First field evidence infection of Culex perexiguus by West Nile virus in Sahara Oasis of Algeria.

BACKGROUND & OBJECTIVES: West Nile virus (WNV) is considered one of the most widely distributed arboviruses in the world which is transmitted by several mosquito species including the Culex genus. Culex pipiens is the major vector of this virus in Europe and USA whereas in African countries, other species such as Cx. perexiguus is considered as an important vector. This paper aimed to study the mosquito species involved in WNV transmission in Aougrout, one of the highly populated Oasis of Timimoun Province in Algeria where human WNV neuroinvasive diseases are prevalent. METHODS: CDC light-traps were installed in animal and human shelters for three nights. Collected mosquitoes were pooled and real-time PCR was performed to detect and identify WNV lineages 1 and 2 in the samples. Results: CDC light-traps collected 270 mosquitoes belonging to three genera. Culex genus was predominant with Cx. perexiguus as main species followed by Aedes and Anopheles genus. A total of 33 pools were tested; one pool containing Cx. perexiguus was found positive for WNV lineage 1. INTERPRETATION & CONCLUSION: This study reports for the first time a WNV natural infection of Culex perexiguus in the study region indicating that species other than Cx. pipiens should be taken into consideration in WNV surveillance, especially in specific environments like Saharan Oasis ecosystem.

[Contribution of nested PCR in the diagnosis of imported malaria in southern Algeria]. / Apport de la PCR nichée dans le diagnostic du paludisme d'importation dans le Sud algérien.

The nested PCR was used to estimate its inputs in malaria diagnosis and in the performance of the microscope operators involved in the surveillance of malaria in remote areas of South Algeria. For the period 2010 to 2015, 112 patients (93 febrile and 19 asymptomatic) coming from sub-Saharan Africa were tested for malaria in the hospital of Tamanrasset. One part of the blood taken from fingertip was used for blood smears and the second part was absorbed in filter paper for molecular diagnosis. Overall, the infection was detected by nested PCR in 63 samples versus 53 by direct examination. In addition, 11 mixed infections and 6 positive asymptomatic cases not detected by microscopy were diagnosed by PCR. Moreover, two negative samples in nested PCR were tested positive by direct examination. The molecular tool is more sensitive than the direct examination in detecting infra-microscopic parasitaemia and mixed infections...

First report of Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Oran, West of Algeria.

The increasing globalisation of trades, human movements and environmental changes facilitate the introduction and the establishment of the invasive Asian tiger mosquito Aedes albopictus outside its native geographical area. Alerted by the complaints about mosquito biting which occurred daytime for the inhabitants of the seaside town Ain Turk (West of Algeria), an entomological survey was conducted in December 2015 to determine the origin of this nuisance. Among the collected mosquitoes, specimens of Ae. albopictus (2 males, 3 females and 3 pupae) were collected. This is the first observation of that invasive mosquito in the west of Algeria which confirms its presence and establishment in Mediterranean Africa.

[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.

[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.

[Demonstration of the Leishmanicidal activity of Stachys guyoniana against Leishmania major in eastern Algeria]. / Mise en évidence de l'activité antileishmanienne de Stachys guyoniana contre Leishmania major dans l'Est algérien.

The authors report the results of the assessment of the Leishmanicidal activity of a plant species of the Lamiaceae family, Stachys guyoniana. This in vitro study used the species endemic in Algeria, Leishmania major. Both the butanolic and ethyl acetate extracts showed strong activity against this strain.

Entomological investigation following the resurgence of human visceral leishmaniasis in southern Algeria.

Visceral and cutaneous leishmaniasis are the main endemic vector born diseases in Algeria. In the Hoggar region (extreme south of the country) human visceral leishmaniasis (HVL) is known to be sporadic but during the last decade the number of cases has increased significantly. In 2010, a peak of HVL cases was registered mostly among children. Therefore an entomological survey and a retrospective study on HVL cases were carried out in order to explore the transmission of the disease. Among the sand fly caught Phlebotomus bergeroti was the most frequent species (68%) followed by Sergentomyia schwetzi (22%). In this work we describe the presence of Phlebotomus (Paraphlebotomus) kazeruni for the first time in the Hoggar region.

