The nucleomorphs associated with secondary plastids of cryptomonads and chlorarachniophytes are the sole examples of organelles with eukaryotic nuclear genomes. Although not as widespread as their prokaryotic equivalents in mitochondria and plastids, nucleomorph genomes share similarities in terms of reduction and compaction. They also differ in several aspects, not least in that they encode proteins that target to the plastid, and so function in a different compartment from that in which they are encoded.Guillardia theta, and the chlorarachniophyte, Bigelowiella natans, have experienced similar evolutionary pressures during their transformation to reduced organelles. We compared the evolutionary rates of genes from nuclear, nucleomorph, and plastid genomes, all of which encode proteins that function in the same cellular compartment, the plastid, and are thus subject to similar selection pressures. Furthermore, we investigated the divergence of nucleomorphs within cryptomonads by comparing G. theta and Rhodomonas salina.Here, we test whether the phylogenetically distinct nucleomorph genomes of the cryptomonad, Chlorarachniophyte nucleomorph genes have accumulated errors at a faster rate than other genomes within the same cell, regardless of the compartment where the gene product functions. In contrast, most nucleomorph genes in cryptomonads have evolved faster than genes in other genomes on average, but genes for plastid-targeted proteins are not overly divergent, and it appears that cryptomonad nucleomorphs are not presently evolving rapidly and have therefore stabilized. Overall, these analyses suggest that the forces at work in the two lineages are different, despite the similarities between the structures of their genomes. While the primary acquisition of the plastid from a free-living cyanobacterium is believed to have occurred only once , plastidThe nucleomorph is often thought of as an anomaly, a rare occurrence, since it is known only in cryptomonads and chlorarachniophytes, but if one considers 'loss or gain' rather than 'presence or absence' then it is perhaps not so anomalous. All lineages that are known to contain secondary plastids  have ancestors that contained a nucleomorph. Depending on the number of secondary endosymbiotic events that took place, which is still contentious ,12-14, tGuillardia theta [Bigelowiella natans [G. theta and B. natans, nucleomorph genomes were completed, all known endosymbiont genomes have been of prokaryotic origin. The best examples of prokaryotic endosymbiont genomes are those of the mitochondrion, once a free-living alpha-proteobacterium, and the chloroplast, once a free-living cyanobacterium [Wolbachia [Buchnera [Wigglesworthia [Blochmania [Buchnera [Cryptomonads and chlorarachniophytes arose from separate endosymbiotic events, and neither host cell nor endosymbiont are very closely related. Yet the nucleomorph genomes of the cryptomonad, ia theta  and the a natans -10 sharea natans ,22. Compolbachia -25, BuchBuchnera , Wigglessworthia  and Blocochmania , the feaochmania -31. Thesochmania ; an effeochmania . The AT Buchnera . A univeBuchnera . The effBuchnera .There is less chromosomal information for eukaryotic obligate intracellular parasites, however certain alveolate and microsporidian genomes show some similar characteristics such as genome compaction , AT biasArabidopsis thaliana serving as an outgroup. Here we test whether the phylogenetically distinct nucleomorph genomes of G. theta and B. natans have experienced similar evolutionary pressures that influenced genome-wide variation in predictable ways and with the same severity and whether these effects are in common to those described in other enslaved nuclei. Proteins from both nucleomorph genomes have been observed to reside on long branches of phylogenetic trees indicating that they are poorly conserved [G. theta and B. natans nucleomorphs encode proteins that are directed to the plastid. Proteins that function in the plastid are presumably subject to similar selection pressures in organisms with nucleomorphs as they are in other algae. We have therefore used plastid proteins encoded in the plastid genome, the nucleomorph, or the nucleus, to examine differences in rates of evolution in the different genomes to determine whether the nucleomorph is evolving at a dissimilar rate to the plastid and nuclear genomes. We also investigate the overall variability of evolutionary rates of nucleomorph-encoded proteins and their homologues in other species to determine if the proteins still encoded within these genomes are generally well conserved, and whether this can shed light on their retention in the nucleomorph. By comparing proteins from the nucleomorph of two cryptomonads, G. theta and Rhodomonas salina, we also investigate whether cryptomonad nucleomorph genomes are diverging at the same rate as their nuclear genomes.With the recent availability of red algal  and greeonserved -45, howeG. theta and B. natans use proteins encoded in the nuclear genome, the nucleomorph genome and the plastid itself. Of the 147 proteins encoded in the G. theta plastid genome [C. merolae and the green plant A. thaliana. Of the 57 proteins encoded in the B. natans plastid genome, 53 are also present in the plastid genome of the green alga C. reinhardtii and A. thaliana. One of these proteins, YCF1 proved to be unalignable and was excluded from the analysis. Since the genomes of all plastids are descendents of the cyanobacterial primary plastid ancestor, these proteins are homologues .The plastids of both d genome  45 are aG. theta, C. merolae and A. thaliana are smaller than the average distances between the nuclear-encoded proteins  between the plastid-encoded proteins of s Figure . The diss Figure . HoweverG. theta and B. natans [G. theta transit peptides have the characteristics of red algal transit peptides [B. natans, of green algal transit peptides [A. thaliana and from the nuclear genomes of C. merolae (for G. theta) or C. reinhardtii (for B. natans). Twenty-four nucleus-encoded plastid-targeted proteins were found in EST data from G. theta for which there were identifiable homologues in C. merolae and A. thaliana, and 45 plastid proteins were identified from B. natans for which there were identifiable homologues from C. reinhardtii and A. thaliana. A G. theta gene encoding an isoform of glycogen (starch) synthase was excluded from the analysis since starch is accumulated in the periplastidial space in this species, but its homologue in C. merolae is active in the cytosol and the its homologues in green algae and plants are active in the plastid [tha4 gene also found in the G. theta nucleomorph. The protein encoded by this gene was longer than the nucleomorph protein, which, in comparison to isoforms from other species appears truncated. It is possible that the nuclear tha4 gene is a recent transfer that has assumed the function of the nucleomorph-encoded protein and that the truncated, nucleomorph copy is in the process of being lost.Nuclear-encoded plastid genes have been transferred from the plastid genome during endosymbiosis resulting in reduced organelle genomes. Nucleus-encoded plastid-targeted proteins of . natans ,47 were . natans . In G. tpeptides , and in  plastid . Also exG. theta, A. thaliana and C. merolae are larger than the plastid-encoded proteins, and are almost identical between the three species  the rates of general substitution are higher in nuclear genomes, or 2) the genes retained in the plastid genome are those under the greatest selection. A combination of both factors may occur. These results for plastid-encoded and nucleus-encoded plastid-targeted proteins are an important indication of the relative distances between the species for which the rates of divergence of the nucleomorph genomes can be compared.G. theta and B. natans each only encode a handful of plastid proteins, and even fewer for periplastidial metabolism. The rest of the genes encode proteins to support the nucleomorph; proteins for transcription, translation, protein folding and degradation and RNA metabolism [Previous phylogenetic observations of nucleomorph-encoded proteins, have led to speculation that the nucleomorph genomes are extraordinarily divergent, however these studies have been made of proteins that do not target to the plastid. The nucleomorph genomes of tabolism ,8. TheseG. theta and nuclear-encoded homologues from A. thaliana &C. merolae are larger than the distances between proteins that are plastid-encoded in all species (red), whereas there is less difference between these protein distances and those of proteins that are nucleus-encoded in all species (blue). However, significantly, the relative distances between taxa are not equal. The distance to G. theta from both A. thaliana and C. merolae is greater than the difference between A. thaliana and C. merolae , which contains 14 further genes encoding proteins targeted to the plastid [The nucleomorph of function . Of thes plastid .tha4, which may have led to the demise of the nucleomorph-encoded gene relatively recently showing the ongoing nature of the process. By extension, it is possible that only the few genes whose proteins are more permissive to mutation can tolerate the high mutation rate of nucleomorph genomes. Selection pressure favouring the successful transfer of genes for proteins under tighter selection for sequence conservation would be stronger. This would suggest that the genes for plastid-targeted proteins remaining in the nucleomorphs would be divergent compared with homologues in other eukaryotes, perhaps as divergent as other nucleomorph proteins on average.Why these plastid-targeted proteins remain encoded in the nucleomorph may be the key to the existence of the genome itself, since almost all other nucleomorph-encoded proteins are for self-maintenance and expression of the genome. A variety of biological explanations have been suggested for the retention of certain core proteins in most chloroplast and mitochondrial genomes ,52, howeG. theta had identifiable homologues in the nuclear genomes of C. merolae and A. thaliana and 17 nucleomorph-encoded plastid-targeted proteins of B. natans had identifiable homologues in the nuclear genomes