Association between genital tract infection and premature rupture of membranes: A retrospective case control study in Tunisia, North Africa.

Premature Rupture of Membranes is responsible for most cases of neonatal death. In most of these cases, the causes of PROM have not been established in Tunisia, although several risk factors have been described. Therefore, we set out to determine the presence of an association between genital infections and PROM among Tunisian women. A case-control study was conducted among 251 womens to detect the presence of association between genital tract infection and Premature Rupture of Membranes.Cases had a premature membranes rupture and the controls had intact membranes or suffering from premature membrane rupture during the latent phase of labour. Data were collected from the medical register including socio-demographic characteristics, obstetrics, and medical history. Association between genital infections and premature rupture of membranes was estimated using the Odds Ratio and 95% CI. One risk factor was identified, including age. There is no association between the presence of Group B streptococcus (OR= 1.08; 95% CI 0.50-2.34), presence of Trichomonasvaginalis (OR= 2.45; 95% CI 0.15-39.83) and presence of Candidiasis (OR= 1.11; 95% CI 0.58-2.14) and premature rupture of membranes. Co-infection was not associated with premature rupture of membranes (OR= 0.43; 95% CI 0.45-6.07). There is no association between genital infections and PROM among pregnant Tunisian women.

[Evaluation of the measurement of capillary glucose concentration versus plasma glucose in the newborn]. / Fiabilité de la mesure de la glycémie capillaire par rapport à la glycémie plasmatique chez le nouveau-né.

BACKGROUND: The reliability of blood glucose monitoring in neonatology is not always confirmed. The aim of this study was to evaluate the reliability of blood glucose measurements made with three different devices in newborns. PATIENTS AND METHODS: The study was prospective, conducted in a medical and neonatal intensive care department over a period of 4 months. Capillary glucose level was measured with three different glucometers and compared with venous glucose level determined using the hexokinase method. An ANOVA and Scheffe test were used for the correlation analysis. RESULTS: Three hundred and nine infants were included, with a mean age of 55h and a mean term of 39 weeks of gestation. Mean blood glucose in the laboratory was 0.62±0.15g/L, 0.71±0.17g/L for Accu-Chek(®) Active, 0.80±0.17g/L for Accu-Chek(®) Performa, and 0.83±0.12g/L for Bionime. An ANOVA showed statistically significant differences between the measurements made by glucometers compared to the reference blood glucose levels, and the Scheffé method showed that glucometers overestimated the real plasma glucose levels. CONCLUSION: None of the devices used in this study was satisfactory. However, an estimation of blood glucose taking into consideration this numerical overestimation would allow early detection of hypoglycemia.

Molecular genotyping of Echinococcus granulosus using formalin-fixed paraffin-embedded preparations from human isolates in unusual tissue sites.

Cystic echinococcosis (CE) caused by Echinococcus granulosus remains a serious problem worldwide for issues relating to public health and the economy. The most predominantly affected sites are the liver and the lungs, but other organs such as the heart, the spleen and the peritoneum can also be infected. Access to cysts from uncommon sites has limited genomic and molecular investigations. In the present study, genotypes of E. granulosus sensu lato were identified from formalin-fixed paraffin-embedded tissues (FF-PETs) implicated in human CE. Tissue samples were obtained from 57 patients with histologically confirmed CE. DNA samples were analysed using Egss 1 polymerase chain reaction (PCR) specific to the mitochondrial 12S rRNA gene of E. granulosus sensu stricto. All cysts were typed as E. granulosus sensu stricto with up to 35% of the liver and 16.6% of lungs being the most frequently infected, and up to 48.4% of samples being from rare sites. No correlation was found between cyst site and either the gender or the age of patients. This study demonstrates the possibility of exploiting atypical cysts using FF-PET samples and highlights the predominance of E. granulosus sensu stricto species in the Tunisian population, even in unusual infection sites.

[Diagnostic performance of graphical anomalies in the detection of large platelets and platelet clumps]. / Performances diagnostiques des anomalies graphiques dans la détection des macroplaquettes et des agrégats plaquettaires.

