WorldWideScience

Sample records for rat-kangaroo bettongia tropica

  1. Locomotion energetics and gait characteristics of a rat-kangaroo, Bettongia penicillata, have some kangaroo-like features.

    Science.gov (United States)

    Webster, K N; Dawson, T J

    2003-09-01

    The locomotory characteristics of kangaroos and wallabies are unusual, with both energetic costs and gait parameters differing from those of quadrupedal running mammals. The kangaroos and wallabies have an evolutionary history of only around 5 million years; their closest relatives, the rat-kangaroos, have a fossil record of more than 26 million years. We examined the locomotory characteristics of a rat-kangaroo, Bettongia penicillata. Locomotory energetics and gait parameters were obtained from animals exercising on a motorised treadmill at speeds from 0.6 m s(-1) to 6.2 m s(-1). Aerobic metabolic costs increased as hopping speed increased, but were significantly different from the costs for a running quadruped; at the fastest speed, the cost of hopping was 50% of the cost of running. Therefore B. penicillata can travel much faster than quadrupedal runners at similar levels of aerobic output. The maximum aerobic output of B. penicillata was 17 times its basal metabolism. Increases in speed during hopping were achieved through increases in stride length, with stride frequency remaining constant. We suggest that these unusual locomotory characteristics are a conservative feature among the hopping marsupials, with an evolutionary history of 20-30 million years.

  2. The high aerobic capacity of a small, marsupial rat-kangaroo (Bettongia penicillata) is matched by the mitochondrial and capillary morphology of its skeletal muscles.

    Science.gov (United States)

    Webster, Koa N; Dawson, Terence J

    2012-09-15

    We examined the structure-function relationships that underlie the aerobic capacities of marsupial mammals that hop. Marsupials have relatively low basal metabolic rates (BMR) and historically were seen as 'low energy' mammals. However, the red kangaroo, Macropus rufus (family Macropodidae), has aerobic capacities equivalent to athletic placentals. It has an extreme aerobic scope (fAS) and its large locomotor muscles feature high mitochondrial and capillary volumes. M. rufus belongs to a modern group of kangaroos and its high fAS is not general for marsupials. However, other hopping marsupials may have elevated aerobic capacities. Bettongia penicillata, a rat-kangaroo (family Potoroidae), is a small (1 kg), active hopper whose fAS is somewhat elevated. We examined the oxygen delivery system in its muscles to ascertain links with hopping. An elevated fAS of 23 provided a relatively high maximal aerobic oxygen consumption ( ) in B. penicillata; associated with this is a skeletal muscle mass of 44% of body mass. Ten muscles were sampled to estimate the total mitochondrial and capillary volume of the locomotor muscles. Values in B. penicillata were similar to those in M. rufus and in athletic placentals. This small hopper had high muscle mitochondrial volume densities (7.1-11.9%) and both a large total capillary volume (6 ml kg(-1) body mass) and total capillary erythrocyte volume (3.2 ml kg(-1)). Apparently, a considerable aerobic capacity is required to achieve the benefits of the extended stride in fast hopping. Of note, the ratio of to total muscle mitochondrial volume in B. penicillata was 4.9 ml O(2) min(-1) ml(-1). Similar values occur in M. rufus and also placental mammals generally, not only athletic species. If such relationships occur in other marsupials, a fundamental structure-function relationship for oxygen delivery to muscles likely originated with or before the earliest mammals.

  3. Characterization of a mutant rat kangaroo cell line with alterations in the cell cycle and DNA repair

    OpenAIRE

    Miyaji, E.N.; Johnson, R.T.; Downes, C.S.; Eveno, E.; Mezzina, M.; Sarasin, A.; Menck, C.F.M.

    2000-01-01

    Using a positive selection system for isolating DNA replication and repair related mutants, we isolated a clone from a rat kangaroo cell line (PtK2) that has increased sensitivity to UV light. Characterization of this clone indicated normal post-replication repair after UV irradiation, and normal removal rates of cyclobutane pyrimidine dimers and pyrimidine(6-4)pyrimidone photoproducts by excision repair. However, this cell line has decreased ability to make early incisions on damaged DNA, po...

  4. Identification of novel Babesia and Theileria genotypes in the endangered marsupials, the woylie (Bettongia penicillata ogilbyi) and boodie (Bettongia lesueur).

    Science.gov (United States)

    Paparini, Andrea; Ryan, Una M; Warren, Kris; McInnes, Linda M; de Tores, Paul; Irwin, Peter J

    2012-05-01

    Piroplasms, which include the genera Theileria and Babesia, are blood-borne parasites transmitted mainly by tick vectors. Relatively little is known about their prevalence and clinical impact in Australian marsupials. In the present study the occurrence and molecular phylogeny of these parasites were studied in both wild and captive marsupials from Western Australia (WA) and Queensland (QLD). Blood samples were screened by microscopy and molecular methods, using PCR and DNA sequencing of the 18S ribosomal RNA gene (18S rDNA). Overall, 7.1% of the blood samples (8/113) were positive for piroplasm 18S rDNA. Theileria and Babesia rDNA was detected in 0.9% (1/113) and 6.2% (7/113) of the animals, respectively. The single Theileria positive was identified in one of three boodies (Bettongia lesueur) screened from a wildlife rehabilitation centre in WA, while all seven Babesia positives were detected in WA in wild captured woylies (Bettongia penicillata ogilbyi). Small intraerythrocytic inclusions were observed in blood films made from six of these individuals. This is the first report of a Babesia sp. in woylies, and Theileria sp. in boodies. Phylogenetic analysis indicated that the woylie-derived Babesia was genetically distinct and most closely related to Babesia occultans, the causative agent of a benign form of cattle babesiosis (genetic similarity 98.4%). The Theileria identified was most closely related to the marsupial-derived species Theileria penicillata from the woylie, Theileria brachyuri from the quokka (Setonix brachyurus), and Theileria sp. from the long-nosed potoroo (Potorous tridactylus). Copyright © 2012 Elsevier Inc. All rights reserved.

  5. Characterization of a mutant rat kangaroo cell line with alterations in the cell cycle and DNA repair

    Directory of Open Access Journals (Sweden)

    Miyaji E.N.

    2000-01-01

    Full Text Available Using a positive selection system for isolating DNA replication and repair related mutants, we isolated a clone from a rat kangaroo cell line (PtK2 that has increased sensitivity to UV light. Characterization of this clone indicated normal post-replication repair after UV irradiation, and normal removal rates of cyclobutane pyrimidine dimers and pyrimidine(6-4pyrimidone photoproducts by excision repair. However, this cell line has decreased ability to make early incisions on damaged DNA, possibly indicating a defect in preferential repair of actively transcribed genes, and a slower cell proliferation rate, including a longer S-phase. This phenotype reinforces the present notion that control of key mechanisms in cell metabolism, such as cell cycle control, repair, transcription and cell death, can be linked.

  6. Experimental Acquisition, Development, and Transmission of Leishmania tropica by Phlebotomus duboscqi

    Science.gov (United States)

    2013-01-01

    ac ta t ropica Experimental acquisition, development, and transmission of Leishmania tropica by Phlebotomus duboscqi Hanafi A. Hanafi, El...August 2012 Accepted 2 September 2012 Available online 10 September 2012 Keywords: Phlebotomus duboscqi Leishmania tropica Transmission Vector...competency a b s t r a c t We report experimental infection and transmission of Leishmania tropica (Wright), by the blood-feeding sand

  7. Biosynthesis of silver nanoparticles by Leishmania tropica | Rahi ...

    African Journals Online (AJOL)

    A novel biosynthesis route for Silver Nanoparticles (Ag-NPs) was attempted in the present study using Leishmania tropica the causative agent of cutaneous leishmaniasis in different countries, particularly in Mediterranean region in Iraq. Silver nanoparticles were successfully synthesized from AgNO3 by reduction of ...

  8. In vitro activity of Arbutus unedo against Leishmania tropica promastigotes.

    Science.gov (United States)

    Kivçak, Bijen; Mert, Tuba; Ertabaklar, Hatice; Balcioğlu, I Cüneyt; Ozensoy Töz, Seray

    2009-01-01

    Pentavalent antimonials are the first choice for the treatment of anthroponotic cutaneous leishmaniasis (ACL) in health centers in Turkey, however in rural areas, traditional plants may be preferred for the treatment of lesions. In recent years a number of papers are published related to the natural products especially plant derivates. Our aim is to investigate the antileishmanial effect of Arbutus unedo which is a wild plant mainly grown in maquis and rocky places of the seabord in South Europe. In the present study, the ethanolic, water and n-hexane extracts from the leaves of Arbutus unedo were originally tested in vitro against Leishmania tropica promastigotes. The ethanol extract of Arbutus unedo leaves at the concentrations of 100, 250, 500 microg/ml were found to be more effective than the other extracts (p:0.000). Our study showed that the ethanolic extract of Arbutus unedo leaves can be a promising antileishmanial agent and further experiments are needed.

  9. The Genetic Relationship between Leishmania aethiopica and Leishmania tropica Revealed by Comparing Microsatellite Profiles.