Anticancer activity of chemically prepared shrimp low molecular weight chitin evaluation with the human monocyte leukaemia cell line, THP-1.

In the present study, anticancer activities of chitin, chitosan and low molecular weight chitin were evaluated using a human tumour cell line, THP-1. A molecular weight-activity relationship and an electrostatic interaction-activity relationship were determined. The cytotoxic effects of chitin and derivatives were also evaluated using a normal human foetal lung fibroblastic cell line, MRC-5 and the specific cytotoxicity of chitin and derivatives to tumour cell lines was demonstrated. The high antitumour effect of low molecular weight of chitin was established.

[Leishmania infantum MON-1 isolated from a golden jackal (Canis aureus) in Grande Kabylie (Algeria)]. / Leishmania infantum MON-1 isolé d'un chacal doré (Canis aureus) en Grande Kabylie (Algérie).

In the north of Algeria, Leishmania infantum is responsible for two clinical forms of leishmaniasis: visceral and cutaneous leishmaniasis, for which dogs are the only proven reservoir host. In this study, the authors report, for the first time, the isolation of L. infantum from a golden jackal (Canis aureus) trapped in the Illoulen ou Malou region (Grande Kabylie). Two isolates were thus obtained from bone marrow and spleen and were identified by starch gel isoenzyme electrophoresis as L. infantum MON-1, the widespread zymodeme in the north of the country. Leishmania parasites have also been detected by polymerase chain reaction (PCR) in the biopsy of the spleen. The golden jackal, a prevalent wild canid in Northern Africa, could play a predominant role in the sylvatic foci of leishmaniasis and in the dissemination of the parasite in this region.

[Spread of Leishmania major to the north of Algeria]. / Extension de Leishmania major au nord de l'Algérie.

Since a long time, Leishmania major and L. infantum foci in Algeria were geographically separated by the mountains of the Tell Atlas which represent a natural barrier. Recently, a new focus of cutaneous leishmaniasis (CL) has emerged in the village of El M'hir, located on the north side of the chain of the Tell Atlas, in the basin of the Soummam. During the period 2004-2010, 152 CL cases have been registered and 12 isolates were obtained from patients who declared never having been outside the village the last years. The identification of the parasites showed that all strains belonged to L major MON-25. Investigations on the reservoir hosts showed the presence of the sand rat (Psammomys obesus), for the first time, in this locality. Five strains isolated from this rodent belonged to L. major MON-25. The sand rat, which is usually observed around the chotts in the Saharan and steppe areas, acts as the main reservoir of L. major in Algeria. Its presence in the new focus of El M'hir is reported for the first time. Entomological surveys carried out in 2009 showed the predominance of two sandfly species: Phlebotomus papatasi and P. perniciosus. The first one is known as a vector of L major in the Algerian Sahara. This study highlights the spread of L. major from the arid zones towards the semi arid areas, particularly in the Soummam valley. Climate changes and desertification observed in the steppe area northern Sahara could play a role in the extension of the disease.

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.

Canine Leishmania infantum enzymatic polymorphism: a review including 1023 strains of the Mediterranean area, with special reference to Algeria.