UNLABELLED: Thrombocytopenia is a current situation for making a blood smear in routine practice in a medical analysis laboratory. Recent automated hematology analyzers enumerate platelets and generate histograms and specific flags. Operators must be aware of the characteristics of their analyzer in order to avoid spurious results in the case where microscopy review is not possible. OBJECTIVE: We evaluated the diagnostic performance of various graphical anomalies in the detection of large platelets and platelet clumps. PATIENTS AND METHODS: Three hundred cases of thrombocytopenia were included in the study on the basis of a platelet count less than 150 × 10(9)/L. This evaluation is expressed by the results of the sensitivity, specificity, positive predictive value and negative predictive value compared to the microscopic review of blood smear. RESULTS: Graphical performances are variable according to microscopic review of blood smears. Indeed, a not fitted curve is the most sensitive change on platelet histogram to the presence of large platelet. A high specificity to the presence of platelet clumps is announced when the platelet curve fails to return to the baseline. Moreover, characteristic findings on the DIFF scattergram are very specific to the presence of platelet clumps. CONCLUSION: A normal platelet histogram can validate with great confidence thrombocytopenia in cases where a blood smear cannot be read immediately.

Biting midges monitoring (Diptera: Ceratopogonidae: Culicoides Latreille) in the governate of Monastir (Tunisia): species composition and molecular investigations.

The results of entomological studies carried out in the governate of Monastir (Tunisia) in 2009-2010 (captures and emergences from muds) focusing on Culicoides species are presented in the present study. Identification of Culicoides at the species level is based on morphological characters, and a molecular study has been carried out based on mitochondrial DNA cytochrome C oxidase I gene (COI) and D1 and D2 domains of the 28S rDNA. The DNA sequences reported here are related to 10 species (on 25 known) of Culicoides described in Tunisia: Culicoides cataneii-gejgelensis, Culicoides circumscriptus, Culicoides imicola, Culicoides jumineri, Culicoides kingi, Culicoides langeroni, Culicoides newsteadi, Culicoides paolae, Culicoides puncticollis and Culicoides sahariensis. DNA sequencing of the COI gene and D1D2 domains discriminated all morphologically determined species. The choice of D1D2 domains considered as a conserved region is informative for Culicoides species identification. The molecular analyses of COI has grouped both C. circumscriptus, C. puncticollis within two clusters and C. newsteadi within five subclusters. However, C. newsteadi shows relatively deep intraspecific divergence using COI sequences.

[Visceral leishmaniasis in immunocompetent adults. About six cases]. / La leishmaniose viscérale de l'adulte immunocompétent. À propos de six cas.

OBJECTIVE: Studying the epidemiological variations of visceral leishmaniasis in Tunisia and proving the importance of parasitological investigations to raise the diagnosis. PATIENTS AND METHODS: Six patients hospitalised during the period between January 1998 and January 2009 at Fattouma Bourguiba Teaching Hospital in Monastir, five men and an only one woman, aged from 26 to 70 years old, originating from the central and eastern regions of the country. Epidemiological, clinical, biological and therapeutic data were obtained from the patient's medical files. RESULTS: The major clinical symptoms were fever, weakness and spleen enlargement. Biological data revealed the presence of anaemia in every case and leucopoenia associated or not associated with thrombopenia in four cases. The diagnosis of visceral leishmaniasis was confirmed by the identification of the parasite in the blood or in the bone marrow. All patients were treated with two courses of antimoniate of meglumine separated by a 6-week interval. The outcome was positive and the patients were cured. CONCLUSION: Visceral leishmaniasis is increasing among adults in Tunisia. Moreover, it is spreading outside its epidemiological area in the north to reach the central and southern regions. It should be raised when fever and spleen enlargement occur. Biological data are hardly specific. Diagnosis is based on finding the parasite in human fluids, mainly by molecular techniques. The rapid establishment of a specific treatment is vital.

