[Intestinal parasitosis among non-permanent resident students in Tunisia: a review of 23 years of monitoring in the department of Parasitology-Mycology at the Rabta Hospital of Tunisia]. / Les parasitoses intestinales chez les étudiants non-residents permanents en Tunisie: Bilan de 23 ans de surveillance au laboratoire de Parasitologie-Mycologie à l'hôpital la Rabta de Tunis.

BACKGROUND: In order to fight digestive parasitism in Tunisia, a national program of surveillance of non-permanent resident students in Tunisia has been found to detect these parasitosis in this target population. OBJECTIVE: To determine the prevalence of intestinal parasitosis among non-permanent resident students in Tunisia, to identify the different parasitic species founded and to show the interest of this screening. METHODS: During a period of 23 years (1990-2012), 7386 parasitological examinations of stools has been made among students essentially from or had visited tropical Africa, Maghreb and Middle-East, at the laboratory of Parasitology-Mycology at the Rabta Hospital of Tunis. RESULTS: The prevalence of intestinal parasitism found was 34.45% (i.e. 2545 infested students). Among the protozoa that have been isolated in the majority of cases (78.75%), amoebae were most frequently found (86.4%) represented mainly by Entamoeba coli and Endolimax nanus in respectively, 25.62 and 23.33% of parasites isolated; while Entamoeba histolytica/dispar, only pathogenic Amoeba was found in 8.05% of the total of parasites isolated. Regarding helminths, found in 21.25% of parasites isolated, Ankylostome was predominant (34.5%) represented by the species of Necator americanus. A single case of Ancylostom duodenale has been isolated. Among the identified parasite species, 38.7% were known parasitic pathogens for humans. CONCLUSIONS: These results note the interest of the control of the non-permanent resident students in Tunisia. The precocious tracking and treatment of affected subjects permits to avoid the introduction and the dissemination of parasites already rare and virulent strains in our country.

[Risk factors for Toxoplasma gondii and immune status of pregnant women: cause and effect?]. / Facteurs de risque pour Toxoplasma gondii et statut immunitaire des femmes parturientes: relation de cause à effet.

BACKGROUND: Human infection with the protozoan parasite Toxoplasma gondii is found world-wide. The differences in prevalence across the countries may be ascribed to variations in hygiene and eating habits. AIM: To identify preventable risk factors for Toxoplasma gondii infection in pregnancy through a transversal study achieved in the laboratory of Parasitology Mycology of Rabta including all pregnant women referred to the laboratory between March 2010 and February 2011 to serological testing for toxoplasmosis methods: The age, the stage of the current pregnancy and exposures to potential risk factors associated with Toxoplasma infection were collected by a questionnaire. An analysis with SPSS and chi-square was used to analyse exposure frequencies for each risk factor variable. RESULTS: A total of 2351 women were included. The median age of the pregnany women was 29.4 years (range, 16 - 48 years). In all, 47.7% of them were seropositive. Seropositivity for toxoplasmosis was significantly associated with eating undercooked meat (p = 0.002) and eating miswashed vegetables (p = 0.003). However, the other factors (contact with cats, cleaning the cat litter box, washing the hands after preparation of raw meat, contact with the ground) were found to be independently associated with seropositivity toxoplasmosis. CONCLUSION: In Tunisia, there is still no prevention program toxoplasmosis well structured. Health education on avoidance of maternal infection is an important aspect of any program for prevention of congenital toxoplasmosis.

Pulsed Field Gel Electrophoresis types of Candida albicans isolates from an intensive care unit in a Tunisian hospital.

Candida albicans is the most important cause of fungal infections in intensive care units. The aim of this work was to compare the profiles of C. albicans in order to specify their genetic polymorphism and to determine the origin of these infections. Thirty-five C. albicans strains were collected from different clinical samples of 12 patients and three health-workers in an intensive care unit (ICU) in Rabta hospital of Tunisia, between August 2007 and April 2008. After digestion with BssHII, the isolates were typed by pulsed field gel electrophoresis (PFGE). The PFGE profiles were analyzed using a visual method, which showed three PFGE types (A, B and C) and the dendrogram generated three clusters (clusters I to III). An average similarity coefficient of 0.83, suggests that isolates are related.

Morphological and molecular differentiation between Culicoides oxystoma and Culicoides kingi (Diptera: Ceratopogonidae) in Tunisia.

