About the presence of Phlebotomus sergenti Parrot, 1917 (Diptera: Psychodidae) in Eastern Sicily, Italy

The note reports the data of a three-year sand fly investigation (1997-99) carried out in Eastern Sicily (Italy) with the aim to study the distribution of Phlebotomus sergenti. The survey involved a densely inhabited area at the foot of Mount Etna and the area of Iblei mounts. A total of 9,095 sand flies, of which 63.4% males, were captured. Five species belonging to the genus Phlebotomus (P. perniciosus, P. perfiliewi, P. neglectus, P. sergenti and P. papatasi) and one to the genus Sergentomyia (S. minuta) were identified. Both the prevalence and distribution of the species were different within the two areas studied. In Mount Etna area, P. perniciosus (77.7%) was the prevalent species followed by S. minuta (19.8%), P. sergenti (2.0%), P. neglectus (0.3%) and P. papatasi (0.2%). While in Iblei mounts region S. minuta (84.5%) showed the highest prevalence, followed by P. perniciosus (14.4%), P. perfiliewi (0.9%) and P. neglectus (0.1%). Here, P. sergenti was a very rare species (< 0.02). P. sergenti was mostly associated to domestic habitats of peri-urban and urban zones located between two and 750 m a.s.l. The density values of P. sergenti, expressed as number of specimens/m2 of sticky trap, were between 0.3 and 5.5 with the highest value in the hilly collecting sites. The low observed abundance of P. sergenti does not allow to draw any prediction on the role that the species could play in the transmission of leishmaniasis in Sicily.     

Entomological and parasitological study on phlebotomine sandflies in central and northern Albania

An entomological survey was carried out in two districts of central (Kruje) and northern (Lezhe) Albania. Six collecting sites, showing a variety of diurnal resting sites, were monitored for adult sandflies from June through October 2002. Flies were collected with CDC miniature light traps, sticky traps and mechanical or hand aspirators in peridomestic sites, in bedrooms and inside cow barns, chicken coops and pigpens. All collecting sites monitored were found positive for sandflies. A total of 849 specimens were caught (29.2% males) belonging to five Phlebotomus species. Phlebotomus neglectus (75.6%) was the most abundant species followed by P. perfiliewi (14.4%), P. papatasi (4.6%), P. tobbi (3.6%) and P. similis (1.8%). The first adult of P. neglectus appeared on June 11 and the last one was collected on October 16. The highest density for this species was observed at the end of July. A total of 111 blood-fed females were caught from the two areas studied. P. neglectus was the only species found blood fed in Lezhe and the same species was prevalent (56.1%) in Kruje followed by P. perfiliewi (30.3%), P. tobbi (10.6%); P. papatasi was represented by only two specimens. Blood meal origin was determined in 45/66 (68.2%) of the females tested from Kruie district. P. neglectus was found fed on four hosts, showing the following feeding patterns: cow (71.4%), dog (117.1%), chicken (5.7%) and human (5.7%); P. perfiliewi was found fed on cow (80.0%) and chicken (20.0%), P. tobbi on cow (50.0%), chicken (25.0%) and dog (25.0%). One specimen of P. papatasi was found fed on cow. When such prevalences were analysed by the available biomass for each host present at the collecting site, P. neglectus resulted to be an opportunistic feeder rather than exhibiting preferences for any specific animal. PCR analysis of 39 P. neglectus from the Lezhe district gave negative results for the presence of Leishmania DNA.     

Entomological survey of sandflies (Diptera, Psychodidae) in the province of L'Aquila (Abruzzo)]

A survey on phlebotomine sandflies (Diptera, Psychodidae) was carried out in the province of L'Aquila, Abruzzo region (Italy). Ten collecting sites at altitudes between 700 and 1,070 m a.s.l., produced a total of 324 specimens (81.1% males), 72.86% being Phlebotomus perniciosus, 3.08% P. mascittii, 0.30% P. papatasi and 23.76% Sergentomyia minuta. The collecting sites at 1,070, 950 and 775 m a.s.l. are the highest levels recorded in Italy for P. perniciosus, P. mascittii and P. papatasi, respectively. P. perfiliewi, the proven vector of cutaneous leishmaniasis (CL) in the Abruzzo region (Maroli et al., 1987) was absent. The probable role of P. perniciosus in the transmission of the sporadic CL cases reported from the area is discussed.     

