[Contribution of Real-Time PCR in the Diagnosis of Cutaneous Leishmaniasis: Experience of the Pasteur Institute of Tunis]. / Contribution de la PCR en temps réel dans le diagnostic biologique de la leishmaniose cutanée : expérience de l'institut Pasteur de Tunis.

The aim of this study was to assess the performance of real-time PCR (qPCR) in the diagnosis of cutaneous leishmaniasis (CL). Culture, direct microscopic examination (DE) and qPCR were performed on dermal exudate samples collected from 235 confirmed CL cases. The qPCR was found to be more sensitive than other diagnostic techniques and was able to correct the diagnosis in 49 patients (20.9%) with negative dermal smears. Median parasitic load (PL) of the 49 dermal exudates with negative DE was lower than that of positive ones in microscopy. This suggests that PL likely impact the sensitivity of microscopy. On the other hand, qPCR was performed on DNA extracts of scraped products collected from the 23 out of 49 archived negative Giemsa-stained slides and showed 11 positive. Parasitic loads in the latter smears were lower than those in corresponding exudates. The results highlight qPCR relevance for the diagnosis of CL and recommend its use directly on dermal exudates collected from CL lesions.

L'objectif de ce travail était d'évaluer la place de la PCR en temps réel (qPCR) dans le diagnostic de la leishmaniose cutanée (LC). Des sucs dermiques prélevés chez 235 cas confirmés de LC ont été traités par examen microscopique direct (ED), culture et qPCR, et les résultats analysés. La qPCR s'est révélée plus sensible que les autres techniques diagnostiques permettant de redresser le diagnostic chez 49 patients (20,9 %). Les charges parasitaires (CP) des 49 sucs dermiques correspondants étaient inférieures à celles de prélèvements microscopiquement positifs suggérant que la CP serait un facteur limitant la sensibilité de l'ED. La qPCR a été également pratiquée sur les produits de grattage de 23 des 49 frottis négatifs. Elle a permis d'en détecter 11 positifs avec des CP inférieures à celles dans les sucs dermiques correspondants. Ces résultats recommandent l'utilisation de la qPCR pour le diagnostic de la LC et privilégient sa réalisation directement sur le suc dermique.

[Management of Toxoplasmic Seroconversion in the Third Trimester of Pregnancy in Tunisia]. / Particularités de la prise en charge en Tunisie des séroconversions toxoplasmiques du troisième trimestre de grossesse.

The purpose of our study was to report the particularities of the management of toxoplasmosis seroconversion cases occurred during the third trimester of pregnancy and diagnosed in the Institute Pasteur of Tunis. The study was conducted in the Laboratory of Parasitology-Mycology of the Institute Pasteur of Tunis between January 2005 and December 2017. A total of 27 cases of toxoplasmosis seroconversion during the third trimester were included. Prenatal diagnosis was performed in five cases. PCR was positive in one case. Pyrimethamine-sulfadoxine was prescribed in one case with positive PCR and in another case as soon as maternal infection was confirmed. Spiramycine was prescribed in 24 cases for the duration of the pregnancy. One woman did not take any treatment because seroconversion was diagnosed just before delivery. Twenty newborns had congenital toxoplasmosis: 19 cases were diagnosed by serology and one case was diagnosed after amniocentesis. Two newborns (10%) were symptomatic at birth. All the newborns had neither clinical nor radiological signs during the follow-up.

L'objectif de notre travail était de rapporter les particularités de la prise en charge d'une série de cas de séroconversion toxoplasmique du troisième trimestre de grossesse diagnostiquée à l'institut Pasteur de Tunis. Il s'agit d'une étude rétrospective colligée au laboratoire de parasitologie-mycologie de l'institut Pasteur de Tunis entre janvier 2005 et décembre 2017, incluant 27 femmes enceintes ayant présenté une séroconversion toxoplasmique au cours du troisième trimestre de la grossesse diagnostiquée au cours de leurs suivis sérologiques. Cinq femmes ont bénéficié d'un diagnostic anténatal. La PCR (polymerase chain reaction) s'est révélée positive dans un seul cas. Deux femmes ont été mises sous pyriméthamine-sulfadoxine dont une avait une PCR positive. La spiramycine a été instituée chez 24 femmes jusqu'à l'accouchement. Une femme n'a reçu aucun traitement, la séroconversion ayant été diagnostiquée la veille de l'accouchement. Le diagnostic de la toxoplasmose congénitale a été retenu chez 20 nouveau-nés (74 %) dont un en anténatal et 19 à la naissance. Deux étaient symptomatiques (10 %) à la naissance. Aucune manifestation clinique ou radiologique n'a été observée au cours de leurs suivis.

