Description of Madathamugadia hiepei n. sp. (nematoda: Splendidofilariinae), a parasite of a South African gecko, and its development in laboratory bred Phlebotomus dubosqi (Diptera: Psychodidae).

Madathamugadia hiepei n. sp., Splendidofilariinae, a parasite of a South African gecko Pachydactylus turneri is described together with its development obtained experimentally in Phlebotomus duboscqi (Diptera: Psychodidae: Phlebotominae). This new species differs from the two small, more highly evolved groups with a short tail and atrophied postcloacal papillae, the first group consisting of two Madagascan species, M. zonosauri and M. hopluri, parasites of the Gerrhosauridae and Iguanidae, and the second containing three species from the Ethiopian Region, M. huambensis, M. versterae and M. bissani, parasites of the Scincidae. It also differs from M. ineichi, the most primitive species of the genus (cuticularised buccal capsule, no atrophy of head papillae and largest number of precloacal papillae), a parasite of the Cordylidae in South Africa. M. hiepei is close to the two species parasitic in the Gekkonidae of the Mediterranean subregion, M. ivaschkini and M. wanjii, all three of which have a post-oesophageal vulva. However, the new species can be distinguished from the Mediterranean parasites by (a) the shorter oesophagus, (b) the number and position of the cloacal papillae and (c) the microfilaria. The three filariae of this group and M. ineichi, the only ones of which aspects of the life-cycles are known, experimentally develop in phlebotomine sand flies.

Molecular systematics of the phlebotomine sandflies of the subgenus Paraphlebotomus (diptera, psychodidae, phlebotomus) based on ITS2 rDNA sequences. Hypotheses Of dispersion and speciation.

Phylogenetic Paraphlebotomus relationships are inferred by a study based on the sequences of ITS2, which has been sequenced in nine Paraphlebotomus species: P. alexandri, P. andrejevi, P. jacusieli, P. kazeruni, P. mireillae, P. mongolensis, P. saevus, P. sergenti and P. similis and in two out-groups species of the subgenus Phlebotomus: P. papatasi and P. duboscqi. Paraphlebotomus alexandri appears as the sister group of all other Paraphlebotomus sandflies. Among the other species, three groupings are clearly highlighted: andrejevi and mongolensis; mireillae and saevus; jacusieli, kazeruni, sergenti and similis. These groupings are related to speculations about the migration of Paraphlebotomus from a centre of dispersion located in the Middle East sometime from the early Eocene to the late Miocene.

Courtship and mating by the sandfly Phlebotomus duboscqi, a vector of zoonotic cutaneous leishmaniasis in the Afrotropical region.

Courtship behaviour of males of the Afrotropical sandfly Phlebotomus duboscqi Neveu-Lemaire (Diptera: Psychodidae) involved mounting the female and clasping her 'waist' with the male coxites placed between the female's thorax and abdomen. This behaviour, which we call 'piggy-backing', was preceded by male wing beating, perhaps involving mate recognition and contact pheromones. It did not seem to be pre- or postcopulatory mate guarding. Piggy-backing was attempted by P. duboscqi males on females of other species (P. papatasi and P. perniciosus) and even on other male P. duboscqi. The majority of female P. duboscqi piggy-backed by males were already inseminated, and most of the courting did not lead to copulation. This, coupled with the presence of a mating plug (semen) in each spermatheca of inseminated females, suggests that female P. duboscqi are monogamous for at least the first gonotrophic cycle. Male courtship with piggy-backing was more intense when females could feed on a hamster than when a hamster was present but the females were denied access to the host. It is suggested that, when a hamster was available to the females, the conditions in the laboratory are similar to those in rodent holes, the natural habitat of P. duboscqi.

Protection of dogs from bites of phlebotomine sandflies by deltamethrin collars for control of canine leishmaniasis.