First report on natural infection of the Phlebotomus tobbi by Leishmania infantum in northwestern Iran

Visceral leishmaniasis (VL) is an important health problem in Ardebil, where it borders Azerbaijan in the northwestern Iran. In spite of the presence of both cutaneous and visceral leishmaniasis (CL and VL) in northwestern Iran, previous researches have consistently revealed the etiologic agent of VL in the region to be Leishmania infantum. This is the first report of natural infection of Phlebotomus tobbi with L. infantum in Bilesavar district in the northern part of Ardebil province bordering Azerbaijan. Polymerase chain reaction (PCR) of kDNA, ITS1-rDNA, and CPB genes of the parasite followed by restriction fragment length polymorphism (RFLP) and gene sequencing analyses revealed presence of L. infantum in six out of 433 tested female sand fly specimens. Although sand flies of P. tobbi were infrequent, two out of 32 (6.25%) females captured in the area were found infected with the parasite. Phlebotomus perfiliewi transcaucasicus, the known vector of VL in the area, were the most dominant species but only four out of 273 (1.47%) tested were infected with L. infantum. This study showed that P. tobbi similar to P. perfiliewi transcaucasicus could play a significant role in the transmission of the L. infantum. However more investigations are needed to demonstrate that L. infantum is the only species circulating in the focus.     

Distribution and altitudinal structuring of phlebotomine sand flies (Diptera: Psychodidae) in southern Anatolia, Turkey: their relation to human cutaneous leishmaniasis.

The two Old World genera, Phlebotomus and Sergentomyia, were both recorded in southern Anatolia in Turkey. Phlebotomus species predominated and comprised about 93% of the entire collection (3,172 specimens). Out of the sixteen species identified, two belonged to the genus Sergentomyia: S. dentata and S. theodori. The remaining fourteen species in the genus Phlebotomus were grouped under four subgenera including some species that are elsewhere known to act as vectors of human cutaneous leishmaniasis. Most of the Phlebotomus were P. tobbi (32.5%), but P. papatasi, P. transcaucasicus, P. halepensis, P. galilaeus, P. sergenti, P. syriacus, P. neglectus, P. simici, P. alexandri, P. similis, P jacusieli, P. perfiliewi, and P. brevis were also identified. There were two associations of sand fly fauna with altitudinal gradient; the first one at relatively higher altitudes and the second one at lower altitudes. The transition between these two assemblages was within the range of 800-1,000 m. It is likely that Adana and Hatay provinces are transitional areas between western and eastern Anatolia. Mountains do not appear to be important geographical barriers for sand fly distribution. We also found that the proven vector P. sergenti is a widely distributed species throughout southern Anatolia and this species, together with its closely related species P. similis, shows sympatry in Konya Province.     

A new stable focus of canine leishmaniasis in northern Italy

A new stable focus of canine leishmaniasis (CanL) was identified in a coastal Adriatic area of the Emilia-Romagna Region in Northern Italy. Following the first clinical cases observed starting from 1998, a seroepidemiological survey was carried out on owned dogs from two communes and on animals housed in dog pounds of the Rimini province. Sixteen out of 612 dogs (2.6%) resulted positive to the IFA test. The 16 positive dogs all came from the two communes, with seroprevalences of 3 and 6%, respectively. The autochthonous origin of the infection was confirmed in all the cases. The parasitological investigation led to the isolation and identification of the parasite as Leishmania infantum Zymodeme MON 1. An entomological survey showed that Phlebotomus perniciosus and P. perfiliewi are present in this area and that P. perfiliewi was very abundant in one collection site. The risk of the establishment of a permanent transmission of the infection in the area, previously considered CanL-free, must be analysed in view of further investigations to be extended also to neighbouring areas.     

