The Late Miocene coldspot of z-coral diversity in the Mediterranean: Patterns and causes

The Late Miocene distribution and diversity of zooxanthellate-like corals in the Mediterranean are analyzed in their paleobiogeographical framework, using our REEFCORAL database. The Late Miocene Mediterranean pool reached 20 z-coral genera. Although this fauna could build flourishing reef ecosystems during the Early Messinian, it was a relict fauna with severely limited speciation that lived on the edge of its ecological requirements in terms of solar energy and temperature range. Most z-coral genera, because they had long stratigraphic ranges and had survived previous extinctions, were able to adapt to the Messinian environments, which were unusual for such biotas. Hence, Porites, the most widespread genus in the region and also the most dominant in ecological assemblages, was the best equipped to cope with the drastic changes related to the Messinian Salinity Crisis.     

Population Genomics of Wall Lizards Reflects the Dynamic History of the Mediterranean Basin

The Mediterranean Basin has experienced extensive change in geology and climate over the past six million years. Yet, the relative importance of key geological events for the distribution and genetic structure of the Mediterranean fauna remains poorly understood. Here, we use population genomic and phylogenomic analyses to establish the evolutionary history and genetic structure of common wall lizards (Podarcis muralis). This species is particularly informative because, in contrast to other Mediterranean lizards, it is widespread across the Iberian, Italian, and Balkan Peninsulas, and in extra-Mediterranean regions. We found strong support for six major lineages within P. muralis, which were largely discordant with the phylogenetic relationship of mitochondrial DNA. The most recent common ancestor of extant P. muralis was likely distributed in the Italian Peninsula, and experienced an “Out-of-Italy” expansion following the Messinian salinity crisis (∼5 Mya), resulting in the differentiation into the extant lineages on the Iberian, Italian, and Balkan Peninsulas. Introgression analysis revealed that both inter- and intraspecific gene flows have been pervasive throughout the evolutionary history of P. muralis. For example, the Southern Italy lineage has a hybrid origin, formed through admixture between the Central Italy lineage and an ancient lineage that was the sister to all other P. muralis. More recent genetic differentiation is associated with the onset of the Quaternary glaciations, which influenced population dynamics and genetic diversity of contemporary lineages. These results demonstrate the pervasive role of Mediterranean geology and climate for the evolutionary history and population genetic structure of extant species.     

The distribution of phlebotomine sandflies in the principal ecological zones of Jordan

A survey of the sandflies (Diptera: Psychodidae) of Jordan increased the number of species known from the country from three to thirteen. Composition of the phlebotomine fauna and the relative abundance of sandfly species differ among the six climatic zones, with a general distinction between the dry desert zones and the more humid Mediterranean zones. Species of Phlebotomus predominate in the former and Sergentomyia in the latter. Within the desert zone some species are restricted to the eastern desert and others to the Jordan Valley in the west.     

Phlebotomus (Legeromyia) multihamatus subg. nov., sp. nov. from Gabon (Diptera: Psychodidae)

During a research project aimed at the study of the Culicinae fauna of Gabon and carried out in the National Park of La Lopé, we captured an unknown sandfly male specimen (genus Phlebotomus) by CDC miniature light trap belonging to a new species for Science. Furthermore, the originality of his genitalia does not allow us to include this species in one of the existing subgenus, thus in this paper we propose the creation of a new subgenus, as Phlebotomus (Legeromyia) multihamatus sp. nov., subg. nov. described from the National Park of La Lopé, through one male captured with CDC miniature light trap. A new species and a new subgenus of sandfly is characterised by a short style with three spines, a paramere wearing a basal hook as well as a basal pouch and the absence of basal lobe on the coxite. The originality of the genitalia of the male gives way to discussion about potential primary homologies between P. multihamatus sp. nov. and Phlebotomus (Abonnencius) fortunatarum, Phlebotomus (Anaphlebotomus) stantoni and Phlebotomus (Euphlebotomus) argentipes, which should be verified for future studies. The discovery of this new species in Gabon must encourage the study of sandflies in this country.     