Asymptomatic intestinal carriage of microsporidia in HIV-positive patients in Tunisia: prevalence, species, and pathogenesis.

Asymptomatic carriage of microsporidia (ACM) has not been described in patients living with HIV (PLHIV) in Tunisia. To determine the prevalence of ACM in PLHIV followed at Tunis la Rabta hospital, describe its clinical features and course, and identify the species involved. This prospective study (2005-2009) included 71 asymptomatic PLHIV compared with 37 PLHIV with diarrhea. One stool sample per patient was examined by microscopy after Weber staining and by PCR. Species identification was confirmed by specific PCR and sequencing. In cases of ACM, a second stool sample was examined in 2010 and a clinical check-up took place in 2013. The prevalence of ACM in asymptomatic PLHIV was 11.3 % (8/71). PCR was more sensitive than microscopy (P = 0.0047). ACM was associated with stage C of HIV infection (P = 0.008) and CD4 T cells <100/µl (P = 0.033). The species involved were E. intestinalis (6 cases) and E. bieneusi (2 cases). Six PLHIV remained asymptomatic with negative stool examinations, but two developed digestive signs. ACM is common among Tunisian PLHIV and it appears to be associated with E. intestinalis.

[Epidemiological, clinical and biological features of infantile visceral leishmaniasis at Kairouan hospital (Tunisia): about 240 cases]. / Profils épidémiologique, clinique et biologique de la leishmaniose viscérale infantile à l'hôpital de Kairouan (Tunisie): à propos de 240 cas.

Visceral leishmaniasis (VL) is an important health problem in Tunisia. It is most common in children under five years of age. The governorate of Kairouan (central Tunisia) is one of the most affected foci. The aim of this study was to update the epidemiological, clinical and biological features of the disease. The study concerned all VL cases admitted in the pediatric department of Kairouan hospital during 10 years (from 2004 to 2013). For every patient included in this study and when available, data such as sex, age, geographical origin and the condition of the patient at admission (clinical and biological findings) were collected. The myelogram results were also exploited as well as results of serology, culture, Real-Time polymerase chain reaction (PCR) and isoenzymatic typing of Leishmania isolates. Two hundred and forty cases were recorded. Rural cases (87.1%) were more prevalent than urban ones (12.9%). Age ranged from 2 months to 13 years (median, 18 months). The female/male sex ratio was 1.03. The diagnosis delays ranged from 1 day to 8 months (median, 15 days). The most common clinical symptoms at admission were splenomegaly (97.9%), fever (79.9%) and hepatomegaly (47.3%). The principal biological disturbances were anemia (91.7%), thrombocytopenia (83.9%) and leucopenia (56.1%). Among the different biological tools used for diagnosis confirmation, PCR was the most sensitive (100%). All 43 typed stocks corresponded to Leishmania (L.) infantum species. Although zymodeme MON-1 was predictably the most frequent (27 cases), L. infantum MON-24 and MON-80 were responsible of no negligible numbers of cases (11 and 5 cases respectively). The present study gave an updated epidemiological, clinical and biological profile of infantile VL in Tunisia. The diagnosis delays were considerably shortened compared to previous reports. However, an even earlier diagnosis of cases is needed to improve the disease prognosis. Real-Time PCR showed to be helpful in VL management.

[Investigation and analysis of an outbreak of cutaneous leishmaniasis in Ksar Ouled Dabbab, Tataouine (Tunisia), 2012-2013]. / Investigation et analyse d'une épidémie de leishmaniose cutanée à Ksar Ouled Dabbab, Tataouine (Tunisie), 2012-2013.

UNLABELLED: Zoonotic cutaneous leishmaniasis (ZCL), due to Leishmania major, and chronic CL, due to L. tropica, are endemic in the governorate (administrative subdivision) of Tataouine (southeast Tunisia). This study aims to analyze epidemiologic, clinical, parasitologic, and entomologic data collected during the 2012-2013 epidemic in Ksar Ouled Dabbab (KOD). METHODS: The KOD health care center diagnosed 164 CL cases from July 2012 to March 2013, identifying 21 Leishmania isolates and local sandflies. RESULTS: The incidence rate of CL in KOD was around 27.5/1000 inhabitants. The age groups most strongly affected were children younger than 10 years (19.9%) and those aged 10-20 years (23.1%). The number of patients with multiple lesions (67.9%) and their location on limbs (71.3%) indicated ZCL. This hypothesis is supported by the high proportions of L. major (81%) and Phlebotomus papatasi found. CONCLUSION: The epidemiologic and clinical profiles of the cases and the predominance of L. major and P. papatasi confirm that the recent outbreak in KOD was related to ZCL. It probably followed recent environmental changes and transmission spread from adjacent neighboring foci.

