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Mosquitoes collected in northern Italy were screened for flavivirus RNA. Positive amplicons were sequenced and found most similar to insect flavivirus (ISF), Usutu virus (USUV) and surprisingly also to Japanese encephalitis virus (JEV). The sequence (167 bp), obtained from one pool of Culex pipiens, was found identical to JEV strains from bats in China. Unfortunately additional sequence data or virus isolations were not obtained in this study. Confirmation of potential introduction of JEV to Italy and other European countries is urgently needed.

This short communication describes a case of human conjunctival dirofilariosis by Dirofilaria immitis. A 51-year-old man from the Emilia-Romagna region of northeastern Italy referred for ocular foreign body sensation in his right eye. A slight swelling of the bulbar conjunctiva was observed. A live, whitish, 10-cm-long nematode was extracted from the subconjunctival space. Histology revealed typical features of a filarioid nematode belonging to the genus Dirofilaria sp. Molecular analysis with polymerase chain reaction confirmed that the extracted nematode was a male of D. immitis. The removal of the parasite leads to complete resolution of symptoms. Ocular human dirofilariosis caused by infection with the filarioid nematode D. immitis is extremely rare, but should be considered in humans living in Italian endemic areas.

Purpose: To investigate the most plausible cause of stillbirth by evaluating clinical records and postmortem examination findings including placental analysis., Methods: A retrospective cohort study concerning 132 stillbirths from 124 pregnancies occurred in the Mother-Infant Department of the University Hospital of Modena, Italy, from January 2000 to December 2004. Collected data were reviewed and classified according to the Gardosi ReCoDe system., Results: A reasonable cause of fetal death was identified in 99/124 pregnancies (79.84%). No associated relevant factors were disclosed in 25 fetuses (20.16%) classified as unexplained stillbirths. A succeeding scrupulous analysis of the placenta and an accurate clinical record review were useful to detect other conditions in 82 cases, including 5 cases of unexplained stillbirth. The major relevant conditions associated to stillbirths were feto-placental infection especially in the early fetal gestation age, under the 24th week of gestation, and placental insufficiency occurred both in early and late gestation age fetuses and mainly associated with a IUGR (<10th customized percentile). The main frequent secondary conditions were represented by placental anomalies including cluster of avascular villi with stromal fibrosis associated to thrombosis in minor and/or major vessel(s). Through the further analysis of the placenta, we were able to reduce the unexplained stillbirth rate from 20.16 to 15%., Conclusion: Accurate fetal autopsy and placental examination related to meticulous clinical collecting data are requisites in the valuation of stillbirth and could play an important role in reduction of unexplained stillbirth rate.

Introduction: The aim of this study was to investigate the incidence of second primary tumors in patients with breast cancer (BC), with particular regard to bidirectional risk for endometrial cancer (EC)., Methods: A total of 7512 and 343 patients with first and second primary BC, respectively, were referenced to the expected number of cases calculated using the standardized incidence ratio (SIR) over the same period, to evaluate the observed and expected ratio between the groups. Data on tamoxifen use were also considered., Results: A total of 499 women with primary BC developed a second tumor. The total SIR, that is, the ratio between observed second primary cancer among patients with BC and the expected primary cancers in the general population, was significantly higher (SIR = 1.23; 95% confidence interval, 1.12-1.34; P = 0.007), particularly for melanoma (2.25), EC (2.15), ovarian cancer (1.74), hematologic malignancies (1.36), and bilateral BC (1.25). A greater risk of BC after thyroid (2.22) and EC (1.62) was also observed. Furthermore, the risk of developing EC was higher in patients treated with tamoxifen (SIR = 2.50 vs 1.34)., Conclusions: Bidirectional risk of endometrial cancer was not exclusively related to tamoxifen use.