Combining climatic projections and dispersal ability: a method for estimating the responses of sandfly vector species to climate change

BACKGROUND: In the Old World, sandfly species of the genus Phlebotomus are known vectors of Leishmania, Bartonella and several viruses. Recent sandfly catches and autochthonous cases of leishmaniasis hint on spreading tendencies of the vectors towards Central Europe. However, studies addressing potential future distribution of sandflies in the light of a changing European climate are missing. METHODOLOGY: Here, we modelled bioclimatic envelopes using MaxEnt for five species with proven or assumed vector competence for Leishmania infantum, which are either predominantly located in (south-) western (Phlebotomus ariasi, P. mascittii and P. perniciosus) or south-eastern Europe (P. neglectus and P. perfiliewi). The determined bioclimatic envelopes were transferred to two climate change scenarios (A1B and B1) for Central Europe (Austria, Germany and Switzerland) using data of the regional climate model COSMO-CLM. We detected the most likely way of natural dispersal ("least-cost path") for each species and hence determined the accessibility of potential future climatically suitable habitats by integrating landscape features, projected changes in climatic suitability and wind speed. RESULTS AND RELEVANCE: Results indicate that the Central European climate will become increasingly suitable especially for those vector species with a current south-western focus of distribution. In general, the highest suitability of Central Europe is projected for all species in the second half of the 21st century, except for P. perfiliewi. Nevertheless, we show that sandflies will hardly be able to occupy their climatically suitable habitats entirely, due to their limited natural dispersal ability. A northward spread of species with south-eastern focus of distribution may be constrained but not completely avoided by the Alps. Our results can be used to install specific monitoring systems to the projected risk zones of potential sandfly establishment. This is urgently needed for adaptation and coping strategies against the emerging spread of sandfly-borne diseases.     

Detection of Wolbachia pipientis, including a new strain containing the wsp gene,in two sister species of Paraphlebotomus sandflies,potential vectors of zoonotic cutaneous leishmaniasis

Individual, naturally occurring Phlebotomus mongolensis and Phlebotomus caucasicus from Iran were screened for infections with the maternally inherited intracellular Rickettsia-like bacterium Wolbachia pipientis via targeting a major surface protein gene (wsp). The main objective of this study was to determine if W. pipientis could be detected in these species. The sandflies were screened using polymerase chain reaction to amplify a fragment of the Wolbachia surface protein gene. The obtained sequences were edited and aligned with database sequences to identify W. pipientis haplotypes. Two strains of Wolbachia were found. Strain Turk 54 (accession {"type":"entrez-nucleotide","attrs":{"text":"EU780683","term_id":"212725653","term_text":"EU780683"}}EU780683) is widespread and has previously been reported in Phlebotomus papatasi and other insects. Strain Turk 07 (accession {"type":"entrez-nucleotide","attrs":{"text":"KC576916","term_id":"499068248","term_text":"KC576916"}}KC576916) is a novel strain, found for first time in the two sister species. A-group strains of W. pipientis occur throughout much of the habitat of these sandflies. It is possible that Wolbachia is transferred via horizontal transmission. Horizontal transfer could shed light on sandfly control because Wolbachia is believed to drive a deleterious gene into sandflies that reduces their natural population density. With regard to our findings in this study, we can conclude that one species of sandfly can be infected with different Wolbachia strains and that different species of sandflies can be infected with a common strain.     

Paratethyan Ostracod immigrants in Italy during the Late Miocene

In this paper the ostracod assemblages recovered from several brackish Late Miocene Italian deposits have been analysed from a palaeobiogeographical perspective. During late Tortonian-early Messinian it is possible to recognize in Italy rich ostracod assemblages characterized by a wide contingent of taxa with central European or Mediterranean affinity, while only few brackish and freshwater ostracods show Paratethyan affinity. The recognized composition of the ostracod assemblages matches the palaeogeographic setting of the palaeo-Mediterranean/Paratethys at that moment. In fact during late Tortonian-early Messinian the palaeo-Mediterranean and Paratethysian domains were divided and, even if the connection via the present Marmara Sea-Strimon Basin was still open, the different salinity between them represented an ecological barrier, preventing faunal exchanges. Since normal aquatic migration was impossible, it must be assumed that the Paratethyan-like taxa entered the palaeo-Mediterranean area via passive dispersal by aquatic birds. On the contrary, the ostracod assemblages from the Italian Lago-Mare deposits show the absolute predominance of Paratethyan taxa, which, according to the known palaeogeographic setting during the late Messinian Lago-Mare event, could actively migrate from the Paratethys domain, colonizing the palaeo-Mediterranean, whose endemic fauna was severely impoverished by the Messinian salinity crisis and the following water dilution.     

