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Microbiology & Humanities Sciences

Team 12: Idir Bitam, CR

 

Team composition

 

  1. Idir BITAM, MCF
  2. Slim BENYACOUB, PU
  3. Nadjet MOUFFOK, PU (Alger)

 

Vectorized bacterial diseases are common in Algeria thus they remain underestimated. These diseases have an epidemiology directly dependent on the geographical location but no comprehensive work has been made in Algeria. The person in charge of this young team project received a formation of medical entomology at the Pasteur Institute and carried out his thesis in Marseilles in the WHO collaborative centre for the vectorized bacterial diseases. Pr MOUFFOK Nadjet received a formation in the field of the infectiology in Marseilles and is an expert in infectious illness in Algeria. Pr BENYACOUB Slim has expertise in the field of the birds. Working in some of the extreme areas in Algeria, he is capturing and identifying birds and then collecting the various ectoparasites.

 

The various elements of work will comprise:

 

Firstly, a collection of the arthropods likely to be vectors of human pathogens. Secondly, at the laboratory of Algiers will be set-up the techniques for the identification of arthropods. Thirdly, the installation of breeding facility for the arthropods representing the most important risks in Algeria. Dr. BITAM will coordinate the collection of the hard ticks on the cattle and the dogs, and on the whole of the territory with correspondents in El Tarf, Sétif and Oran. As for soft ticks, no work was completed for the moment. A work in collaboration with Dr. Jean François TRAPE will be set up to collect the soft ticks in the rodents' burrows initially at the Moroccan border then in other places. A coordination will be done with people of El Tarf (Pr. BENYACOUB) for the collection of the soft ticks. Dr. Bitam will collect in collaboration with the veterinary services and departments of health in Oran and Algiers, the rodents and other wild mammals in zones suspect of plague like the rural areas. As for the collections of lice, the young team is of connection with the services of the cribs to collect the lice of hair and with the mental health departments and the Algerian Red Crescent to collect the lice of the body of the homeless individuals. Concerning the clinical part, Pr MOUFFOK will collect blood and skin biopsies of patients with a presomptive diagnosis of Rickettsioses, Bartonelloses and Borrelioses. These samples will be conveyed initially at the Institute Pasteur d' Algérie where they will be treated (first tests serologic and PCR), in the second time the positive samples will be sent to corresponding Unit IRD for the confirmation of the cases.

 

  • Arthropods Identification:

The identification of the arthropods will be done through the usual taxonomic keys as long as the establishment of techniques of molecular biology will not have been carried out. The molecular identification will be done initially in collaboration with the team of Marseilles.

 

  • Repertory of the bacteria present in the arthropods in Algeria

We began this work of repertory on the hard ticks, among which we already identified three rickettsia, in four species of ticks with the first description of the relation between Rickettsia aeschlimannii and Hyalomma specific aegyptium tick of tortoise (Bitam et al., 2006a. Bitam et al., 2009). We identified a Lyme disease group Borrelia with unknown pathogenicity  of the type Borrelia lusitaniae, we will continue to thus supplement the repertory in this field. In addition, we will systematically seek for Coxiella, Ehrlichia, Anaplasma and Bartonella. For the moment our search for Bartonella in the ticks appeared unfruitful. For the fleas, we already collected those of the hedgehogs in which we identified Rickettsia felis and we made the first description of the relation between Rickettsia felis and Archeopsylla erinacei, chip specific of the hedgehog (Bitam et al., 2006b). We will go on our work on Bartonella including one species specific to Algeria (Bartonella eldjazairii) which is under description (four sequences of the new genotype are published in GenBank). The fleas are used as extremely important indicator to detect the cases of plague, in particular on the wild rodents. A work of monitoring was completed in the localities where there were the epidemics (Oran, Mascara and Ain Temouchent and recently in Laghouat), the results of this work show the presence of the infectious agent in the fleas collected in these zones (Bitam et al., 2006c. Bitam et al., 2010). For the lice, the samples which we collected for the moment did not allow to highlight disease-causing agents directly on the lice, but it is known that the lice can transmit the recurring borrelioses, the typhus and the trench fever due to Bartonella quintana. For the soft ticks, overall their disease-causing agents are ignored in the whole world. They are known however in Africa as being the vectors of a frequent disease in Sahelian Africa, which is the recurring borrelioses with ticks (Trappe JF et al., 2007). This disease described forever in Algeria but its presence at the Moroccan border lets suppose that it could emerge in Algeria. Coxiella burnetii, the agent of the fever Q is also found in the soft ticks. This disease appears frequent in Algeria, in particular according to work which was completed in Sétif (Lacheheb et al., 2009). Other disease-causing agents are sought by more general techniques of amplification making it possible to discover new micro-organisms.

 

  • Evaluation of infection in humans

The person in charge of the young team drew up a network with infectious disease physicians in Algeria, in particular in Oran. Dr. Nadjet Mouffok is associated with the project; she described the Mediterranean Spotted Fever in Algeria and showed that it was an extremely frequent infection in Oran including the fatal form (Mouffok et al., 2009). A work of evaluation of the factors of gravity in the Mediterranean Spotted Fever is in hand with the team of Marseilles. In addition, the team of infectiologists of Laghouat (Algeria of the south) is interested by a work on the zoonoses following an alarm on a problem of plague and this team is of agreement to begin a collaboration with the teams of Marseilles and Algiers for the monitoring of these pathologies. It is to be announced that cases of human plague were diagnosed in Algiers and were confirmed in Marseilles, and a publication is currently in hand (subjected to Emerging infectious diseases). The monitoring can be carried out, according to the same methods as those adopted by Dr. Mouffok with taking away carried out in agreement with the local ethics committees among all feverish patients (fever of unexplained etiology) to study, with serological microarrays, the all pathogens identified in the arthropods prevalent in Algeria. This work will make it possible to evaluate the public health problems which the bacterial diseases transmitted by the ticks in Algeria represent.