In order to optimize resources, an assay allowing the detection of several etiological agents would be useful. This paper describes a polymerase chain reaction (PCR)-based assay for the parallel detection of different pathogens in mosquitoes (Diptera: Culicidae). The assay combines the analysis of both RNA and DNA, hence allowing the detection of both flaviviruses (Flaviviridae: Flavivirus) and filarial nematodes (Spirurida: Onchocercidae). The sensitivity of diagnostic assay for both flavivirus and nematode infections was tested by means of spiked samples and resulted adequate for the purpose of screening mosquito samples. The assay was implemented to check pathogen infections in 637 mosquitoes field-collected in Romagna, northern Italy. The integrity of RNA extracted from mosquito pools was checked by control amplifications targeting the 18 S rRNA of mosquitoes. Control amplifications were successful in 118 out of 119 pools. Flavivirus RNA was not found in any of these 118 pools, whereas DNA of Dirofilaria immitis (Leidy) was detected in one pool of Aedes albopictus (Skuse).

Infections with Spargana, the larvae of Spirometra spp., are rarely found in humans in Europe. So far only six cases have been discussed in the literature, four from Italy and two from France. We here report a new case in a 50-year-old workman, a freshwater sports fisherman from Bologna in Northern Italy. The infection manifested as a subcutaneous nodule in the thigh, 2x3 cm in size, slightly painful when compressed, somewhat mobile under the skin, present for 9 months, with recurrent periods of local itchiness associated with redness and slight oedema. The lump was removed surgically. Histological sections of the biopsied material revealed the presence of a sparganum. Drinking water contaminated by copepods, containing procercoid larvae of this parasite, seems to be the medium of infection.

The authors report on eleven new human cases of anisakiasis occurring in Italy, and emphasize the importance of the infection in clinical medicine, histopathology and public health. For ten of these cases, the diagnosis was based on histological findings: an eosinophilic granuloma associated with a larva of Anisakis sp. For one of them, the larva was removed from the stomach by endoscopy. Nine of the subjects were from Apulia and two from Molise (regions of southern and central Italy, respectively). Ten of them were surgically treated, and in one case the endoscopical extraction of the parasite resolved the situation. In two cases, the gastric wall was affected, in three the intestinal wall, in a further three the omentum, in one the spleen, and in the final two the mesentery and the epiploic appendix. In all the cases, the parasite was discovered unexpectedly during surgical treatment of the patients for supposed illnesses, which had originally been misdiagnosed. In three cases, the patients were also affected by cancer. Human anisakiasis must be taken into account in the differential diagnosis of acute, abdominal synddromes in subjects who have ingested raw fish or squid a few hours to a few days before the onset of symptoms. Histopathologists should consider the possibility of this parasitic infection when confronted with an eosinophilic granuloma of the digestive tract, mesentery or peritoneum. The incidence of anisakiasis in Italy is probably higher than reported, as some cases might not be diagnosed and others might heal spontaneously.

Histoplasmosis, which is highly endemic in the United States, is rare in Europe, usually imported but sometimes autochthonous. In Africa, histoplasmosis capsulati coexists with "African histoplasmosis", a characteristic skin infection caused by H. capsulatum var. duboisii. Histoplamosis due to H. capsulatum is one of the 12 secondary infections listed in the surveillance definitions of AIDS. We report the case of a 36-year-old black man with acquired immunodeficiency syndrome (AIDS) who was living in Italy but originally came from Ghana. Histoplasmosis was disseminated with fever and cutaneous manifestations. The diagnosis was demonstrated morphologically based on the presence of yeast, observed by light microscopy, in skin lesions and by identification of H. capsulatum var. capsulatum DNA by nested PCR from a paraffin sample. No clinical reports of histoplamosis capsulati in Ghana have been published until now. The present case stresses the role of immigration of subjects from outside Europe who have been infected in their native country.