Limits of a rapid identification of common Mediterranean sandflies using polymerase chain reaction-restriction fragment length polymorphism

A total of 131 phlebotomine Algerian sandflies have been processed in the present study. They belong to the species Phlebotomus bergeroti, Phlebotomus alexandri, Phlebotomus sergenti, Phlebotomus chabaudi, Phlebotomus riouxi, Phlebotomus perniciosus, Phlebotomus longicuspis, Phlebotomus perfiliewi, Phlebotomus ariasi, Phlebotomus chadlii, Sergentomyia fallax, Sergentomyia minuta, Sergentomyia antennata, Sergentomyia schwetzi, Sergentomyia clydei, Sergentomyia christophersi and Grassomyia dreyfussi. They have been characterised by sequencing of a part of the cytochrome b (cyt b), t RNA serine and NADH1 on the one hand and of the cytochrome C oxidase I of the mitochondrial DNA (mtDNA) on the other hand. Our study highlights two sympatric populations within P. sergenti in the area of its type-locality and new haplotypes of P. perniciosus and P. longicuspis without recording the specimens called lcx previously found in North Africa. We tried to use a polymerase chain reaction-restriction fragment length polymorphism method based on a combined double digestion of each marker. These method is not interesting to identify sandflies all over the Mediterranean Basin.     

Bionomics of phlebotomine sandflies in the Galilee focus of cutaneous leishmaniasis in northern Israel

The bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied for three years (2001-2003) in the Galilee focus of cutaneous leishmaniasis in northern Israel, where the causative Leishmania tropica (Kinetoplastida: Trypanosomatidae) is transmitted by Phlebotomus (Adlerius) arabicus Theodor and Phlebotomus (Paraphlebotomus) sergenti Parrot, comprising 22% and 8%, respectively, of the local sandfly fauna sampled by light traps. The predominant species overall was Phlebotomus (Larroussius) tobbi Adler & Theodor (51%) with lesser numbers of Phlebotomus (Adlerius) simici Theodor (11%), Phlebotomus (Larroussius) syriacus Adler & Theodor (5%), Phlebotomus (Larroussius) perfiliewi Perfil'ev (3%) and Phlebotomus (Phlebotomus) papatasi Scopoli (0.05%). Sandfly adult populations were prevalent from April to November and peaked between June and August, being more abundant through the summer in irrigated habitats, such as gardens and orchards, than in open grassland. Of the two cutaneous leishmaniasis vectors, P. sergenti preferred boulder mounds located at the outskirts of settlements, whereas P. arabicus was more abundant overall and near houses in particular. Females of all these sandfly species displayed a peak of activity after sunset (20.00-22.00 hours), whereas activity of males persisted longer through the night. Another slight increase in activity was noted before dawn (02.00-04.00 hours). Phlebotomus arabicus appears to be the main vector of L. tropica in the Galilee focus, due to its denser populations, more endophily and preference for peridomestic habitats than shown by P. sergenti in northern Israel.     

Latest Messinian “Lago-Mare” Lymnocardiinae from Italy: Close relations with the Pontian fauna from the Dacic Basin

Rich hypo- to mesohaline molluscan assemblages characterising the latest Messinian “Lago-Mare” biofacies, composed of prosobranch gastropods (Neritidae, Thiaridae, Melanopsidae, Hydrobiidae) and bivalves of the families Cardiidae (subfamily Lymnocardiinae) and Dreissenidae, are widespread in shallow water basins characterized by low salinities within the Mediterranean realm, during the post-evaporitic phase in the time-span 5.5-5.3 Ma. Several genera and species are recorded in the Italian uppermost Messinian sediments. While the gastropods show endemic character being linked to continental water-systems, Lymnocardiinae and Dreissenidae have strong Paratethyan affinity. New records of significant species of Lymnocardiinae from the uppermost Messinian sediments of Tuscany, Marches and Sicily and the systematic review of the old literature data point out close relations of the Italian fauna with that from the Pontian sediments of the Dacic Basin. The palaeobiogeographical data referred to Messinian and Pontian Lymnocardiinae suggest that the Aegean Basin could be an intermediate basin from whence the Pontian Paratethyan-type fauna migrated into the Mediterranean area during the latest Messinian. Since the ecology of Lymnocardiinae is mainly tied to oligo- and mesohaline water, spreading of suitable habitats in depositional systems of marginal settings characterized by increasing freshwater influx after the “salinity crisis” favoured their dispersal into the Mediterranean area from the Paratethys realm.     