    Science.gov (United States)

    Krayter, Lena; Schnur, Lionel F; Schönian, Gabriele

    2015-01-01

    Leishmania (Leishmania) aethiopica and L. (L.) tropica cause cutaneous leishmaniases and appear to be related. L. aethiopica is geographically restricted to Ethiopia and Kenya; L. tropica is widely dispersed from the Eastern Mediterranean, through the Middle East into eastern India and in north, east and south Africa. Their phylogenetic inter-relationship is only partially revealed. Some studies indicate a close relationship. Here, eight strains of L. aethiopica were characterized genetically and compared with 156 strains of L. tropica from most of the latter species' geographical range to discern the closeness. Twelve unlinked microsatellite markers previously used to genotype strains of L. tropica were successfully applied to the eight strains of L. aethiopica and their microsatellite profiles were compared to those of 156 strains of L. tropica from various geographical locations that were isolated from human cases of cutaneous and visceral leishmaniasis, hyraxes and sand fly vectors. All the microsatellite profiles were subjected to various analytical algorithms: Bayesian statistics, distance-based and factorial correspondence analysis, revealing: (i) the species L. aethiopica, though geographically restricted, is genetically very heterogeneous; (ii) the strains of L. aethiopica formed a distinct genetic cluster; and (iii) strains of L. aethiopica are closely related to strains of L. tropica and more so to the African ones, although, by factorial correspondence analysis, clearly separate from them. The successful application of the 12 microsatellite markers, originally considered species-specific for the species L. tropica, to strains of L. aethiopica confirmed the close relationship between these two species. The Bayesian and distance-based methods clustered the strains of L. aethiopica among African strains of L. tropica, while the factorial correspondence analysis indicated a clear separation between the two species. There was no correlation between

  10. Evolutionary history of Leishmania killicki (synonymous Leishmania tropica) and taxonomic implications.

    Science.gov (United States)

    Chaara, Dhekra; Ravel, Christophe; Bañuls, Anne- Laure; Haouas, Najoua; Lami, Patrick; Talignani, Loïc; El Baidouri, Fouad; Jaouadi, Kaouther; Harrat, Zoubir; Dedet, Jean-Pierre; Babba, Hamouda; Pratlong, Francine

    2015-04-01

    The taxonomic status of Leishmania (L.) killicki, a parasite that causes chronic cutaneous leishmaniasis, is not well defined yet. Indeed, some researchers suggested that this taxon could be included in the L. tropica complex, whereas others considered it as a distinct phylogenetic complex. To try to solve this taxonomic issue we carried out a detailed study on the evolutionary history of L. killicki relative to L. tropica. Thirty-five L. killicki and 25 L. tropica strains isolated from humans and originating from several countries were characterized using the MultiLocus Enzyme Electrophoresis (MLEE) and the MultiLocus Sequence Typing (MLST) approaches. The results of the genetic and phylogenetic analyses strongly support the hypothesis that L. killicki belongs to the L. tropica complex. Our data suggest that L. killicki emerged from a single founder event and that it evolved independently from L. tropica. However, they do not validate the hypothesis that L. killicki is a distinct complex. Therefore, we suggest naming this taxon L. killicki (synonymous L. tropica) until further epidemiological and phylogenetic studies justify the L. killicki denomination. This study provides taxonomic and phylogenetic information on L. killicki and improves our knowledge on the evolutionary history of this taxon.

  11. Draft genome sequence of Paraburkholderia tropica Ppe8 strain, a sugarcane endophytic diazotrophic bacterium.

    Science.gov (United States)

    Silva, Paula Renata Alves da; Simões-Araújo, Jean Luiz; Vidal, Márcia Soares; Cruz, Leonardo Magalhães; Souza, Emanuel Maltempi de; Baldani, José Ivo

    Paraburkholderia tropica (syn Burkholderia tropica) are nitrogen-fixing bacteria commonly found in sugarcane. The Paraburkholderia tropica strain Ppe8 is part of the sugarcane inoculant consortium that has a beneficial effect on yield. Here, we report a draft genome sequence of this strain elucidating the mechanisms involved in its interaction mainly with Poaceae. A genome size of approximately 8.75Mb containing 7844 protein coding genes distributed in 526 subsystems was de novo assembled with ABySS and annotated by RAST. Genes related to the nitrogen fixation process, the secretion systems (I, II, III, IV, and VI), and related to a variety of metabolic traits, such as metabolism of carbohydrates, amino acids, vitamins, and proteins, were detected, suggesting a broad metabolic capacity and possible adaptation to plant association. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  12. Identification of Leishmania tropica from micro-foci of cutaneous leishmaniasis in the Kenyan Rift Valley.

    Science.gov (United States)

    Odiwuor, Samwel; Muia, Alfred; Magiri, Charles; Maes, Ilse; Kirigi, George; Dujardin, Jean-Claude; Wasunna, Monique; Mbuchi, Margaret; Auwera, Gert Van der

    2012-07-01

    We performed diagnosis and species identification of parasites in lesion samples from suspected cutaneous leishmaniasis patients in four villages, three of which are in a known Leishmania tropica endemic region in Kenya. Samples were analyzed both by microscopy and PCR for Leishmania, and typed by an assay using four ribosomal DNA-based species-identification PCRs. The lesions were demonstrated to be caused by L. tropica, which confirms the re-emergence of cutaneous leishmaniasis from this species after a period of reduced incidence in the endemic zone. Our report highlights the importance of an intervention and sustained Leishmania control program.

  13. Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology.

    Science.gov (United States)

    Baneth, Gad; Yasur-Landau, Daniel; Gilad, Matan; Nachum-Biala, Yaarit

    2017-03-13

    Infection and clinical disease associated with Leishmania major and Leishmania tropica, two common agents of human cutaneous leishmaniosis, have rarely been reported in dogs. This study describes dogs infected with these Leishmania spp. prevalent in the Middle East and North Africa, and compares the serological response of dogs infected with Leishmania infantum, L. major or L. tropica to whole promastigote antigen enzyme-linked immunosorbent assay (ELISA) of each species and to rK39 dipstick. Leishmania major infection in a 5-month-old male dog was associated with alopecic and ulcerative periocular and limb skin lesions which responded to allopurinol treatment. Infection was detected by skin and blood polymerase chain reaction (PCR) and confirmed by DNA sequencing but the dog was seronegative. Leishmania tropica infection was detected in a 3-month-old female dog co-infected with Babesia vogeli and Anaplasma platys and with no skin lesions. PCR and DNA sequencing of the blood and parasite culture were positive for L. tropica. Sera from 11 dogs infected with L. infantum, L. major or L. tropica were reactive with all three Leishmania spp. antigens except for sera from a dog with L. major infection. No significant differences were found between reactivity of dog sera to the antigen of the infecting species, or to the other Leishmania spp. antigens. Sera from dogs infected with L. infantum and L. tropica were positive with the rK39 antigen kit, while dogs with L. major infection were seronegative. Skin lesions in L. major infected dogs from this study and previous reports (n = 2) were ulcerative and located on the muzzle, feet and foot pads and not associated with generalized lymphadenomegaly and splenomegaly. In previous L. tropica infections, skin lesions were proliferative mucocutaneous in young dogs (n = 2), or associated with widespread dermatitis, lymphadenomegaly and splenomegaly in older dogs with similarity to L. infantum infection (n = 2). This

  14. Multilocus Microsatellite Typing reveals intra-focal genetic diversity among strains of Leishmania tropica in Chichaoua Province, Morocco.

    Science.gov (United States)

    Krayter, Lena; Alam, Mohammad Zahangir; Rhajaoui, Mohamed; Schnur, Lionel F; Schönian, Gabriele

    2014-12-01

    In Morocco, cutaneous leishmaniasis (CL) caused by Leishmania (L.) tropica is a major public health threat. Strains of this species have been shown to display considerable serological, biochemical, molecular biological and genetic heterogeneity; and Multilocus Enzyme Electrophoresis (MLEE), has shown that in many countries including Morocco heterogenic variants of L. tropica can co-exist in single geographical foci. Here, the microsatellite profiles discerned by MLMT of nine Moroccan strains of L. tropica isolated in 2000 from human cases of CL from Chichaoua Province were compared to those of nine Moroccan strains of L. tropica isolated between 1988 and 1990 from human cases of CL from Marrakech Province, and also to those of 147 strains of L. tropica isolated at different times from different worldwide geographical locations within the range of distribution of the species. Several programs, each employing a different algorithm, were used for population genetic analysis. The strains from each of the two Moroccan foci separated into two phylogenetic clusters independent of their geographical origin. Genetic diversity and heterogeneity existed in both foci, which are geographically close to each other. This intra-focal distribution of genetic variants of L. tropica is not considered owing to in situ mutation. Rather, it is proposed to be explained by the importation of pre-existing variants of L. tropica into Morocco. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  15. Development of a direct species-specific PCR assay for differential diagnosis of Leishmania tropica

    Czech Academy of Sciences Publication Activity Database

    Jirků, Milan; Zemanová, Eva; Al-Jawabreh, A.; Schönian, G.; Lukeš, Julius

    2006-01-01

    Roč. 55, č. 1 (2006), s. 75-79 ISSN 0732-8893 Grant - others:European Comission(EU) QLK2-CT-2001-01810 Institutional research plan: CEZ:AV0Z60220518 Keywords : Kinetoplastida * Leishmania tropica * PCR assay Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.553, year: 2006

  16. Mapping the genes for susceptibility and response to Leishmania tropica in mouse

    Czech Academy of Sciences Publication Activity Database

    Sohrabi, Yahya; Havelková, Helena; Kobets, Tetyana; Šíma, Matyáš; Volkova, Valeriya; Grekov, Igor; Jarošíková, T.; Kurey, Irina; Vojtíšková, Jarmila; Svobodová, M.; Demant, P.; Lipoldová, Marie

    2013-01-01

    Roč. 7, č. 7 (2013), s. 1-17 ISSN 1935-2735 R&D Projects: GA ČR GA310/08/1697; GA MŠk LH12049 Institutional support: RVO:68378050 Keywords : Leishmania tropica * gene controlling susceptibility * host-parasite interactions * leishmaniasis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.489, year: 2013

  17. Investigation of Double-Band Electrophoretic Pattern of ITS-rDNA Region in Iranian Isolates of Leishmania Tropica

    Directory of Open Access Journals (Sweden)

    MA Ghatee

    2013-06-01

    Full Text Available Background: Leishmania tropica is a genetically divergent species. Amplification of entire internal tran­scribed spacer (ITS region of L. tropica isolates obtained from Bam district, one of the well known focus of anthroponotic cutaneous leishmaniasis ACL( in Iran, revealed a double-band pat­tern in agarose gel electrophoresis. This study explains how this pattern occurs.Methods: Twenty seven L. tropica smear preparations were collected from Bam district, south east Iran, and eight L. major and one L. infantum smear preparations were gathered from Shiraz, south west Iran. Furthermore one L. major and one L. infantum cultured standard strains were tested using entire ITS-PCR to survey their electrophoretic pattern. The ITS sequences of L. tropica, L. major, and L. infantum already deposited in GenBank were analyzed. Analysis of GenBank sequences of L. tropica revealed two groups of sequences based on length size, one group having a 100 bp gap. Therefore, a new re­verse primer namely LITS-MG was designed to exclude this gap in PCR products.Results: Whole ITS fragment amplification resulted in a double-band pattern in all L. tropica cases, while a sharp single band was observed for L. infantum and L. major isolates. This result was correspond­ing to the result obtained from in silico analysis of GenBank sequences. Use of LITS-MG primer was expectedly resulted in a single band including ITS1, 5.8s and partial ITS2 product for L. tropica which is appropriate for following molecular studies such as sequencing or restriction analysis.Conclusion: Sequences analysis of GenBank L. tropica sequences and following practical laboratory tests revealed at least two alleles in L. tropica which were confirmed in Bam isolates. This especial double-band pattern is because of a 100 bp fragment difference within ITS-rDNA alleles

  18. [Infecting glial cells with antimony resistant Leishmania tropica: A new ex-vivo model].