Geographical distribution updating of Tunisian leishmaniasis foci: about the isoenzymatic analysis of 694 strains.

Over a period of ten years, a series of 694 Leishmania strains from Tunisian leishmaniasis foci were isolated and identified by isoenzymatic analysis. Strains were obtained from human cutaneous and visceral leishmaniasis in immunocompetent subjects, visceral leishmaniasis in imunocompromised individuals and from dogs with visceral leishmaniasis. Two classically dermotropic species, Leishmania (L.) major and Leishmania killicki were found. L. major with the single zymodeme MON-25 was the most isolated in cutaneous leishmaniasis foci of the Centre and South of Tunisia with a recent northern extension. L. killicki zymodeme MON-8 was sporadically found both in its classical microfocus of Tataouine in southeastern Tunisia as well as in some new foci in Southwestern, Central and Northern Tunisia. Leishmania infantum with its three zymodemes MON-1, MON-24 and MON-80 was isolated from both visceral and cutaneous human cases. The majority of L. infantum strains were found in the Northern part of the country; however, some strains were reported for the first time in the Southern part. L. infantum MON-1 was the only zymodeme isolated from canine leishmaniasis.

[Retrospective study of the distribution and the fertility of hydatid cysts in the child in Tunisia]. / Étude rétrospective de la distribution et de la fertilité des kystes hydatiques chez l'enfant en Tunisie.

PURPOSE OF THE STUDY: Our aim was to study the distribution and the fertility of the hydatid cysts in function of the age and the sex of patients and to identify the strain(s) responsible(s) of the children hydatidosis. PATIENTS AND METHODS: We have analyzed a total of 241 cysts coming from 195 children aged 2 to 16 years operated in the CHU F. Bourguiba of Monastir during the period from November 1999 to December 2009. For each cyst, the localization and the fertility of the métacestode as well as age, sex and origin of the patient are listed. Identification of strains was carried out by PCR/RFLP and has targeted the ribosomal gene ITS1. RESULTS: The lung was the primary localization of cyst (61.8%) followed by the liver (34.85%). The greatest number of cases is observed in the age groups 4-9 years (138 cases) where children's infection is more frequent in the male than in the female sex. The fertility of the cyst was independent of its site or its size and no incidence of age of children was detected. The G1 sheep strain is responsible for the contamination of children. CONCLUSION: The cystic echinococcosis described as a young adult disease may actually observed at any age and remains a serious problem of public health in Tunisia.

Usefulness of a PCR-based method in the detection and species identification of Leishmania from clinical samples.

The aim of this study is to assess the usefulness of a simple, low-cost method for the detection and species identification of Leishmania isolated by in vitro culture or detected directly from clinical samples. A total of 110 samples were used in this study. Among these, 21 were human and canine peripheral bloods, 63 skin lesion material samples, eight reference strains and 18 Leishmania culture. Detection of Leishmania DNA with PCR using primers designed to amplify the internal transcribed spacer 1 (ITS1) region of the rRNA gene proved sufficiently sensitive at the level of 0.1 parasites per PCR reaction. Furthermore, followed by single-strand conformational polymorphism (SSCP), the PCR-ITS1 allowed the species identification of Leishmania. The inter-specific polymorphism of Leishmania was first validated on reference strains, and then this method was applied on clinical samples and culture. Typing identified all human and canine visceral leishmaniasis samples (21 samples) as L. infantum, 95.23% of the cutaneous leishmaniasis samples as L. major and 3.17% as L. killicki and 1.58% as L. infantum. A scheme of the PCR diagnosis procedure for the detection and identification of Leishmania parasites is proposed in this study.

[Evaluation of the immunoblotting for the detection of immunoglobulin G Toxoplasma antibodies in immunocompetent patients]. / Apport de l'immunoblot dans la détection des IgG anti-toxoplasmiques chez le sujet immunocompétent.