Dog collars made of PVC plastic impregnated with the pyrethroid insecticide deltamethrin at 40 mg/g were investigated for their protective efficacy against phlebotomine sandflies. Collared dogs were kept separately (two untreated control dogs lived together) in outdoor enclosures, each with a kennel, in the Cévennes, southern France. To measure sandfly mortality and anti-feeding effects due to the deltamethrin-impregnated collars worn continuously by the dogs for up to 8 months, each dog was periodically sedated and exposed for 2h to 150-200 laboratory-reared Phlebotomus perniciosus females (plus c. 25 males) inside a net (1.2 m square, 1.8 m high) indoors. After dogs were removed from the nets, allowed to recover and returned to their kennels, any dead sandflies were collected from inside the net and counted. Surviving flies were kept overnight, then scored according to whether they were still alive or dead, unfed or blood-fed. From tests 2, 3, 4, 13, 20, 26 and 34 weeks after the dogs began wearing collars, the overall numbers of blood-fed female sandflies recaptured were 75 from two dogs with collars, compared with 1911 from two collarless dogs. Thus, for every 100 flies which fed on collarless dogs, only 4 fed on collared dogs, i.e. the collars protected dogs from 96% of the bites and this activity was maintained for up to 34 weeks. During the same period, the percentage of recaptured female sandflies that had fed on collared dogs was 0-12% compared to 55-95% on collarless dogs. Immediately after dogs were taken out of the nets, 21-60% of flies confined with the collared dogs were found dead, compared to 0-12% with the controls. It is concluded that, at least in the Mediterranean subregion, this insecticidal collar would protect a dog from the majority of sandfly bites and retain a killing effect for a complete sandfly season. Moreover, it seems likely that the use of collars on all dogs in a focus of Leishmania infantum would reduce contact between sandfly vectors and canine reservoir hosts sufficiently to diminish the risk of infection for humans as well as dogs.

A laboratory model of canine leishmaniasis: the inoculation of dogs with Leishmania infantum promastigotes from midguts of experimentally infected phlebotomine sandflies.

Twenty-five dogs (beagles) were infected with Leishmania infantum by the intradermal inoculation of an estimated 5-8,000 metacyclic promastigotes harvested from the midguts of 320 experimentally infected P. perniciosus. Details are given of the methods of infecting the flies and harvesting the parasites. All dogs developed small, self-healing chancres at the sites of inoculation. Parasites were isolated from lymph nodes, bone marrow or spleen of 21 dogs, 12 of which developed signs of disease and raised IFAT litres to Leishmania. Nine of the 21 remained healthy over a five-year observation period. Six of the nine were shown to have a cell mediated immune response to Leishmania. No parasites were isolated from four of the 25 dogs, two of which had a demonstrable cell mediated immunity and another had low transitory IFAT titres. The fourth had chancres at the sites of inoculation. The results show that dogs can be readily infected with promastigotes from the midguts of sandflies. However, a high proportion develop a cell mediated immunity and show on signs of disease. It is suggested that serological surveys of dogs for canine leishmaniasis reveal neither the true prevalence of infection nor the intensity of transmission. The efficacy of controlling human visceral leishmaniasis caused by L. infantum by destroying seropositive dogs is questioned.

The armature in the genital atrium as a new taxonomic character distinguishing females of Phlebotomus papatasi and P. duboscqi (Diptera: Psychodidae).

Descriptions are given of armatures in the genital atria of females of the two morphologically similar sandfly species, Phlebotomus papatasi and P. duboscqi. The species are distinguishable by the size and shape of the armature, the grouping of the spines in the armature and the length and shape of the spines. These characters have been shown to separate females of other closely related species of phlebotomine sandflies.

Phlebotomine sandflies of Kenya (Diptera: Psychodidae). IV. The armature in the genital atrium of female Larroussius as a means of identification.

Descriptions are given of armatures in the genital atria of the six known Kenyan species of phlebotomine sandflies of the subgenus, Larroussius, namely Phlebotomus aculeatus, P. elgonensis, P. guggisbergi, P. longipes, P. orientalis and P. pedifer. Phlebotomus aculeatus, P. longipes and P. pedifer can be recognized by the shapes of their armatures. Differences in the length and arrangement of the spines in the armature of P. elgonensis and P. longipes are diagnostic features. The distinguishing feature of P. guggisbergi is a wide variation in spine thickness. The most notable feature of P. orientalis is the angle at which the spines lie. The appearance of the base of the spermathecal duct remains the method of choice for the identification of all six but, if this feature is not well displayed in dissected females, they can be distinguished by the armature. It is suggested that descriptions of new species should include an illustration of the armature in the genital atrium.