BACKGROUND: Culicoides kingi and Culicoides oxystoma belong to the Schultzei group of biting midges. These two species are vectors of disease in livestock of economic importance. As described in the literature, morphological identification for discrimination between them is still unclear. However, species-specific identification is necessary to solve taxonomic challenges between species and to understand their roles in disease transmission and epidemiology. This study aims to develop accurate tools to discriminate C. oxystoma from C. kingi using traditional morphometry and polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) assays for use in developing countries. METHODS: Specimens were collected from the region of Kairouan in central Tunisia. A total of 446 C. oxystoma/C. kingi individuals were identified using traditional morphometric analyses combined with PCR-RFLP of the cytochrome c oxidase subunit I gene. Thirteen morphometric measurements were performed from the head, wings, and abdomen of slide-mounted specimens, and six ratios were calculated between these measurements. Multivariate analyses of the morphometric measurements were explored to identify which variables could lead to accurate species identification. RESULTS: Four variables, namely antennae, wings, spermathecae, and palpus length, were suitable morphometric characteristics to differentiate between the species. Digestion with the SspI restriction enzyme of the PCR product led to good discriminative ability. Molecular procedures and phylogenetic analysis confirmed the efficiency of this simple and rapid PCR-RFLP method. CONCLUSIONS: This study highlights for the first time in Tunisia the presence of C. oxystoma and its discrimination from C. kingi using abdominal measurements and the PCR-RFLP method. This approach could be applied in future epidemiological studies at the national and international levels.

[Keratomycosis in the area of Tunis: epidemiological data, diagnostic and therapeutic modalities]. / Les kératites fongiques dans la région de Tunis: caractéristiques épidémiologiques, modalités diagnostiques et thérapeutiques.

Fungal keratitis is a serious disease involving the visual prognosis. This pathology is not well known in Tunisia. The aim of our study is to determine epidemiological data and clinical and mycological characteristics of fungal keratitis in the area of Tunis (North of Tunisia) and discuss its therapeutic modalities. This is a retrospective study including 19 cases of fungal keratitis collected over a period of 11 years (January 1998-December 2008). The diagnosis of keratomycosis was based on clinical and mycological data. Mycological examination interested corneal scraping including direct examination and culture. The cases of fungal keratitis concerned 13 men and 6 women with a mean age of 48.7 years. The most common risk factors was corneal trauma (47.4%). The mean delay between the first ophthalmic signs and consultation was 17.7 days. Most frequently fungal isolated fungi were Candida albicans (6 cases), followed by Aspergillus spp (5 cases) and Fusarium spp (4 cases). All patients received topical and systemical antifungal therapy. The evolution was favourable in six cases. Three patients retained corneal scars. The surgery was necessary in 7 cases, consisting of a penetrating keratoplasty (5 cases), an enucleating (1 case) and amniotic membrane transplantation (1 case). In conclusion, despite the improvement of diagnosis and treatment of fungal keratitis, its prognosis remains pejorative. This prognosis depends on early diagnosis and choice of antifungal therapy.

[Subcutaneous dirofilariasis of the upper lip in Tunisia]. / Dirofilariose sous-cutanée en Tunisie : une observation à localisation labiale.

INTRODUCTION: Dirofilariasis is a rare anthroponotic disease caused by Dirofilaria, the principal reservoir of which is the dog. The first case of subcutaneous dirofilariasis in Tunisia was reported in 1990. CASE REPORT: We report a case involving a 40-year-old woman living in northeastern Tunisia who presented with a subcutaneous lesion of the upper lip resembling a sebaceous cyst. Excisional biopsy released a worm identified on morphologic examination as Dirofilaria repens. Only excision allows simultaneous diagnosis and treatment. CONCLUSION: This is the 14th case of dirofilariasis reported in Tunisia. This parasitic disease is probably underestimated in our country. It should be considered for all subcutaneous nodules, regardless of localisation.

[Acanthamoeba keratitis: diagnosis to consider]. / La kératite amibienne: il faut y penser.