UNLABELLED: The serological tests commonly used for the diagnosis of toxoplasmosis raise the problem of the interpretation of the borderline immunoglobulin G (IgG) levels and discordant results between various tests. OBJECTIVE: The purpose of our study was to evaluate the contribution of the immunoblotting in the detection of specific IgG in acquired toxoplasmosis of immunocompetent patients especially when levels are equivocal or discordant in enzyme-linked immunosorbent assay (Elisa) and indirect fluorescent antigen test (IFAT). MATERIAL AND METHODS: [corrected] We tested three groups of sera. The first included 87 positive sera, the second 33 negative sera, and the last one 29 equivocal sera. RESULTS: Results obtained with the first and the second group of sera led us to identify the bands 30kDa and 32kDa as markers of the toxoplasmic infection. The simultaneous presence of both bands showed a sensitivity of 91.5%, a specificity of 96.9%, a VPP of 98.7%, a VPN of 74.4% and a Youden's index of 0.88. Our findings suggest that the presence of these two bands is a reliable criterion for the confirmation of the presence of anti-toxoplasmic IgG in the corresponding serum. The immunoblot allowed us to ascertain serological status of 27 (93.1%) patients from the third group in which results were discrepant or equivocal in Elisa and/or in IFAT. CONCLUSION: Immunoblot is a useful serological test for detection of very low or equivocal titers.

Use of PCR, IFAT and in vitro culture in the detection of Leishmania infantum infection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia.

The aim of this study was to assess the use of parasitological, serological and molecular methods for the detection of Leishmania infection in blood of 67 dogs and to investigate the prevalence of canine leishmaniasis (CanL) in Kairouan (central Tunisia), an area known to be of reduced endemicity and has not been studied since 1973. Veterinarians clinically examined all dogs, and the titer of anti-Leishmania antibodies was determined by indirect immune-fluorescence antibody test. The presence of Leishmania was performed by PCR and in vitro culture. IFAT was positive in 12% of dogs and promastigote form of the parasite was isolated by in vitro culture from only 4.5% of them. However, DNA of Leishmania was detected by PCR in 20.9% of dogs. PCR was more sensitive than IFAT (p = 0.004) and in vitro culture (p < 10(-5)). A prevalence of 21% was found in Kairouan, which is significant high (p < 10(-3)) when compared to that of thirty years ago. This state is in correlation with the increase in other Mediterranean countries. Furthermore, 50% of positive dogs were asymptomatic. Preventive measures must be taken against these dogs as for symptomatic ones since their role in the transmission of the infection to vectors has been proven.

The phlebotomine fauna (Diptera: Psychodidae) of the eastern coast of Tunisia.

To identify the phlebotomine sand fly populations of the eastern coast of Tunisia, an entomological survey was carried out between September and October 2005 at 71 sites located in three districts. CDC light traps and sticky papers were used to collect a total of 2,138 phlebotomine sand flies representing nine species. The predominant species occurring on the eastern coast of Tunisia are, in order of abundance, Phlebotomus longicuspis Nitzulescu, 1930 (40%); Phlebotomus papatasi Scopoli, 1786 (21%); Sergentomyia minuta Parroti Adler & Theodor, 1927 (19%); Phlebotomus perniciosus Newstead, 1911 (9.5%); Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 (9%); Sergentomyia fallax Parrot, 1921 (1.5%); Sergentomyia dreyfussi Parrot, 1933 (0.23%); Phlebotomus langeroni Nitzulescu, 1930 (0.05%); and Phlebotomus perfiliewi Parrot, 1930 (0.05%). Species involved in the transmission of Leishmania, namely P. papatasi and P. perniciosus, represent 31% of the total number of flies captured. In the central sites (district of Monastir), P. longicuspis predominates, P. perniciosus predominates in the northern sites (district of Sousse) and P. papatasi in the southern sites (district of Mahdia), which is consistent with the distribution of Leishmania infantum and L. major in this region. Analysis of the degree of presence (D) revealed that Phlebotomus papatasi was the most common species and showed the broadest distribution (D = 95%), followed by P. longicuspis and S. minuta parroti (D = 90%) and P. perniciosus and P. chabaudi (D = 86% and 68% respectively).