of C. reinhardtii and A. thaliana.To test these hypotheses, we first compared the relative distances of nucleomorph-encoded plastid-targeted proteins to nucleus-encoded plastid-targeted and plastid-encoded proteins Figure . FifteenG. theta and nuclear-encoded homologues from A. thaliana and C. merolae are larger than plastid-encoded proteins, but similar to nucleus-encoded plastid-targeted proteins. The distances between the three species are not equal. As for the plastid-encoded proteins, the distance to A. thaliana from both G. theta and C. merolae is much greater than the difference between G. theta and C. merolae  and plastid-encoded proteins (37%) fail the relative rate test in B. natans in which cases B. natans is typically the most rapidly evolving peptide. In G. theta, nucleomorph-encoded plastid-targeted proteins fail the relative rate test more frequently than those encoded in the plastid or nucleus, but the difference is not nearly as pronounced as in B. natans. In fact, nucleomorph encoded plastid-targeted proteins in G. theta only fail the relative rate test 11% more frequently than nuclear-encoded plastid-targeted proteins in which G. theta is the most rapidly evolving taxon. Interestingly, of the 17% of the plastid-encoded peptides that fail the relative rate test, G. theta is not the most rapidly evolving ingroup. This may indicate that the plastid of C. merolae is evolving at an accelerated rate compared to that of G. theta.Relative rate tests can be used to measure the degree of divergence of two genes from an equally distant outgroup ,54. Relag RRTree  and wereOverall, the rate of evolution of plastid proteins encoded in the nucleomorph of cryptomonads is in line with those encoded in the nucleus, despite the fact that other nucleomorph-encoded proteins are generally evolving at a higher rate. In chlorarachniophytes, however, the nucleomorph-encoded plastid-targeted proteins are evolving much faster than those encoded in the nucleus , which provides one of the first indications that the mode of evolution in these two genomes is fundamentally different.Thalassiosira pseudonana, and the distance of these proteins compared to an extant free-living plastid relative; the cyanobacterium Synechocystis PCC 6803. This analysis showed that plastid proteins that are encoded in the nucleomorph of either G. theta or B. natans  from G. theta and R. salina are actually less than the distances between nucleus-encoded proteins  and Rhodomonas salina (CCMP 1319) and also previously published data from B. natans [B. natans where several lateral gene transfers have been identified [DQ383756-DQ383799. Proteins were also identified from the coding sequences of the ongoing sequencing project of the plastid genomes of Bigelowiella natans and the plastid genomes of G. theta [Arabidopsis thaliana [Cyanidioschyzon merolae [Odontella sinesis [Thalassiosira pseudonana [A. thaliana [C. merolae [Synechocystis sp. PCC 6803, and the nucleomorph genomes of G. theta [B. natans (DQ158856 – DQ158858). When multiple isoforms existed in the algal or plant nucleus and it was not obvious which isoform was the orthologue, the distances for all isoforms were calculated and the isoforms with the closest distance to the cryptomonad or chlorarachniophyte was used, providing that the same isoforms from the algae and plant were also closest to each other. Alternatively, in a few cases, a neighbour-joining phylogenetic tree was constructed to determine groups of isoforms. In a minority of cases for nucleomorph-encoded plastid proteins in B. natans where there were multiple paralogues in both A. thaliana and C. reinhardtii, the nearest Arabidopsis paralogue to B. natans was not nearest to the C. reinhardtii paralogue closest to B. natans. In these cases the paralogue closest to B. natans in pair-wise distance (using Dayhoff) was chosen. If it was not possible to determine which isoform was the likely original paralogue then that protein was excluded from the analysis. For analyses with nucleomorph-encoded non-plastid proteins a subset of proteins involved in transcription, translation  and protein folding for which homologues could be identified in A. thaliana and C. reinhardtii or C. merolae, was used.Proteins representing known plastid functions from other eukaryotes and cyanobacteria, were used to search ongoing EST projects from the cryptomonads . natans ,47, resuentified , only nuG. theta , Arabidothaliana , Cyanidi merolae  and Odon sinesis . Homologeudonana , A. thalthaliana , C. mero merolae . ProteinGuillardia theta were used to search a database of Rhodomonas salina (CCMP 1319) ESTs using tBLASTn. The GC content of the transcripts was calculated and compared to the GC content of the G. theta nucleomorph and nuclear genome and also to R. salina proteins identified as being nuclear-encoded, plastid-targeted. R. salina transcripts with homologues in the G. theta nucleomorph with coding regions of 28% GC content or less were determined to be nucleomorph encoded.Proteins encoded in the nucleomorph genome of G. theta distances were also calculated with the Dayhoff substutution matrix and nine rates catagories (eight variable and one invariable), to test for saturation [see Protein alignments were made using Clustal X  and refition see .C. reinhardtii as an ingroup and A. thaliana as an outgroup for B. natans datasets. C. merolae was used as an ingroup and A. thaliana as an outgroup for G. theta datasets. The test was used to compare the evolutionary rate of individual genes from each of the three genomes of B. natans and G. theta to its compartment specific homologue in the genomes of C. reinhardtii and C. merolae. Since a failure of a relative rate test does not indicate which taxon is evolving more rapidly, we compare failures where G. theta or B. natans is the most rapidly evolving ingroup.Relative rate tests were performed using the RRTREE program  using C.EST, Expressed Sequence TagAll authors contributed to the experimental concept and design, the generation of EST data, data acquisition, analysis, statistics, and manuscript preparation. All authors read and approved the final manuscript.G. theta (GT), the red algae C. merolae (CM) and the plant A. thaliana (AT) calculated with the Dayhoff substitution matrix and nine catagories of gamma corrections (eight variable +1 invariable).Supplementary Figure 1. Radar graph of average distances of plastid-, nucleus-, and nucleomorph-encoded plastid proteins, and nucleomorph-encoded non-plastid proteins of the cryptophyte Click here for file


Anopheles maculipennis complex are effective malaria vectors in Europe and the Caspian Sea region in northern Iran, where malaria has been re-introduced since 1994. The current study has been designed in order to provide further evidence on the status of species composition and to identify more accurately the members of the maculipennis complex in northern Iran.Members of The second internal transcribed spacer of ribosomal DNA (rDNA-ITS2) was sequenced in 28 out of 235 specimens that were collected in the five provinces of East Azerbayjan, Ardebil, Guilan, Mazandaran and Khorassan in Iran.maculipennis complex could be easily clustered into three groups: the An. atroparvus – Anopheles labranchiae group; the paraphyletic group of An. maculipennis, An. messeae, An. persiensis; and An. sacharovi as the third group.The length of the ITS2 ranged from 283 to 302 bp with a GC content of 49.33 – 54.76%. No intra-specific variations were observed. Construction of phylogenetic tree based on the ITS2 sequence revealed that the six Iranian members of the An. maculipennis complex including An. atroparvus, An. messae and An. labranchiae, as shown as new records in northern Iran, is somehow alarming. A better understanding of the epidemiology of malaria on both sides of the Caspian Sea may be provided by applying the molecular techniques to the correct identification of species complexes, to the detection of Plasmodium composition in Anopheles vectors and to the status of insecticide resistance by looking to related genes.Detection of three species of the  Anopheles maculipennis, the historic vector of malaria in Europe and the Middle East was the first sibling species complex to be discovered among mosquitoes , An. messeae [GenBank: AY050639], and An. atroparvus [GenBank: AY050640] in Iran by using RAPD, SSR and ITS2 sequences. Recently, Sedaghat et al. [An. maculipennis complex from Iran, including An. maculipennis, An. sacharovi, and An. persiensis. The sequence of An. persiensis was identified in 2002 by Djadid and Romi from Rasht (Guilan province) and Amol city in Mazandran province, and indeed it was described later by Sedaghat et al. [et al. [An. maculipennis complex from the north-west and central regions of Iran, only found An. maculipennis and An. sacharovi. However, in study, for the first time, the presence of six species of this complex  are reported in northern Iran based on the sequence of rDNA-ITS2. The three species of An. messeae, An. atroparvus and An. labranchiae have not been reported before in Iran.This study has been designed in order to provide molecular evidence and to verify the real composition of the nce 1986  and latence 1986 . Howeverh et al.  and Mano [et al.  claimed . Djadid , working [et al. , workingAn. maculipennis complex varied in length from 283 bp in An. maculipennis up to 302 bp in An. sacharovi. This is in the range of ITS2 length in other examined Anopheles species; 363–369 bp in Anopheles nunestovari [Anopheles quadrimaculatus complex [An. maculipennis complex 305 – 310 bp [An. maculipennis complex about 280–300 bp [The ITS2 sequences of six Palearctic species of estovari , 287–329 complex , in the – 310 bp  and in s0–300 bp . Howeveret al. [An. maculipennis complex inferred by maximum parsimony analysis of the PAUP programme and neighbour joining and maximum likelihood analysis of the PHYLIP program. All the trees obtained were almost identical in topology although the relationships among the three species i.e. An. maculipennis, An. messeae and An. melanoon, remained unresolved. Perhaps due to the differentiation of these species from neighbouring taxa within a brief evolutionary time-frame that dispensed insufficient