INTRODUCTION: Acanthamoeba keratitis is a rare but severe disease that can cause blindness. The objective of this study is to call attention to its severity and conditions of development and to emphasize the importance of early treatment. CASE REPORT: We report a case of a 27-year-old woman. After wearing soft contact lenses for a month, she consulted for redness, pain and poor visual acuity in her left eye. The ophthalmological examination showed an epithelial corneal abscess measuring 2 mm in diameter in the left eye and very tiny epithelial corneal abscesses in the right eye. Microscopic examination and culture of samples from the contact lenses and the contact lens solution found Acanthamoeba spp. Rapid anti-amoeba treatment led to disappearance of the corneal abscesses and improvement of her vision. CONCLUSION: This case confirmed that the prognosis of Acanthamoeba keratitis depends on rapid diagnosis and early treatment.

Mucormycosis: four case reports.

INTRODUCTION: Mucormycosis is a rare and invasive fungal infection, but frequently fatal when it occurs. It commonly affects patients with diabetes mellitus. The aim of this study is to assess the clinical presentation, radiological findings, management and prognosis of mucormycosis. METHODS: This retrospective study was conducted in the department of infectious diseases of Rabta hospital between January 1988 and December 2004 and included patients hospitalized for mucormycosis confirmed by mycological and/or histological findings. RESULTS: the study is about four diabetic patients with mucormycosis (3 men and a woman). Three of them had diabetic ketoacidosis at the time of diagnosis. The infection was sinusal in 2 cases and rhinocerebral in the the other two cases. Treatment consisted in systemic amphotericin B combined with surgical debridement in 3 cases. A fatal outcome was noted in 2 cases. CONCLUSION: Mucormycosis remains a severe infectious disease in diabetic patients. Early diagnosis and treatment is mandatory for a successful management of this infection.

[Confirmation by PCR of the existence in Tunisia of Entamoeba histolytica and Entamoeba dispar]. / Confirmation par l'amplification genique de l'existence en Tunisie des deux especes : Entamoeba histolytica ET entamoeba dispar.

AIM: Amibias are illness in Tunisia diagnosed until now on the sole basis of the morphological aspects of the parasite. Our aim is to report the first Tunisian results concerning the molecular identification of E. histolytica/E. dispar, METHODS: 25 stools presenting cysts and/or vegetative shapes of E. histolytica/E. dispar were gathered at the "Laboratoire de Parasitologie Hôpital La Rabta Tunis" between 2001and 2004 for PCR. The stools came from 24 subjects, one of them having two samples: 9 Tunisian patients, 5 adressed to the hospital services for abdominal pains or diarrheas and 4 adressed for a systemic tracking (food manipulation), and 15 foreign students for which a tracking is done each fall. RESULTS: The identification showed thus for the Tunisian patients the presence of : E. histolytica alone for a patient (food manipulator) 11%. E. histolytica associated to E. dispar for two patients 22%. E. dispar alone for six patients 67%. Nearly similar results has been obtained for foreign student's samples: E. histolytica alone in one case (7%), E. histolytica associated to E. dispar in four cases (26%) and E. dispar alone in 10 cases (67%). CONCLUSION: These results show therefore the existence in Tunisia the two species E. histolytica and E. dispar for symptomatic or non symptomatic patients. The distinction between the two species is very important on the therapeutic level as well as the epidemiologic and public health level.

Blood meal analysis of culicoides (Diptera: ceratopogonidae) in central Tunisia.

To evaluate the host preferences of Culicoides species (Diptera: Ceratopogonidae) in Central Tunisia, we identified the source of blood meals of field collected specimens by sequencing of the cytochrome b (cyt b) mitochondrial locus and Prepronociceptine single copy nuclear gene. The study includes the most common and abundant livestock associated species of biting midges in Tunisia: C. imicola, C. jumineri, C. newsteadi, C. paolae, C. cataneii, C. circumscriptus, C. kingi, C. pseudojumineri, C. submaritimus, C. langeroni, C. jumineri var and some unidentified C. species. Analysis of cyt b PCR products from 182 field collected blood-engorged females' midges revealed that 92% of them fed solely on mammalian species, 1.6% on birds, 2.4% on insects and 0.8% on reptiles. The blast results identified the blood origin of biting midges to the species level with exact or nearly exact matches (≥98%). The results confirm the presence of several Culicoides species, including proven vectors in Central Tunisia. Blood meal analyses show that these species will indeed feed on bigger mammals, thereby highlighting the risk that these viruses will be able to spread in Tunisia.

Plasmodium falciparum causing hemophagocytic syndrome after allogeneic blood stem cell transplantation.