Bionomics of phlebotomine sand flies (Diptera: Psychodidae) in the province of Al-Baha, Saudi Arabia

The bionomics of phlebotomine sand flies (Diptera: Psychodidae) were studied for two successive years (January 1996-December 1997) at 12 collecting stations representing six sectors of the province of Al-Baha, Saudi Arabia. The predominant species was Phlebotomus bergeroti (41.7%), followed by lesser numbers of Phlebotomus sergenti (11%), Phlebotomus arabicus (10.6%), Sergentomyia tiberiadis (10.5%), Phlebotomus papatasi (10.2%), Sergentomyia antennata (9.6%), Phlebotomus alexandri (3%), Phlebotomus orientalis (2.3%) and Sergentomyia clydei (1.1%). The distribution of the collected species including species that are elsewhere known to act as vectors of human cutaneous leishmaniasis were distributed across different altitudes in Al-Baha. P. bergeroti, P. papatasi and P. arabicus were more abundant indoors; however, P. sergenti was more abundant outdoors. Sand fly populations exhibited three patterns of seasonal abundance in terms of their monthly activity. P. bergeroti, P. sergenti and P. arabicus were found to be naturally infected with Leishmania-like flagellates at an infection rate of 0.2%.     

Phylogenetic analysis of Phlebotomus species belonging to the subgenus Larroussius (Diptera, psychodidae) by ITS2 rDNA sequences

In the genealogy of Phlebotomus (Diptera: Psychodidae), morphological analyses have indicated that the subgenus Larroussius is a monophyletic group which is most closely related to the subgenera Transphlebotomus and Adlerius. We conducted a phylogenetic analysis of the relationships among six representative species of the subgenus Larroussius and one species representatitive of the Phlebotomus subgenus, assessing sequences of the Second Internal Transcribed Spacer (ITS2) of the ribosomal RNA (rRNA). Three of the species (P. perniciosus, P. ariasi and P. perfiliewi perfiliewi) were collected in different parts of the Mediterranean area. The trees estimated from parsimony and neighbour-joining analyses supported the monophyly of the Larroussius subgenus inferred from the morphological analysis. According to our data, P. ariasi may be a sister group to the rest of the Larroussius subgenus, although additional sequence data are needed to confirm this observation. Our results suggest that P. perniciosus and P. longicuspis are distinct species, in spite of the fact that there are only slight morphological differences. The strict congruence between the phylogeny of the Larroussius subgenus inferred from the ITS2 sequences and that based on morphological studies further confirmed the ability of the spacer sequence to identify recently-derived affiliations.     

Free amino acids in wild-caught Phlebotomus perniciosus and P. perfiliewi (Diptera: Psychodidae).

Free amino acids occurring in wild Phlebotomus perniciosus and P. perfiliewi were analyzed using high performance liquid chromatography. For both species the total concentration of amino acids, per unit weight, is over three times greater in females than in males. Qualitative inter-sex differences in amino acid compositions were also discovered: the most significant involving higher proportions of taurine and gamma-aminobutyric acid in females and higher proportions of arginine and glutamic acid in males. Although amino acid composition is consistent between females of both species, there are significant differences between males. The most important difference between males is an increase in tyrosine levels, at the expense of taurine, in P. perniciosus (as compared with P. perfiliewi). It is suggested that the intra- and inter-specific variation in amino acid composition identified in these studies could be due to ecological factors, such as differences in the sandflies natural amino acid sources (honeydew and/or plants), although a physiological explanation is not ruled out.     

Leishmaniasis in Sardinia. III. Soil analysis of a breeding site of three species of sandflies

Soil analysis was carried out on samples obtained from a breeding-site of the sandflies Phlebotomus perniciosus Newstead, P. perfiliewi perfiliewi Parrot and Sergentomyia minuta (Rondani) in Sardinia. Soil texture, pH, CaCo3, organic matter and water content showed no correlation with the number of sandflies that emerged from the places where soil samples were taken. It appears that pre-imaginal stages of these phlebotomine sandflies are associated with a relatively stable, cool and humid environment protected from sunshine and rain, rich in clay and in organic nitrogen.     

