypes, age) in the observed pattern of ordination of the 189 patients in the symptom categories.

Molecular and phylogenetic analysis of the filarial nematode Micipsella numidica from the hare Lepus europaeus in Italy.

The genus Micipsella comprises three species of filariae to date identified in lagomorphs only, whereas the other genera belonging to the subfamily Splendidofilariinae are described as parasites of birds, reptiles and mammals. In the present study seven specimens of Micipsella numidica (Seurat, 1917), collected from the hare Lepus europaeus in Italy, were characterized genetically by molecular amplification of the mitochondrial genes (12S rDNA; cox1) and the 5S rDNA gene spacer region. Phylogenetic trees inferred using available sequences from filariae and those identified in this study evidenced a close relationship between M. numidica and Splendidofilariinae of other mammals and reptiles (Rumenfilaria andersoni and Madathamugadia hiepei). The present findings, apart from adding new data about the hosts in Italy, support the taxonomic position of M. numidica and highlight the substantial biological and molecular differences existing between Splendidofilariinae and other Onchocercidae. The study also contributes to our knowledge of the molecular/genetic diagnosis of filarial parasites of veterinary and medical concern in any vertebrate or invertebrate host.

Strongyloides stercoralis infection in allogeneic stem cell transplant: a case report and review of the literature.

Strongyloides stercoralis infections may be documented in low-endemicity areas, particularly in immigrants from endemic areas. The case of a patient from Bangladesh, an immigrant to Italy who developed a S. stercoralis infection after allogeneic stem cell transplant, is described, and 7 further cases are reviewed. Because of the atypical clinical presentation, the low predictive role of the eosinophil count, and the low sensitivity of the microbiological tests, diagnosis of strongyloidiasis is a challenging problem. When a case of S. stercoralis infection is suspected, previous exposure may be the only clue to guide the diagnostic approach.

Chagas disease in Italy: breaking an epidemiological silence.

Chagas disease, a neglected tropical disease that due to population movements is no longer limited to Latin America, threatens a wide spectrum of people(travellers, migrants, blood or organ recipients,newborns, adoptees) also in non-endemic countries where it is generally underdiagnosed. In Italy, the available epidemiological data about Chagas disease have been very limited up to now, although the country is second in Europe only to Spain in the number of residents from Latin American. Among 867 at-risk subjectsscreened between 1998 and 2010, the Centre for Tropical Diseases in Negrar (Verona) and the Infectious and Tropical Diseases Unit, University of Florence found 4.2% patients with positive serology for Chagas disease (83.4% of them migrants, 13.8% adoptees).No cases of Chagas disease were identified in blood donors or HIV-positive patients of Latin American origin. Among 214 Latin American pregnant women,three were infected (resulting in abortion in one case).In 2005 a case of acute Chagas disease was recorded in an Italian traveller. Based on our observations, we believe that a wider assessment of the epidemiological situation is urgently required in our country and public health measures preventing transmission and improving access to diagnosis and treatment should be implemented.

[Acute abdomen in renal transplant recipients. Epidemiology and treatment in not referral transplantation centers]. / L'addome acuto nel paziente trapiantato renale. Considerazioni epidemiologiche, diagnostiche e terapeutiche in DEA non dedicati.

The incidence of gastrointestinal complications in renal transplant recipients is relatively high while about 10% is related to acute abdomen. Data concerning gastrointestinal (GI) complications were reported in literature mainly from referral center studies. A multicenter retrospectively survey was performed in Lazio, Italy, in order to evaluate the incidence of acute abdomen in renal transplant recipients observed to the emergency departments of not referral transplantation centers. Clinical and demographic findings regarding 14 patients who experienced acute abdomen between February 2005 and Dicember 2008 have been collected. The following data was investigated: etiology, diagnostic workup, duration of symptoms, elapsed time between admission and emergency operation if performed, morbility and mortality. The severity of disease at presentation was assessed by mean of the Acute Physiology and Chronic Health Evaluation score (APACHE II). Acute abdomen was due to pancreatitis in three patients (23.1%); to cholecystitis in three (23.1%); to acute diverticolitis with colon perforation in two patients (15.4%); to acute appendicitis in two (15.4%) and to intestinal obstruction in 2 patients (15.4%). Small bowel perforation was observed in two patients (15.4%) which one case, upon pathological examination, showed malignant lymphoma. The mean APACHE II score was 14.0 ± 5.9. Ten patients (71.4%) were submitted to surgery. Overall mortality and morbidity were 35% and 42% respectively. Statistical analysis showed admission APACHE II score (p<0.01), duration of symptoms (p<0.05), and total time elapsed between the onset of symptoms and treatment (p<0.04) as factors significantly related to mortality.

