I. Chelbi

An entomological investigation was carried out in 2014 at two sites located in Central Tunisia, one irrigated and another non-irrigated situated in arid bio-geographical areas. Sand flies of the subgenus Larroussius namely Phlebotomus perfiliewi, Phlebotomus perniciosus, and Phlebotomus longicuspis are the most abundant sand fly species in the irrigated site. However, in the non-irrigated site, Phlebotomus papatasi of the Phlebotomus genus is the most abundant species. A total of 3191 sand flies were collected and pooled with up to 30 specimens per pool based on sex, trapping location and collection date, were tested for the presence of phleboviruses by nested reverse transcriptase polymerase chain reaction in the polymerase gene and sequenced. Of a total of 117 pools, 4 were positive, yielding a minimum infection rate of sand flies with phleboviruses of 0.12%. Phylogenetic analysis performed using partial nucleotide and amino acid sequence in the polymerase gene showed that these phleboviruses belonged to four different clusters corresponding to Toscana virus (TOSV), Saddaguia virus (SADV), Sandfly Fever Sicilian Virus (SFSV) and Utique virus (UTIV). This study provides more evidence that the abundance of P. perfiliewi is associated with the development of irrigation in arid bio-geographical areas of Central Tunisia which may have led to the emergence of phleboviruses. We report the first detection of TOSV from sand flies collected from Central Tunisia.

Several outbreaks of human West Nile virus (WNV) infections were reported in Tunisia during the last two decades. Serological studies on humans as well as on equine showed intensive circulation of WNV in Tunisia. However, no virus screening of mosquitoes for WNV has been performed in Tunisia. In the present study, we collected mosquito samples from Central Tunisia to be examined for the presence of flaviviruses. A total of 102 Culex pipiens mosquitoes were collected in September 2014 from Central Tunisia. Mosquitoes were pooled according to the collection site, date and sex with a maximum of 5 specimens per pool and tested for the presence of flaviviruses by conventional reverse transcription heminested PCR and by a specific West Nile virus real time reverse transcription PCR. Of a total of 21 pools tested, 7 were positive for WNV and no other flavivirus could be evidenced in mosquito pools. In addition, WNV was isolated on Vero cells. Phylogenetic analysis showed that recent Tunisian WNV strains belong to lineage 1 WNV and are closely related to the Tunisian strain 1997 (PAH 001). This is the first detection and isolation of WNV from mosquitoes in Tunisia. Some areas of Tunisia are at high risk for human WNV infections. WNV is likely to cause future sporadic and foreseeable outbreaks. Therefore, it is of major epidemiological importance to set up an entomological surveillance as an early alert system. Timely detection of WNV should prompt vector control to prevent future outbreaks. In addition, education of people to protect themselves from mosquito bites is of major epidemiological importance as preventive measure against WNV infection.

An entomological investigation was performed in 2004 in the region of Rmilia, home to well-known zoonotic cutaneous leishmaniasis, located in Central Tunisia. The authors confirm the presence of Sergentomyia (Sintonius) clydei (Sinton, 1928). Thus, the sand fly fauna list of Tunisia includes 17 species.

The objective of this study was to evaluate the systemic insecticidal activity of an imidacloprid-treated rodent oral bait, against Phlebotomus papatasi Scopoli, 1786 vector of Leishmania major Yakimoff & Schokhor, 1914 (Kinetoplastida: Trypanosomatidae), etiologic agent of zoonotic cutaneous leishmaniasis (ZCL). Shaw's gerbil Meriones shawi Duvernoy, 1842 (Rodentia, Gerbillidae) were treated with imidacloprid-treated bait (0.05%). In the laboratory, effects on adult and larval of Phlebotomus papatasi fed on treated M. Shawi and on its faeces were studied. The effectiveness of this approach was tested under field conditions. In the laboratory, 100% of P. Papatasi were killed within 24 hours after blood feeding on Meriones shawi treated up to four weeks prior with a single application of imidacloprid (0.05%) bait. In addition, none of the P. Papatasi larvae that consumed feces from M. shawi treated with the imidacloprid bait survived to pupation. In the field, application of the ...