We describe a case of Plasmodium falciparum infection in a 25-year-old male patient with a myelodysplastic syndrome, who underwent allogeneic peripheral blood stem cell transplantation (PBSCT) in September 2003. Conditioning regimen consisted of total body irradiation (10 Gy) and cyclophosphamide 60 mg/kg for 2 days. A dose of 4 x 10(6) CD34+ cells/kg was transfused. Engraftment was well documented on day 17 post-transplantation. Spiking fevers occurred on days 19 and 21, associated with a pancytopenia, hepatosplenomegaly and neurological signs. P. falciparum parasites were found on the peripheral blood smear (parasitemia = 23%). Marrow aspiration showed P. falciparum parasites and proliferation of mature histiocytes with hemophagocytosis. Quinine 10 mg/kg i.v. three times a day for 10 consecutive days was given. The fever subsided within 3 days, and pancytopenia vanished in 14 days. Parasitemia cleared in 6 days. The patient left the unit on day 46 with no further complications. The screening of donors showed that infection was acquired from two blood units (from a single donor) given 5 days before transplantation. We report the first case of profound hemophagocytosis in immunosuppressed patient with malaria of high parasitemia after a bone marrow transplant.

[Efficacy of doxycycline in thoracic actinomycosis: a case report]. / Efficacite de la doxycycline dans l'actinomycose thoracique: a propos d'une observation.

The authors present a case of thoracic actinomycosis in a 32 year-old man with a history of cervico-facial actinomycosis. Diagnosis was made by bacterial examination of purulent drainage from sternal wound showing typical sulfures granules. One year course of doxycycline gives clinical and radiological improvement.

[Toxocariasis: under-estimated disease. Report of 9 cases]. / La toxocarose: une pathologie sous-estimée. A propos de 9 cas.

Toxocariasis or visceral larva migrans is a parasitosis due to the migration in the human organism of animal ascarid larvae. Its importance is under-estimated and the reported tunisian cases are rare. We report nine cases of toxocariasis, noted between January of 2000 to March of 2002, in the laboratory of parasitology-mycology in La Rabta Hospital. They were six children and three adults. The clinical forms are varied: ophthalmologic form (5 cases), general syndrome (1 case), oedema (1 case) and hypereosinophilia (2 cases), and confirmed serologically by the presence of anti Toxocara canis antibody using ELISA test. Visceral larva migrans should be kept in mind to avoid severe forms such as ophthalmologic forms.

[Bronchopulmonary aspergillosis: study of 17 cases]. / L'aspergillose broncho-pulmonaire: etude de 17 cas.

Aspergillosis is a fungic infection depending on the local or general physiologic and immunologic state of the host. We report the result of retrospective five year study (1995-1999) about 17 cases in the laboratory of Parasitology-Mycology of Rabta hospital in Tunis. Six aspergillomas were observed, they occurred after a pulmonary tuberculosis, two cases of allergic broncho-pulmonary aspergillosis described in two asthmatic patients, nine cases of invasive pulmonary aspergillosis complicating two cancers, one leukaemia, six chronic granulomatous disease. Aspergillus fumigatus is the most frequent species (67%). The clinical and biological characteristic of those will be studied, and compared with those of the literature.

First detection of Leishmania infantum (Kinetoplastida: Trypanosomatidae) in Culicoides spp. (Diptera: Ceratopogonidae).

BACKGROUND: Culicoides (Diptera: Ceratopogonidae) species are known to be the vectors of Bluetongue virus and African Horses Sickness virus (AHSV) in different areas of the world. Nevertheless, other researchers have hypothesized that these arthropods could be involved in the transmission of other pathogens such as Schmallenberg virus, Plasmodium and Leishmania parasites. Identification of the Culicoides' potential vector competence is crucial in understanding the worldwide Culicoides/Leishmania life cycle. FINDINGS: Blood fed and parous females of biting midges Culicoides spp. were collected between 2009 and 2010 in Central Tunisia. DNA was extracted from individual blood fed Culicoides and used as a template in a genus-specific PCR. Leishmania DNA was detected in 14 Culicoides imicola specimens and one Culicoides circumscriptus. In a second step, parasite identification was performed based on a single copy Topo-isomerase II gene specific amplification and sequencing. Leishmania infantum was identified in two infected Culicoides spp. CONCLUSION: This is the first report of Leishmania DNA detection from naturally infected wild caught Culicoides spp. Our finding supports the assumption that Culicoides spp. are a potential vector for L. infantum.