DNA hybridizations on squash-blotted sandflies to identify both Phlebotomus papatasi and infecting Leishmania major

Epidemiological field studies on leishmaniasis have been hampered by the laborious, and often inefficient, methods used to assess the rates of infection of sandfly vectors (Diptera; Phlebotominae) by species of the causative disease organisms, protozoal parasites of the genus Leishmania (Kinetoplastida; Trypanosomatidae). We report the rapid and accurate identification of both sandfly vector (Phlebotomus (Phlebotomus) papatasi (Scopoli] and infecting Leishmania major Yakimov & Schokov by DNA hybridizations to squash-blotted sandflies. Large numbers of whole (infected) sandflies can be quickly squashed on to nylon hybridization filters and (following standard procedures) the filter-bound DNA can be hybridized sequentially to cloned, multicopy genomic sequences that are specific for species of Leishmania (kinetoplast DNA) or for the sandfly (ribosomal (r) DNA). Our sandfly probe consists of a 3.2 kb fragment of the intergenic 'non-transcribed' spacer of rDNA of P. papatasi that we have detected only in this species: it is present in all six geographically isolated populations tested (from Tunisia through to India) but cannot be detected in the morphologically similar P. (Phlebotomus) duboscqi Neveu-Lemaire, the vector of Leishmania major south of the Sahara; it also cannot be detected in Phlebotomus species of the subgenera Larroussius and Paraphlebotomus that together with P. papatasi are the dominant man-biting sandflies in north African foci of zoonotic cutaneous leishmaniasis, where (as in many arid regions of western Asia) P. papatasi is believed to be the sole vector of L. major.     

CAPA-gene products in the haematophagous sandfly Phlebotomus papatasi (Scopoli) – vector for leishmaniasis disease

Sandflies (Phlebotominae, Nematocera, Diptera) are responsible for transmission of leishmaniasis and other protozoan-borne diseases in humans, and these insects depend on the regulation of water balance to cope with the sudden and enormous intake of blood over a very short time period. The sandfly inventory of neuropeptides, including those that regulate diuretic processes, is completely unknown. Direct MALDI-TOF/TOF mass spectrometric analysis of dissected ganglia of Phlebotomus papatasi, combined with a data-mining of sandfly genome ‘contigs’, was used to identify native CAPA-peptides, a peptide class associated with the regulation of diuresis in other hematophagous insects. The CAPA-peptides identified in this study include two CAPA-PVKs, differentially processed CAPA-PK, and an additional CAPA precursor peptide. The mass spectrometric analysis of different parts of the neuroendocrine system of the sandfly indicate that it represents the first insect which accumulates CAPA-PVKs exclusively in hormone release sites of abdominal ganglia and CAPA-PK (nearly) exclusively in the corpora cardiaca. Additionally, sandflies feature the smallest abdominal ganglia (∼35 μm) where CAPA-peptides could be detected so far. The small size of the abdominal ganglia does not appear to affect the development of the median neurosecretory system as it obviously does in another comparably small insect species, Nasonia vitripennis, in which no capa-gene expression was found. Rather, immunocytochemical analyses confirm that the general architecture in sandflies appears identical to that of much larger mosquitoes.     

Hourly Activity and Natural Infection of Sandflies (Diptera: Psychodidae) Captured from the Aphotic Zone of a Cave,Minas Gerais State,Brazil

Sandflies are holometabolous insects that are of great epidemiological importance in the neotropical region as vectors of leishmaniases. Caves are ecotopes that significantly differ from external environments and, among the insects that live or visit their internal area and adjacent environment, sandflies are commonly found. Based on this context, the objective of this work was to examine the period of activity of sandflies in the cave environment in the aphotic zone. Thus, four sandfly captures were conducted, one in each season of the year, in a cave where studies on the bioecological aspects of sandfly fauna have been conducted since 2008. In this same study, we have also noticed the presence of flagellates in some captured females. Catches were carried out for 24 hours using a Shannon trap, light bait, and cave walls were actively searched. We collected a total of 638 sandflies, representing 11 species. The most abundant species and with more intense period of activity were, in descending order: Lu. cavernicola (62%), Ev. spelunca (16%) and Ev. sallesi (14%). A total of 69 females were dissected to check for natural infection, and in five specimens we found living flagellated forms: two Ev. spelunca, two Ev. sallesi and one Sc. sordellii. This study shows that the activity of some species caught in the aphotic zone of the cave, especially Lu. cavernicola, differs from what has already been reported in previous sandfly captures, which are almost always conducted at night and during twilight. The existence of sandflies that were naturally infected with flagellates and the lack of awareness regarding the behaviour of sandflies in cave environments are strong indicators of the need for further study on this group of insects in this ecotope, as a safety measure to protect the visitors of such environment.     