Mucosal leishmaniasis of the lip: an emerging clinical form in Tunisia.

BACKGROUND: Mucosal leishmaniasis (ML), which mostly occurs in the New World, is mainly associated with Leishmania braziliensis. Primary lip ML is very rare in the Mediterranean basin and particulary in Tunisia despite the endemicity of both cutaneous and visceral leishmaniasis in this area. OBJECTIVES: To highlight a recent emergence of primary lip ML in Tunisia, to describe its epidemiological and clinical features and to identify the causative Leishmania species. METHODS: Epidemiological, clinical and therapeutic data of 10 cases presenting a ML of the lip were collected. Diagnosis confirmation of leishmaniasis was obtained by microscopic examination of Leishmania parasites in Giemsa stained smears of the lesion sampling and in cutaneous biopsies. Polymerase chain reaction (PCR) detecting Leishmania DNA directly from dermal scraping was also performed for diagnosis and species identification. RESULTS: Seven men and three women with lip ML were diagnosed during the last 6 years (2008-2013). The mean age was 29.7 years. Clinical presentation was characterized by an infiltrated and ulcerated plaque leading to macrocheilitis involving the upper lip in eight cases and the lower lip in two cases. Mean diagnosis delay was 6.9 months. PCR identified L. infantum in seven cases and L. major in two cases. Seven patients received intramuscular injections of meglumine antimoniate (MA) and three patients received both MA intralesional injections of MA and cryotherapy. A clinical remission was rapidly observed in all cases (on average in 2.2 months). CONCLUSIONS: Primary lip ML is emerging in Tunisia. Macrocheilitis of the upper lip is the main clinical presentation. PCR revealed more sensitive than direct examination in the diagnosis of such form (P < 0.01). Leishmania infantum was the most identified species (7 cases) while L major was involved in only two lesions. A benign local evolution and a rapid recovery were observed in all cases after MA treatment.

Identification of soluble and membrane antigenic markers of acquired toxoplasmosis by immunoblot.

The overall performance of quantitative assays in the detection of anti-Toxoplasma IgG is satisfactory, but discrepancies between assays are not uncommon especially when IgG concentrations are close to the limit of detection of the tests. The purpose of our study was to identify soluble and membrane antigens extracted from Toxoplasma gondii tachyzoites by immunoblot to select the most relevant antigenic bands to be used for qualitative serodiagnosis of acquired toxoplasmosis. We selected five relevant bands (98, 36, 33, 32 and 21 kDa) with soluble antigens and four relevant bands (42, 35, 32 and 30 kDa) with membrane antigens which gave high sensitivity and/or specificity in immunodiagnosis. The association on the same blot of at least three of the five relevant bands in the soluble antigen immunoblot showed the highest sensitivity/specificity (97.4%/99.0%, respectively). Our results indicate that immunoblot using soluble tachyzoite extract with simultaneous detection of at least three of the five bands (98, 36, 33, 32 and 21 kDa) represents a valuable test for serodiagnosis of acquired toxoplasmosis and should be further evaluated as a confirmatory test for sera which give discrepant results in quantitative assays.

Phlebotomus (Paraphlebotomus) riouxi: a synonym of Phlebotomus chabaudi without any proven vectorial role in Tunisia and Algeria.