Endoparasites of red fox (Vulpes vulpes) in central Italy.

A parasitologic study on 129 red foxes (Vulpes vulpes) from Tuscany (central Italy) was carried out in 2004-2006. Five intestinal species were found at necropsy: Dipylidium caninum (prevalence 57.3%), Mesocestoides lineatus (45.4%), Uncinaria stenocephala (39.1%), Toxocara canis (9.1%), and Toxascaris leonina (5.4%). Other parasites not associated with the intestine included Crenosoma vulpis (14.7%), Capillaria aerophila (7.0%), Angiostrongylus vasorum (7.0%), and filarial parasites (17.8%). Coprologic tests were less sensitive and less specific in identifying parasites than direct examinations at necropsy. Trichinella larvae were not found in muscles submitted to artificial digestion. By immunologic assay, antigens of Echinococcus spp. were detected in fecal samples of 20 foxes, but results could not be confirmed by fecal examination or molecular tests.

Morphology and genetics of a Babesia isolate from Capreolus capreolus.

A Babesia isolate that was morphologically distinct from Babesia capreoli and very similar to B. divergens was found in the blood of a roe deer (Capreolus capreolus) found dead in central Italy. Sequences corresponding to the full coding region of the 18S ribosomal RNA (rRNA) gene were identical to a sequence reported for Babesia divergens from a reindeer (Rangifer tarandus) and 99.9% and 99.8% similar to those reported for B. capreoli and bovine origin B. divergens, respectively.

Aedes albopictus and Culex pipiens implicated as natural vectors of Dirofilaria repens in central Italy.

To identify the natural vectors of Dirofilaria repens Railliet et Henry, entomological samplings were carried out in four sites within the Lazio region, foci of canine subcutaneous dirofilariasis. Collections were made in 2002-2003 by means of dog-baited and miniature Centers for Disease Control and Prevention traps as well as on humans. Microscopy identified 1576 attracted mosquito females as belonging to six species, but molecular diagnostics detected filarial DNA only in Culex pipiens L. and Aedes albopictus (Skuse, 1894). Dirofilaria immitis Leidy DNA, D. repens DNA, or both were found in the head and thorax of both mosquitoes. The simultaneous presence of vectors showing diurnal and nocturnal activity patterns is of concern for animal and human health. The finding of D. immitis DNA in mosquitoes in areas where only D. repens was been recovered in dogs also demonstrates that this filarial parasite circulates among carnivores (wild or domesticated pets).

Animal babesiosis: an emerging zoonosis also in Italy?

In Italy, babesiosis is widespread in several Central and Southern Regions, but few data are available on its presence in most Italian areas. In 2004 a project was financed by the MIUR to investigate on the babesiosis epidemiology in vertebrate and invertebrate hosts, and on the transmission risk for humans in Central and Northern Regions of the country. Microscopy and/or molecular tools were applied to blood samples of wild animals, livestock and pets, and to 1,677 ticks collected on animals or in the environment, with the aim of detect babesial parasites. Moreover, serological tests were used to evaluate the circulation of these protozoa among animals and people at risk. Microscopy identified as positive 5.0% of the animals, mostly living in Central Regions, but also in Northern areas considered Babesia-free. Serology evidenced the same general trend. PCR detected "piroplasm" DNA in 13.8% of the animals, and sequencing identified babesial parasites in 101/233 samples. The ticks were identified as belonging to 12 species, mostly represented by Ixodes ricinus, Rhipicephalus sanguineus and Dermacentor marginatus. Molecular analyses evidenced babesial parasites in 3.8% of them; in Rh. sanguineus was also demonstrated the vertical transmission of Babesia canis canis. To date 30 human sera have been analysed: 3 showed antibodies to B. microti. Animal babesiosis is largely present among pets, wild and farm animals, whereas goats seem refractory to the infection. In wild ungulates have been found the B. divergens-like, and the Babesia EU1 strains (reported in Italy in humans). Our findings evidenced the low reliability of microscopy in epidemiological studies, and the need of new/improved immunological tests to face diagnostic problems. The monitoring of infected areas and infection rates, joined to appropriate control programs, seems necessary to avoid the transmission of babesiosis to humans.