differences to support these individuals' lineages [An. maculipennis complex have been reported from Romania and Russia, namely An. daciae, An. artemievi and An. beklemishevi [An. beklemishevi specimens from Russia by their ITS2 ribosomal DNA sequences to amend and to specify the phylogenetic tree of the An. maculipennis species complex [An. persinsis, and that An. messeae and An. atroparvus are not as close as demonstrated by Kampen [An. beklemishevi is in a closer relationship to the Nearctic rather than to the Palearctic sibling species, which is in concordance with the demonstration of final phylogenetic tree drawn from this study by including An. beklemishevi sequence (GenBank: AY593958) to other sequences  collected during the suspected hibernation period (October-March) showed that this species will go into hibernation with gonotrophic dissociation. In this case, all dissected mosquitoes have shown no dilatation proving that the female adult mosquitoes over winter as nuliparus and the last blood they took will be used for producing the fat body allowing the female to start the next generation in the beginning of next seasonal activity. However, it remains un-clear how these ecological data fit with the presence of one or more species within An. maculipennis complex in northern Iran, and what is the role of other anophelines in this region  in malaria transmission and since re-introduction of malaria in northern Iran.With regards to the presence of six members of Anopheles culicifacies, Anopheles gambiae, Anopheles fluviatilis [Beklemishev  listed eviatilis ,55-59. TAn. sacharovi, which is responsible for the majority of Plasmodium vivax transmissions in the Asian part of Turkey [An. labranchiae, formerly was the main vector of malaria in Italy [An. atroparvus is the most efficient vector in Britain [An. maculipennis, An. messeae and An. sacharovi are capable of transmitting malaria; however, they exhibit different vector capacities [An. maculipennis complex including An. atroparvus, An. messae and An. labranchiae, as new records in northern Iran, is a case for concern because of their potential in malaria transmission and more important, the extent of their geographical distribution towards southern territory of the An. maculipennis complex.Nowadays, the most important vectors are considered to be f Turkey , and An.in Italy . An. atr Britain . An. macpacities ,60,61. IAn. maculipennis complex is distributed over an area that covers central Iran to the Caspian Sea region and up to Sweden, an international effort is required to prevent the re-introduction of malaria to those areas where "anophelism without malaria" prevails.A better understanding on the epidemiology of malaria on both sides of the Caspian Sea may be provided by applying the molecular techniques to enable the correct identification of species complexes, the detection of plasmodium composition in anopheles vectors and the status of insecticide resistance by looking at related genes. In addition, it is worth remembering that as the


Lepilemur and their phylogenetic relationships is disputed and controversial. In order to establish their evolutionary relationships, a comparative cytogenetic and molecular study was performed. We sequenced the complete mitochondrial cytochrome b gene (1140 bp) from 68 individuals representing all eight sportive lemur species and most major populations, and compared the results with those obtained from cytogenetic studies derived from 99 specimens.The number of species within the Malagasy genus Lepilemur ankaranensis, L. dorsalis, L. edwardsi, L. leucopus, L. microdon, L. mustelinus, L. ruficaudatus and L. septentrionalis on species level. Additionally, within L. ruficaudatus large genetic differences were observed among different geographic populations. L. dorsalis from Sahamalaza Peninsula and from the Ambanja/Nosy Be region are paraphyletic, with the latter forming a sister group to L. ankaranensis.Interspecific genetic variation, diagnostic characters and significantly supported phylogenetic relationships were obtained from the mitochondrial sequence data and are in agreement with cytogenetic information. The results confirm the distinctiveness of L. ruficaudatus and L. dorsalis. Based on molecular data we propose to recognize the sportive lemur populations from north of the Tsiribihina River, south of the Betsiboka River, and from the Sahamalaza Peninsula, as distinct species.Our results support the classification of the eight major sportive lemur taxa as independent species. Moreover, our data indicate further cryptic speciation events within  Lepilemur, are small nocturnal primates endemic to the island of Madagascar. They are amongst the most widely distributed lemurs, occurring in almost all natural evergreen or deciduous forest formations on the island . Intra-specific differences range from 0.00–7.63%. Interestingly, the observed differences of 5.88–7.63% between the three major populations of L. ruficaudatus, which are separated by wide rivers or large distances, and those found between L. dorsalis from Sahamalaza Peninsula and from Ambanja/Nosy Be (5.18–5.88%) exceed those detected between L. ankaranensis and the two L. dorsalis populations, indicating speciation events in L. ruficaudatus and L. dorsalis. Further details about pairwise differences within and between species are reported in Complete mitochondrial cytochrome b gene sequences (1140 bp) were generated from 68 individuals representing all currently recognized species. Among the 68 sequences we detected 39 haplotypes. Uncorrected pairwise differences within the genus range from 0.00–16.82%, with overlapping intra- and inter-specific differences . Although most inter-specific differences are in the range of 7.37–16.82%, there are also two exceptions which show that differences between species can be much lower , or to those detected between the two widely recognized species L. septentrionalis and L. ankaranensis (82 characters).Based on the 1140 bp of the mitochondrial cytochrome b gene, a population aggregation analysis was performed with 373 positions serving as diagnostic characters see . With 16L. mustelinus was the first species to split off. The remaining species diverged into two subgroups, of which one contained L. ankaranensis, L. dorsalis, L. edwardsi, L. microdon and L. septentrionalis, and the other L. leucopus and L. ruficaudatus. Within L. ruficaudatus three very distinct clades were observed, corresponding to different geographic locations. In the other subgroup a major split occurred between L. microdon/L. edwardsi and the remaining species, which later separated into L. septentrionalis and a group consisting of L. dorsalis and L. ankaranensis. L. dorsalis is further divided into two subclades, with one containing individuals from Ambanja and Nosy Be, and the other those from Sahamalaza. Moreover, L. dorsalis emerges as paraphyletic, with specimens from Nosy Be/Ambanja forming a sister clade to L. ankaranensis.Phylogenetic trees reconstructed on the basis of different algorithms and models of sequence evolution produced identical tree topologies, with mainly significantly supported branching patterns Fig. . Based oLepilemur species.As in many nocturnal mammals, pelage colouration among sportive lemurs is not a suitable characteristic for distinguishing among taxa, since it is not well defined and consistent within taxa. In contrast, cytogenetics or molecular methods such as sequencing of marker genes offer powerful tools that lead to important insights into the diversity and phylogeny of several Malagasy lemur genera ,19,28,29Lepilemur can be divided into the eight traditionally recognized species, L. ankaranensis, L. dorsalis, L. edwardsi, L. leucopus, L. microdon, L. mustelinus, L. ruficaudatus and L. septentrionalis. These differ by several chromosomal rearrangements (3 to 19), with the exception of L. septentrionalis and L. ankaranensis that only one rearrangement separates [L. ruficaudatus populations and the two L. dorsalis populations, no chromosomal differences were detected.Based on cytogenetic data, the genus eparates -9 , as well as from comparable data from other lemur genera, e.g. Microcebus [Geogale aurita, Echinops telfairi, Hypogeomys antimena, Mungotictis decemlineata, Propithecus verreauxi verreauxi, Microcebus berthae; Mahavavy: Eulemur fulvus rufus, Galida elegans occidentalis; Betsiboka: Oryzorictes talpoides, Avahi occidentalis; Sambirano: Phaner furcifer parienti and possibly Avahi unicolor).Besides the high genetic differences observed among the eight jahamena ,17,25,31crocebus ,18. ThusL. dorsalis populations of Ambanja and Nosy Be are closely related and form one clade, indicating both their common origin and a recent isolation of the island of Nosy Be. The Nosy Be/Ambanja L. dorsalis and L. ankaranensis together form a sister group to L. dorsalis from Sahamalaza, indicating a paraphyly of L. dorsalis. These phylogenetic and biogeographic patterns are also confirmed by RAPD analysis [Propithecus (Indriidae) in that P. v. coquereli appears to cluster with P. tattersalli instead with other subspecies of P. verreauxi [L. ankaranensis and Nosy Be/Ambaja L. dorsalis , and heSkull , north-east side of the Mahavavy du Sud River, Fokotany Ambatomahavavy, Firaisana Antongomena-Bevary, Fivondronona Mitsinjo, Province Mahajanga, Madagascar.Pelage coloration is considerably variable in the expression of the colours, possibly as a function of the age of individuals. Depending on light conditions (daylight or flashlight at night) the impression of colours may change subjectively. However, some constant characters are present, though variably expressed. The face is essentially grey and the ears are protruding and rounded. Sometimes there is the impression of a "facemask", in that there may be a darker diffuse, patch of hair in the middle of the forehead. Above the eyes darker coloured but diffuse stripes may run upwards to join in the middle of the head. These confluent stripes continue as one darker and distinct stripe along the back. The stripe is especially well expressed until it reaches the middle of the back, and then continues less prominently to the tail. On the back, the animals are essentially grey and