    Science.gov (United States)

    Zorbozan, Orçun; Harman, Mehmet; Evren, Vedat; Erdoğan, Mümin Alper; Kılavuz, Aslı; Tunalı, Varol; Çavuş, İbrahim; Yılmaz, Özlem; Özbilgin, Ahmet; Turgay, Nevin

    2018-01-01

    Leishmaniasis is a vector-borne zoonotic disease that shows different clinical features like cutaneous, mucocutaneous, visceral and viscerotropic forms. The protocols used in the treatment of leishmaniasis are toxic and have many limitations during administration. One of the limitations of treatment is the resistance against the protocols in practice. There is also a need to define new treatment options especially for resistant patients. Ex-vivo models using primary cell cultures may be a good source for evaluating new drug options in patients with antimony resistance, in addition to in-vitro and in-vivo studies. In this study, it was aimed to define a new ex-vivo culture model to evaluate treatment options in patients with cutaneous leishmaniasis who did not respond to treatment. In our experimental model of ex-vivo infection, Leishmania tropica promastigotes isolated from a case previously diagnosed with cutaneous leishmaniasis were used. The primary astroglial cell culture used for the ex-vivo model was prepared from 2-3 days old neonatal Sprague Dawley rat brains under sterile conditions by the modification McCarthy's method. The astroglia cells, which reached sufficient density, were infected with antimony resistant L.tropica promastigotes. After 24 hours of incubation, the supernatant on the cells were collected, the cell culture plate was dried at room temperature, then fixed with methyl alcohol and stained with Giemsa to search for L.tropica amastigotes. Amastigotes were intensely observed in glia cells in primary cell cultures infected with L.tropica promastigotes. No promastigotes were seen on Giemsa stained preparations of the precipitates prepared from the bottom sediment after the centrifugation of the liquid medium removed from the infected plates. In this study, promastigotes from a cutaneous leishmaniasis patient unable to respond to pentavalent antimony therapy were shown to infect rat glia cells and converted to amastigote form. This amastigote

  19. Disseminated Leishmaniasis Caused by Leishmania Tropica in a Puppy from Karaj, Central Iran

    Directory of Open Access Journals (Sweden)

    M Mohebali

    2011-06-01

    Full Text Available A 5-month old puppy with muco-cutaneous lesions in the chin, around lips and eyes was exam­ined physically and microscopically for leishmaniasis. Muco-cutaneous lesions containing a large num­ber of amastigotes of Leishmania spp. were observed. Amastigotes were also detected in liver and spleen of the puppy. The animal was positive with Dipstick rK39 kit and high level of anti-Leishmania antibodies was detected by direct agglutination test (DAT. DNA, Using PCR-RFLP technique extracted from cultured Leishmania promastigotes and L. tropica was identified. This is the first report of concurrent mucosal and visceral involvement of L. tropica in a puppy from Iran.

  20. Detection of Leishmania donovani and L. tropica in ethiopian wild rodents

    Czech Academy of Sciences Publication Activity Database

    Kassahun, A.; Sádlová, J.; Dvořák, V.; Košťálová, T.; Rohoušová, I.; Frynta, D.; Aghová, Tatiana; Yasur-Landau, D.; Lemma, W.; Hailu, A.; Baneth, G.; Warburg, A.; Volf, P.; Votýpka, J.

    2015-01-01

    Roč. 145, May 2015 (2015), s. 39-44 ISSN 0001-706X R&D Projects: GA ČR GAP506/10/0983 EU Projects: European Commission(XE) 261504 - EDENEXT Institutional support: RVO:68081766 Keywords : Leishmania donovani * Leishmania tropica * Phlebotomine sand fly * Rodents * kDNA * ITS1 Subject RIV: EG - Zoology Impact factor: 2.380, year: 2015

  1. Cloning, expression, purification and characterization of Leishmania tropica PDI-2 protein

    Directory of Open Access Journals (Sweden)

    Ali Dina

    2016-01-01

    Full Text Available In Leishmania species, protein disulfide isomerase (PDI is an essential enzyme that catalyzes thiol-disulfide interchange. The present work describes the isolation, cloning, sequencing and expression of the pdI-2 gene. Initially, the gene was amplified from L. tropica genomic DNA by PCR using specific primers before cloning into the expression vector pET-15b. The construct pET/pdI-2 was transformed into BL21(DE3 cells and induced for the protein expression. SDS-PAGE and western blot analysis showed that the expressed protein is about 51 kDa. Cloned gene sequence analysis revealed that the deduced amino acid sequence showed significant homology with those of several parasites PDIs. Finally, recombinant protein was purified with a metal-chelating affinity column. The putative protein was confirmed as a thiol - disulfide oxidoreductase by detecting its activity in an oxidoreductase assay. Assay result of assay suggested that the PDI-2 protein is required for both oxidation and reduction of disulfide bonds in vitro. Antibodies reactive with this 51 kDa protein were detected by Western blot analysis in sera from human infected with L. tropica. This work describes for the first time the enzymatic activity of recombinant L. tropica PDI-2 protein and suggests a role for this protein as an antigen for the detection of leishmaniasis infection.

  2. Variability of Cutaneous Leishmaniasis Lesions Is Not Associated with Genetic Diversity of Leishmania tropica in Khyber Pakhtunkhwa Province of Pakistan.

    Science.gov (United States)

    Khan, Nazma Habib; Llewellyn, Martin S; Schönian, Gabriele; Sutherland, Colin J

    2017-11-01

    Leishmania tropica is the causative agent of cutaneous leishmaniasis in Pakistan. Here, intraspecific diversity of L. tropica from northern Pakistan was investigated using multilocus microsatellite typing. Fourteen polymorphic microsatellite markers were typed in 34 recently collected L. tropica isolates from Pakistan along with 158 archival strains of diverse Afro-Eurasian origins. Previously published profiles for 145 strains of L. tropica originating from different regions of Africa, Central Asia, Iran, and Middle East were included for comparison. Six consistently well-supported genetic groups were resolved: 1) Asia, 2) Morroco A, 3) Namibia and Kenya A, 4) Kenya B/Tunisia and Galilee, 5) Morocco B, and 6) Middle East. Strains from northern Pakistan were assigned to Asian cluster except for three that were placed in a geographically distant genetic group; Morocco A. Lesion variability among these Pakistani strains was not associated with specific L. tropica genetic profile. Pakistani strains showed little genetic differentiation from strains of Iraq, Afghanistan, and Syria (F ST = 0.00-0.06); displayed evidence of modest genetic flow with India (F ST = 0.14). Furthermore, genetic structuring within these isolates was not geographically defined. Pak-Afghan cluster was in significant linkage disequilibrium (I A = 1.43), had low genetic diversity, and displayed comparatively higher heterozygosity (F IS = -0.62). Patterns of genetic diversity observed suggest dominance of a minimally diverse clonal lineage within northern Pakistan. This is surprising as a wide clinical spectrum was observed in patients, suggesting the importance of host and other factors. Further genotyping studies of L. tropica isolates displaying different clinical phenotypes are required to validate this potentially important observation.

  3. Genome sequencing reveals complex secondary metabolome in themarine actinomycete Salinispora tropica

    Energy Technology Data Exchange (ETDEWEB)

    Udwary, Daniel W.; Zeigler, Lisa; Asolkar, Ratnakar; Singan,Vasanth; Lapidus, Alla; Fenical, William; Jensen, Paul R.; Moore, BradleyS.

    2007-05-01

    Recent fermentation studies have identified actinomycetes ofthe marine-dwelling genus Salinispora as prolific natural productproducers. To further evaluate their biosynthetic potential, we analyzedall identifiable secondary natural product gene clusters from therecently sequenced 5,184,724 bp S. tropica CNB-440 circular genome. Ouranalysis shows that biosynthetic potential meets or exceeds that shown byprevious Streptomyces genome sequences as well as other naturalproduct-producing actinomycetes. The S. tropica genome features ninepolyketide synthase systems of every known formally classified family,non-ribosomal peptide synthetases and several hybrid clusters. While afew clusters appear to encode molecules previously identified inStreptomyces species,the majority of the 15 biosynthetic loci are novel.Specific chemical information about putative and observed natural productmolecules is presented and discussed. In addition, our bioinformaticanalysis was critical for the structure elucidation of the novelpolyenemacrolactam salinilactam A. This study demonstrates the potentialfor genomic analysis to complement and strengthen traditional naturalproduct isolation studies and firmly establishes the genus Salinispora asa rich source of novel drug-like molecules.

