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Wild-caught Phlebotomus ariasi Tonnoir permitted to feed on dogs infected with Leishmania infantum Nicolle were marked with fluorescent powder and released into their natural habitat in an uninhabited area of the Cévennes in southern France. Over a period of 29 days after release, 253 females were recaptured with CDC miniature light traps or by active search at night with portable UV lamps. The ovaries and infections in the alimentary tract were then examined. The females oviposited 6 nights after in infecting blood meal. Second blood meals were never taken during the maturation of eggs. During the first ovarian cycle, midgut infections with promastigotes were only moderately heavy. The intensity of infection increased markedly during the second ovarian cycle and, in the third ovarian cycle, the first pharynx infected with paramastigotes was seen (on day 19). From day 19 to day 29, 76% of the flies had pharyngeal infections. Three out of 19 sand flies with pharyngeal infections recaptured during this period had metacyclic promastigotes in their mouthparts. The long time required for parasites to reach the proboscis in completely natural conditions suggests that their presence in the mouthparts is not a prerequisite for transmission by bite. It is more likely that transmission is most commonly by the regurgitation of metacyclic promastigotes from the thoracic midgut following damage to the stomodaeal valve by chitinase produced by the parasite during its development in the gut of the fly. Nevertheless, it is reasonable to assume that the bite of a fly with metacyclic promastigotes in the proboscis (or salivary glands) would also be infective.

Phlebotomus mariae was first described in the South of Morocco in 1974. More specimens were found again recently in the High Atlas, showing that this species can be considered as a valid one, placed in the sub-genus Larroussius. The female remains to be found.

The development of Leishmania spp. in their vectors is a complex process involving behavioural, morphological and biochemical changes. The process differs between Leishmania species. Recent information on the developmental process and its regulation, summarised in this report, relates to possible separation of amastigotes into two separate developmental lines, development of infective forms of promastigote, attachment of certain, possibly altruistic promastigotes to the gut wall, and the possibility of genetic exchange.

In a Moroccan focus of cutaneous leishmaniasis caused by Leishmania tropica, 7,907 female sandflies captured with CDC traps were dissected from summer to autumn 1989. Among species of the genus Phlebotomus, only P. sergenti harbored promastigotes. Eighty-nine strains belonging to the complex L. tropica were isolated. The frequency of vector infection was zero in June, rose to 1.3% in August, and reached 9.9% in October, which indicates that the period of high risk is at the end of the hot season. Out of 89 strains isolated, 74 were completely typed and corresponded to the following four zymodemes: MON-102 (one strain), MON-107 (56 strains), MON-122 (two strains), and MON-123 (15 strains). Only the first two were observed in humans. The distribution of zymodemes MON-102 and MON-107 was very different in humans, dogs, and the vector. In one of the sites surveyed, which was strongly dominated by MON-107, the absence of human cases involving this zymodeme suggests the existence of a wild reservoir.

The dispersal of Phlebotomus ariasi was studied in mark-release-recapture experiments in the summer of 1980 in a valley on the north-eastern slopes of the Oiselette range in the Cévennes mountains, in the commune of Roquedur, Gard, 50 km north of Montpellier, France. More than 5,000 specimens of P. ariasi were marked with fluorescent powders and released in 9 batches at 3 different places. Seven batches were engorged females and two were unengorged females and males. From 1-29 days after release, 497 marked sandflies (approximately 9%) were recaptured by active searches with UV lamps or in 58 CDC light traps set up in groups of 4 or 5 at 12 recapture stations. Females released engorged generally remained within 250 m of the release point for the first eight days while the bloodmeal was being digested after which there was a tendency to disperse to distances greater than 350 m presumably in a search for oviposition sites or another bloodmeal. The furthest distance to which a female released engorged was shown to move was 925 m; it was caught 12 days after release. Some of the females released unfed quickly moved away from release points, sometimes to distances of 1,000 m or more. One of these was caught 68.5 hrs after release at a station 2,200 m from the release point. Male sandflies tended to stay near the point of release and were not recaptured at distances greater than 600 m. There was no evidence that the movement of the sandflies was assisted by wind. Observations on the dispersal of female sandflies confirm that leishmaniasis can be more widely spread than generally assumed by the movements of the vector.

Engorged sandflies were collected using 58 CDC light traps set up nightly for 5 weeks at 12 stations in the commune of Roquedur, Gard, France, in the Cévennes focus of leishmaniasis. Of 782 engorged females, 593 were Phlebotomus ariasi, 3 were P. mascittii and 186 were not identified. Using the precipitin ring test and counter-current immuno-electrophoresis, the bloodmeals were tested with antisera to man, leporids, rodents, canids, mustelids, equids, suids, bovids, birds, lizards and amphibia. Of 530 identified bloodmeals, 2 were avian blood, 26 were mixed meals of mammalian blood and 25 were meals taken from unidentified mammals. Of the 477 other sandflies, 211 (44.2%) had fed on canids, 107 (22.4%) on man, 70 (14.7%) on bovids (presumed to be cattle at one station and goats elsewhere), 33 (6.9%) on leporids (probably mostly domesticated rabbits), 26 (5.5%) on mustelids (probably badgers), 17 (3.6%) on horses and 13 (2.7%) on rodents (probably brown rat). The proportions of feeds on different mammals varied according to their availability at each station. When both man and dog were equally available, the dog was preferred. The finding that, away from human habitation, P. ariasi commonly feeds on mustelids suggests the need for a reappraisal of animals of this family as possible reservoirs of leishmaniasis in the Cévennes.