reddish grey. The middle part of the back, especially, may show considerable reddish colouration that extends onto both shoulders and the upper and lower arms. The thigh and lower limb in general are less reddish than the upper part of the body. The ventral pelage is light to darker grey. The tail is variably coloured between grey with some red influence to deep rusty red with negligible grey influence.L. randrianasoli and L. ruficaudatus, from all other sportive lemurs in karyotype , which has supported our fieldwork for 12 years.The taxon occurs between the Betsiboka and Mahavavy du Sud rivers. The southern extension of the taxon across the Mahavavy du Sud River is unknown, and needs further research.Tissue and DNA from one individual stored at the Gene Bank of Primates, German Primate Centre, Germany (GBP 941).Andramasay , Province Toliary, Madagascar.Measurements of five males and four females from the type locality Andramasay are listed in L. aeeclis and L. ruficaudatus, from all other sportive lemurs in karyotype .Sahamalaza Peninsula , Province Mahajanga, Madagascar. Hypodigm: 6 animals from Ankarafa , 25 km southeast of the type locality, for which morphometric measurements have been taken.Pelage coloration is variable, possibly also depending on age of individuals. Depending on light conditions (daylight or flashlight at night) the impression of colours may change subjectively. However, some constant characters are present, though variably expressed. The face is essentially grey. The forehead and the hairline around the ears are red-brown with sometimes darker diffuse patches. A dark diffuse line runs from the middle of the upper skull down the spine, ending in the middle or at the lower part of the back, but is never present on the tail. The dorsal pelage, including shoulders and the upper and lower arms, is predominantly red-brown, whereas the thigh and lower limbs in general are less reddish than the upper part of the body. The ventral pelage is generally grey to creamy. The coloration of the tail is red-brown to deep brown. Measurements of 2 males and of 4 females from Ankarafa are listed in L. dorsalis and L. leucopus, from all other sportive lemurs in diploid chromosome number  data, e.g. from nuclear DNA loci, are required to confirm the distinct species status of the populations. Furthermore, behavioural and morphological studies will provide independent data to delimit the species status of the three new forms. The discovery of three new possible primate species however, shows how diverse Madagascar's fauna is and how limited our knowledge currently still is. Ongoing work in the field and in the laboratory is urgently required to describe the complete diversity of sportive lemurs, especially of the different populations representing L. leucopus, L. microdon and L. mustelinus, and of other Malagasy lemurs. Greater knowledge of ecological, evolutionary and biogeographic patterns and processes will provide the necessary basis for protecting Madagascar's unique biodiversity.The combination of cytogenetic and molecular approaches reported here revealed important information about the diversity and evolution of the sportive lemurs. Phylogenetic relationships obtained from the mitochondrial sequence data are completely resolved and significantly supported, so that they most likely display the real evolutionary relationships among the analysed species and populations. Both methods were able to confirm the species status of the eight traditionally recognized species, while mitochondrial sequencing indicated in addition further cryptic speciation events in Lepilemur species were collected during field surveys in Madagascar  or GM2 [L. ruficaudatus in the Kirindy forest. However, morphometric data were collected by different people under various conditions, and hence we expect low inter-observer reliability. We therefore did not use morphometric data for species delimitation, but rather use them as a tool to present a picture of the taxa. Immediately after recovery from anaesthesia, animals were released in their respective capture areas.Samples from 68 individuals representing all currently recognized car Fig. . Skin bi) or GM2 . A part ) or GM2 . From so) or GM2 . For morA total of 99 sportive lemur individuals with at least one specimen per locality was cytogenetically analysed during studies performed from 1975 till 2005 ,37-39. CDNA from the biopsies was extracted using a standard proteinase K digestion, followed by a phenol chloroform extraction  with minThe complete mitochondrial cytochrome b gene was amplified via PCR using the oligonucleotide primers CYT-LEP-L: 5'-AATGATATGAAAAACCATCGTTGTA-3' and CYT-LEP-H: 5'-GGCTTACAAGGCCGGGGTAA-3'. Standard, wax-mediated hot-start PCRs were carried out for 40 cycles, each with a denaturation step at 94°C for 60 s, annealing at 60°C for 60 s, and extension at 72°C for 90 s, followed by a final extension step at 72°C for 5 min. Aliquots of the PCR amplifications were checked by agarose gel electrophoresis. Subsequently, PCR products were cleaned using the Qiagen PCR Purification Kit and sequenced on an ABI 3100-Avant sequencer using the BigDye Terminator Cycle Sequencing Kit (Applied Biosystems), primers as indicated above and the internal primers CYT-LEP-L400: 5'-TGAGGACAAATATCATTCTGAGG-3' and CYT-LEP-H545: 5'-TGGAGTGCGAAGAATCGGGT-3'. The respective sequences were deposited in GenBank and are available under the accession numbers DQ108990-DQ109034 and DQ234881-DQ234900.L. ruficaudatus and one L. dorsalis. As outgroup for phylogenetic tree reconstructions, we selected Phaner furcifer because it displays the most similar orthologous sequence of all Malagasy lemurs to Lepilemur [Sequences were easily aligned by eye due to the lack of insertions or deletions, and were checked for their potential to be correctly transcribed in order to eliminate data set contaminations with pseudogenes. For a comprehensive evaluation of the sequence data, we expanded our data set with orthologous sequences, already deposited at GenBank, from two epilemur . FurtherUncorrected pairwise differences within and between species and major populations were calculated with PAUP 4.0b10  and DnaSA population aggregation analysis (PAA) was performed according to the diagnostic character framework described in . AccordiPhylogenetic tree reconstructions were carried out with the maximum-parsimony (MP), neighbor-joining (NJ) and maximum-likelihood (ML) algorithms as implemented in PAUP or TREEPUZZLE 5.0 . For MP Lepilemur. She did the field work for L. microdon, L. mustelinus, L. edwardsi and L. aeeclis, took morphometric measurements and did a part of the cytogenetic study in the Institut Pasteur de Madagascar (IPM) in Antananarivo.NA participated in the design of the study and the concept of the revision work on Lepilemur specific primers, generated several sequences, performed the statistical analysis of the phylogenetic data and wrote a draft version of the MS in collaboration with YR.JLF designed Lepilemur specific primers, generated several sequences, performed the statistical analysis of the phylogenetic data and wrote the draft and final version of the MS in collaboration with DZ.ChR designed L. ruficaudatus south of the Morondava River. He also sampled L. ruficaudatus in the Kirindy Forest and took morphometric measurements in collaboration with RH. DZ generated sequences from these individuals, did the statistical analysis of the morphometric data and wrote the draft and final version of the MS in collaboration with ChR.DZ organised, lead and conducted the expedition to collect data and samples of L. aeeclis sp. nov. in the field, participated in the design and drafting of the MS, critically revised it several times and approved the final version.UT conceived, planned, organised, lead and conducted the expeditions to collect the samples, data and pictures of Lepilemur, in collaboration with YR and NA. He organized and conceptualized the field work of NA, participated in the collection of samples of L. edwardsi and participated in a revision of a draft version of the MS.ClR participated in the design of the study and the concept of the revision work on Lepilemur. She did the field work for L. septentrionalis, L. ankaranensis, L. dorsalis in Ambanja and part of Nosy Be.IR participated in the design of the study and the concept of the revision work on L. randrianasoli and L. ruficaudatus and provided morphometric data and tissue samples.JG sampled BM organized financed and conceptualized part of the field work on on Nosy Be (with CL), in Sahamalaza Peninsula (with TH and IR), on the ridge of Andrafiamena (with IR) and in Ankarana (with IR). At several of these sites, he collected samples and animal measurements.L. ruficaudatus south of the Tsiribihina River and took morphometric measurements in collaboration with DZ and he took photographs of L. sahamalazensis sp. nov. in the field.RH sampled Lepilemur specific primers and generated several sequences.LW designed L. aeeclis sp. nov., and found the skull of the animal of the type series. He considerably facilitated field work and data collection.AZ participated in the expeditions to collect data on CL did field work on Nosy Be.TH did field work on Sahamalaza Peninsula.EZ organized, financed and conceptualized part of the field work and revised the MS.UR organized, financed and conceptualized part of the field work and revised the MS.MC conducted part of the field work including capturing, measuring and sampling the Sahamalaza population, and revised the MS.Lepilemur.JT participated in the design of the study and the concept of the revision work on IT provided systematic input and checked the MS.Lepilemur, wrote a draft version of the MS, critically revised it several times and approved its final version.YR participated in the design of the study and the concept of the revision work on A table showing details on studied sportive lemur individuals.Click here for fileA table showing diploid number (2N) and chromosomal rearrangements among species and populations .Click here for fileA table showing minimum and maximum uncorrected pairwise genetic differences (in %) within and among analysed species and populations based on complete mitochondrial cytochrome b sequence data.Click here for fileA table showing number of diagnostic characters as obtained from population aggregation analysis (PAA).Click here for fileL. aeeclis syntype skullA table showing morphometric measurements for the Click here for fileL. randrianasoli and L. aeeclis in comparison with L. ruficaudatus.A table showing morphometric measurements of Click here for fileL. sahamalazensis in comparison to L. dorsalis.A table showing morphometric measurements for Click here for fileA table showing classification of sportive lemurs.Click here for file