Occurrence of the G3 Indian buffalo strain of Echinococcus granulosus in cattle.

During a survey carried out to define the occurrence of Echinococcus granulosus in cattle bred in the province of Rieti (Central Italy), molecular diagnostics (PCR amplification and sequencing of a partial region of the mitochondrial CO1 gene) showed that 6/10 positive bovines harboured hydatid cysts (No.=16) genetically identical (95.8-100%) to the Indian buffalo genotype G3. As far the location of the 16 cysts, 11 of them were found in the lungs of three animals, whereas 5 cysts were in the liver of three parasitized hosts. The occurrence of genotype G3 in 60% of parasitized bovines living in an area never studied before provides more definite evidence about the existence of the strain in this region, and proves that cattle have to be considered a non-accidental host.

[Double ileo-ileal and ileo-cecocolic intussusception due to submucous lipoma: case report]. / Doppia invaginazione ileo-ileale e ileo-ciecocolica da lipoma sottomucoso del tenue: case report.

Intestinal intussusception is a rare pathology in adults and counts for approximately 5-16% of all invaginations, according to literature. We report the case of a 54 years-old male affected by a double ileo-ileal and ileocecocolic invagination secondary to a submucosal intestinal lipoma. Because of a bowel obstruction symptomathology, the diagnosis was based on abdomen X-rays, abdominal ultrasound and multislice CT scan that showed a probable pre-operative diagnosis of intestinal intussusception. The diagnosis of double invagination was confirmed intraoperatively and was therefore performed a right colectomy extended to a long tract of ileum. Opening the specimen demonstrated the evidence of a submucosal lipoma on the head of the intussusceptum. After the description of this case, we proceed with review of the literature of adult intestinal invagination.

[Modern orientation in parotid surgery]. / Moderni orientamenti nella chirurgia della parotide.

The choice of the surgical approach is very relevant in parotid surgery because of the extreme anatomic variability of the parotideal district and the functional importance of the branches of facial nerve. The aim of this paper is to prove how an accurate knowledge of the anatomy and a careful execution of surgery permit radicality in case of tumors and respect of nerves and other structures of the region. We studied anatomy of the different intra- and periparotideal structures to compare two different approaches in parotidectomy, proving how the orthograde technique is safer and more effective in preventing hyatrogenous nerve injuries and other major complications.

[Multidetector CT urography: diagnostic role in the evaluation of patients with non-traumatic hematuria]. / Urografia-TC multidetettore: ruolo diagnostico nella valutazione del paziente con ematuria non traumatica.

Blood in the urine (hematuria) can originate from any site along the urinary tract and may be the only sign of renal or vesical malignancy. Therefore, literature recommends for the evaluation of any case of macroscopic or microscopic hematuria. Our aim was to define the diagnostic role of multidetector CT urography (MDCTu) in the evaluation of this symptom through the analysis of 181 consecutive patients from January 2003 to March 2006.

Natural vectors of dirofilariasis in rural and urban areas of the Tuscan region, central Italy.

Entomological investigations by means of dog- and human-baited traps were carried out in summers 2000-2002 in urban and rural areas of the Tuscan region in central Italy. The aim of the study was to define the mosquito species involved in the transmission of Dirofilaria nematodes and to assess the risk that their presence might represent for animal and human health. Nocturnal fieldwork on host-seeking activity and feeding preferences was followed by microscopic identification of the mosquito species attracted and by molecular identification of Dirofilaria parasites in mosquitoes. In total, 3,611 mosquito females belonging to 12 species, largely represented by Culex pipiens L. and Aedes caspius (Pallas), were caught. Some females of each species collected fed on the dogs, indicating their possible role as an intermediate host, but filarial DNA was found only in Cx. pipiens, Anopheles maculipennis s.l. (Meigen), and Coquillettidia richiardii (Ficalbi). In rural environments, the DNA evidence indicated the presence of infective larvae of Dirofilaria immitis, whereas in urban areas, infective larvae of Dirofilaria repens were present. The role of Cx. pipiens as a vector for heartworm disease and subcutaneous infections in natural and artificial environments was confirmed, whereas Ae. caspius seemed refractory to the infection. The different role of the collected species is discussed. The vector competence of An. maculipennis and Cq. richiardii needs further investigation, because the importance of these species poorly represented, and the role of species such as Aedes albopictus (Skuse), characterized by a dominant diurnal activity pattern, has to be evaluated.