  4. Burkholderia tropica una bacteria con gran potencial parasu uso en la agricultura

    Directory of Open Access Journals (Sweden)

    Hernando José Bolívar-Anillo

    2016-01-01

    Full Text Available El género Burkholderia con más de 90 especies reportadas hasta la fecha, se encuentra dividido en dos grupos mayores filogenéticamente distantes. El primer grupo se encuentra constituido por especies patógenas donde destacan los patógenos oportunistas referidos como el complejo Burkholderia cepacia (Bcc;el otro grupo está conformado por especies no patógenas con habilidades para la promoción del crecimiento vegetal y la rizoremediación. Burkholderia tropica es unabacteria con capacidad de fijar nitrógeno; aislada de la rizósfera, rizoplano, tallo y la raíz de plantas de maíz y caña de azúcar. Además de su capacidad diazotrofa, B. tropica presenta características que permiten catalogarla como una bacteria promotora del crecimiento vegetal, por su capacidad de producir sideróforos, solubilizar fosfatos, producir exo-heteropolisacáridos, además de utilizarse como biocontrol para algunos fitoparásitos, lo que la convierte en una bacteria prometedora para su aplicación en el sector agrícola.

  5. Phlebotomus sergenti in a Cutaneous Leishmaniasis Focus in Azilal Province (High Atlas, Morocco): Molecular Detection and Genotyping of Leishmania tropica, and Feeding Behavior

    OpenAIRE

    Ajaoud , Malika; Es-Sette , Nargys; Charrel , Rémi N.; Laamrani-Idrissi , Abderahmane; Nhammi , Haddou; Riyad , Myriam; Lemrani , Meryem

    2015-01-01

    International audience; Background Phlebotomus (Paraphlebotomus) sergenti is at least one of the confirmed vectors for the transmission of cutaneous leishmaniasis caused by Leishmania tropica and distributed widely in Morocco. This form of leishmaniasis is considered largely as anthroponotic, although dogs were found infected with Leishmania tropica, suggestive of zoonosis in some rural areas. Methodology and Findings This survey aimed at (i) studying the presence of Leishmania in field caugh...

  6. Molecular detection of Leishmania infantum and Leishmania tropica in rodent species from endemic cutaneous leishmaniasis areas in Morocco.

    Science.gov (United States)

    Echchakery, Mohamed; Chicharro, Carmen; Boussaa, Samia; Nieto, Javier; Carrillo, Eugenia; Sheila, Ortega; Moreno, Javier; Boumezzough, Ali

    2017-10-02

    Leishmaniasis remains a major public health problem in African nations, including Morocco, where little is known about the vertebrate reservoirs involved in the causal parasites' transmission cycles. The present study investigates the role of rodent species as potential reservoirs of Leishmania spp. in central Morocco, where both L. tropica and L. infantum have been reported. Rodents were caught from 22 sites in central Morocco, by using Sherman metal traps, and identified morphologically. For each specimen, genomic DNA was extracted from different tissues using the Speed Tools DNA extraction Kit. Then, samples were PCR-analyzed, targeting the SSU rRNA gene to detect Leishmania spp. DNA, followed by amplification of the internal transcribed spacer 1 (ITS1) and its sequencing to identify the species. A total of 197 rodents belonging to ten species were captured and identified: Rattus rattus (40.61%), Mus musculus (25.38%), Apodemus sylvaticus (8.63%), Mus spretus (7.11%), Meriones shawi (5.58%), Rattus norvegicus (4.57%), Meriones libycus (3.05%), Mastomys erythroleucus (2.03%), Gerbillus campestris (2.03%) and Lemniscomys barbarus (1.01%). Molecular analysis revealed the presence of Leishmania species in 18 specimens: six R. rattus (out of 80 captured; 7.5%), 11 M. musculus (out of 50 captured; 22%), and one R. norvegicus (out of 9 captured; 11.11%). To the best of our knowledge, L. infantum and L. tropica were identified in rodent species for the first time in Morocco. These findings suggest that rodent species may be involved in L. infantum and L. tropica transmission cycles in this country but that further studies are needed to confirm their role as reservoirs of Leishmania species in Morocco.

  7. Phlebotomus sergenti in a Cutaneous Leishmaniasis Focus in Azilal Province (High Atlas, Morocco): Molecular Detection and Genotyping of Leishmania tropica, and Feeding Behavior

    Science.gov (United States)

    Ajaoud, Malika; Es-Sette, Nargys; Charrel, Rémi N; Laamrani-Idrissi, Abderahmane; Nhammi, Haddou; Riyad, Myriam; Lemrani, Meryem

    2015-01-01

    Background Phlebotomus (Paraphlebotomus) sergenti is at least one of the confirmed vectors for the transmission of cutaneous leishmaniasis caused by Leishmania tropica and distributed widely in Morocco. This form of leishmaniasis is considered largely as anthroponotic, although dogs were found infected with Leishmania tropica, suggestive of zoonosis in some rural areas. Methodology and Findings This survey aimed at (i) studying the presence of Leishmania in field caught Phlebotomus sergenti, (ii) investigating genetic diversity within Leishmania tropica and (iii) identifying the host-blood feeding preferences of Phlebotomus sergenti. A total of 4,407 sand flies were collected in three rural areas of Azilal province, using CDC miniature light traps. Samples collected were found to consist of 13 species: Phlebotomus spp. and 3 Sergentomyia spp. The most abundant species was Phlebotomus sergenti, accounting for 45.75 % of the total. 965 female Phlebotomus sergenti were screened for the presence of Leishmania by ITS1-PCR-RFLP, giving a positive rate of 5.7% (55/965), all being identified as Leishmania tropica. Nucleotide heterogeneity of PCR-amplified ITS1-5.8S rRNA gene-ITS2 was noted. Analyses of 31 sequences obtained segregated them into 16 haplotypes, of which 7 contain superimposed peaks at certain nucleotide positions, suggestive of heterozygosity. Phlebotomus sergenti collected were found to feed on a large variety of vertebrate hosts, as determined by Cytochrome b sequencing of the DNA from the blood meals of 64 engorged females. Conclusion Our findings supported the notion that Phlebotomus sergenti is the primary vector of Leishmania tropica in this focus, and that the latter is genetically very heterogeneous. Furthermore, our results might be suggestive of a certain level of heterozygosity in Leishmania tropica population. This finding, as well as the feeding of the vectors on different animals are of interest for further investigation. PMID:25826399

  8. Phlebotomus sergenti in a cutaneous leishmaniasis focus in Azilal province (High Atlas, Morocco: molecular detection and genotyping of Leishmania tropica, and feeding behavior.

    Directory of Open Access Journals (Sweden)

    Malika Ajaoud

    2015-03-01

    Full Text Available Phlebotomus (Paraphlebotomus sergenti is at least one of the confirmed vectors for the transmission of cutaneous leishmaniasis caused by Leishmania tropica and distributed widely in Morocco. This form of leishmaniasis is considered largely as anthroponotic, although dogs were found infected with Leishmania tropica, suggestive of zoonosis in some rural areas.This survey aimed at (i studying the presence of Leishmania in field caught Phlebotomus sergenti, (ii investigating genetic diversity within Leishmania tropica and (iii identifying the host-blood feeding preferences of Phlebotomus sergenti. A total of 4,407 sand flies were collected in three rural areas of Azilal province, using CDC miniature light traps. Samples collected were found to consist of 13 species: Phlebotomus spp. and 3 Sergentomyia spp. The most abundant species was Phlebotomus sergenti, accounting for 45.75 % of the total. 965 female Phlebotomus sergenti were screened for the presence of Leishmania by ITS1-PCR-RFLP, giving a positive rate of 5.7% (55/965, all being identified as Leishmania tropica. Nucleotide heterogeneity of PCR-amplified ITS1-5.8S rRNA gene-ITS2 was noted. Analyses of 31 sequences obtained segregated them into 16 haplotypes, of which 7 contain superimposed peaks at certain nucleotide positions, suggestive of heterozygosity. Phlebotomus sergenti collected were found to feed on a large variety of vertebrate hosts, as determined by Cytochrome b sequencing of the DNA from the blood meals of 64 engorged females.Our findings supported the notion that Phlebotomus sergenti is the primary vector of Leishmania tropica in this focus, and that the latter is genetically very heterogeneous. Furthermore, our results might be suggestive of a certain level of heterozygosity in Leishmania tropica population. This finding, as well as the feeding of the vectors on different animals are of interest for further investigation.

  9. Genetic, serological and biochemical characterization of Leishmania tropica from foci in northern Palestine and discovery of zymodeme MON-307

    Directory of Open Access Journals (Sweden)

    Azmi Kifaya

    2012-06-01

    Full Text Available Abstract Background Many cases of cutaneous leishmaniasis (CL have been recorded in the Jenin District based on their clinical appearance. Here, their parasites have been characterized in depth. Methods Leishmanial parasites isolated from 12 human cases of CL from the Jenin District were cultured as promastigotes, whose DNA was extracted. The ITS1 sequence and the 7SL RNA gene were analysed as was the kinetoplast minicircle DNA (kDNA sequence. Excreted factor (EF serotyping and multilocus enzyme electrophoresis (MLEE were also applied. Results This extensive characterization identified the strains as Leishmania tropica of two very distinct sub-types that parallel the two sub-groups discerned by multilocus microsatellite typing (MLMT done previously. A high degree of congruity was displayed among the results generated by the different analytical methods that had examined various cellular components and exposed intra-specific heterogeneity among the 12 strains. Three of the ten strains subjected to MLEE constituted a new zymodeme, zymodeme MON-307, and seven belonged to the known zymodeme MON-137. Ten of the 15 enzymes in the profile of zymodeme MON-307 displayed different electrophoretic mobilities compared with the enzyme profile of the zymodeme MON-137. The closest profile to that of zymodeme MON-307 was that of the zymodeme MON-76 known from Syria. Strains of the zymodeme MON-307 were EF sub-serotype A2 and those of the zymodeme MON-137 were either A9 or A9B4. The sub-serotype B4 component appears, so far, to be unique to some strains of L. tropica of zymodeme MON-137. Strains of the zymodeme MON-137 displayed a distinctive fragment of 417 bp that was absent in those of zymodeme MON-307 when their kDNA was digested with the endonuclease RsaI. kDNA-RFLP after digestion with the endonuclease MboI facilitated a further level of differentiation that partially coincided with the geographical distribution of the human cases from which the strains

  10. [Ecoepidemiology of leishmaniases in Syria. 2--Presence, in dogs, of Leishmania infantum Nicolle and Leishmania tropica (Wright) (Kinetoplastida-Trypanonomatidae)].