Dr. Heather F. Henry joined the Center for Risk Analysis (CRIS) on 10 July as a program administrator for the Superfund Basic Research Program (SBRP). She received a B.S. in Biology from the University of Rochester, which included a year of study in plant-derived medicine at universities in Australia and Ecuador. In 2004, she completed a Ph.D. from the University of Cincinnati, where she was an SBRP trainee participating in numerous multidisciplinary research projects. Henry has recently returned from a Fulbright Postdoctoral Fellowship in Australia studying the role of arbuscular mycorrhizal fungi on arsenate acquisition by native grasses growing on gold mine tailings.http://www-apps.niehs.nih.gov/sbrp/products/products4.cfm.Henry’s primary role has been to initiate the SBRP’s new Individual Research Project Program, which is designed to address specific issues that complement the multiproject research programs, meet high-priority research needs of the SBRP, or tackle issues of emerging concern. The first request for information (RFA), released in November, encouraged the development of innovative approaches to remediate contaminated sediments. Henry’s most recent activity has been to organize the upcoming web-based seminar series “Nanotechnology—Applications and Implications for Superfund,” which will highlight the potential of nanotechnology to support characterization and remediation of hazardous waste sites as well as explore the potential risks of this new class of compounds. For more information on the series, please see Dr. Daniel Shaughnessy joined the Susceptibility and Population Health Branch (SPHB) on 16 May. He received a BM degree from the Eastman School of Music in Rochester, New York. He completed an MPH and a Ph.D. from the University of North Carolina, Chapel Hill, in 2000 and 2002 respectively.His research interests are in the molecular mechanisms of antimutagenic compounds, in the role of diet in modulating cancer risk, and in developing short-term markers of effect related to dietary exposures. He has also studied the genotoxic effects of disinfection by-products in drinking water. As a graduate student, Shaughnessy worked with David DeMarini at the U.S. EPA’s Environmental Carcinogenesis Division. He joined Jack Taylor’s lab at NIEHS as a postdoctoral fellow in 2002, where he conducted a human controlled feeding study on the effects of fried meat on DNA damage and the possible inhibition of that damage by other dietary compounds. Results from this work demonstrate that protective dietary components—cruciferous vegetables, yogurt, and chlorophyllin—significantly inhibited DNA damage induced by the fried meat.http://www.gei.nih.gov/exposurebiology/index.asp.Shaughnessy is managing a portfolio of grants related to DNA damage, DNA repair, and mutagenesis. He is also involved in the Exposure Biology Program within the Genes and Environment Initiative, participating in the formulation and implementation of two RFAs aimed at stimulating research to improve measures of early biological responses to common environmental stressors. For more information on the Genes and Environment Initiative, please see ContactsHeather Henry, Ph.D. |henryh@niehs.nih.govDaniel Shaughnessy, Ph.D. |shaughn1@niehs.nih.gov


PLoS Medicine, volume 4, issue 3, doi: 10.1371/journal.pmed.0040101:In http://pubmlst.org/neisseria) developed by Keith Jolley and Man-Suen Chan and sited at the University of Oxford. The development of this site has been funded by the Wellcome Trust and European Union. We are grateful for support and contributions of E. Arnold, F. Binka, S. Droz, I. Ehrhard, B. Genton, and M. Tanner. In the Navrongo Health Research Center, we greatly appreciate the assistance of A. Bugri, S. Abudulai, and A. Wahab in the laboratory; all nurses and health workers in the War Memorial Hospital, Navrongo and the Health Centers of the KND; C. Tindana with all field workers and drivers for excellent work in the field; and T. Tei and M. Bugase for logistic support. We acknowledge the use of the Navrongo Demographic Surveillance System database, and we thank all study participants for their trust and contribution.”The text of the Acknowledgments section was incomplete. It should have read: “We acknowledge the use of the Neisseria Multi Locus Sequence Typing Web site (


AbstractCalomeraaulicoides  is (re-) established (stat. rest.) as this taxon can be found sympatrically and parapatrically together with Calomeralittoraliswinkleri . Morphological character states are discussed to identify Cicindelajavetii Chaudoir, 1861 and C.herbacea Klug, 1832. Calomeraaphrodisia  is recorded for the first time from Israel. The presence of Calomeraaulica  and Grammognathaeuphratica  is confirmed by new records. At least five taxa are PageBreakthreatened or extinct in Israel. For one of these species, Israel has a national responsibility for the conservation as the main part of the distribution range is within this country.The tiger beetles of the southern Levant  and adjacent regions of the neighboring countries Lebanon, Syria, Iraq, Saudi Arabia and Egypt are reviewed in terms of species taxonomy, ecological and distributional traits and conservation biology. An illustrated dichotomous identification key from the species of this region is provided. Based on the classical identification key, we developed a digital identification application for smartphones and tablets. The species status of https://doi.org/10.3897/zookeys.734.21989.suppl1. See also disclaimer of warranties.Availability: The application TIGER BEETLE ID for Android devices can be freely downloaded at  Cicindela’ devoted exclusively to this group, and highlights the public interest in these animals which belong to the best known insects  In the faunistic part of their work erlooked . Therefo(2) An identification key which includes the species of the adjacent countries would be useful. To meet the need of many enthusiasts and laypersons, for example in Facebook groups which discuss entomology, we present a field guide for mobile devices, such as smartphones and tablets cf. . MoreoveCicindelalittoralis group in the southern Levant, need to be revised.(3) Open systematic questions, for example, the systematic rank of the two parapatric and sympatric “subspecies” of the (4) Finally, as claimed by both taxonomists and conservationists e.g. , we incoWe define the southern Levant as a section of Southwest Asia comprised of the Sinai Peninsula (Egypt), Israel , and Jordan. Species known from surrounding regions in Egypt, Lebanon, Syria, Iraq and Saudi-Arabia are also considered. For an overview of the study area see Fig. Geadephaga. In many other systematic compilations of Adephaga, tiger beetles are ranked as a subfamily of Carabidae . However, the taxonomy of these genus group names is not consistently used in the literature, especially experts from the PageBreakPageBreakNew World and many professional biologists still adopt the broad and conservative definition of the genus Cicindela . We studied approximately 2,000 specimens stored in the following collections:CAL Working collection Assmann, Lüneburg, Germany (part of ZSM)CGD Working collection Gebert, Dresden, GermanyCOQ Working collection Orbach, Qiryat Tiv’on, Israel CSW Working collection Starke, Warendorf, Germany CSH Working collection Schnitter, Halle/S., GermanyCWB Working collection Wrase, Berlin, Germany (part of ZSM)SMNHTAU Steinhard Museum of Natural History, National Collections, Tel Aviv University, Tel Aviv, IsraelNHMP Muséum National d’Histoire Naturelle, Entomology Department, Paris, FranceZISP Zoological Institute of the Academy of Sciences, St. Petersburg, RussiaZSM Zoological State Collection Munich (Zoologische Staatssammlung München), München, GermanyWe received information from colleagues about few records from the following collections:SDEI Senckenberg German Entomological Institute (Deutsches Entomologisches Institut), Müncheberg, GermanyZISPColeoptera Department, Laboratory of Insect Taxonomy, Zoological Institute of the Russian Academy of Sciences, St. Petersburg, RussiaPageBreakFor detailed explanations about measurements, photography, traits and distributional data see other publications about the carabid beetles of the southern Levant ; 2015b.As few biologists and citizen scientists work on tiger beetles in the Middle East, our data do not allow for the estimation of a trend for all species. Thus, we used the approach of If large parts of the distribution range (>67 %) of a taxon are in the study region we derive the national responsibility of the given states for the preservation of the given taxa cf. .We developed two tools for the identification of the tiger beetles in the southern Levant:(a) A “classical”, dichotomous identification key  containi(b) An Android application for mobile phones and tablets . Text and figures are adopted from the “classical” identification key.In general, tiger beetles differ from all other ground beetles in the position of antennae which insert on the frons of head, between the bases of mandibles Fig. . In othePageBreakTiger beetles are agile, usually diurnal beetles with a head (including the eyes) wider than the pronotum; long, thin legs and long, sickle-shaped mandibles with long, simple teeth along the inner edge of the mandibles Figs , 3. All The habitus photographs Figs –38 proviPageBreakPageBreakPageBreakFor ease of orientation, the numbering schemes of the species found in the identification key and in the species accounts are identical. Species which are known from adjacent countries, but not from the southern Levant itself are given in parentheses.Cephalotadeserticola  for Israel. However, its