Leishmaniasis vectors (Diptera, Psychodidae) in 3 different environments of western Sicily and 2 of the Aeolian Islands

Some cases of visceral leishmaniasis in children and in adults have been diagnosed in the last fifteen years in Western Sicily and an increasing number of cases of canine leishmaniasis has been recorded, mainly in the province of Palermo. Such a human leishmaniasis recrudescence, after the antimalaria campaign had reduced the transmission, and the increase of canine leishmaniasis in the city of Palermo and its suburbs suggest that transmission in urban areas may be possible. Using the oiled paper method, a survey of phlebotomine sandflies was carried out during June and July 1988 both in the centre and suburbs of Palermo, and during July and August '88 in the Madonie mountains (at 800 to 1200 meters above sea level) and in two Eolian Islands: Lipari and Filicudi. Details of collected species are given. In both surveys, the oiled paper method did not turn out to be selective towards any given species of sandflies.     

Trapping sand flies (Diptera: Psychodidae) in the Emilia-Romagna region of northern Italy.

The efficiency and practicality of two trapping methods for adult Phlebotomine sand flies in two areas of the Emilia-Romagna region (Italy) were evaluated. Suction traps (CO2) and sticky traps (ST) were used to collect sand flies every two weeks, from June to September, 1999, from 16:00 to 07:00. Two CO2 traps were activated at the same time for each area (one with light and one without light), whereas 38 (four with light and 34 without lights) and 48 (four with light and 44 without) sticky traps were activated in Borghi and Longiano, respectively. An Index of Apparent Abundance (IAA) was calculated for each trap type and area. A total of 2,253 sand flies was trapped over the four-month period, with 1,765 collected from Borghi and 488 from Longiano. Phlebotomus perfiliewi was the most abundant species collected, comprising 99.6% and 84.6% of the total flies trapped in Borghi and Longiano, respectively. Other species were also collected within the two areas (Phlebotomus perniciosus and Phlebotomus mascittii) but were not considered for further analyses due to low catches. Significantly more specimens were caught using CO2 than sticky traps and the addition of a light source also improved the catches, however, a significantly greater number of female specimens were collected by a CO2 trap without a light source. Phlebotomus perfiliewi thus appears to show a photophobic reaction in the case of females when trapped using CO2/light attractants.     

Intraspecific variation in the cuticular hydrocarbons of the sandfly Phlebotomus perfiliewi from Italy

The cuticular hydrocarbons of Phlebotomus perfiliewi Parrot from representative localities on both sides of Italy show contrasts which support the view of Ward et al., (1981) that there are distinguishable populations of this sandfly on the Adriatic and Tyrrhenian coasts. It is suggested that the Appennine mountains form a barrier between the Adriatic and Tyrrhenian populations. A third distinct hydrocarbon type of P. perfiliewi was detected in Calabria, southern Italy, where contact between the Adriatic and Tyrrhenian types of flies is most likely to occur. P. perfiliewi from a trans-Appennine pass were also of the Calabrian hydrocarbon type. These hydrocarbon contrasts between allopatric populations of P. perfiliewi may result from clinical variation across the Appennines, associated with different host preferences, or they may represent apomictic populations with differential mate recognition systems.     

Sand fly (Diptera: Psychodidae) distribution in the endemic and non‐endemic foci of visceral leishmaniasis in northwestern Iran

An entomological study was conducted from June to September, 2010 in rural regions of Azarbayjan‐e‐sharqi, Azarbayjan‐e‐qarbi, and Ardabil provinces in northwestern Iran to determine sand fly fauna, diversity, and distribution in different habitats and altitudes using both sticky papers and light traps. Geographical distribution of sand flies and the similarity of populations in different locations were analyzed ecologically based on the Shannon‐Wiener Index and Jacard Coefficient, respectively. A total of 3,982 specimens was collected and sixteen species recorded. They belonged to the genera Phlebotomus [subgenus Phlebotomus (P. papatasi), Paraphlebotomus (P. sergenti, P. mongolensis, P. caucasicus, P. jacusieli), Larroussius (P. major s.l., P. tobbi, P. perfiliewi transcaucasicus, P. kandelakii) and Adlerius (P. halepensis, P. brevis, P. longiductus, P. balcanicus)], and Sergentomyia [subgenus Sergentomyia (S. sintoni, S. dentata and S. theodori)]. P. papatasi was the predominant species in all the locations except Bileh Savar, Macu, and Meshkin Shahr, followed by P. perfiliewi transcaucasicus and P. kandelakii. The latter species were caught from different habitats and altitudes with higher frequency than other species of the subgenus Larroussius. The lowest abundance belonged to P. jacusieli. The predominant species of subgenus Adlerius was P. halepensis. Data analysis showed that Meshkin Shahr and Bileh Savar had high and low diversities of sand fly distribution, respectively. Meshkin Shahr and Sarab districts had the highest similarity. Both are located in the foothills of Sabalan Mountain, with high diversity and richness.     