Population structure and geographical subdivision of the Leishmania major vector Phlebotomus papatasi as revealed by microsatellite variation

Abstract Multi‐locus microsatellite typing (MLMT) has been employed to infer the population structure of Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae) sandflies and assign individuals to populations. Phlebotomus papatasi sandflies were collected from 35 sites in 15 countries. A total of 188 P. papatasi individuals were typed using five microsatellite loci, resulting in 113 different genotypes. Unique microsatellite signatures were observed for some of the populations analysed. Comparable results were obtained when the data were analysed with Bayesian model and distance‐based methods. Bayesian statistic‐based analyses split the dataset into two distinct genetic clusters, A and B, with further substructuring within each. Population A consisted of five subpopulations representing large numbers of alleles that were correlated with the geographical origins of the sandflies. Cluster B comprised individuals collected in the Middle East and the northern Mediterranean area. The subpopulations B1 and B2 did not, however, show any further correlation to geographical origin. The genetic differentiation between subpopulations was supported by F statistics showing statistically significant (Bonferroni‐corrected P < 0.005) values of 0.221 between B2 and B1 and 0.816 between A5 and A4. Identification of the genetic structure of P. papatasi populations is important for understanding the patterns of dispersal of this species and to developing strategies for sandfly control.     

Ecology of cutaneous leishmaniasis in Sinai: linking parasites,vectors and hosts

Cutaneous leishmaniasis (CL) is a neglected clinical form of public health importance that is quite prevalent in the northern and eastern parts of Egypt. A comprehensive study over seven years (January 2005-December 2011) was conducted to track CL transmission with respect to both sandfly vectors and animal reservoirs. The study identified six sandfly species collected from different districts in North Sinai: Phlebotomus papatasi, Phlebotomus kazeruni, Phlebotomus sergenti, Phlebotomus alexandri, Sergentomyia antennata and Sergentomyia clydei. Leishmania (-)-like flagellates were identified in 15 P. papatasi individuals (0.5% of 3,008 dissected females). Rodent populations were sampled in the same districts where sandflies were collected and eight species were identified: Rattus norvegicus (n = 39), Rattus rattus frugivorous (n = 13), Rattus rattus alexandrinus (n = 4), Gerbillus pyramidum floweri (n = 38), Gerbillus andersoni (n = 28), Mus musculus (n = 5), Meriones sacramenti (n = 22) and Meriones crassus (n = 10). Thirty-two rodents were found to be positive for Leishmania infection (20.12% of 159 examined rodents). Only Leishmania major was isolated and identified in 100% of the parasite samples. The diversity of both the vector and rodent populations was examined using diversity indices and clustering approaches.     

Biogeography and evolution of Abies (Pinaceae) in the Mediterranean Basin: the roles of long‐term climatic change and glacial refugia

Aim The genus Abies exemplifies plant diversification related to long‐term climatic, geological and evolutionary changes. Today, the Mediterranean firs comprise nine species, one natural hybrid and several varieties. Here I summarize current knowledge concerning the origin and evolution of the genus Abies in the Mediterranean Basin and propose a comprehensive hypothesis to explain the isolation and speciation pattern of Mediterranean firs. Location The Mediterranean Basin. Methods The literature on Abies was reviewed, focusing on the morphology, fossil records, molecular ecology, phytosociology and biogeography of the genus in the Mediterranean Basin. Results Abies fossils from the western Mediterranean indicate a wide Tertiary circum‐Mediterranean distribution of the Abies ancestor. Palaeogeographical data also suggest a single eastern Mediterranean Tertiary ancestor. Following the Miocene to Pliocene climate crisis and marine transgressions, the ancestor of the northern Mediterranean firs is hypothesized to have separated into two eastern groups, one on the Balkan Peninsula and the other in Asia Minor. However, land bridges may have permitted gene flow at times. A southward migration of A. alba to refugia, where older fir species may have remained isolated since the Miocene, could explain recent findings indicating that morphologically distant species are more closely related than expected based on such morphological classification. Main conclusions The Abies genus appears to have undergone significant morphological differentiation that does not necessarily imply reproductive isolation. That is, long‐term Mediterranean Basin dryness along a south‐eastern to north‐western gradient may have caused an initial Miocene–Pliocene speciation sequence. Pleistocene glacial cycles probably forced migrations to occur, leading to repeated contact between fir species in glacial refugia.     