    Science.gov (United States)

    Dereure, J; Rioux, J A; Khiami, A; Pratlong, F; Périères, J; Martini, A

    1991-01-01

    In an ecoepidemiological study in the Syrian foci of human visceral (VL) and cutaneous (CL) leishmaniasis, domestic dogs infected with Leishmania infantum MON-1 and L. tropica MON-76 were found for the first time. In Syria canine leishmaniasis caused by L. infantum, occurs in the humid and sub-humid western belt from the coastal zone to the nearby mountain ranges. Sporadis cases of human VL occur in this area. Canine CL, caused by L. tropica was found in the semi-arid zone in a village with a high human infection rate. The infection causes small papules or crusted ulcers on the nose or muzzle. These observations are relevant to control. In human VL foci it is recommended not to control the dog population, unless this is linked with other activities, such as rabies control. Control of dogs in the CL areas is not justified because of the small size of the lesions and the rarity of the parasites. Indeed the role of the dog as the "true" reservoir host is questionable. Human CL infection is best controlled by active case detection and specific treatment.

  11. Distribution pattern of anthroponotic cutaneous leishmaniasis caused by Leishmania tropica in Western Afghanistan during 2013-2014.

    Science.gov (United States)

    Fakhar, Mahdi; Karamian, Mehdi; Ghatee, Mohammad Amin; Taylor, Walter Robert; Pazoki Ghohe, Hossein; Rasooli, Sayed Abobakar

    2017-12-01

    Anthroponotic cutaneous leishmaniasis (ACL), caused by Leishmania tropica, is the main cause of cutaneous leishmaniasis (CL) in the Herat province, Western Afghanistan. We investigated the role of environmental factors on ACL distribution in Herat. Epidemiological data from 2457 patients were retrieved from the local WHO sub-office. Shapefile layers of districts, cities, villages, land cover, soil type and digital elevation model (DEM) of the Herat province were used to assess, by logistic regression modelling, the effects of land cover, soil types, elevation, and proximity to the Harirud river on the distribution of ACL. The key determinants of distribution were: (i) close proximity to the Harirud river, (ii) elevation between 700 and 1200m, (iii) intensive and intermittent irrigated cultivated land, and (iv) Haplocalcids with Torriorthents and Torrifluvents soil types. No ACL cases were found below 700m, and a few cases were present at >1200m in irrigated areas around the Harirud river. These findings suggest that moist soil and the humidity from irrigated areas found between 700 and 1200m provide suitable breeding sites of Phlebotomus sergenti, the main sandfly vector of L. tropica in Afghanistan. The effect of elevation also explains the predominance of ACL over ZCL in this region. The present study showed that distribution of ACL is strongly associated with environmental factors in West Afghanistan where the political and socio-economic conditions may also affect the epidemiology of CL. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Epidemiological analysis of Leishmania tropica strains and giemsa-stained smears from Syrian and Turkish leishmaniasis patients using multilocus microsatellite typing (MLMT.

    Directory of Open Access Journals (Sweden)

    Mehmet Karakuş

    2017-04-01

    Full Text Available Turkey is located in an important geographical location, in terms of the epidemiology of vector-borne diseases, linking Asia and Europe. Cutaneous leishmaniasis (CL is one of the endemic diseases in a Turkey and according to the Ministry Health of Turkey, 45% of CL patients originate from Şanlıurfa province located in southeastern Turkey. Herein, the epidemiological status of CL, caused by L. tropica, in Turkey was examined using multilocus microsatellite typing (MLMT of strains obtained from Turkish and Syrian patients. A total of 38 cryopreserved strains and 20 Giemsa-stained smears were included in the present study. MLMT was performed using 12 highly specific microsatellite markers. Delta K (ΔK calculation and Bayesian statistics were used to determine the population structure. Three main populations (POP A, B and C were identified and further examination revealed the presence of three subpopulations for POP B and C. Combined analysis was performed using the data of previously typed L. tropica strains and Mediterranean and Şanlıurfa populations were identified. This finding suggests that the epidemiological status of L. tropica is more complicated than expected when compared to previous studies. A new population, comprised of Syrian L. tropica samples, was reported for the first time in Turkey, and the data presented here will provide new epidemiological information for further studies.

  13. Les leishmanioses cutanées à Leishmania major et à Leishmania tropica au Maroc: aspects épidémio-cliniques comparatifs de 268 cas

    Science.gov (United States)

    Chiheb, Soumia; Slaoui, Widad; Mouttaqui, Tarik; Riyad, Meriem; Benchikhi, Hakima

    2014-01-01

    Introduction Depuis 1995, le Maroc a connu une réactivation des foyers de leishmanioses cutanées (LC) à L. major et une nouvelle répartition géographique des foyers à L. tropica. Le but de cette étude est de comparer les aspects épidémio-cliniques associés aux LC potentiellement dûes à L. major et à L. tropica. Méthodes Une étude rétrospective a colligé 268 cas de LC au service de dermatologie du CHU Ibn Rochd de Casablanca entre Janvier 1995 et Septembre 2010. Les données étaient analysées par Epi info version 3.5.1. Le test X2 était appliqué (Différence significative = p< 0,05). Résultats Deux cent soixante-huit cas de LC ont été colligés, dont 160 femmes et 108 hommes. Ils ont été répartis en 123 patients originaires des foyers à L.major et 145 patients originaires des foyers à L. tropica. L'aspect ulcéronodulaire, ulcérovégétant ou végétant était retrouvé dans 58 cas (47,2%) des cas de LC à L. major versus 24 cas (16,7%) dans la L.C à L. tropica. L'aspect papulonodulaire était retrouvé dans 84 cas (58%) de LC à L. tropica contre 41 cas (33,3%) de LC à L. major. Conclusion Dans la LC à L. major, l'atteinte des membres et les aspects cliniques végétant ou ulcéro-végétant restent toujours prédominants. Dans la L.C à L. tropica, l'atteinte papulonodulaire unique du visage reste prédominante mais des formes ulcéronodulaires, végétantes ou ulcérovégétantes existent également dans les foyers récents à L. tropica, prêtant à confusion cliniquement avec des LC à L. major. PMID:25810796

  14. Genetics of host response to Leishmania tropica in mice - different control of skin pathology, chemokine reaction, and iInvasion into spleen and liver

    Czech Academy of Sciences Publication Activity Database

    Kobets, Tetyana; Havelková, Helena; Grekov, Igor; Volkova, Valeriya; Vojtíšková, Jarmila; Slapničková, Martina; Kurey, Irina; Sohrabi, Yahya; Svobodová, M.; Demant, P.; Lipoldová, Marie

    2012-01-01

    Roč. 6, č. 6 (2012), e1667 ISSN 1935-2735 R&D Projects: GA ČR GA310/08/1697; GA ČR GD310/08/H077; GA MŠk(CZ) LC06009 Institutional support: RVO:68378050 Keywords : leishmaniasis * Leishmania tropica * Leishmania major Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.569, year: 2012

  15. Habitats of the sandfly vectors of Leishmania tropica and L. major in a mixed focus of cutaneous leishmaniasis in southeast Tunisia.

    Science.gov (United States)

    Tabbabi, Ahmed; Ghrab, Jamila; Aoun, Karim; Ready, Paul Donald; Bouratbine, Aïda

    2011-08-01

    From 2009 to 2010, 3129 sandflies were caught in CDC light traps placed in various habitats in Ghomrassen, Tataouine governorate, southeast Tunisia, a mixed focus of human cutaneous leishmaniasis caused by Leishmania tropica and Leishmania major. Species diversity was quantified in anthropogenic, semi-anthropogenic and semi-natural locations. Sandflies were identified according to morphological characters and also by the comparative sequence analysis of a fragment of the mitochondrial cytochrome b gene to distinguish between two putative local vectors of L. tropica, namely Phlebotomus chabaudi and Phlebotomus riouxi. The lowest sandfly diversities were found in L. major sites, where the incriminated vector P. papatasi predominated in the burrows of the rodent reservoir hosts (Meriones) as well as inside and outside houses of human cases. In L. tropica sites, the incriminated peri-domestic vector Phlebotomus sergenti was the most abundant species inside houses, whereas P. riouxi or P. chabaudi was the dominant species in the semi-natural rocky habitats favoured by the putative rodent reservoir, Ctenodactylus gundi. All specimens of P. chabaudi identified molecularly had the diagnostic cytochrome b characters of P. riouxi, indicating either that the latter represents only a geographical variant of P. chabaudi or that these two species may sometimes hybridize. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Prophylactic immunization against experimental leishmaniasis. III. Protection against fatal Leishmania tropica infection induced by irradiated promastigotes involves Lyt-1+2- T cells that do not mediate cutaneous DTH

    International Nuclear Information System (INIS)

    Liew, F.Y.; Howard, J.G.; Hale, C.