[The pharyngeal dentition of the females of Phlebotomus duboscqi Neveu-Lemaire, 1906 and of Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae)]. / La denture pharyngienne des femelles de Phlebotomus duboscqui Neveu-Lemaire, 1906 et de Phlebotomus papatasi (Scopoli, 1786) (Diptera:Psychodidae).

Phlebotomus duboscqi and P. papatasi are morphologically closely related species. Scanning electron microscopy showed constant differences between pharyngeal teeth of the females.

Phlebotomine sandflies of Kenya (Diptera: Psychodidae). III. The identification and distribution of species of the subgenus Larroussius.

The six species of phlebotomine sandflies of the subgenus Larroussius recorded in Kenya are Phlebotomus aculeatus, P. elgonensis, P. guggisbergi, P. longipes, P. orientalis and P. pedifer. Five are proven vectors of leishmaniasis in that country or elsewhere. Males of all six can be identified by the morphology of the aedeagus or the number and position of the hairs on the inner surface of the coxite. Additional features separating some of the species are the sizes of the palpal and antennal segments. The females have usually been considered difficult or impossible to distinguish. A comparison of the base of the spermathecal ducts is made and it is shown that all six can be identified by this character alone. A map of Kenya is given, showing the presently known distribution of the six Larroussius species. Further surveys are necessary in parts of the country that have not been systematically surveyed.

Anthroponotic cutaneous leishmaniasis in Kabul, Afghanistan: the low susceptibility of Phlebotomus papatasi to Leishmania tropica.

Females of a laboratory colony of Phlebotomus papatasi from Kabul were fed through a membrane on a high dose of amastigotes of Leishmania tropica from the same place. Less than one-third of the flies became infected and only 6% developed heavy infections of the stomodaeal valve. From these observations, and previous studies by other workers, it is concluded that P. papatasi probably plays little or no part in the transmission of anthroponotic cutaneous leishmaniasis in Kabul. As in Saudi Arabia and Morocco, the most likely vector in Afghanistan is P. sergenti.

Phlebotomine sandflies of Kenya (Diptera: Psychodidae). I. The validity of Phlebotomus (Larroussius) elgonensis Ngoka, Madel and Mutinga.

Sandflies collected in Kitum cave on the Kenyan side of Mount Elgon were identified as Phlebotomus (Larroussius) elgonensis Ngoka, Madel and Mutinga, a species synonymized with P. (L.) aculeatus Lewis, Minter and Ashford, by D. J. Lewis. A comparison of the flies from Kitum cave with P. aculeatus from Lake Elmentaita, Kenya, shows marked differences in the lengths of: (a) the spermathecal ducts of the female; (b) the style, coxite, genital pump and aedeagus of the male; and (c) the palpal segments of both sexes. Other notable differences are in the shape of the base of the spermathecal ducts; the position of spines on, and the shape of, the tip of the aedeagus; and the appearance of the pharyngeal armatures of both sexes. Phlebotomus elgonensis is redescribed and it is concluded that it is sufficiently different from P. aculeatus to deserve recognition as a valid species.

Epidemiology of canine leishmaniasis: prevalence, incidence and basic reproduction number calculated from a cross-sectional serological survey on the island of Gozo, Malta.

Assessment of the resilience of canine leishmaniasis to control or, more ambitiously, the effort needed to eradicate infection, requires an estimate of the basic case reproduction number (R0). This paper applies the theoretical results of Hasibeder, Dye & Carpenter (1992) to data from a cross-sectional survey on the Maltese island of Gozo in which dogs of known age, sex and occupation (pet, guard etc) were subjected to three different serological tests for the presence of specific antibody (IFAT, DAT and ELISA). Difficulties in interpreting these test results, and hence of determining the proportion of dogs infected, present the main obstacle to estimating R0: estimates are critically dependent on the choice of threshold separating seropositives from seronegatives. The data do, however, allow a robust comparative analysis of risk which shows that the force of infection experienced by working dogs is about three times higher than that of pet dogs, a degree of non-homogeneous contact which actually has little effect on estimates of R0. We suggest a cautious point estimate of R0 congruent to 11, and comment briefly on its significance for leishmaniasis control.