Phlebotomus (Paraphlebotomus) riouxi Depaquit, Léger & Killick-Kendrick (Diptera: Psychodidae) was described as a typological species based on a few morphological characters distinguishing it from Phlebotomus (Paraphlebotomus) chabaudi Croset, Abonnenc & Rioux. The naming of P. riouxi coincided with its incrimination as a rural vector of Leishmania tropica Wright (junior synonym: Leishmania killicki Rioux, Lanotte & Pratlong) in Tataouine governorate, an arid region of southern Tunisia. The current report finds insufficient evidence to incriminate either phlebotomine sandfly as a vector of L. tropica in North Africa. Phlebotomus riouxi was found not to have the characteristics of a phylogenetic or biological species, and therefore it is synonymized with P. chabaudi. Both taxa were recorded together for the first time in Tunisia, in Tataouine, where three of 12 males showed intermediate morphology and both sexes of each taxon were not characterized by specific lineages of the nuclear gene elongation factor-1α or the mitochondrial gene cytochrome b, for which a long 3' terminal fragment is recommended for phlebotomine phylogenetics. This case study indicates that the eco-epidemiology of leishmaniasis should focus more on identifying key components of vectorial transmission that are susceptible to interventions for disease control, rather than on defining sibling species of vectors.

[Toxoplasmic infections in pregnancy: about 94 cases diagnosed at the Pasteur Institute of Tunis]. / Infections toxoplasmiques au cours de la grossesse: à propos de 94 cas diagnostiqués à l'institut Pasteur de Tunis.

OBJECTIVES: The objectives of this study were to estimate the risk of maternal-fetal transmission of toxoplasmosis and its consequences on the fetus and to emphasize the importance of follow-up of newborns in Tunisia. PATIENTS AND METHODS: It was a retrospective study of 94 cases of pergravidic toxoplasmic seroconversion who were diagnosed and followed in the Laboratory of Parasitology of Pasteur Institute of Tunis between 2005 and 2010. RESULTS: In our series, amniocentesis was performed for 60 parturients. Among the amniotic fluid tested, research of toxoplasmosis DNA by PCR was positive in 12 cases (12/60, 20 %). Twenty-six cases of congenital toxoplasmosis were diagnosed with 14 postnatal cases. The rate of maternal-fetal transmission of Toxoplasma gondii was 27.6 % (26/94). This risk increases with gestational age, from 19 % at seroconversion of the 1st quarter to 29.4 % in the 2nd quarter and 44.4 % in the 3rd trimester. Monitoring of newborns with congenital toxoplasmosis showed that only 3 children were symptomatic. There were 2 cases of toxoplasmic chorioretinitis and a case of brain damage. Under serological monitoring of newborns, 21 cases were lost to follow-up and monitoring was stopped for 29 after decrease of anti-toxoplasmic IgG. DISCUSSION AND CONCLUSION: The prenatal diagnosis allowed to decrease the severe forms of congenital toxoplasmosis in Tunisia. Nevertheless, it is always necessary to raise the problem of the significant number of newborn children whose follow-up is incomplete.

[Contribution of Leishmania identification using polymerase chain reaction--restriction fragment length polymerase for epidemiological studies of cutaneous leishmaniasis in Tunisia]. / Place de l'identification des leishmanies par la polymerase chain reaction--restriction fragment length polymerase dans l'étude de l'épidémiologie des leishmanioses cutanées en Tunisie.

AIM: Three forms of cutaneous leishmaniasis (CL) are endemic in Tunisia. The identification of the causative species is useful to complete epidemiological data and to manage the cases. The aim of this study is to assess PCR-RFLP technique in the identification of Leishmania species responsible of CL in Tunisia and to compare the results of this technique to those of isoenzyme analysis. PATIENTS AND METHODS: Sixty-one CL lesions were sampled. Dermal samples were tested by culture on NNN medium and analyzed by PCR-RFLP assay targeting the ITS1 region of ribosomal DNA. Species identification was performed by both iso-enzymatic typing for positive cultures and analysis of restriction profiles after enzymatic digestion by HaeIII of the obtained amplicons. RESULTS: Thirty-eight (62%) samples were positive by culture. The iso-enzymatic typing of 32 isolates identified 3 L. infantum, 23 L. major MON-25 and 6 L. tropica MON-8. Sixty samples were positive by PCR. The PCR-RFLP digestion profiles of the 56 PCR products identified 12 L. infantum, 38 L. major and 6 L. tropica. The results of both techniques were concordant in the 32 strains identified by both techniques. Species identification correlated with the geographical distribution of CL forms endemic in Tunisia. CONCLUSION: Results of PCR-RFLP revealed highly concordant with those of isoenzyme electrophoresis. Thanks to its simplicity, rapidity and ability to be performed directly on biological samples, this technique appears as an interesting alternative for the identification of Leishmania strains responsible of CL in Tunisia.

[Evaluation and contribution of OptiMAL-IT® test for the diagnosis of imported malaria in Tunisia]. / Évaluation et apport du test OptiMAL-IT® dans le diagnostic du paludisme d'importation en Tunisie.