A case of persistent infestation of the scalp due to Dermanyssus gallinae.--Infestation due to Dermanyssus gallinae, the common red mite of poultry, in a country-woman aged 69 years from Crevalcore (Emilia-Romagna region, Northern Italy), is described. The case was unusual either for its location, the scalp, and for its persistence, 9 months. Specimens of the mite were also found in the henhouse adjacent to the patient's house. Apparently the woman recovered by means of daily washing of camomile tea.

Aims: Sixty new cases of human dirofilariasis due to Dirofilaria repens, occurring in Italy between 1990 and 1999, are presented. This is the most extensive case study of this zoonosis reported worldwide by a single study group. The aim is to utilize this large experience to characterize the different histopathological findings in the parasitic lesions in man., Methods and Results: Diagnosis was performed on histological sections of the nematode enclosed in the nodules excised at biopsy or surgery. The nematode was located in the subcutaneous tissue (49 cases), the epididymis (two cases), the spermatic cord (two cases), the lung (two cases), the breast (two cases), the omentum (two cases) and under the conjunctival tissue (one case). The majority of cases (46) were from Piedmont; the remainder were from Emilia-Romagna, Sardinia, Sicily, Tuscany, Apulia and Lombardy. The histopathological features of the lesions are described and the clinical and epidemiological aspects of the zoonosis are discussed. The prevalence in Italy in general and in the area of Piedmont in particular, comprising the provinces of Alessandria, Asti, Novara and Vercelli, which is one of the most severely affected areas of the world, is emphasized. The evident increase in the number of cases in the last few years is a clear indication that it is an emergent zoonosis., Conclusions: We recommend that each and every case observed be recorded, to enable the true extent of human dirofilariasis in Italy to be assessed, and that a reference centre be set up in the area to collate the data. The importance of the histopathologist's role in the diagnosis is stressed.

No clinical reports of blastomycosis in Italy have been published until now. We here report two cases of histologically diagnosed, unexpected cutaneous involvement in patients, aged 78 and 52 years, living in North Italy and never having been abroad. The histological differential diagnosis between blastomycosis and other fungal pathogens is discussed. Even in the absence of culture the present cases can confidently be considered as genuine examples of Blastomyces dermatitidis infection in Italy.

Objectives: The main objective of the study was to evaluate if speculoscopy, a magnified chemiluminescent examination, combined with a Pap smear, could improve the detection of early cervical lesions compared with the Pap smear alone., Setting: Pap tests and speculoscopies were performed in two family planning centres located in the surrounding areas of Modena. Colposcopic investigations and biopsies of the uterine cervix were performed in a second level centre (Gynaecological Prevention Centre of Modena Policlinico). Histological specimens were analysed at the Section of Pathological Anatomy of Modena Policlinico., Subjects: The study population comprised 1000 women aged 25-64 years, invited to undergo a Pap smear every three years in accordance with the screening programme for cervical cancer started in Modena in 1997., Methods: Midwives performed the Pap smear and speculoscopy in succession. Women with a positive Pap test and/or positive speculoscopy underwent colposcopy and, if colposcopic findings were positive, targeted punch biopsies were performed., Results: A total of 1000 patients were subjected to cytology and speculoscopy examinations. Among these women, 10 had abnormal Pap smear findings whereas 144 had an abnormal speculoscopic pattern. Only three of 59 patients with a histological diagnosis of cervical intraepithelial neoplasia grade I (CIN I)/human papillomavirus and only three of seven patients with CIN II/CIN III had a positive Pap test., Conclusions: Speculoscopy combined with a Pap test can significantly increase the detection of cervical lesions when included in a screening programme.

Eight new cases of Human Subcutaneous Dirofilariasis due to Dirofilaria repens in subjects aged 14 to 83 years, 5 men and 3 women, are recorded. The patients were living in Piedmont (5 cases), Tuscany (1), Friuli-Venetia Julia (1) and Apulia (1). The nematodes were immature in 6 cases (2 males and 4 females) and adults in 2 (both females). Microfilariae in the uterus of one of them were detected.