The initial establishment of sandfly colonies.

The initiation of laboratory colonies of phlebotomine sandflies is far more difficult than the maintenance of already established colonies. Based on our experience of setting up more than two dozen colonies, we present an account of the methods which we have found most successful. They include: techniques for collection and feeding wild-caught females: transportation in the field: optimum conditions for oviposition: choice and preparation of larval-rearing pots: larval food: handling adults: and the influence of cage size for sandfly copulation in the laboratory.

The identification of female sandflies of the subgenus Larroussius by the morphology of the spermathecal ducts.

The complete spermathecae of 13 species of Larroussius were dissected and examined. The base of the duct of 4 species (aculeatus, guggisbergi, tobbi, wui) is bell-shaped: they are easily distinguished by differences in shape. Five other species (perniciosus, pedifer, longicuspis, perfiliewi, galilaeus) have lateral structures at the base of the duct: slight differences between perniciosus, pedifer and longicuspis distinguish these species: the structures of perfiliewi and galilaeus are the same. Species of a third group (neglectus, syriacus) have a common duct with no lateral structures: they are distinguishable by minor differences. Two species (ariasi, orientalis) from a fourth group with expanded distal parts of the duct; the size and shape of the expanded parts easily separates the species. The study shows a great variation in the morphology of the base of the spermathecal ducts of Larroussius and confirms the value of this feature in identifying otherwise indistinguishable females of the subgenus.

[Comparison between Phlebotomus duboscqi Neveu-Lemaire, 1906, and Phlebotomus papatasi (Scopoli, 1786): morphological and isoenzymatic studies]. / Comparaison entre Phlebotomus duboscqi Neveu-Lemaire, 1906 et Phlebotomus papatasi (Scopoli, 1786): études morphologique et isoenzymatique.

Phlebotomus duboscqi et P.papatasi are morphologically closely related. Both species can be vectors of zoonotic cutaneous leishmaniasis in the Old World. In some African areas these two species are sympatric. Until now the females of the two species could not be differentiated. The authors investigated morphological characters and isoenzymatic characterization. Constant differences were observed. Moreover three interbreeding attempts were unsuccessful. These two species seemed to be considered as genuine species.

Isoenzyme characterization of Leishmania from man, dog and sandflies in the Maltese islands.

As a part of a general survey on leishmaniases and sandflies of the Maltese islands, 22 Leishmania stocks were isolated from human visceral (1) and cutaneous (1) cases, dogs (16) and sandflies (4). They were characterized by the analysis of 15 enzymes. The commonest Mediterranean L. infantum zymodeme, MON 1, was found to cause human and canine visceral leishmaniasis; L. infantum MON 78, which has so far been isolated only in Malta, was the agent of human cutaneous leishmaniasis. Both zymodemes were isolated from the same sandfly species, Phlebotomus perniciosus.

Absence of outer caudal setae on all larval instars of Phlebotomus tobbi from the Ionian Greek islands.

Larval instars 2, 3 and 4 of Phlebotomus tobbi Adler, Theodor & Lourie from the Greek islands of Corfu and Zakynthos were found to have two caudal setae instead of the four usually present on these instars of Phlebotomus larvae. In a scanning electron microscope comparison with larvae of P. papatasi (Scopoli), a sensillum was seen in place of each outer seta of P. tobbi larvae, suggesting secondary loss of the setae. As the larvae of less than a tenth of the species and subspecies of the genus Phlebotomus have been seen and described, it cannot be assumed that those of P. tobbi are unique in having only two caudal setae. However, four caudal setae in late instars can no longer be considered as a constant character of the genus Phlebotomus. Observations on the larvae of P. tobbi also raise the question of the unknown function of the caudal setae.