[Epidemiology of Leishmania (L.) infantum, L. major and L. killicki in Tunisia: results and analysis of the identification of 226 human and canine isolates]. / Epidémiologie de Leishmania (L.) infantum, L. major et L. killicki en Tunisie: résultats et analyse de l'identification de 226 isolats humains et canins et revue de la littérature.

The epidemiological situation of leishmaniasis in Tunisia is characterised by the co-existence in a very limited territory (165,000 km2, Sahara included), of 4 clinical forms: the infantile visceral leishmaniasis (VL) and 3 cutaneous leishmaniasis (CL) forms (sporadic, zoonotic and chronic). In addition to the useful epidemiological data, identification of the causative parasitic species is essential to determine the geographic distribution of each form and to select appropriate therapeutic procedure and suitable control measures. 226 Leishmania isolates, 135 human's coming from 59 VL cases and 76 CL cases and 91 canine's were identified by the isoenzyme electrophoresis reference technique. Results confirm the endemicity of the 4 forms mentioned above. The sporadic CL, confined to the North of the country is principally caused by L. infantum MON-24 (72.2%). VL which has reached the southern ridge in the central area of Tunisia, in the governorate of Kairouan (36 typed isolates), presents an unusual high proportion of L. infantum MON-24. In fact, this zymodeme, rather dermotropic is responsible for 47.2% of the cases vs 13% in the other regions of the country where L. infantum MON-1 remains predominant with 78.3% of typed isolates, the difference being statistically significant (P < 0.01). A third zymodeme, L. infantum MON-80 is sporadically pointed out during VL or in sporadic CL. Despite the high number of canine isolates (n=91) coming from 6 governorates, only the zymodeme L. infantum MON-1 was identified, letting hypothetic the reservoir of the 2 other zymodemes of the species identified in humans. Those absences may be related to cross infections, with a low sensitivity to L. infantum MON-24 leading to a selection of MON-1 at the time of culture passages. Hence it is important to develop molecular tools of direct identification on initial biological samples without going through cultures. Zoonotic CL remains the predominant cutaneous form in the central and southern area of Tunisia. However L. killicki, agent of the chronic CL, is confirming its presence out of its original focus of Tataouine in the southern-east of the country in both zoonotic CL and VL areas.

Pediatric visceral leishmaniasis diagnosis in Tunisia: comparative study between optimised PCR assays and parasitological methods.

There has been a steady increase of visceral leishmaniasis during the past 20 years in Tunisia. In this study, we assess the value of two optimised PCR versus those of classical methods for the diagnosis of human visceral leishmaniasis. 106 samples were collected from 53 cases of pediatric visceral leishmaniasis. Peripheral blood and bone marrow samples were analysed both by parasitological methods (direct examination, leukocytoconcentration (LCC) and culture) and by PCR methods with two primer pair (R221/R332 and Lei 70L/Lei 70R). We diagnosed visceral leishmaniasis in all patients: 44 cases were diagnosed by culture (83%), 42 by direct examination of bone marrow (79%), 17 by LCC (32%), and 53 positive cases with both PCR assays (R221/R332 and/or Lei 70L/Lei 70R) (100 %). Regarding each PCR assay, for blood samples, the difference between the sensitivities of PCR Lei 70L/Lei 70R (86,8%) and PCR R221/R332 (17 %) is statistically significant with p-value 0.025. For bone marrow, the sensitivities of the two PCR methods were respectively 96,2% (Lei 70L/Lei 70R) and 75,5% (R221/R332). On the whole, PCR Lei 70L/Lei 70R was more effective than PCR R221/R332 and conventional methods for the two biological samples. Moreover, the requirement of less invasive sample using blood has the advantage of being repeatable for screening and for post therapeutic monitoring.