distribution range stretches from western Iran to Central Asia and China (SMNHTAU (including the recently transferred collection of Ptashkovsky), we treat the published records for C.deserticola as misidentifications .nd China  and therions cf. , and do Cylinderapygmaea , 31. Calomeracaucasica , the Salpingophora species 32. S.bellana , 33. S.hanseatica  and 34. S.rueppelii , and the Hypaetha species 35. H.schmidti  and 36. H.copulata . These species are recorded from the adjacent countries , and thus their occurrence in the southern Levant is unlikely. In many cases, the photographs from The following species are also not incorporated in the key: 30. ies e.g. , but exchttps://doi.org/10.3897/zookeys.734.21989.suppl1. On most devices, the app requires less than 150 Mega bytes (MB) of storage.The above presented key for the tiger beetles of the southern Levant and adjacent territories is also available as a stand-alone application (app) for portable Android devices  of 21 or higher is recommended), and can be downloaded from PageBreaklevel. The species list is probably be more helpful for experienced users, while beginners should start with the identification key. Figure After the loading screen, the users will first see a short morphological definition of cicindelids and drawings of the external morphology with key terms indicated (see above). The next screen leads to the dichotomous identification key and to the species list. All photographs and most of the drawings have a zoom function which enables viewing at a higher resolution. Each species name is linked to the species’ accounts with information about habitat, distribution and conservation status (shortened version of the species accounts given below). Here too, a habitus photograph which can be enlarged allows for better orientation and helps to verify identification to species All species are macropterous and flight active. If the species are not recorded from the southern Levant, or if not enough data about the populations during the last decades are available, no conservation information is given.Taxon classificationAnimaliaColeopteraCicindelidae1.Anthrocnemum). Around the Dead Sea and in the Arava Valley in salty wetlands and in date palm plantations (own observations). Nocturnal. Attracted by light.In salty habitats, on the Mediterranean coast in marshlands , adults are active until approximately November (own observations). The number of eggs laid per females in a laboratory experiment ranges from 3 to 25 , which iFrom southern Spain, Morocco and Sardinia to Central Asia , 2017.CGD).Mediterranean Sea coast of the Sinai Peninsula (eastwards to El-Arish) and close to Haifa (Atlit); Red Sea coast of the Sinai Peninsula (incl. near Eilat); in the Dead Sea area  and in the Arava Valley . PutchkoGrammognatha Motschulsky, 1850 is frequently ranked as a subgenus of Megacephala Latreille, 1802, but see armenica  which occurs westwards to Iran. The dark form also occurs on the Mediterranean Sea coast, though it is rare.Endangered in Israel. The species is sensitive to disturbances  . The popPageBreakTaxon classificationAnimaliaColeopteraCicindelidae2.Chaudoir, 1861Open habitats with dwarf shrubs and bare ground, mostly on loamy soils (own observations and Chikatunov pers. comm.), also in quarries. Israeli records from about 1000 m a.s.l. upwards, in Lebanon up to about 2200 m a.s.l. Diurnal.Adults found mainly in May .The species is most likely extinct in Israel, as there have been no new records in the last two decades despite intensive searches on the sites from which the species was previously known. In most cases, the relevant habitats have been destroyed. Populations still exist on the Syrian side of the Hermon, as specimens have been collected there as recently as 2007 (<Syria Occ. Bludan / 40 km west of Damascus / 1700–2300m Antilebano(n) / leg. A. Wrzecionko / 5.5.2007> and same locality, but < … 2200m / Skoupý leg.>; Taxon classificationAnimaliaColeopteraCicindelidae3.Klug, 1832Unknown. Diurnal.CAL, CGS).End of March to July .May Sandy beaches . Larvae PageBreakMain activity period of adults from June to August .Along the coasts of the Mediterranean Sea from Aegean Islands to Syria  .PageBreakNo records.rouxi   .This species is sensitive to disturbances caused by touristic activities on beaches. Even relatively extensive tourism can reduce the activity of adult beetles, and can prevent the development of larvae .Taxon classificationAnimaliaColeopteraCicindelidae6.On river banks and next to freshwater ponds with sparse vegetation on sandy, sometimes cohesive soil . In deseIn Turkey, adults from the end of May to the beginning of September .SDEI: <Cyprus, Baudi>  Therefore, the species is listed correctly by from southeastern Europe to central Asia and India, southwards to Turkey and Syria . Austin SMNHTAU, and the species is mentioned neither by Coleoptera  is found on the Arabian Peninsula In riverbeds with gravel banks and stones, or on sandy ground close to water .PageBreakUnknown.Northeast Africa and the Arabian Peninsula . Horn Mainly in salty habitats, such as sea shores and marshlands with salt crusts, or rocky habitats ; 2000. DPageBreakOn the Sinai Peninsula from February until October , in the From Senegal through northern Africa and Greece to the Middle East and Pakistan .SMNHTAU). Now also recent records from Jordan: “JOR-at-Tafila, Hammam Afra, Hot Springs, 08.05.2010” (CGD), “Dead Sea, Wadi ‘Atun, N Wadi Mujib, same date” (CSH).In northern and southern Sinai along the coasts of the Mediterranean and of the Red Sea, and along the Suez Canal. In Israel in the Dead Sea region . RittnerCalomera species, especially C.aulica, C.diania, C.littoralis and C.aulicoides C.aulica, which can occur on both coastal and inland habitats, C.diania is an exclusive inland species End of February to August Rocky habitats in the littoral zone . Larval May to August .PageBreakFrom Sicily and Greece to Turkey and Syria .ZISP. No recent records from Israel, but suitable habitats still exist in northern Israel (e.g. close to Akko and to Hadera).The first verifiable record from Israel was found by the cicindelid expert A. Putchkov (pers. comm.). He saw an old specimen from northern Israel (label information: <Izrael: Khaifa env.>), together with an old record from Syria ; both specimens are preserved in Declining in Turkey due to touristic activities on the beaches . As the Three subspecies are known, with the nominate form occurring in Turkey and in Syria .PageBreakTaxon classificationAnimaliaColeopteraCicindelidae11.Anthrocnemum species and by Tamarixtetragyna. Also found in river mouths and in freshwater habitats  and C.aulicoides, from where they mention also C.littoralis. We do not know of any verifiable record for Jordan.Along the Mediterranean coast from the mouth of Nahal Betzet  to the Gaza strip . It probwinkleri can be differentiated from the other subspecies of littoralis using the form of the copulatory piece of the median lobe of the aedeagus  and winkleri. These populations are most probably transitional, and likely are hybrid populations. Nonetheless, they are described as a separate subspecies, mandli Mandl, 1934 , stat. rest.On sandy and stony banks close to freshwater , especially in wadis. Also found in salty habitats close to the Dead Sea. Diurnal.Throughout the year (records from February to December)  , which occurs in Sicily (In the past most authors ranked this taxon as a subspecies (or even as a form with a rank below the subspecies) of up) e.g. . Howevering e.g. , and theing e.g. . Pesarining e.g. : 10 are,n Sicily , but is littoraliswinkleri and aulicoides occur. They live in sympatry in the north of Israel , while further southwest they are (at least) parapatric with a distance of about 20 km (Tamra – Eilabun) between populations. Although both taxa are flight-active, no intermediate forms are known . Calomeraaulicoides has a clear and easily accessible character for differentiation from C.littoralis. While the latter one usually has three teeth on the inner side of the left mandible, C.aulicoides has four of them Habroderanilotica.Sandy and stony river banks and on the shore of lakes . See alsIn Africa throughout most of the year .From Senegal to Sudan and Ethiopia . ProbablNo record.Described from Ambukol (= Ambukohl) which belongs today to Sudan .Taxon classificationAnimaliaColeopteraCicindelidae14.C.fimbriata  . ChikatuTaxon classificationAnimaliaColeopteraCicindelidae15. [sic]On seashores and in marshland habitats. Diurnal and nocturnal. Attracted by light .PageBreakMay to September .From southern Spain to the Arabian Peninsula and Northeast Africa .SMNHTAU, CAL).Only in southern Sinai . No recoCephalotalittorea and C.tibialis have long been confused  occurs south of the distribution range of C.littorea s.str. , which occurs along the coasts of the western Mediterranean, is currently ranked as a subspecies of C.littorea, it is probably a valid species. Cephalotalittorea s.str. and goudotii do not occur parapatrically as their ranges are separated from each other by a gap which is partially filled by the distribution range of tibialis. Moreover, the differences in the median lobe of the aedeagus  may support the species status of both goudotii and littorea .Although the taxon The correct spelling of the author name is Forskål  and not Taxon classificationAnimaliaColeopteraCicindelidae16.Shorelines of salt lakes and ponds , sandy bFebruary to September .From Tunisia to Egypt .along the Mediterranean coast of the Sinai Peninsula . No recoC.littorea.Only the nominate subspecies in the southern Levant, the other two subspecies in northern Africa and on Cyprus . See alsTaxon classificationAnimaliaColeopteraCicindelidae17.On salty habitats which have very sparse vascular plant vegetation. Often found on salty crusts of lagoons and ponds behind the coastal dunes , but canPageBreakIn Italy from June to October with activity maximum in June and July .A Mediterranean species from Spain and Algeria to Turkey .PageBreak could not examine specimens from there or from anywhere else on the peninsula. C.circumdata from Sinai are preserved in the Frey collection, and only few tiger beetle individuals from Alfieri’s collection are found