The genus Leishmania in Italy

Seventy-four Leishmania isolates collected in Italy from six different Regions where leishmaniases are endemic, have been typed. Parasites have been isolated from: man (VL and CL), dog, black rat (Rattus rattus), fox (Vulpes vulpes) and geckoes (Tarentola mauritanica and Cyrtodactylus kotschyi). The isolates have been characterized by starch-gel electrophoresis for 9-16 enzymes whose mobility was compared with that of international reference strains for L. infantum, L. tropica, L. major, L. donovani, L. aethiopica and L. tarentolae. The results obtained have shown that the genus Leishmania in Italy is represented by five zymodemes which may be grouped into two taxa: L. infantum s.l. (L. infantum s.st., L. infantum NH130 variant, L. infantum NH140 variant and L. infantum GOT, MDH, NH variant), agent of mammalian leishmaniases (including human leishmaniases), and L. tarentolae, parasite of geckoes. At the moment, the absence of L. tropica in Italy as agent of CL has been revealed. Through the analysis of epidemiological data obtained from the foci where Leishmania parasites were isolated two zymodemes only, L. infantum s.st. and L. infantum NH140 variant, show to be widely distributed. However, L. infantum s.st. appears to be prevalent in Thyrrenean foci which are characterized by VL cases and by high density of Phlebotomus perniciosus, and L. infantum NH140 variant is present in Adriatic areas where CL is diffuse and P. perfiliewi is the probable vector.     

Effects of indomethacin, luteinizing hormone (LH), prostaglandin E(2) (PGE(2)), trilostane, mifepristone, ethamoxytriphetol (MER-25) on secretion of prostaglandin E (PGE), prostaglandin F(2alpha) (PGF(2alpha)) and progesterone by ovine corpora lutea of pregnancy or the estrous cycle

Two experiments were conducted to determine the luteotropin of pregnancy in sheep and to examine autocrine and paracrine roles of progesterone and estradiol-17 beta on progesterone secretion by the ovine corpus luteum (CL). Secretion of progesterone per unit mass by day-8 or day-11 CL of the estrous cycle was similar to day-90 CL of pregnancy (P >/= 0.05). In experiment 1, secretion of progesterone in vitro by slices of CL from ewes on day-8 of the estrous cycle was increased (P /= 0.05) while PGE(2) increased (P /= 0.05) detectable quantities of PGF(2alpha) or PGE while day-90 ovine CL of pregnancy secreted PGE (P /= 0.05). Trilostane, mifepristone, or MER-25 did not affect secretion of progesterone, PGE, or PGF(2alpha) by day-11 CL of the estrous cycle or day-90 CL of pregnancy (P >/= 0.05). It is concluded that PGE(2), not LH, is the luteotropin at day-90 of pregnancy in sheep and that progesterone does not modify the response to luteotropins. Thus, we found no evidence for an autocrine or paracrine role for progesterone or estradiol-17 36 on luteal secretion of progesterone, PGE or PGF(2alpha).     