In the Cévennes, the distribution of canine leishmaniasis and, therefore, of human visceral leishmaniasis is usually on the middle slopes of hillsides. From previous work, it was assumed that the reason is that the same places are the preferred habitats of the sandfly vector, Phlebotomus ariasi Tonnoir, 1921. An investigation was made throughout one season (13.05.1977 to 7.10.1977) of the comparative densities of sandflies at different altitudes on two opposite sides of a selected hill (le massif d'Oiselette) in the department of Gard. Oiled papers, changed every 15 days, were distributed at 22 points at altitudes from 165 to 600 m above sea level. A total of 5,050 sandflies were collected and analysed according to altitude and time of year. Of the sandflies collected, 86,2% were P. ariasi, 0,3% were P. perniciosus Newstead, 1911, 0,1% were P. mascittii Grassi, 1908 and 13,4% were Sergentomyia minuta (Rondani, 1843). P. ariasi was present at all altitudes with maximum densities between 300 and 500 m. This species was present from mid May until late September, with a maximum density at the end of July. The results support the notion that, in the Cévennes, inhabitants of farms and villages on the slopes of hillsides at altitudes of 300-500 m are at the greatest risk of acquiring leishmaniasis because these places are the preferred habitat of the sandfly vector, P. ariasi.

Surveys of the phlebotomine fauna in a focus of zoonotic cutaneous leishmaniasis (ZCL) in the Al-Hassa oasis, Eastern Province, Saudi Arabia, revealed only one species of Phlebotomus (P. papatasi) and three of Sergentomyia (S. antennata, S. clydei and S. fallax). 11 specimens of P. papatasi from six sites in the oasis were found with promastigotes in the midgut. An isolate from one of the sandflies was typed by the examination of isoenzymes and identified as Leishmania major, zymodeme LON-4 (= Montpellier zymodeme 26), the principal zymodeme of L. major isolated from patients with ZCL in the oasis. Three isolates from leishmanial lesions at sites of the bites of wild caught specimens of P. papatasi were also identified as the same zymodeme of L. major as the isolate from the sandfly. The findings show that P. papatasi is the vector of ZCL in the Al-Hassa oasis and probably in other ecologically similar foci in the Kingdom.

In the analysis of the ecology of foci of leishmaniasis in the South of France, a comparison was made of two methods of sampling the vector: the technique of oiled paper traps and of hand-catching off human bait. The techniques, used in parallel in a rural site (Cévennes) and in an urban site (built up areas in Marseilles) gave comparable results. In the Cévennes focus of visceral leishmaniasis, Phlebotomus ariasi Tonnoir, 1921 is the dominant sandfly; in the urban focus, it is replaced by Phlebotomus perniciosus Newstead, 1911. The analysis of captures shows that, in the South of France, in general, the method of sampling by oiled papers can be used instead of man-biting catches. The former method is easier and permits work over a wide area at many places simultaneously.

By use of mark-release-recapture methods, evidence was obtained of the distance of the dispersion of Phlebotomus ariasi Tonnoir, 1021. Female flies, collected in a valley in the Cévennes mountains (at Roquedur, Gard), were marked with fluorescnt powders, released an then recaptured by searching with ultra-violet lamps. The majority of the flies recapture were among ones given a blood meal shortly before release. Recaptures were made from the second day after release. A number of females migrated with blood in the midgut; their ovaries were up to and including stage V. The distance between the extreme points of recapture was about 1 km, and the maximum recorded distance between points of release and recapture was 750 m. The epidemiological interest of these observations is that, at least in the Cévennes foci, the sandfly as well as the dog must now be considered as capable spreading the pathogenic agent of visceral leishmaniasis.

The systematic inventory and annual following of Phlebotomus population is established by the authors in a transmission area of visceral (human and canine) and cutaneous (human) leishmaniasis in the Yemen Arab Republic (province of Taez). Seven species of Phlebotomus and nine species of Sergentomyia are thus identified. Among them, four are considered as potential vectors: on the one hand, P. orientalis (s.g. Larroussius) and, probably P. arabicus (s.g. Adlerius) for L. infantum and L. donovani, in the other, P. sergenti and P. saevus (s.g. Paraphlebotomus) for L. tropica.

As part of a study of visceral leishmaniasis in the Cévennes in southern France, an infection was transmitted from dog to dog by the bite of a single sandfly, Phlebotomus ariasi Tonnoir, 1921. The role of this species as a vector, suspected from earlier studies, is, therefore, confirmed. Twenty female sandflies, which had engorged on a naturally infected dog 23 +/- 2 days previously were put with a healthy dog which, after an incubation period of 15 months, developed viscero-cutaneous leishmaniasis. In dissections of the sandflies immediately after contact with the experimental dog, it was found that only one had engorged. All of 17 sandflies dissected had midgut infections: 10 of these also had infections in the pharynx. The engorged specimen additionally had parasites in the proboscis, a condition thought by many workers to be necessary for the parasite to be transmitted by bite.