Human infections due to nematode helminths nowadays: epidemiology and diagnostic tools.

An update on the epidemiology of intestinal and tissue roundworms, in the world as well as in Italy, is presented. This analysis indicates that the most important changes happened in developed countries, where two main phenomena have been registered. The improvements occurred in social, economic, hygienic and sanitary conditions resulted in decrease of intestinal geohelminth prevalence but, besides, the changes happened at level of other factors, like environment, climate and human behaviour, promote more and more the opportunity for infections due to animal nematode species, which in people induce tissue syndromes. Therefore, the easier circulation of zoonoses among humans constitutes the main remark, which has to be strongly taken into account in countries like Italy, in that it is an emerging sanitary problem, important both for the number of people involved, and for the difficulties induced by their diagnosis. The general trend of "traditional" and "new" nematode infections that can affect humans is analysed, changes in the distribution patterns are tentatively explained, and available diagnostic tools are summarised.

[Intraoperative quick parathyroid hormone assay in thyroid surgery: preliminary experience on 36 cases]. / Dosaggio intraoperatorio dell'ormone paratiroideo in chirurgia tiroidea: esperienza preliminare su 36 casi.

Through the analysis of our preliminary experience on 36 consecutive cases of thyroid surgery, we assessed the feasibility of the rapid intraoperative assay of the intact parathyroid hormone (iPTH) as predictive risk factor of hypocalcemia versus seric calcium level on the first post-operative day to select the patients eligible to an early discharge. Furthermore, we managed to determine if iPTH level during thyroid surgery could point out the cases in which parathyroid autotransplantation is necessary, as the macroscopic evaluation of the parathyroid gland's viability is inadequate.

[The actual role of bilateral neck exploration in the treatment of primary hyperparathyroidism]. / Il ruolo attuale della esplorazione bilaterale del collo nel trattamento dell'iperparatiroidismo primario.

Surgery is the only curative treatment for primary hyperparathyroidism (pHPT). Surgical exploration is recommended for all patients with biochemically documented pHPT and signs or symptoms of the disease. Some patients are asymptomatic, others have subtle symptoms that disappear after parathyroid surgery. Felix Mandl successfully performed the first parathyroidectomy in 1925, using a bilateral neck exploration (BNE) with examination of all four glands and this remained the procedure of choice for pHPT into the 1990s. As over 80% of pHPT cases are due to a single parathyroid adenoma, many authors have questioned the need of BNE and have proposed directed unilateral approaches, termed "mini-invasive parathyroidectomies". The aim of this report is to define which is the actual role of the conventional surgical approach to pHPT.

Epilepsy, cysticercosis, and toxocariasis: a population-based case-control study in rural Bolivia.

OBJECTIVE: To assess the relationship between epilepsy and infection with Taenia solium and Toxocara canis with a case-control study, in the rural area of the Cordillera Province, Bolivia. METHODS: A preliminary two-phase door-to-door prevalence survey determined the prevalence of epilepsy and identified cases and control subjects. At least two control subjects per case were selected, matching on sex, age, and community of residence. Cases and control subjects were assessed serologically for antibodies against T. canis by ELISA and against T. solium by enzyme-linked immunoelectrotransfer blot (EITB). RESULTS: The prevalence survey found 130 confirmed cases of epilepsy, of which 113 were eligible for the case-control study (59 partial seizures and 54 generalized seizures). Two hundred thirty-three control subjects were selected. Multivariable analysis for a matched case-control study was carried out. There was an association between EITB positivity for T. solium and epilepsy with an OR of 1.85 (95% CI 0.99 to 3.4) for all cases. A stronger association was found in those with partial epilepsy with a late onset of disease (15 years and older), where the OR was 3.66 (95% CI 1.10 to 12.10). A positive association was also found with T. canis for all cases with an OR of 2.70 (95% CI 1.41 to 5.19). This increased for those with late-onset partial epilepsy to an OR of 18.22 (95% CI 2.10 to 158.10). CONCLUSION: This finding suggests that both neurocysticercosis and toxocariasis may in part explain the higher prevalence of epilepsy, particularly partial epilepsy, in developing countries.