[Cutaneous leishmaniasis caused by Leishmania infantum MON-24 in Tunisia: extension of the focus to the center of the country]. / La leishmaniose cutanée due a Leishmania infan- tum MON-24 en Tunisie: extension du foyer vers le centre du pays.

Three clinico-epidemiological forms of cutaneous leishmaniasis (CL) were described in Tunisia: the zoonotic CL (ZCL) epidemic which occurred in the centre of the country caused by Leishmania major MON-25, the chronic CL (CCL) In the south-east of the country caused by Leishmania killicki MON-8 and the sporadic CL In the North (SCL) caused by Leishmania infantum MON-24. The latter form, described in 1991, prevails in northern Tunisia with approximately thirty cases per year. Its vector, unknown for a long time could be according to the last publications, Phlebotomus perfiliewi or Phlebotomus langeroni; however, its reservoir remains unknown until now. The systematic isoenzymatic characterization permits to identify a great number of strains improving then knowledge on the eco-epidemiology of the disease. Indeed, changes were noted in the geographical distribution of these clinical forms: extension of the ZCL to the North and South, extension of the CCL to North and the SCL to the centre. We report in this note the first mention of L. infantum MON-24 in the two provinces of the centre of Tunisia: Kairouan and Sidi Bouzid, confirming the extension of the SCL to the Centre.

Isoenzymatic variability of Leishmania infantum in Tunisia concerning 254 human strains.

The different clinical forms of leishmaniasis are the result of both the immunological status of individuals and the species of the parasite causing the infection. In Mediterranean countries, the Leishmania infantum complex groups zymodemes which are responsible for visceral, cutaneous and exceptionally cutaneomucosal or mucosal leishmaniasis. We report in this study a synthesis concerning 254 cases of L. infantum that have been characterized at the "Laboratoire de Parasitologie" of the Rabta Hospital. The strains were isolated from human cases of visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) by culture on NNN medium: 156 VL cases and 98 CL cases. The isoenzymatic characterization revealed three zymodemes of L. infantum. * L. infantum MON 1, a common zymodeme of VL,occurred in 154 cases (61%): 147 VL (95%) and 7 CL (5%). All CL cases were from the northern provinces, six of them occurring during an epidemic disease in 2001. * L. infantum MON 24, a common zymodeme of CL in the north, occurred in 98 cases (38.5%): 91 CL (93%) and 7 VL (7%). The seven VL cases were immunocompetent children aged from 8 months to 9 years and native of northern Tunisia. Two of the CL cases were from central regions of the country. This is the first time that cases from these regions are reported. * L. infantum MON 80, an uncommon zymodeme in Tunisia, occurred in two VL cases (0.5%): two children aged 7 and 5. The small number of strains of this zymodeme does not allow understanding of its epidemiological role. The results of this study indicate a low enzymatic variability of L. infantum in the country. However, our study includes only human strains and should be extended to animal ones (dogs, rodents and sand flies). This would lead to a better understanding of the epidemiology of leishmaniasis in Tunisia.

Post-surgical follow-up (by ELISA and immunoblotting) of cured versus non-cured cystic echinococcosis in young patients.

The study was designed to determine comparatively the prognostic value of immunoblotting and ELISA in the serological follow-up of young cystic echinococcosis (CE) patients exhibiting either a cured or a progredient (non-cured) course of disease after treatment. A total of 54 patients (mean age 9 years, range from 3 to 15 years) with surgically, radiologically and/or histologically proven CE were studied for a period up to 60 months after surgery. Additionally, some of the patients underwent chemotherapy. Based on the clinical course and outcome, as well as on imaging findings, patients were clustered into 2 groups of either cured (CCE), or non-cured (NCCE) CE patients. ELISA showed a high rate of seropositivity 4 to 5 years post-surgery for both CCE (57.1%) and NCCE (100%) patients, the difference found between the two groups was statistically not significant. Immunoblotting based upon recognition of AgB subcomponents (8 and 16 kDa bands) showed a decrease of respective antibody reactivities after 4 years post-surgery. Only sera from 14.3% of CCE patients recognized the subcomponents of AgB after 4 years, while none (0%) of these sera was still reactive at 5 years post-surgery. At variance, immunoblotting remained positive for AgB subcomponents in 100% of the NCCE cases as tested between 4 and 5 years after surgical treatment. Immunoblotting therefore proved to be a useful approach for monitoring post-surgical follow-ups of human CCE and NCCE in young patients when based upon the recognition of AgB subcomponents.