in Basel . We do not know of any verifiable record from the Sinai. As all other populations are known from areas with a typical Mediterranean climate, we believe that C.circumdata is not found in the Sinai (cf. No verified population. – The occurrence in El Tor (southern Sinai) has been questioned by inai cf. .Cephalotacircumdata has several subspecies which are mainly characterized by the elytral pale patterns. However, Taxon classificationAnimaliaColeopteraCicindelidae18.Saline habitats with sparse vegetation and salt crusts during summer. Diurnal and nocturnal. Attracted by light .Spring, records from February to June .Israel, Syria to Iran .In the Dead Sea region of Israel . We do nC.zarudniana , Cephalotavartianorum differs from C.zarudniana by slightly slender habitus as well as shape of the median lobe of aedeagus, and in the complete lack of white setae on the genae.While in older publications this taxon is listed as a subspecies of Critically endangered in Israel. Israel has a national responsibility for the worldwide conservation of the taxon.COQ, CAL) and from 2000 in Syria . All other 34 entries in the database Gebert date back to the late 1980s and 1990s. Intensive searches, including use of light traps at night, of several sites in Israel such as the Enot Tsukim Reserve (= Enot Zuqim = Enot Zukim = Einot Zukim = En Fescha) from where populations have been previously recorded, have yielded no new records. The lowering of the water table and changes in land use in the Dead Sea region have strongly impacted many habitats, both freshwater and saltwater. Therefore, at further studies of the Dead Sea region, both on the Israeli and the Jordanian side, are needed to establish wether or not populations still exist.Tiger beetles of coastal habitats are often sensitive to touristic use of beaches . Most ofTaxon classificationAnimaliaColeopteraCicindelidae19.In open forests and in grasslands, mainly on sandy ground in higher elevations .Spring species .Turkey and Syria .No records.PageBreakTaxon classificationAnimaliaColeopteraCicindelidae20.In the southern Levant found exclusively in sandy shore habitats along the Mediterranean Sea. Larval development takes place a few centimeters above the mean sea level, just above where most of the waves break , though Adults from the beginning of May until mid-November . Few aduvaldenbergi subspecies  is patchily located in a small area along the Mediterranean coast from western Egypt to northern Israel (The nominate form is found from south-eastern Europe (Romania and Ukraine) and Asia Minor to Central Asia and China. The n Israel .In Egypt around Abu Qir, Maadia and Ras el Bar . In Isravaldenbergi from Ma’agan Michael as a subspecies. It is characterized by the excessive pale elytral pattern . As the entire world population of this taxon is located in Israel (the majority of the known populations) and in Egypt, these countries bear the responsibility for the worldwide preservation of this subspecies.Critically endangered in Israel. Tiger beetles of coastal habitats tend to be very sensitive to touristic use of beaches . Most ofTaxon classificationAnimaliaColeopteraCicindelidae21.Calomeraaulica, C.alboguttata and Myriochilamelancholica Margins of both stagnant and running freshwater bodies, including artificial water reservoirs, and in salty habitats . DiurnalMarch to December .PageBreakFrom southern Europe to southern Africa and from the Cape Verde Islands to China .Widespread in Sinai, Israel and Jordan .Not endangered, it is abundant even in habitats strongly influenced by human activity .Taxon classificationAnimaliaColeopteraCicindelidae23.Unknown.Unknown.From Turkey and Syria to China .No record.Taxon classificationAnimaliaColeopteraCicindelidae24.Semi-desert and savanna habitats .Unknown.Southern Sahel zone from Mauritania and Senegal to Sudan . Listed No record.Taxon classificationAnimaliaColeopteraCicindelidae25.Sandy seashores .SMNHTAU are also specimens collected in September (own observation).Recorded in Egypt from May to August . In SMNHFrom Egypt east of the Nile to the Arabian Desert , also found in tropical Africa .SMNHTAU records also originate Eurytopic species found on sea shores, in saltmarshes, in dune depressions, on river banks, in oases and in palm plantations, not restricted to coastal habitats .Throughout most of the year, from February to December .From Morocco to Israel .Numerous records from the Mediterranean coast in Israel and northern Sinai, southwards to the Negev and central Sinai .Not threatened. A widespread species which also can be found in highly disturbed habitats.Taxon classificationAnimaliaColeopteraCicindelidae27.On sparsely vegetated escarpments along rivers .Poorly known, records from April and May, but may have a longer activity period .From Turkey to Iran .No record.PageBreakL.hilariola from L.flexuosa.A table found in Taxon classificationAnimaliaColeopteraCicindelidae28.C.fischeri , Jordan, Lebanon, Syria (without the easternmost parts) and northern Saudi Arabia. In the investigated study region, the southern Levant, there are ied e.g. . The incCicindelajavetii, C.herbacea; Calomeraaulicoides, C.littoralis, and C.aulica). However, the majority of the species can be identified correctly under field conditions, by examining live individuals using basic magnification tools. In such circumstances the Android application may be more useful rather than the classic key. We hope that both identification tools will be useful in a range of contexts, such as PageBreakeducation at levels, academic research, the activities of citizen scientists and in practical conservation work like surveying.Under laboratory conditions, all species can be reliably identified using the “classical” identification key, including those requiring the dissection of the male genitalia  must be taken into consideration, and in some cases may need to be developed, as established terms in Hebrew and in Arabic are in many cases lacking.Our application for mobile Android devices, TIGER BEETLES ID, can serve as a starting point for the development of additional tools, with the translation of the app’s text into both Hebrew and Arabic being greatly desired. Moreover, a simple version of the identification application is possible by deleting those alternatives considering the Calomeraaphrodisia for Israel has now been documented in the form of an old specimen in ZISP. The occurrence of two species is confirmed by new records from Jordan . It is likely that additional species which occur in Jordan have not yet been recorded , especially as the oases in the eastern part of the country have, to our knowledge, not yet been sampled.As far as we know, the first record of PageBreakPageBreakPageBreakSMNHTAU which are listed by Calomeraaulicoides for the northern Hula Valley, Calomeralittoralis in eastern Lower Galilee, Calomeraaulica on the Mediterranean Sea coast). The material found in the collection of the Steinhardt Museum at the Tel Aviv University (SMNHTAU) can be used to help bridging the gap between taxonomy and nature conservation biology. Thus the knowledge of the distribution of tiger beetles can be deepened, and the decline of many tiger beetle species can be investigated. This is of particular importance in regions such as the southern Levant, where species diversity has not yet been thoroughly studied in terms of taxonomy and systematics , as we did not find individuals in any of our collecting trips, including nocturnal light trapping methods. Specifically in Enot Zuqim, a decline of the phytodiversity in this nature reserve has been reported ((a) reported .PageBreakCicindelajavetii in the Golan Heights has been destroyed, and no further records since the 1990s are known from Israel. Due to the high power of dispersal of the species, with all individuals being fully winged and flight-active, (re-) colonization of suitable habitats in the southern Levant is possible.(b) The known habitat of Cylinderacontortavaldenbergi, as its entire distribution range is confined to these two countries. However, it is unclear if populations of this taxon still exist, or if C.contortavaldenbergi is extinct. The definition of national responsibility is important in the assessment of national conservation priorities as well as in decision making about inclusion in international conventions on species conservation. The larval development of Cylinderacontortavaldenbergi occurs in close proximity to sea water line on beaches, a habitat which is often extremely disturbed by tourism and recreational activities such as swimming facilities and off-road vehicles which compress the soil and sand and destroy the habitat of the species. For Habroscelimorphadorsalis s.str. Say , an endangered species covered by the U.S. Endangered Species Act  Israel and Egypt are responsible for the world-wide preservation of cies Act  which incies Act . We suggWe hope that our identification tools and the species’ accounts describing the ecology and conservation biology of the tiger beetles of the southern Levant will encourage further work on tiger beetles in the Middle East and enhance the conservation and preservation of these attractive insects.


The present study attempts to examine the longitudinal impact of a curriculum-based positive youth development program, entitled the Project P.A.T.H.S. , on adolescent problem behavior in Hong Kong. Using a longitudinal randomized group design, six waves of data were collected from 19 experimental schools  in which students participated in the Project P.A.T.H.S. and 24 control schools . At each wave, students responded to questions asking about their current problem behaviors, including delinquency and use of different types of drugs, and their intentions of engaging in such behaviors in the future. Results based on individual growth curve modeling generally showed that the participants displayed lower levels of substance abuse and delinquent behavior than did the control students. Participants who regarded the program to be helpful also showed lower levels of problem behavior than did the control students. The present findings suggest that the Project P.A.T.H.S. is effective in preventing adolescent problem behavior in the junior secondary school years.