Mosquitoes as vectors of Setaria labiatopapillosa.

An infected bovine-baited trap was utilised in summer 1994 to catch possible intermediate hosts of S. labiatopapillosa in northeastern Italy. Collections were made for 21 nights from 8.00 p.m. to 6.00 a.m. every 2 h and, after 12 September, every 30 min. Among the 16,159 mosquitoes sampled, 11,052 were freshly blood-fed. Most of the unfed females and a representative sample of those which had fed were identified as follows: Culex pipiens, Aedes caspius, A. vexans, Culiseta annulata, Anopheles maculipennis s.l., A. claviger and Coquillettidia richiardii. In spite of having the highest relative density, C. pipiens is the species which fed the least frequently and showed lower susceptibility and efficiency. The 2 Aedes species appear to act as vectors, particularly A. caspius, which proved to be the most efficient vector (K.I. = 0.8). A. claviger and A. maculipennis contribute to S. labiatopapillosa transmission, but their scarce presence reduces their epidemiological relevance. The other species identified showed a complete refractoriness to the infection. The risks for veterinary and medical health are discussed.

Epidemiological evidence for a differential effect of hookworm species, Ancylostoma duodenale or Necator americanus, on iron status of children.

BACKGROUND: The hookworms, Ancylostoma duodenale and Necator americanus, cause significant gastrointestinal blood loss. In clinical studies, greater blood losses have been reported with A. duodenale. However, there has been no evidence that endemic A. duodenale infection has greater impact than N. americanus infection on the iron status of populations. METHODS: In a sample of 525 school children in Pemba Island, Tanzania, we compared the degree of anaemia and iron deficiency associated with the two hookworm species at the individual and community (i.e. school) levels. Multiple regression was used to control for infection intensities and other child characteristics. RESULTS: In the 492 children with hookworm positive faecal cultures, haemoglobin and ferritin concentrations decreased with increasing proportions of A. duodenale. Among children with only N. americanus larvae, the prevalence of anaemia was 60.5 % and the prevalence of ferritin <12 microg/l was 33.1%, while in children with > or =50% A. duodenale larvae, the respective prevalences were 80.6% and 58.9%. When children were grouped by the prevalence of A. duodenale at the school level, children from high prevalence (> or =20%) schools had significantly worse iron deficiency and anaemia than children from low prevalence schools. CONCLUSIONS: The species of hookworm being transmitted in a community influences the burden of iron deficiency anaemia in the community, and should be considered in prioritizing and planning programmes for hookworm and anaemia control.

PIP: Hookworms infect 1.3 billion people annually. Iron-deficiency anemia caused or exacerbated by intestinal blood loss is the major feature of hookworm infection, with such loss caused by the feeding of hookworms upon the intestinal mucosa. The hookworms Ancylostoma duodenale and Necator americanus cause major gastrointestinal blood loss. While relatively greater blood loss has been reported in clinical studies due to infection with A. duodenale, there has been no evidence that endemic A. duodenale infection has a greater impact than N. americanus infection upon the iron status of populations. The authors compared the degree of anemia and iron deficiency associated with these 2 hookworm species at the individual and community levels using a sample of 525 school children in grades 1-4 in Pemba Island, Tanzania, and controlling for infection intensities and other child characteristics through multiple regression. In the 492 children with hookworm-positive fecal cultures, hemoglobin and ferritin concentrations decreased with increasing proportions of A. duodenale. Among children with only N. americanus larvae, the prevalence of anemia was 60.5% and the prevalence of ferritin under 12 mcg/l was 33.1%. In children with 50% or higher proportions of A. duodenale larvae, the respective prevalences were 80.6% and 58.9%. Children from schools with high prevalences of A. duodenale infection had significantly worse iron deficiency and anemia than did children from low prevalence schools.