Incidence of spontaneous ovulation and development of the corpus luteum in non-mated dromedary camels (Camelus dromedarius)

The occurrence of spontaneous ovulation in dromedaries was examined in two separate studies including 20 non-lactating, barren and 12 lactating dromedaries, respectively. Lactating camels were milked twice a day with an automatic bucket milking machine. Ovarian activity was monitored by repeated ultrasonography. Blood samples for progesterone were collected daily or two to three times a week. To compare CL development after spontaneous and induced ovulations, ovulation was induced by a GnRH analogue in eight lactating dromedaries. Spontaneous ovulation was observed in one non-lactating camel (1 of 20 camels, 5%; 1 of 70 follicular waves, 1.4%), whereas, spontaneous ovulation was detected more frequently in lactating dromedaries (5 of 12 camels, 41.7%; 13 of 91 follicular waves, 14.3%). In one lactating camel, spontaneous ovulation occurred repeatedly for nine times. There was a significant effect of type of ovulation (spontaneous versus induced, P < 0.05) and day (P < 0.001) on serum progesterone concentration. Mean serum progesterone levels and total progesterone production (AUC) were higher after induced ovulation. Luteal diameter and serum progesterone concentration were positively correlated (r = 0.71, P < 0.001), but there was a significant difference between morphological and functional development of the CL. In dromedaries, morphological development starts earlier, morphological regression starts later and last longer than functional development and regression of the CL. Compared to induced ovulation, functional development of the CL after spontaneous ovulation might be altered but the morphological development is not affected.     

The Parallel Analysis of Phase Sensitive Inversion Recovery (PSIR) and Double Inversion Recovery (DIR) Images Significantly Improves the Detection of Cortical Lesions in Multiple Sclerosis (MS) since Clinical Onset

BackgroundDouble inversion recovery (DIR) detects only a minority (<20%) of cortical lesions (CL) in multiple sclerosis (MS). Phase-sensitive inversion recovery (PSIR) was suggested to be substantially superior to DIR in the detection of cortical lesions (CL). These two sequences might be complementary.ObjectivesTo analyze CL frequency and type in MS patients having different disease duration and disability, including patients at clinical onset, and to discern more correctly the artifacts, by combining DIR and PSIR images.ResultsPSIR disclosed CL in 100% of the patients and was capable of identifying more than four times lesions (455.5%, p<0.00001), especially IC (mean numbers: 36.5 in CIS/eRRMS, 45.0 in RRMS and 52.3 in SPMS) and LC (mean numbers: 10.9 in CIS/eRRMS, 20.1 in RRMS and 25.3 in SPMS), compared to DIR (p<0.00001). CL number was significantly higher in SPMS compared to RRMS (p<0.0001). Artifacts were more accurately identified by comparing the two sequences.ConclusionsOur study confirms the higher ability of PSIR in disclosing and classifying CL. The presence of CL in all CIS patients further points out the relevance of cortical pathology in MS. Whether the parallel analysis of DIR and PSIR images may be useful for diagnostic purposes, especially when a diagnosis of MS is suspected but not confirmed by routine MRI, needs to be evaluated in larger patient series. The analysis of the cortex by DIR and PSIR may also allow a better stratification of the patients for prognostic and counseling purposes, as well as for their inclusion in clinical studies.     

Effect of progesterone from induced corpus luteum on the characteristics of a dominant follicle in dromedary camels (Camelus dromedarius)

The present study was carried out to elucidate the effect of progesterone (P4) from the induced corpus luteum (CL) on the characteristics of the dominant follicle (DF) in dromedary camels (Camelus dromedarius). Ovarian follicular and induced CL dynamics were monitored by transrectal ultrasonography in eight camels during the peak breeding season. The characteristics of the DF were monitored daily from the day of emergence into a wave, until it appeared to lose its dominance and the DF of a subsequent wave grew to a diameter of 13-17 mm. At this stage ovulation was induced by hCG and the DF was monitored every 8 h for 48 h. After ovulation, CL dynamics and follicular development (emergence of a new wave, growth and mature phase of the selected DF) were monitored daily. Blood samples were collected during each ultrasound examination to study the P4 profile in these animals. The CL developed to a maximum size (22.55 ± 3.24 mm) with a peak concentration of P4 (4.60 ± 2.57 ng/ml) 7 days after ovulation. The size of the CL was positively correlated with the P4 concentration (r = 0.612) during the different stages of the CL dynamics. The presence of CL did not affect the linear growth rate, duration of growth and mature phases of the DF. The development of the DF to its maximum size during its mature phase and inter-wave interval were not affected by the P4 secreted by the induced CL. In conclusion, there is no evidence from this study to suggest that P4 from induced CL altered the characteristics of a DF in dromedary camels.