Anopheles in relation to malaria transmission in Addis Zemen, Ethiopia.Malaria is a public health problem in Ethiopia and its transmission is generally unstable and seasonal. For the selection of the most appropriate vector control measures, knowledge on the ecology of the vector is necessary at a local level. Therefore, the objectives of this study were to document the species composition, breeding habitat characteristics and occurrence of anopheline larva in Sheni stream and the vectorial role of the prevailing Plasmodium and source of blood meals.Immature anophelines were sampled from breeding habitats and characteristics, such as water temperature, turbidity, water current, water pH and other variables, of the habitats were measured from October 2011 to February 2012. Adult anophelines were sampled inside human dwellings using space spray and Center for Disease Control light traps. Artificial pit shelters and clay pots were also used for outdoor adult collections. Anophelines collected were identified using morphological key. The enzyme-linked immunosorbent assay was applied to detect circumsporozoite proteins of Anopheles larvae were collected and identified morphologically. Five anopheline species were found: An. gambiae (s.l.), An. cinereus, An. demeilloni, An. christi and An. pretoriensis. Anopheles gambiae (s.l.) existed in most of the habitats investigated. Only the former three species were captured in the adult collections. Sun-lit Sheni stream, rain pools, hoof prints, drainage and irrigation canals were found to be habitats of larvae. Anopheles gambiae (s.l.) larvae were most abundantly sampled from sand mining and natural sand pools of Sheni stream. Multiple regression analysis showed that clear, permanent and temporary habitats devoid of mats of algae were the best predictors of An. gambiae (s.l.) larval abundance. It is also the responsible malaria vector in the study area and exhibits anthropophilic and endophagic behaviour.A total of 6258 An. gambiae (s.l.) was found in Addis Zemen throughout the study period from both adult and larval collections. Sheni stream is the main larval habitat responsible for the occurrence of anopheline larvae during the dry season of the study area when other breeding sites perish.The malaria vector  Malaria is one of the main public health problems globally and is endemic in 91 countries of the world. Its incidence is estimated to be 212 million cases in 2015; of these, 90% of the cases occur in Africa . After 1 h, wells were washed three times with PBS-Tween 20. Finally, the absorbance at 414 nm was determined with microplate reader 30 min after the addition of 100 μl of ABTS peroxidase substrate. Each blood-meal sample was considered positive if the absorbance value exceeded the mean plus three standard deviations of the mean of three negative controls and also by observing color change (green color).Blood meals of r et al.  at the EAnopheles mosquitoes were tested for sporozoite infection using ELISA as described in Wirtzet al.  were caught using CDC light traps and pyrethrum spray sheet collection (Table An. gambiae (s.l.) and An. cinereus were obtained from 44 houses inspected using PSC method. Anopheles gambiae (s.l.) was more predominant in human dwellings than An. cinereus and An. demeilloni. The mean daily density in CDC light traps (number of anopeline species/trap/night) was 0.87 for An. gambiae (s.l.), 0.19 for An. cinereus and 0.21 for An. demeilloni. The density of An. gambiae (s.l.) varied between the months; the highest density was in October whereas the lowest was in December . Anopheles gambiae (s.l.) was collected only in February from pit shelters. Clay pots were not productive, with only one An. cinereus collected throughout the sampling period.Attempts to collect An. gambiae (s.l.) indicated the source as both human and bovine (Table An. gambiae (s.l.), 47 An. cinereus and 47 An. demeilloni, were tested to detect circum-sporozoite proteins of Plasmodium falciparum and P. vivax by ELISA method, however, none were positive.Blood meals of 29 freshly fed anophelines, of which the majority was sampled from indoor locations, were tested to determine their source of blood meals. The small number of blood meals of ne Table . The othAn. gambiae (s.l.) (presumably An. arabiensis), the principal vector of malaria in the country [An. cinereus, An. demeilloni, An. christiand and An. pretoriensis) is ascertained from both larval and adult sampling. Larvae of An. gambiae (s.l.) were the second most abundant in almost all habitats, including sun-lit pools formed at the bed and edges of Sheni stream, rain pools, hoof prints and drainage canals. All the types of habitats reported here have been previously documented in Ethiopia as well as elsewhere in Africa [An. gambiae (s.l.) and An. cinereus was higher here than other breeding sites. This is inconsistent with previous study in Eretria [An. gambiae (s.l.) as the water is clear and sunlit. This observation is similar to the findings noted by Keneaet al. [This study provided baseline information for the species composition of anophelines, types of larval breeding habitats and adjoining characteristics as well as some entomological indicators in Addis Zemen in relation to malaria transmission. The presence of  country , and foun Africa , 17–19.  Eretria , 19. Saneaet al.  in ZiwayAn. gambiae (s.l.), An. cinereus abundantly breeds in Sheni stream, while the contrary was noted for An. demeilloni, An. christi and An. pretoriensis. All these species are regarded as highland mosquitoes, except for An. gambiae (s.l.) and An. pretoriensis whose distribution extends to the lowlands [Like lowlands , 21.An. gambiae (s.l.). This agrees with recent findings in Ethiopia which indicated that An. arabiensis breeds in clear, temporary and often sun-lit pools of water [An. gambiae (s.l.) larval density with habitats devoid of mats of green algae reported here may be due to the exposure of habitats with muddy substrate to sunlight provide favorable conditions for the survival of bacteria from which larvae get their nutrients [An. gambiae (s.l.) is similar to the findings of Keneaet al. [Anopheles gambiae (s.l.) is positively associated with habitats located between 0 and 100 m from human dwellings, which is one of the strong predictors for indoor Anopheles abundance [An. cinereus.Multiple regression analysis revealed that clear and sun-lit temporary habitats are positively associated with the abundance of of water , 21. Thiof water –24. In cof water . The posutrients , 27. Theeaet al. . This maeaet al. –24. Anopbundance . Change Anopheles collections contained more An. gambiae (s.l.) than the other species, the majority of which were captured from CDC light traps indoors indicating possible host-seeking behaviour, although they may have been attracted by light from the trap. This might be consistent with its anthropophilic and endophagic behaviorur noted by other investigators [An. gambiae (s.l.) tested for blood meal analysis was very low, the few positive reactions exhibited both the zoophilic and anthropophilic behaviour of the vector, which is typical of its biting behaviour [Although the number is small, adult tigators , 29. Thetigators , which wehaviour  and is sehaviour , 30, 31.Anopheles cinereus and An. demeilloni were the second greatest in adult sampling, after to An. gambiae (s.l.), and the majority of these tested for blood meal analysis showed a preference towards cattle feeding showing their zoophilic and poor anthropophilic behaviorur. This is in agreement with the study conducted in highlands of western Kenya [rn Kenya . These srn Kenya .An. cinereus was caught resting in clay pots in outdoor collection, showing very little attraction to man-made habitats. However, in western Kenya, more mosquitoes were captured from clay pots (37%) than pit shelters [Only one shelters .An. gambiae (s.l.), An. cinereus and An. demeilloni were present in collections throughout the study period. Moreover, larvae of the former two species continued to survive in Sheni stream during the dry months, showing the importance of this stream in providing suitable condition for the survival of the two species, particularly to An. gambiae (s.l.), during the period when other breeding sites perish. The presence of An. gambiae (s.l.) in both stages during the entire study period indicates that active transmission of malaria might take place throughout the entire year. Therefore, further study on the prevalence of malaria in conjunction with anophelines is required to better describe the disease.Adults of An. gambiae (s.l.) and other Anopheles species. In addition, this study indicates that the presence of the principal malaria vector, An. gambiae (s.l.), in the country in the study area from both larval and adult collections. Since the collection of adult mosquitoes was low, owing to the brief period of study and the dry season, a more detailed and year-round investigation is required to gather appropriate and relevant entomological indices of transmission towards contributing knowledge-based strategy for effective vector control management.The present study demonstrated the preferred anopheline larval habitats and best predictor environmental factors for larval abundance. Sheni stream, present in the study area, plays an important role in maintaining


Because of a technical error, an incorrect version of the article “We Run This City: Impact of a Community–School Fitness Program on Obesity, Health, and Fitness” appeared on our website. The PDF version of the article was correct as published.Major corrections were to add the middle initial for author Meredith A. Goodwin and to correct the degree for Barbara A. Clint to BA. Other differences were several editorial changes to the article.https://www.cdc.gov/pcd/issues/2018/16_0471.htm. We regret any confusion or inconvenience this error may have caused.The article was corrected on May 4, 2018, and appears online at


Micrargus herbigradus-species group in Poland. The analysis is based on a thorough literature survey, our own materials, and verification of some older collections. We give new diagnostic drawings and review the characters that are useful in identification of species within the group. Three species are present in Poland: M. herbigradus , M. apertus  and M. georgescuae Millidge, 1976. The latter is recorded for the first time in the country, and we add numerous new localities for the two former species. Micrargus herbigradus is common and widespread in Poland, living in various habitats, with only a slight preference to forests. In contrast, M. apertus is widely distributed but rarely found, while its affinity to forests is the highest within the group. The records of this species are most numerous in lowland forests (up to c. 300 m a.s.l), but it can also be found at higher altitudes. M. georgescuae is found only in montane habitats, both in the Sudetes and the Carpathian Mountains, from above 650 m a.s.l. The adults of all three species occur the whole year round, but seem to be most abundant in May and June.We review the known information on the distribution and habitat affinities of the The online version of this article (10.2478/s11756-018-0026-5) contains supplementary material, which is available to authorized users. Micrargus herbigradus-group contains four Central European species: M. alpinus Rëlys and Weiss, 1997, M. apertus , M. georgescuae Millidge, 1976 and M. herbigradus . Two species from this group, M. apertus and M. herbigradus, have been recorded in Poland. Here we add one more species to the list of the Polish fauna, i.e. M. georgescuae. Micrargus subaequalis , which is also present in Poland, belongs to a separate species group.The Micragus species, their actual distribution in Poland was uncertain. Some authors suggested the presence of more than two herbigradus-group members , but to the typical features of a habitat type, e.g. all lowland alder forests or different mire types were recognized as ‘very moist’. The data in the literature were obviously of different quality or credibility, thus selected records were excluded from some counts. The exact number of records used in each analysis is given in the results.For the analysis of phenology as a unit we have used a single record of a species in one plot, per year (i.e. records from the same plot and date coming from two different years were counted separately). We have divided the records into those coming from the first half of each month (1st–15th) and those from the second half (16th–end). For data from pitfall traps we have assigned the record to the respective period by taking the middle date of the trap exposure time.Micrargus herbigradus-group can be identified by comparing their embolus, size and form of the adjacent lamellae, and the shape of the process  that accompanies the embolus. Micrargus georgescuae has a conspicuously wide basal part of the embolus  were collected in several mountain ranges of the Carpathians – the Beskid Wyspowy, the Tatra Mountains, the Gorce ; in the Orawa-Nowy Targ Basin, at high altitudes; and in the Sudetes – the Stołowe Mountains and the Giant Mountains  squares previously. The localities are scattered around the country, from the lowlands to the mountains , even though these ranges have been very intensively surveyed recently. It is also absent in the Slovak part of this mountain massif . Similarly, M. georgescuae was not found in the Izera Mountains (the Western Sudetes) that are close to the other mountain ranges where the species occurs. These mountains have also been intensively surveyed during the last decade.Our data update information on the distribution of the s Thaler , but alss Thaler , Czechias Thaler  and Slovs Thaler . It is as Thaler , namely s Thaler  and in Rs Thaler . The spes Thaler , the Hrus Thaler  or the Gs Thaler , but alss Thaler , the Cens Thaler  and the s Thaler . MicrargMicrargus apertus is a species that is rarely found. Considering the arrangement of its localities in other countries, e.g. in Germany , although sometimes the species were observed together . There is a clear affinity of Micrargus herbigradus was sometimes described as a silvicolous species  by Tomek