    1984-01-01

    Protective immunity against fatal L. tropica infection in genetically vulnerable BALB/c mice can be induced by prophylactic immunization with irradiated promastigotes even when heat-killed. Such immunity is adoptively transferable transiently into intact or durably into sub-lethally irradiated (200 or 550 rad) syngeneic recipients by splenic T but not B cells. The effector T cells are of the Lyt-1 + 2 - phenotype, devoid of demonstrable cytotoxic activity. The immune splenic T cell population expresses specific helper activity for antibody synthesis. A causal role for helper T cells in this capacity, however, seems unlikely, because it was shown that antibody does not determine the protective immunity against L. tropica. The immunized donors show no detectable cutaneous DTH or its early memory recall in response to live or killed promastigotes or a soluble L. tropica antigen preparation. Spleen, lymph node, and peritoneal exudate cells from protectively immunized donors similarly fail to transfer DTH locally or systemically. These cells also lack demonstrable suppressive activity against the expression or induction of DTH to L. tropica. Thus, protection against L. tropica induced by prophylactic i.v. immunization with irradiated promastigotes appears to be conferred by Lyt-1 + 2 - T cells that are distinguishable from T cells mediating either both DTH and T help, or cytotoxicity

  17. Leishmania tropica isolates from non-healed and healed patients in Iran: A molecular typing and phylogenetic analysis.

    Science.gov (United States)

    Bamorovat, Mehdi; Sharifi, Iraj; Mohammadi, Mohammad Ali; Eybpoosh, Sana; Nasibi, Saeid; Aflatoonian, Mohammad Reza; Khosravi, Ahmad

    2018-03-01

    The precise identification of the parasite species causing leishmaniasis is essential for selecting proper treatment modality. The present study aims to compare the nucleotide variations of the ITS1, 7SL RNA, and Hsp70 sequences between non-healed and healed anthroponotic cutaneous leishmaniasis (ACL) patients in major foci in Iran. A case-control study was carried out from September 2015 to October 2016 in the cities of Kerman and Bam, in the southeast of Iran. Randomly selected skin-scraping lesions of 40 patients (20 non-healed and 20 healed) were examined and the organisms were grown in a culture medium. Promastigotes were collected by centrifugation and kept for further molecular examinations. The extracted DNA was amplified and sequenced. After global sequence alignment with BioEdit software, maximum likelihood phylogenetic analysis was performed in PhyML for typing of Leishmania isolates. Nucleotide composition of each genetic region was also compared between non-healed and healed patients. Our results showed that all isolates belonged to the Leishmania tropica complex, with their genetic composition in the ITS1 region being different among non-healed and healed patients. 7SL RNA and Hsp70 regions were genetically identical between both groups. Variability in nucleotide patterns observed between both groups in the ITS1 region may serve to encourage future research on the function of these polymorphisms and may improve our understanding of the role of parasite genome properties on patients' response to Leishmania treatment. Our results also do not support future use of 7SL RNA and Hsp70 regions of the parasite for comparative genomic analyses. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Predicting the risk of an endemic focus of Leishmania tropica becoming established in South-Western Europe through the presence of its main vector, Phlebotomus sergenti Parrot, 1917.

    Science.gov (United States)

    Barón, S D; Morillas-Márquez, F; Morales-Yuste, M; Díaz-Sáez, V; Gállego, M; Molina, R; Martín-Sánchez, J

    2013-09-01

    The aim of the study was the construction of risk maps for exposure to Phlebotomus sergenti, the main vector of Leishmania tropica, with a view to identifying hot spots for the potential establishment of this parasite in the southwest of Europe. Data were collected on the presence/absence of this vector and the ecological and climatic characteristics of 662 sampling sites located in the southeast, centre and northeast of the Iberian Peninsula (south-western Europe). The environmental factors associated with the distribution of P. sergenti were determined. The best predictors for the presence of this dipteran were ‘altitude’, ‘land use’, ‘land surface temperature’, ‘aspect’, ‘adjacent land cover’, ‘absence of vegetation in wall’ and the ‘absence of PVC pipes in the drainage holes of retaining walls’. Risk maps for exposure to the vector were drawn up based on these variables. The validation of the predictive risk model confirmed its usefulness in the detection of areas with a high risk of P. sergenti being present. These locations represent potential hot spots for an autochthonous focus of L. tropica becoming established. The risk maps produced for P. sergenti presence revealed several areas in the centre and south of the Iberian Peninsula to be the most prone to this process, which would make it possible for the disease to enter south-western Europe.

  19. Fire, hypogeous fungi and mycophagous marsupials

    Science.gov (United States)

    James M. Trappe; Andrew W. Claridge; Ari Jumpponen

    2005-01-01

    In their interesting research on post-fire foraging behaviour of northern bettongs (Bettongia tropica) in tropical Queensland, Australia, Vernes et al. (2004) used forage-diggings of their study animals to locate plots for estimating biomass of hypogeous fungi on prescribed-burnt sites in comparison with unburnt control sites. They concluded that...

  20. ANALISIS KELAYAKAN TEKNIS DAN FINANSIAL DARI INVESTASI PENGADAAN TRAY DRYER BERBAHAN BAKAR BIOMASSA PADA USAHA ARANG TEMPURUNG KELAPA BERBASIS EKSPOR (Studi Kasus di Tropica Nucifera Industry – Yogyakarta

    Directory of Open Access Journals (Sweden)

    Fanny Widadie

    2013-11-01

    Full Text Available The purpose of the study is to assess the technical and financial feasibility of procurement investment Tray Dryer fueled biomass to operations of coconut shell charcoal briquette in PT. Tropica Nucifera Industry. The result showed that investment-Tray Dryer on cocnut shell charcoal briquette bussiness was highly feasibile both the technical and financial aspects. The technical aspect was seen that the quantity-production capabilities of coconut shell charcoal briquettes using a dryer-Tray Dryer could be increased 15 percent compared the previous consdition-used of solar energy. While the quality of the charcoal briquettes produced from Tray Dryer was a much butter quality, moisture content (6.39%; ash (2.65%; carbon-bound (85.83%, compared to using sunlight, water content (15.89; ash (2.57% and carbon-bound (77.32%.The financial aspects of procurement Tray Dryer investment that its value was more feasible than previous, nothing Tray Dryer. Coconut shell charcoal briquet enterprises without using Tray Dryer was feasible to be done with NPV 1,058,928.12, IRR 63%, Payback Period 1 year and 9 moths, Net B/C 2.25 and analysis of swicthcing value reached 20 percent. And after the business using Tray Drayer, the financial analysis was much more feasible with NPV 2,285,500,498.12, IRR 113%, Payback Period 11 months, Net B/C 3.5 and analysis of switching value reached 50 percent.

  1. Comparative jaw muscle anatomy in kangaroos, wallabies, and rat-kangaroos (marsupialia: macropodoidea).

    Science.gov (United States)

    Warburton, Natalie Marina

    2009-06-01

    The jaw muscles were studied in seven genera of macropodoid marsupials with diets ranging from mainly fungi in Potorous to grass in Macropus. Relative size, attachments, and lamination within the jaw adductor muscles varied between macropodoid species. Among macropodine species, the jaw adductor muscle proportions vary with feeding type. The relative mass of the masseter is roughly consistent, but grazers and mixed-feeders (Macropus and Lagostrophus) had relatively larger medial pterygoids and smaller temporalis muscles than the browsers (Dendrolagus, Dorcopsulus, and Setonix). Grazing macropods show similar jaw muscle proportions to "ungulate-grinding" type placental mammals. The internal architecture of the jaw muscles also varies between grazing and browsing macropods, most significantly, the anatomy of the medial pterygoid muscle. Potoroines have distinctly different jaw muscle proportions to macropodines. The masseter muscle group, in particular, the superficial masseter is enlarged, while the temporalis group is relatively reduced. Lagostrophus fasciatus is anatomically distinct from other macropods with respect to its masticatory muscle anatomy, including enlarged superficial medial pterygoid and deep temporalis muscles, an anteriorly inflected masseteric process, and the shape of the mandibular condyle. The enlarged triangular pterygoid process of the sphenoid bone, in particular, is distinctive of Lagsotrophus. (c) 2009 Wiley-Liss, Inc.

  2. Pesquisas sobre a imunidade da Framboesia tropica no homem: observações feitas em 33 superinoculações e 7 reinoculações

    Directory of Open Access Journals (Sweden)

    F. Nery Guimarães

    1946-03-01

    em germes. 5 Existe na framboesia trópica uma verdadeira imunidade além de uma simples resistência á superinoculação devido a presença da infecção ativa ou latente. Com efeito, pacientes tratados em determinado período da molestia e curados clinica e sorologicamente, mostraram resistência parcial á reinoculação, reagindo de modo semelhante a outros do mesmo período de molestia e não tratados. 6 A imunidade na framboesia tropica se manifesta seja como uma resistência á superinoculação ou reinoculação seja como uma modificação da lesão boubatica inicial, seja, finalmente, como uma resistência á generalização da doença. 7 Os resultados das esperiências sugerem que as diferentes manifestações cutaneas da molestia são condicionadas até certo ponto pelo estado imunitario do organismo infectado. 8 Os diferentes gráus de imunidade, encontrados na framboesia trópica, estão até certo ponto relacionados com o tempo de doença. Porém, são atingidos mais ou menos ràpidamente, segundo o organismo infectado e, talvez segundo a virulência do treponema, do mesmo modo como os chamados "secundarismo" e 'terciarismo" da doença.The following conclusions are drawn from the results of the 40 experiments described above: 1 From the 2nd to the 8th month there is a great resistance to super-infection. a Such resistance seems to be independent of the presence of cutaneous lesions and occurred even in cases with only the initial lesion. b Within this period, resistance disappears with treatment: patients treated and reinoculated acquire yaws in the normal time. This fact suggests that this resistance is conditioned by the presence of an active or latent infection, being not a true immunity (an immunity in its classical conception. c When homovirus is used, resistance lasted about one year of the disease. 2 From the 10th month to the 4th year of the disease, there is a partial resistance, which manifests itself by two modes, according to the nature of

  3. Selection and evaluation of reference genes for RT-qPCR expression studies on Burkholderia tropica strain Ppe8, a sugarcane-associated diazotrophic bacterium grown with different carbon sources or sugarcane juice.