Fish otoliths from the pre-evaporitic (Early Messinian) sediments of northern Italy: their stratigraphic and palaeobiogeographic significance

The study of otolith assemblages from the pre-evaporitic Messinian deposits allows the reconstruction of a fauna of 79 taxa of which 35 could be identified at the specific level. Three of these are new: Diaphus rubus, Myctophum coppa, and Uranoscopus ciabatta. The assemblages reflect mainly a neritic environment influenced by the oceanic realm. Analysis of the global present-day geographic distribution of 42 of the recognised Messinian genera indicates that 88% of these are still living in the Mediterranean, 98% in the Atlantic and 78% in the Indo-Pacific realm. These results are in good agreement with the evolutionary trends documented for the Oligocene and Miocene teleost fauna, specifically an increase in percentage of genera inhabiting the modern Mediterranean, a very high percentage of Atlantic and Indo-Pacific genera, and a slight fall of the importance of present-day Indo-Pacific genera from the Rupelian up to the Late Miocene. Analysing the composition of the Early Messinian fauna at the level of nominal species indicates that about 53% of the species represented in the assemblages are still living in the Recent Mediterranean, and that a significant number of these were already present in the Tortonian. It is interesting that these species are mainly neritic. This seems to confirm that the close affinity of the fossil assemblage with the present-day Mediterranean neritic fauna, which was already recorded at the genus level for the Rupelian fauna, persists during the Neogene and continues until the Pleistocene.     

Evolution and biogeography of the endemic Roucela complex (Campanulaceae: Campanula) in the Eastern Mediterranean

At the intersection of geological activity, climatic fluctuations, and human pressure, the Mediterranean Basin – a hotspot of biodiversity – provides an ideal setting for studying endemism, evolution, and biogeography. Here, we focus on the Roucela complex (Campanula subgenus Roucela), a group of 13 bellflower species found primarily in the eastern Mediterranean Basin. Plastid and low‐copy nuclear markers were employed to reconstruct evolutionary relationships and estimate divergence times within the Roucela complex using both concatenation and species tree analyses. Niche modeling, ancestral range estimation, and diversification analyses were conducted to provide further insights into patterns of endemism and diversification through time. Diversification of the Roucela clade appears to have been primarily the result of vicariance driven by the breakup of an ancient landmass. We found geologic events such as the formation of the mid‐Aegean trench and the Messinian Salinity Crisis to be historically important in the evolutionary history of this group. Contrary to numerous past studies, the onset of the Mediterranean climate has not promoted diversification in the Roucela complex and, in fact, may be negatively affecting these species. This study highlights the diversity and complexity of historical processes driving plant evolution in the Mediterranean Basin.     

Sandfly (Diptera, Psychodidae, Phlebotominae) fauna of South-Western Pakistan. 1. Diagnostic morphology of Phlebotomus papatasi (Scopoli), Ph. bergeroti (Parrot) and Ph. salehi (Mesghali)

A survey was conducted to study the morphology of the sandfly fauna in South-Western Pakistan (Balochistan). During the revision of different genera of sandflies the specimens of Phlebotomus papatasi (Scopoli) (N = 720), Ph. bergeroti Parrot (N = 30) and Ph. salehi Mesghali (N = 70) were encountered in various localities. These localities appear to be new records of the subgenus in the literature to date. Ph. bergeroti is reported for the first time from Pakistan and Ph. salehi from Balochistan as well. Characters of these three Pakistanese Phlebotomus are compared with the published data of these species from other countries. Keys for the identification of Pakistanese Phlebotomus are also constructed. Two female Ph. papatasi collected from indoors out of 132 female flies (1.5%) were found positive with flagellate infection in pharynx and midgut. The possible vectorial role of these flies is also discussed. Further surveys are necessary in parts of the country that have not been systematically surveyed.