 

  • The forward programme

First part: Reinforcement of the networks with the clinicians. Collect arthropods and micro mammalian. Identification of the arthropods.

Second part: Installation of the molecular techniques for the identification of all the pathogens detected in these arthropods and micro mammals. Installation of serological techniques for the diagnosis of the patients hospitalized in our collaborators' department in Oran and Algiers. Molecular diagnosis starting from the cutaneous biopsies on the patients. Organization of a workshop of formation intended for the health professionals in the field of bacterial research in infectious illness to Algeria; this workshop will be organized at the Algeria Pasteur Institute in collaboration with the URMITE.

Third part: Consolidation and creation of an official network of monitoring of the emergent infectious diseases in Algeria, and possibly creation with the local health authorities of a micro association of health monitoring in Algeria which will include all partners of this multi-field. One day organization international which will make it possible to expose our results obtained at the time of this project in front of an assistance including/understanding the local and foreign scientists. This will also make it possible to make exchanges and the development of the various techniques of diagnosis between the various researchers present and guests. The scientific publications will be made in international, peer-reviewed journals. According to the results obtained and the organization, the young team will have an international extension between Algiers and Marseilles.

This project consists of a partnership between various specialists in various fields including medical entomology, medical bacteriology, veterinary surgeons, the clinicians and nurses, and biologists. This project is organized in four sectors sets of themes. Each one of them corresponding to a working group. In each work group, the tasks are shared among the participants of the various fields. To be done, a domestic network split in areas will be installed and of the persons in charge will be indicated in each area at the risk where they will collect data of all the participants in all the fields (entomologist, veterinary surgeons, doctors, biologists, epidemiologists, agronomists) whose tasks will be done as follows:

  • Biologists and ecologists: they will study the effect of the vegetation and the micro_climate on the density of the ticks and fleas.
  • The clinicians, doctors and nurses will collect clinical data and blood samples and cutaneous biopsies.
  • The veterinary surgeons and entomologists will facilitate animal studies and animal facilities.

 

Fields of action

 

The first sector will be devoted to the study of the bioecology of the ticks, lice and fleas. The role of the ticks and fleas was proven by the various research tasks on the presence of the bacteria in the ticks, fleas and lice (Beaucournu et al., 1985; Parola et al., 2003, 2001, 2005; Raoult et al., 2001; Perez-Eid et al. 1998; Houhamdi et al., 2005; Davoust et al., 2006; Rolain JM et al., 2003; Loftis et al., 2006; Trap door et al., 2007; Zhioua, 1999).

  • The regular collection of the ticks and fleas will be carried out with the regional partners and will be carried out by taking account of the development cycle of the ticks.

The second sector will study:

  • The prevalence of the infection of the ticks by Borrelia, Rickettsia and Ehrlichia, Coxiella and Bartonella, will be determined by methods of Immunofluorescence and PCR.
  • The pathogenic ones will be identified on the level of the species by using targets for Borrelia (intergenic spacer rrf-rrl and the gene ospA), targets for Rickettsia (primers comprising a fraction of citrate gene synthase RP CS. 877p and RP CS.1258n and Omp have Rr190.70p and Rr190.701n) and using URB1 and URB2 primers to amplify ftsZ gene to seek for Bartonella.

The third sector will ensure the eco-epidemiologic studies of the diseases transmitted by the ticks and fleas: an important sum of data was accumulated in Europe on the animal species which were prone to studies of determination of hosts but no data to was now obtained Mediterranean countries.

  • We project to carry out a series of captures of micro-mammals, birds and lizards. A larval xenodiagnosis will be carried out in Algeria; it consists in nourishing larvae of not-infected ticks (coming from the breeding available to the level of the laboratory two species of hard ticks: Rhipicephalus sanguineus and Ixodes ricinus) on animals captured then to analyze the ticks after their blood meal to seek the pathogenic ones.
  • Then collection of skin biopsies from the patients. The culture will be carried out in a laboratory in Marseilles.

The fourth sector will ensure the determination of the frequencies of the infections to Rickettsia, Borrelia, and of the clinical demonstrations in Algeria, which they are human or animal. A questionnaire for the epidemiologic and clinical data-gathering was developed and will be widely distributed to the doctors in the various areas of the study. The answers will be centralized to the level of the laboratory where they will be analyzed. All the serums and skin biopsies will be collected by the clinicians within a clinical network already set up. Then will be carried out an evaluation of the risk of development of a clinical disease following an exposure to infected ticks and fleas and the exploration of the relation between the behavior of the patient and the infection and the disease. The improvement of the knowledge of the public and the doctors of the local risk factors is necessary to implement the precautionary measures, since no vaccine is currently available against the pathogenic species transmitted by the ticks and fleas.

 

AWAITED RESULTS:

  • List of the ticks and fleas as potential vectors of infectious disease in Algeria,
  • Cartography of the distribution of these ticks and fleas according to the bioclimatic zones,
  • Bioecologic data concerning these ticks and fleas: cycles, activities, hosts
  • Identification of the ticks and fleas playing a vector part, - Insulation of pathogenic at the ticks and fleas and constitution of a reference collection,
  • Identification of the reservoirs of unquestionable pathogenic transmitted by the ticks and fleas.
  • Creation of a serothèque for Rickettsioses, Bartonelloses, Ehrlichioses, Borrelioses. 

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