    Science.gov (United States)

    da Silva, Paula Renata Alves; Vidal, Marcia Soares; de Paula Soares, Cleiton; Polese, Valéria; Simões-Araújo, Jean Luís; Baldani, José Ivo

    2016-11-01

    Among the members of the genus Burkholderia, Burkholderia tropica has the ability to fix nitrogen and promote sugarcane plant growth as well as act as a biological control agent. There is little information about how this bacterium metabolizes carbohydrates as well as those carbon sources found in the sugarcane juice that accumulates in stems during plant growth. Reverse transcription quantitative PCR (RT-qPCR) can be used to evaluate changes in gene expression during bacterial growth on different carbon sources. Here we tested the expression of six reference genes, lpxC, gyrB, recA, rpoA, rpoB, and rpoD, when cells were grown with glucose, fructose, sucrose, mannitol, aconitic acid, and sugarcane juice as carbon sources. The lpxC, gyrB, and recA were selected as the most stable reference genes based on geNorm and NormFinder software analyses. Validation of these three reference genes during strain Ppe8 growth on the same carbon sources showed that genes involved in glycogen biosynthesis (glgA, glgB, glgC) and trehalose biosynthesis (treY and treZ) were highly expressed when Ppe8 was grown in aconitic acid relative to other carbon sources, while otsA expression (trehalose biosynthesis) was reduced with all carbon sources. In addition, the expression level of the ORF_6066 (gluconolactonase) gene was reduced on sugarcane juice. The results confirmed the stability of the three selected reference genes (lpxC, gyrB, and recA) during the RT-qPCR and also their robustness by evaluating the relative expression of genes involved in glycogen and trehalose biosynthesis when strain Ppe8 was grown on different carbon sources and sugarcane juice.

  4. Effects of Simultaneous Radiofrequency Radiation and Chemical Exposure of Mammalian Cells. Volume 2

    Science.gov (United States)

    1988-07-01

    chromosome - - - - - - -I aberrations and sister chromatid exchanges (SCE). Yao (1982) exposed rat kangaroo RH5 and RH1l6 cells to 2.45 GHz radiation, and...control was reported in chromosome aberrations. Yac (1982) investigated the cytogenetic consequences of chronic microwave exposure on rat kangaroo RH5...was said to be 280C. The cells were exposed both as conidia, which are "rather inactive metabolically ," and also after DNA replication had been

  5. Luteolin as a potent anti-leishmanial agent against intracellular ...

    African Journals Online (AJOL)

    intraperitoneally to L. tropica KWH23 amastigotes-infected albino mice at a dose of ≥ 3 mg/kg for 5 ... Journal Citation Reports/Science Edition, Directory of Open Access Journals ..... tropica field isolate against buparvaquone in comparison.

  6. Molecular Markers for Breast Cancer Susceptibility.

    Science.gov (United States)

    1998-09-01

    experiments have allowed insight into the nature of Laboratory of Biochemistry and Metabolism , Bethesda, Mary- interions bewe the strom and the nathe...rat kangaroo cells. These investigators further demonstrated a direct correlation between the number of fluorescent spots in the interphase nuclei and...overall length of human chromosomes was greatest after treatment with 5-azacytidine following a fluorodeoxyuridine (FUdR)/uridine metabolic block

  7. 78 FR 27253 - Endangered Species; Marine Mammals; Receipt of Applications for Permit

    Science.gov (United States)

    2013-05-09

    ... specimens of African wild dog (Lycaon pictus) for the purpose of enhancement of the survival of the species... bettong (Bettongia penicillata) from Aqua Zoo Friesland, Netherlands, for the purpose of enhancement of... from a ranch in the Republic of South Africa, for the purpose of enhancement of the survival of the...

  8. The Installation Restoration Program Toxicology Guide. Volume 3

    Science.gov (United States)

    1987-06-01

    exchanges were observed in rat- kangaroo cells treated with p,p’-DDT at 10 Ug/mL/24 hours. The p,p’-isomer accounts for most ot thi toxicity attributed to...ity of degra..ative enzymes (2000,1991). In mammals, including man, DDT is metabolized by 2 pathways. It is convertud to a slight extent to DDE which...sympathetic discharge were observed to accompany the tremors and convulsions. These were thought to be due to changes in the metabolism of brain

  9. Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused by Leishmania donovani in Turkey

    NARCIS (Netherlands)

    Özbilgin, Ahmet; Harman, Mehmet; Karakuş, Mehmet; Bart, Aldert; Töz, Seray; Kurt, Özgür; Çavuş, İbrahim; Polat, Erdal; Gündüz, Cumhur; van Gool, Tom; Özbel, Yusuf

    2017-01-01

    In Turkey, the main causative agents are Leishmania tropica (L. tropica) and Leishmania infantum (L. infantum) for cutaneous leishmaniasis (CL) and L. infantum for visceral leishmaniasis (VL). In this study, we investigated leishmaniasis cases caused by L. donovani and established animal models for

  10. Ancistrotanzanine C and related 5,1 '- and 7,3 '-coupled naphthylisoquinoline alkaloids from Ancistrocladus tanzaniensis

    DEFF Research Database (Denmark)

    Bringmann, G.; Dreyer, M.; Faber, J.H.

    2004-01-01

    from this newly discovered plant species. The structural elucidation was achieved by chemical, spectroscopic, and chiroptical methods. The biological activities of the alkaloids against the pathogens causing malaria tropica, leishmaniasis, Chagas' disease, and African sleeping sickness were evaluated....

  11. Community monitoring of carbon stocks for REDD+

    DEFF Research Database (Denmark)

    Brofeldt, Søren; Theilade, Ida; Burgess, Neil David

    2014-01-01

    Reducing emissions from deforestation and forest degradation in developing countries, and the role of conservation, sustainable management of forests, and enhancement of forest carbon stocks in developing countries (REDD+) is a potentially powerful international policy mechanism that many tropica...

  12. Distribution and abundance of copepods in the pollution gradient zones of Bombay Harbour-Thana Creek-Bassein Creek, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah, Neelam

    . Eucalanus subcrassus and Paracalanus aculeatus were more abundant in the outer zone while A. tropica was very common in the interior region. Hyposaline species, Pseudodiaptomus binghami malayalus was recorded from the interior locations particularly during...

  13. Quantifying the key factors that create road flooding : research project capsule.

    Science.gov (United States)

    2012-02-01

    Road fl ooding is a serious operational hazard for low-lying areas of south Louisiana. This is : especially true for emergency evacuation routes, which must be accessible for coastal residents : evacuating ahead of an approaching hurricane or tropica...

  14. The current status of phlebotomine sandflies (Diptera: Psychodidae) in Tunisia and their role on Leishmania transmission: A review

    OpenAIRE

    Ahmed Tabbabi; Sajida Sboui; Jabeur Daaboub

    2017-01-01

    In Tunisia, the epidemiological situation of leishmaniasis is characterized by the coexistence in a rather circumscribed territory (165000 km2, including the Sahara) of 4 forms of leishmaniasis caused by 3 species: Leishmania infantum, Leishmania major and Leishmania tropica (L. tropica) (synonymous Leishmania killicki). One of the factors determining the clinical, epidemiological and immunological diversity of leishmanioses is certainly the existence of a vector-parasite specificity of of...

  15. Radioimmunoassay studies on repair of ultraviolet damaged DNA in cultured animal cells

    International Nuclear Information System (INIS)

    Yatani, Ryuichi; Tohgo, Yukihiro; Kunishima, Nobuyoshi.

    1975-01-01

    UV (ultraviolet) damaged DNA and its repair of various cultured animal cells were observed by radioimmunoassay using anti-serum against the UV irradiation induced heat-degenerated DNA. There is some difference among the cells of used animals according to their DNA repairabilities. The cells were divided into four groups according to the existence or strength of their repairabilities. 1) excision repair type: cells of men and chimpanzees. 2) photoreactivation type: cells derived from Tachydromus tachydromoides and chicks. 3) photoreactivation with excision repair: cells of rats, kangaroos and mosquitos. 4) non-excision repair type: cells of mice, Meriones and rats. Animal cells have plural types of repair. Main types of repair will differ according to the kind of animals. (Ichikawa, K.)

  16. Damage and repair in mammalian cells after exposure to non-ionizing radiations. 1

    International Nuclear Information System (INIS)

    Harm, H.

    1978-01-01

    Cornea cells of the rat kangaroo or 'potoroo' (Potorous tridactylus) were exposed to far-UV (254 or 302 nm) radiation, with or without subsequent illumination by near-UV or visible light. The DNA of these cells was extracted and tested for the presence of photoproducts binding yeast photoreactivating enzyme (PRE). The effects on repair kinetics of the transforming DNA indicate that in UV-irradiated potoroo cornea cells up to approximately 90% of photorepairable DNA damage can be photorepaired within 15 min. However, the extent of cellular photorepair depends appreciably on experimental parameters during photoreactivating treatment, including the spectral composition of photoreactivating light. Apparently superposition of damage by the photoreactivating treatment itself is the critical factor. This may explain experimental discrepancies existing in different laboratories studying photorepair in UV-irradiated cells of placental mammals. (Auth.)

  17. Induced recovery from near- and far-UV damage in cultured marsupial cells

    International Nuclear Information System (INIS)

    Hoy, D.A.

    1982-01-01

    Colony-forming ability of cultured rat kangaroo cells (Ptk-2) decreases with increasing exposure to light from a daylight fluorescent lamp. After reaching a minimum, survival increases with further exposure, forming a V-shaped survival curve. This increase is inhibited by low concentrations of the protein synthesis inhibitor cycloheximide. The resulting V-shaped survival curve was characterized with respect to several experimental parameters; it appears to be due to an induced repair system dependent on protein synthesis. These results suggest that induced repair, dependent on protein synthesis, in response to fluorescent, near UV, and far UV light damage, is a major repair pathway in Ptk-2 cells, and provides a characterizable system that can elucidate induced repair mechanisms in other mammalian cells

  18. Identification and phylogenetic relationship of Iranian strains of various Leishmania species isolated from cutaneous and visceral cases of leishmaniasis based on N-acetylglucosamine-1-phosphate transferase gene.