The rapid test OptiMAL-IT® was evaluated in the diagnosis and the screening of imported malaria in Tunisia in comparison with microscopic techniques. This prospective study focused on 500 individuals recruited from September 2010 to September 2012 in laboratory of Parasitology of Pasteur Institute of Tunis. They include 192 patients with clinical manifestations suggestive of malaria and 308 students originating from endemic areas. Microscopy of thick-and-thin blood smears and OptiMAL-IT® test were systematically performed on blood samples of all participants. Sixty individuals revealed infected by Plasmodium (12%). Positivity rates were respectively 20.3% in patients (44 cases) and 5.2% among asymptomatic students (16 cases) (p<0.01). The sensitivity and specificity of the OptiMAL-IT® test were respectively 88.6% and 100%. The concordance kappa was 0.92. The sensitivity and specificity during the screening of asymptomatic subjects were respectively 68.8% and 98.3% with a concordance of 0.67.

[Toxoplasmosis mother-to-child screening: study of cases followed in the Pasteur Institute of Tunis (2007-2010)]. / Dépistage de la toxoplasmose materno-foetale : étude des cas suivis à l'Institut Pasteur de Tunis (2007-2010).

Toxoplasmosis when occurring during pregnancy can be transmitted to the fetus and lead to congenital toxoplasmosis (CT). Therefore, pregnant women are a risk group, for which it is necessary to determine the serologic profile. The objective of this study is to determine the serologic profile of toxoplasmosis in pregnant women followed at the Parasitology Laboratory of the Pasteur Institute in Tunis, to establish the prevalence of toxoplasmic infections during pregnancy and the incidence of the CT, noting the difficulties faced in the interpretation of serological results. This is a retrospective study concerning 2833 toxoplasmic serologies practiced on 2070 pregnant women, followed at the Parasitology-Mycology Laboratory of the Pasteur Institute of Tunis, between 2007 and 2010. Serological diagnosis of toxoplasmosis was done by ELISA (Enzyme Linked Immunosorbent Assay) for the detection of Immunoglobulin (Ig) G and M and the study of toxoplasmosis IgG avidity. Prenatal diagnosis was performed for 58 women by amniotic fluid sampling. Toxoplasma gondii was detected by Polymerase Chain Reaction (PCR). At birth, the diagnosis of congenital toxoplasmosis was established based on serology. The toxoplasmic serologies carried out have shown that 45.6% of the pregnant women were formerly immunized while 49.6% had a negative serology. A toxoplasmosis primary infection acquired during pregnancy was detected in 79 cases (3.8%). Among them, 33% had a true seroconversion while 67% had a recent toxoplasmosis infection in view of the positivity of IgG and IgM on the first sample with a low index of avidity (IA). For 21 parturients whose serology showed the presence of IgG, IgM and an intermediate or high IA. Among the 58 parturients in whom prenatal diagnosis was performed, PCR was positive in four cases. After birth, six cases of congenital toxoplasmosis were detected by serology.

[Comparative characterization of skin lesions observed in the three endemic varieties of cutaneous leishmaniasis in Tunisia]. / Caractérisation comparative des trois formes de leishmaniose cutanée endémiques en Tunisie.

BACKGROUND: The recent spread in the geographical distribution of the three forms of cutaneous leishmaniasis (CL) endemic in Tunisia has resulted in the coexistence of more than one species of Leishmania (L.) in some foci, rendering characterization on the basis of geographical criteria alone more difficult. The aim of the study was to establish clinical criteria associated with these noso-geographic forms, namely sporadic CL (SCL) due to L. infantum, zoonotic CL (ZCL) due to L. major and chronic CL (CCL) due to L. tropica. PATIENTS AND METHODS: One hundred and twelve patients with biologically confirmed CL were involved in the study. Leishmania species was systematically identified by iso-enzyme analysis and/or PCR-RFLP. Details of the number, the location, the morphological aspect and the month of outbreak of the lesions were noted for each patient. RESULTS: SCL lesions appeared later than ZCL lesions (53.8% of cases appeared from December onwards vs. 23.6%, P<0.001). ZCL lesions were often multiple (75%) and situated on the limbs (84.7%, P<0.001), whereas SCL lesions were single (92.3%, P<0.001) and located on the face (84.6%, P<0.001). CCL lesions were also single (78.6%) and located on the face (71.4%). The classical ulcerous presentation with scabs was mainly observed in ZCL patients (69.4%) and the erythematous presentation was described more frequently in SCL patients (75%; P<0.001). CONCLUSION: The number, site, morphological aspect and month of outbreak of lesions could be considered as useful criteria that help differentiate between the three noso-geographical forms of CL prevailing in Tunisia. Such characterization is useful for the individual management of patients and for optimizing the combat against the disease.