[Molecular and morphological characterisation of two closely related species belonging to the subgenus Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 et P. riouxi Depaquit, Killick-Kendrick & Léger, 1998 (Diptera: Psychodidae)]. / Caractérisation moléculaire et morphologique de deux espèces affines de Paraphlebotomus: Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 et P. riouxi Depaquit, Killick-Kendrick & Léger, 1998 (Diptera: Psychodidae).

In 1998, the description of Phlebotomus riouxi emphasised the difficulty to differentiate the female from the closely related P. chabaudi, a suspected vector of Leishmania killicki in several foci in Tunisia. In order to be able to distinguish the females of these two species, we started a molecular study based on 37 Algerian and Tunisian specimens. The alignment of the sequences of the cytochrome b of the mitochondrial DNA and their analysis using Neighbor-Joining, maximum likelihood and maximum parsimony shows the individualisation of two species including an intraspecific variability. Following a morphological approach, it is not possible to distinguish the females on the basis of their spermathecae. A new character is proposed: the presence of anterolateral teeth of the pharyngeal armature for P. chabaudi, never observed in P. riouxi. However, a molecular typing is necessary at the present time for a sure identification of the females.

Leishmaniasis in central and southern Tunisia: current geographical distribution of zymodemes.

The authors report the identification of Leishmania strains isolated from the Centre and the South of Tunisia. 266 strains were isolated between 1998 and 2006 from human (n=221 strains) and dogs (n=45 strains) hosts. The isoenzymatic identification exhibits the presence of in total five zymodemes belonging to three Leishmanio complexes: Leishmania infantum, L. major and L. killicki. All strains isolated from human and canine visceral leishmaniasis belonged to L. infantum. zymodeme MON-1 was the only one isolated from canine visceral leishmaniasis. However, it is predominant in human visceral leishmaniasis beside zymodeme MON-24 which was detected in two provinces of the Centre (Monastir and Kairouan) and zymodeme MON-80 isolated for the first time in Kairouan province. Three complexes are responsible for human cutaneous leishmaniasis: L. major MON-25 is the parasite the most frequently found in its classic foci in the Centre and the South of the country. L. infantum MON-24 was isolated for the first time in a small locality of Sfax (southern Tunisia) showing the appearance of a new focus of L. infantum. L. killicki was isolated in its original focus of Tataouine and in two new foci of the central part of the country (Sidi Bouzid and Kairouan).

Increase of canine leishmaniasis in a previously low-endemicity area in Tunisia.

An epidemiological study of canine leishmaniasis (CanL) was carried out in nine districts of Sfax, in the southern central part of Tunisia. Sera from 250 dogs were tested by two serological methods: the indirect immunofluorescence antibody test and the counter-immunoelectrophoresis. Seven to eight months later, before the next season of transmission, seropositive dogs from the first test were re-examined and a second sampling was performed. Infection status was assessed by serology and by other methods. PCR, in vitro culture and direct examination were applied on blood and other samples (bone marrow, liver, lymph node, spleen and cutaneous biopsies). The seroprevalence of the infection in dogs was 6%. Infection was then confirmed by at least one other method. The PCR is the method which agreed most with serology, all seropositive dogs were found PCR-positive. The sensitivity of the direct examination and the culture was only 33% and 55% respectively as compared with serology. A similar value of seroprevalence has been observed previously in Sousse, in the northern central part of Tunisia. The present report suggests a significant increase of CanL in the Sfax area and confirms that the disease is continuing to move southwards in Tunisia.