For some Dirofilaria spp. of zoonotic importance, such as D.immitis, D.tenuis, D.ursi, reviews of human cases were often published. For D. repens there is no published information giving a picture of the importance of the problem and of the geographical distribution of the parasite in the world. From a study of the international literature and from direct experience (our workgroup observed 75 cases in Italy) we have ascertained that the condition associated with D. repens is the most frequent and the most widely distributed in the world, of the dirofilariases of medical relevance: comprehensively 397 cases were recorded in 30 countries, exclusively in the Old World, with endemic foci in Southern and Eastern Europe, Asia Minor, Central Asia and Sri Lanka. Apparently, the most affected country is Italy with a total of 168 cases. The most common localisations are the subcutaneous (normally nodular) and the submucosal (nodular or not). The most affected areas are the head, the thoracic wall and the upper limbs. However, 15 cases with internal localisations are recorded (9 of them being pulmonary) which were always diagnosed erroneously as malignant neoplasiae. Clinical diagnosis for the other localisations was also generally wrong with the exception of the subconjunctival cases, where because of the transparency of the bulbar conjunctiva, it is possible to see the parasite directly. The correct diagnosis is usually histological, based on the identification of the nematode using the morphological characters of the species to differentiate it from the other dirofilariae or other erratic nematodes. It is possible that the zoonosis in man is more frequent than shown by the literature: many cases are not diagnosed, viz. not published, others recover spontaneously without medical intervention.

A case of subcutaneous human dirofilariasis associated with Dirofilaria repens in the arm of a 9-year-old child resident in Campobasso is described. It is the first report of the nematode in the Molise region. Blood samples taken from dogs in the same region, confirmed the presence of D. repens microfilariae in the natural reservoir, few km from the clinical case.

Eight new cases of Human Dirofilariasis in people living in Northern Italy were recorded: 3 from the province of Asti and 2 from Alessandria (Piedmont), 1 from Corsico (Miland, Lombardy) and 2 from Ravenna (Emilia-Romagna). All the patients were adults, 42 to 66 years old, 4 men and 4 women. The localization of the nematode in all the cases was subcutaneous. Signs and symptoms were always mild and consisted in the growth of a 1 to 2 cm large nodule associated with occasional localized itching and oedema, sometimes with reddening. In one case, the sting of a non-specified winged insect was recorded in the anamnesis, in the same spot where the nodule developed. In another case, the nematode came out spontaneously from the lesion. The aetiological agent was always identified as Dirofilaria repens. These 8 new cases confirm the comparatively frequent presence of Human Dirofilariasis in regions of Northern Italy, namely in the Po Valley.

Two new cases of human subcutaneous dirofilariasis in Northern Italy are described. The first one occurred in a 56 year-old man living in Venezia Lido and was localised in the frontal region; the second one in a 52 year-old woman living in Modena and was localised in the sacrococcygeal region. Both lesions were associated with Dirofilaria repens. In the same geographical areas other cases of this zoonosis had occurred in the past.

Since 1971, at the Laboratory of Parasitology, Faculty of Veterinary Medicine, University of Bologna, 30 cases of human dirofilariasis were observed out of the 101 cases reported in Italy. The causative agent was identified in nearly all the cases as Dirofilaria repens (Nematoda, Filarioidea) a parasite relatively common in the dog in Italy. 23 cases were localized in the subcutaneous tissue, 4 in the submucosa and 3 in the lungs. In the majority of the cases the diagnosis had been misinterpreted, the lesions being first seen as malignant or benign tumours or foreign-body granuloma. The parasite was histologically recognized. In 3 cases the parasite could be extracted intact from the nodule. The authors believe that human dirofilariasis is more frequent than reported by the relevant scientific literature in that many cases are likely to recover spontaneously while others would remain undetected.

The presence of E. gregorii, a species recently identified by Hugot in France in man, is signaled by the authors in Italy and in the Central African Republic. The diagnosis was based on the differential morphological characters reported by Hugot and Tourte-Schaefer in 1985.