Genotype identification of Enterocytozoon bieneusi isolates from stool samples of HIV-infected Tunisian patients.

The microsporidian species Enterocytozoon bieneusi is a major cause of chronic diarrhea and malabsorption in patients with AIDS. Genotyping was performed on seven E. bieneusi strains for the first time in Tunisia. All the strains were isolated from stool samples of humans with immunodeficiency virus (HIV) infection. Analysis of the ribosomal RNA gene internal transcribed spacer (rDNA ITS) allowed the identification of three distinct genotypes previously described in other studies. Genotypes D and B were characterized in four and two respectively. The Peruvian genotype (Peru 8) was detected in the last isolate. These results indicate a genetic diversity in E. bieneusi strains from HIV Tunisian patients and suggest the coexistence of both zoonotic and anthroponotic route of transmission.

[Contribution of PCR for detection and identification of intestinal microsporidia in HIV-infected patients]. / Apport de la PCR dans la recherche et l'identification des microsporidies intestinales chez les sujets infectés par le VIH.

AIM: Intestinal microsporidiosis are among the most frequent opportunistic diseases in immunocompromised subjects. This study aimed to evaluate the contribution of PCR for a better detection and species identification of microsporidia in stool specimens of HIV-infected patients. PATIENTS AND METHODS: Stool samples obtained from 119 HIV-infected Tunisian subjects were screened for intestinal microsporidiosis by light microscopy using Weber's modified Trichrome stain and by a PCR method using universal primers V1/PMP2 which amplified a common fragment of the small subunit rRNA gene of microsporidia. The obtained PCR products were then sequenced using an ABI PRISM 377 DNA sequencer. RESULTS: The results showed a better sensitivity of PCR in the detection of microsporidia with an infection rate of 14.3% significantly higher than that of 6.7% obtained by light microscopy (p=0.03). As previously described, intestinal microsporidiosis was associated with low CD4 cell counts; 23.9% infection rate in patients having CD4 cell count under 200/mm(3) against 5.6% in patients with higher CD4 cell count (p=0.008). The sequencing of 15 out of the 17 positive PCR products has confirmed in all cases the species identified based on the PCR fragment size i.e., 250pb for Enterocytozoon bieneusi (seven cases) and about 270pb for Encephalitozoon intestinalis (nine cases); one case revealed a double infection. CONCLUSION: PCR proved to be more effective than classical Trichrome stain for the diagnosis of intestinal microsporidiosis. Moreover, the ability of PCR to identify the species involved could also be useful for cases management.

[Ileocecal ameboma: a case diagnosed by pathology and molecular biology]. / Amoebome iléocaecal: un cas diagnostique par l'anatomie pathologique et la biologie moléculaire.

Colonic ameboma is a rare benign inflammatory tumor due to the infection by Entamoeba histolytica and poses frequently the problem of colon cancer. We report a case of a 52 year-old patient who presented a cecal amoeboma revealed by a painful mass in the right iliac fossa. Radiologic and endoscopic examinations depicted a parietal thickening of the right colon and the cecum. A presumptive diagnosis of colon cancer was firstly discussed. Confirmation of ameboma was made on pathological examination, PCR and serology.

[Cryptosporidium infection in patients with major histocompatibility complex class II deficiency syndrome in Tunisia: description of five cases]. / La cryptosporidiose chez les enfants atteints de déficits immunitaires primitifs par défaut d'expression de protéines du complexe majeur d'histocompatibilité classe II en Tunisie : à propos de 5 observations.