In Tunisia, Phlebotomus (Larroussius) perfiliewi Parrot, 1939, is highly suspected to be involved in the transmission of sporadic cutaneous leishmaniasis. The geographical distribution of this sandfly species is limited to the northern part of Tunisia and more specifically to the humid, sub-humid and semi-arid bio-geographical areas. To date, the geographical distribution of P. perfiliewi and subsequently of the sporadic cutaneous leishmaniasis extends to the arid bio-geographical areas located in central Tunisia. This extension is related to the intense development of irrigation systems in these areas, which may lead to the establishment of a new ecological niche suitable for the development of P. perfiliewi.

An entomological investigation was performed in 2004 in the region of Rmilia, home to well-known zoonotic cutaneous leishmaniasis, located in Central Tunisia. The authors confirm the presence of Sergentomyia (Sintonius) clydei (Sinton, 1928). Thus, the sand fly fauna list of Tunisia includes 17 species.

The ability of the sand fly Phlebotomus papatasi to transmit Leishmania major, the etiologic agent of zoonotic cutaneous leishmaniasis, to Meriones shawi, the natural reservoir host of this parasite, was studied under laboratory conditions. Sand flies became infected with L. major after feeding on a lesion of needle-inoculated M. shawi. Moreover, P. papatasi, previously infected with L. major, transmitted the parasite to M. shawi by bite during a second bloodmeal. Two months after the blood-meal, the animal developed a lesion on its ears. Xenodiagnosis was performed on the infected animal. The infectivity of M. shawi to P. papatasi lasted for five months, period corresponding to winter season in North Africa. We have thus demonstrated the transmission of L. major by P. papatasi to M. shawi under laboratory conditions. Our results show that reservoir hosts surviving winter time are the main source of infection for P. papatasi during the following season, and subsequently they play a major role in the persistence and transmission of L. major between transmission cycles.

ABSTRACT Details on the productivity and developmental times of a colony of Phlebotomus papatasi Scopoli (Diptera: Psychodidae) over 14 generations are reported and compared with findings of previous studies. The average productivity (percentage of eggs laid that were reared to adults) over six generations at 26-27 and at 29 -30 degrees C was 44.08 and 59.53%, respectively. The maximum productivity was 69.5%. The average developmental time over six generations at 26-27 and at 29 -30 degrees C was 35 and 26 d, respectively. The minimum developmental time from egg to adults was 25 d. The Tunisian strain of P. papatasi can reproduce autogenously or anautogenously, depending on the availability of a suitable bloodmeal source.

Phlebotomus papatasi (Diptera: Psychodidae) is the Old World sand fly vector of zoonotic cutaneous leishmaniasis caused by Leishmania major (Trypanosomatidae: Kinetoplastida), a debilitating and disfiguring protist parasitic disease prevalent throughout southern Mediterranean countries, the Middle East, as well as southern and eastern European countries, where it is regarded as a serious public health problem. Little is known of the mating ecology of P. papatasi, and, in particular, the role (if any) of pheromones is not known. In this laboratory- and field-based study, we have shown that a male-produced sex pheromone exists in P. papatasi. Young female P. papatasi are attracted to the headspace volatiles of small groups of males, males and females together, but not females alone. Males were not attracted to males, females, or mixed groups of males and females in the laboratory. Larger groups of males or males and females together were repellent in the laboratory study. Field experiments showed that Centers for Disease Control (CDC) light traps baited with small groups of males and females together were attractive to females, but not males. CDC traps baited with large groups of males and females together caught significantly fewer females and males than the control traps; however, the proportion of females caught compared with males overall was much higher than with CDC traps baited with small numbers of males and females. These results suggest that females may be attracted in preference to males to the vicinity of the baited traps and are highly sensitive to the concentration of male pheromone. It also suggests that P. papatasi mating behavior is fundamentally different from that of Lutzomyia longipalpis, where large mating aggregations of males and females occur.

ABSTRACT The population density of Phlebotomus papatasi Scopoli (Diptera: Psychodidae), vector of Leishmania major Yakimoff & Schokhor (Kinetoplastida: Trypanosomatidae), the etiologic agent of zoonotic cutaneous leishmaniasis (ZCL), was assessed May-November 2005 in central Tunisia by using sticky traps. The densities of P. papatasi were found to peak in early spring and again in the autumn. The lowest densities were observed in August. Prevalence of ZCL in the governorate of Sidi Bouzid peaks in December, 3 to 3.5 mo after the fall sand fly population peak, suggesting a close temporal association with the abundance of P. papatasi.