This bibliographic review reports the isoenzyme polymorphism of 1023 Leishmania infantum strains isolated from dogs that have been characterized by multilocus enzyme electrophoresis in the Leishmania Reference Centre of Montpellier, or in other laboratories, to which this typification technique has already been transferred. Between 1981 and 2010, a total of 12 zymodemes were identified around the Mediterranean basin: MON-1, MON-24, MON-34, MON-72, MON-77, MON-80, MON-98, MON-105, MON-108, MON-199, MON-199 var NP1130 and MON-281, of which 6 were present in Algeria. The zymodeme MON-1 was predominant (86.5% of the strains). The dog was confirmed as the main reservoir of L. infantum MON-1, while the reservoir of the other zymodemes has not yet been identified. The enzymatic polymorphism is relatively high in Algeria and in Spain in contrast to other Mediterranean countries. The reasons for this polymorphism are discussed.

Increase in the prevalence of canine leishmaniasis in urban Algiers (Algeria) following the 2003 earthquake.

Between 2005 and 2008, a serological survey for leishmanial infection was conducted among dogs from urban and peri-urban Algiers, with the focus on the new, densely populated areas that were built after the 2003 earthquake. Serum samples were collected from 1810 animals and tested for the presence of leishmanial antibodies by IFAT, ELISA and western blotting. The overall seroprevalence recorded was 25.1%. Of the seropositive dogs, 58.8% showed no clinical signs of the disease, 25.8% had a few, minor signs and the remaining 15.4% showed more severe illness. The major clinical signs of infection were weight loss, skin lesions and lymphadenopathy. Although seropositive dogs were found in all of the boroughs (daïras) of Algiers, seroprevalences were highest in the western part of the city (i.e. in the boroughs of Bouzaréah, Chéraga and Zéralda), ranging from 23.0% to 44.5%. Statistical analysis showed a relationship between seropositivity for leishmanial infection and the dog's age and lifestyle (i.e. whether the dog lived outside and/or in areas with dense vegetation). Only two zymodemes were identified amongst the 50 isolates investigated: MON-1 (88%) and MON-281 (12%). The latter zymodeme has not been previously found in Algeria, sandflies or dogs.

[Malaria in Algerian Sahara]. / Le paludisme au Sahara algérien.

Thanks to the malaria eradication campaign launched in Algeria in 1968, the number of malaria cases fell down significantly from 95,424 cases in 1960 to 30 cases in 1978. At that time the northern part of the country was declared free of Plasmodium falciparum. Only few cases belonging to P. vivax persisted in residual foci in the middle part of the country. In the beginning of the eighties, the south of the country was marked by an increase of imported malaria cases. The resurgence of the disease in the oases coincided with the opening of the Trans-Saharan road and the booming trade with the neighbouring southern countries. Several authors insisted on the risk of introduction of malaria or its exotic potential vectors in Algeria via this new road. Now, the totality of malaria autochthonous cases in Algeria are located in the south of the country where 300 cases were declared during the period (1980-2007). The recent outbreak recorded in 2007 at the borders with Mall and the introduction of Anopheles gambiae into the Algerian territory show the vulnerability of this area to malaria which is probably emphasized by the local environmental changes. The authors assess the evolution of malaria in the Sahara region and draw up the distribution of the anopheles in this area.

[Imported dracunculiasis: four cases confirmed in the south of Algeria]. / La dracunculose d'importation: quatre cas confirmés dans le sud algérien.

The authors report for the first time, in the south of Algeria, four imported cases of dracunculiasis in Malian Touaregs. The patients came from Mali. They remained two months in Tamanrasset before settling in Illizi where they have been working illegally for more than nine months. In August 2007, because of infected lesions appearing on their legs a month earlier, the patients decided to go to the Illizi hospital for treatment The diagnosis of dracunculiasis was made by direct observation of worms emerging from lesions and by microscopic examination of larvae. The patients were then isolated and treated. The Ministry of Health immediately took measures to avoid local transmission of the disease, never seen in the region. The measures consisted in increasing awareness campaign among medical and paramedical staff about dracunculiasis and in pointing out the importance of detection of new cases among nomadic populations. The risk of emergence of a new dracunculiasis indigenous focus in the south of Algeria should be seriously considered, especially as the number of immigrants from endemic countries is increasing considerably

[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.