BACKGROUND: In Tunisia, Cryptosporidium is frequently identified in diarrheic stools of children and immunocompromised patients. The infection is usually self-limited in immunocompetent populations, but can be severe and life-threatening in immunocompromised individuals. Cryptosporidiosis is well-documented in patients with the human immunodeficiency virus; however, few data are available concerning children with primary immunodeficiencies (PIDs). PATIENTS AND METHODS: A retrospective study was conducted on 5 cryptosporidiosis cases diagnosed in 11 children with PIDs. Cryptosporidium was systematically investigated when patients presented chronic diarrhea. Stool samples were examined for the parasite oocysts by modified Ziehl-Neelsen staining, and DNA was systematically extracted for a nested polymerase chain reaction (PCR). The species were identified by the analysis of restriction patterns. Epidemiological and clinicobiological data were presented for each patient. RESULTS: All cryptosporidiosis cases presented a CMH class II deficiency syndrome. Chronic diarrhea was associated with failure to thrive in all cases. PCR provided the diagnosis in all patients, while Ziehl-Neelsen staining revealed Cryptosporidium oocysts in only 3 cases. Species identification yielded Cryptosporidium hominis in 2 cases, Cryptosporidium meleagridis in 1 case, and Cryptosporidium parvum in 1 case; a C. hominis/C. meleagridis co-infection was observed in the last case. C. hominis was isolated in children from rural areas, suggesting that the infection could have been contracted in the hospital and thus a probability of nosocomial transmission. One of the C. hominis carriers developed sclerosing cholangitis with a high parasite load. CONCLUSION: Cryptosporidiosis with serious clinical symptoms is observed in PID patients, particularly those with CMH class II deficiency syndrome. Early, regular, and repeated screening, improved by PCR, is recommended in this group of patients. The predominance of C. hominis, the anthropophilic species, in children from rural areas should emphasize hygiene measures in care centers where PID cases are treated.

[Congenital toxoplasmosis following infection occurring late in pregnancy]. / Toxoplasmose congénitale faisant suite à une primo-infection maternelle en fin de grossesse.

Monthly serological screening of non immune pregnant women is recommended for prevention of congenital toxoplasmosis. However, this screening is often interrupted before delivery. We report a case of congenital toxoplasmosis following infection occurring late in pregnancy. This documented case highlights the need for a final routine serological test, 2-3 weeks post-partum for all seronegative pregnant women. In fact, the screening of congenital toxoplasmosis cases allows the early administration of specific treatment that avoids later severe complications such as chorioretinitis.

[Microsporidia and cryptosporidia coinfection in an HIV-infected newborn]. / Co-infection par des microsporidies et des cryptosporidies chez un nouveau-né infecté par le VIH.

Microsporidiosis and cryptosporidiosis are emerging opportunistic infections responsible for intestinal manifestations that are often severe in immunocompromised patients. A case of microsporidiosis-cryptosporidiosis coinfection is reported in an HIV-infected newborn. The patient was a 17-day-old female, exclusively breastfed and with no contact with animals. Microsporidiosis and cryptosporidiosis were diagnosed after systematic screening in stool samples using both specific staining and PCR. Two species of microsporidia, Encephalitozoon intestinalis and Enterocytozoon bieneusi, and Cryptosporidium hominis were identified. The contamination of the newborn probably resulted from direct human-to-human transmission during close contact with the mother (who had diarrhea and refused stool sampling). This report highlights the usefulness of the screening of intestinal microsporidiosis and cryptosporidiosis in HIV-infected subjects for better management.

Phlebotomus sergenti (Parrot, 1917) identified as Leishmania killicki host in Ghardaïa, south Algeria.

Since 2005, an outbreak of human cutaneous leishmaniasis (CL) in Ghardaïa, south Algeria, was studied and one output of these investigations was the identification of two Leishmania species, Leishmania major and Leishmania killicki, as the CL causative agents. In the present study, we were curious to focus on sand fly fauna present in this area and detection of Leishmania-positive sand fly females. Sand flies (3717) were collected during two seasons using sticky papers and CDC light traps in urban, rural and sylvatic sites. Twelve Phlebotomus species were identified. Phlebotomus papatasi was dominant in the urban site while Phlebotomus sergenti and Phlebotomus riouxi/chabaudi were dominant in the sylvatic site. Out of 74 P. sergenti females captured by CDC light traps in the sylvatic site populated by Ghardaïas' Gundi (Massoutiera mzabi), three ones were hosting Leishmania promastigotes. PCR-RFLP and sequencing of seven single-copy coding DNA sequences identified the promastigotes as L. killicki. Furthermore, laboratory experiments revealed that L. killicki isolate sampled from a CL patient inhabiting the studied region develop well in P. sergenti females. Our findings strongly suggest that the human cutaneous leishmaniases caused by L. killicki is a zoonotic disease with P. sergenti sand flies acting as hosts and vectors and gundi rodents as reservoirs.