    Science.gov (United States)

    Hajjaran, Homa; Mohebali, Mehdi; Teimouri, Aref; Oshaghi, Mohammad Ali; Mirjalali, Hamed; Kazemi-Rad, Elham; Shiee, Mohammad Reza; Naddaf, Saied Reza

    2014-08-01

    The identity of Iranian Leishmania species has been resolved to some extent by some genetic markers. In this study, based on N-acetylglucosamine-1-phosphate transferase (nagt) gene, we further elucidated the identity and phylogeny of the prevalent species in this country. DNAs of 121 isolates belonging to cutaneous leishmaniasis (CL) patients, canine visceral leishmaniasis (CVL) cases, and Rhombomys opimus rodents were amplified by targeting a partial sequence of nagt gene. All the amplicons were analyzed with restriction fragment length polymorphism (RFLP) using Acc1 enzyme, and 49 amplicons representing different reservoir hosts were sequenced and aligned with similar sequences from GenBank database. The RFLP analysis revealed that 41 CL patients were infected Leishmania tropica and 36 with Leishmania major. Among 10 CVL isolates, 6 were identified as Leishmania infantum and 4 as L. tropica. Amongst 34 rodents' isolates, 11 and 23 isolates exhibited patterns similar to those of L. major, and L. tropica/Leishmania turanica, respectively. The sequencing results from all CL patients, CVL cases, and 4 reservoir rodents were in agreement with RFLP analysis and showed 99-100% homologies with the registered species of L. major, L. tropica, and L. infantum from Turkey, Tunisia, Iraq and Israel. Of the 7 rodent isolates exhibiting RFLP patterns similar to L. tropica/L. turanica, 3 exhibited the highest homologies (99-100%) with L. turanica and 4 with Leishmania gerbilli. The 49 nagt DNA sequences were grouped into five clusters representing L. major, L. tropica, L. infantum, L. turanica and L. gerbilli species, encompassing 19 haplotypes. No correlation was observed between intraspecies divergence and geographic distribution of haplotypes. The L. tropica haplotypes exhibited more homologies with those of L. infantum than L. major (97.2% vs. 96.9%), a probable indication to the potential ability of L. tropica to visceralize. Characterization of Iranian Leishmania isolates

  19. Short Communication Reduced seabird night strikes and mortality in ...

    African Journals Online (AJOL)

    Birds killed were broad-billed prions Pachyptila vittata (41%), common diving petrels Pelecanoides urinatrix (23%), and storm petrels (Pelagodroma marina and Fregetta grallaria/tropica 36%). All these species are listed as Least Concern globally, and the numbers killed per year are <0.1% of the island populations.

  20. Cutaneous leishmaniasis: An emerging infection in a non-endemic area and a brief update

    Directory of Open Access Journals (Sweden)

    Rastogi V

    2007-01-01

    Full Text Available We report here the emergence of a new focus of cutaneous leishmaniasis (CL due to Leishmania tropica (L. tropica in the Ajmer city of Rajasthan, India, a previously non-endemic area. Between January-February 2006, 13 new indigenously acquired cases of CL were diagnosed among the patients attending the Skin and STD department, JLN Hospital, Ajmer. The diagnosis was based on clinical presentation, demonstration of amastigotes (LT bodies in Giemsa stained smear of the lesion and response to intralesional / local anti-leishmanial drug therapy. In addition, culture of the promastigote forms of L. tropica from the lesion was successfully attempted in four of the smear negatives cases. By retrospective analysis, 23 new indigenous cases of CL have been diagnosed in the same setting during the period January 2004 - December 2005, based on clinical and therapeutic response alone. There was no clear-cut history of sandfly bite and travel outside the district or state to endemic area in any of the cases. However, all of them came from a common residential area (famous dargah of Ajmer and the peak incidence was seen in January, four months after the famous Urs fair of Ajmer, the location was urban and the lesions were characteristic of L. tropica. Therefore, the disease is suspected to be anthroponotic. These features are suggestive of a common mode of transmission, source and/or vector signalling introduction of this infection into a non-endemic area.

  1. Molecular epidemiological study of cutaneous leishmaniasis in the east north of Iran

    Directory of Open Access Journals (Sweden)

    Kazem Hassanpour

    2014-09-01

    Full Text Available Objective: To identify and study the epidemiology of Leishmania species isolated from patients with leishmaniasis by PCR method in Sabzevar, Khorasan Razavi Province, Iran. Methods: Aspirated samples from the lesions of 86 patients with confirmed leishmaniasis used for direct smear preparation. The samples are prepared and cultured in the Novy-MacNealNicolle and Roswell Park Memorial Institute-1640 culture mediums. After extraction of DNA by using phenol-chloroform, parasite kintoplast DNA gene amplification was done by using PCR. The electrophoresis pattern of each species was compared with standard species of Leishmania tropica (L. tropica and Leishmania major (L. major. Results: Results of PCR patterns of kintoplast DNA gene suggested that two types of L. tropica and L. major isolated from leishmaniasis patients in Sabzevar and their findings indicated that both species of parasites, L. tropica and L. major are prevalent in Sabzevar. Conclusions: According to this information, Sabzevar can be divided in two focal points of dry and dry-wet in terms of leishmaniasis, while it was previously known as the dry focal point. Likewise a significant relationship observed between Leishmania species to time distribution, type of lesion, amount of parasites and infection districts.

  2. 1608-IJBCS-Article-André T Kabré

    African Journals Online (AJOL)

    hp

    Eisenia fetida (Oligochaeta) and Oreo- chromis mossambicus (Cichlidae) in. South Africa. Acta Tropica, 63: 1-14. Chiayvareejja S, Rittibhonbhum N, Hong- promyart M,Wiriyachitra P. 1997. Toxic- ity of the Thai piscicidal plant, Maesa ramentacea, to freshwater fishes in ponds. Aquaculture, 158: 229-234. Davies W. 1983.

  3. Phylogenetic position of Leishmania isolates from Khyber Pakhtunkhwa province of Pakistan.

    Science.gov (United States)

    Khan, Nazma Habib; Messenger, Louisa A; Wahid, Sobia; Sutherland, Colin J

    2016-08-01

    Several species of the genus Leishmania are causative agents of cutaneous leishmaniasis in Pakistan. This study aimed to determine phylogenetic placement of Leishmania species causing cutaneous leishmaniasis in Khyber Pakhtunkhwa province, Pakistan (34 Leishmania tropica, 3 Leishmania infantum), in-relation to species from other geographical areas using gene sequences encoding cytochrome b (cytb) and internal transcribed spacer 2 (its2). Based on cytochrome b sequence analysis, L. tropica strains from Pakistan and other geographical regions were differentiated into two genotype groups, A and B. Within the province, five distinct L. tropica genotypes were recognized; two in group A, three in group B. Two L. infantum isolates from the province were closely associated with both Afro-Eurasian and American species of the Leishmania donovani complex, including Leishmania chagasi, L. infantum and L. donovani from Sudan and Ethiopia; while a third L. infantum isolate could not be differentiated from visceralizing Kenyan and Indian L. donovani. We observed apposite phylogenetic placement of CL-causing L. tropica and L. infantum from Khyber Pakhtunkhwa. Affinities ascribed to Leishmania spp. From the region are valuable in tracing potential importation of leishmaniasis. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  4. Ongoing epidemic of cutaneous leishmaniasis among Syrian refugees, Lebanon.

    Science.gov (United States)

    Saroufim, Maya; Charafeddine, Khalil; Issa, Grace; Khalifeh, Haifaa; Habib, Robert H; Berry, Atika; Ghosn, Nada; Rady, Alissar; Khalifeh, Ibrahim

    2014-10-01

    In September 2012, a cutaneous leishmaniasis outbreak began among Syrian refugees in Lebanon. For 948 patients in whom leishmaniasis was not confirmed, we obtained samples for microscopic confirmation and molecular speciation. We identified Leishmania tropica in 85% and L. major in 15% of patients. After 3 months of megulamine antimonite therapy, patients initial cure rate was 82%.

  5. In Vitro Inhibitory Effect of Berberis vulgaris (Berberidaceae) and Its Main Component, Berberine against Different Leishmania Species.

    Science.gov (United States)

    Mahmoudvand, Hossein; Sharififar, Fariba; Sharifi, Iraj; Ezatpour, Behrouz; Fasihi Harandi, Majid; Makki, Mahsa Sadat; Zia-Ali, Naser; Jahanbakhsh, Sareh

    2014-03-01

    Leishmaniasis has been identified as a major public health problem in tropical and sub-tropical countries. The present study was aimed to investigate antileishmanial effects of various extracts of Berberis vulgaris also its active compoenent, berberine against Leishmania tropica and L. infantum species on in vitro experiments. In this study in vitro antileishmanial activity of various extracts of B. vulgaris also its active compoenent, berberine against promastigote and amastigote stages of L. tropica and L. infantum was evaluated, using MTT assay and in a macrophage model, respectively. Furthermore, infectivity rate and cytotoxicity effects of B. vulgaris and berberine in murine macrophage cells were investigated. The findings of optical density (OD) and IC50 indicated that B. vulgaris particulary berberine significantly (P<0.05) inhibited the growth rate of promastigote stage of L.tropica and L.infantum in comparison to meglumine antimoniate (MA). In addition, B. vulgaris and berberine significantly (P<0.05) decreased the mean number of amastigotes in each macrophage as compared with positive control. In the evaluation of cytotoxicity effects, it could be observed that berberine as compared with B. vulgaris exhibited more cytotoxicity against murine macrophages. Results also showed that when parasites were pre-incubated with B. vulgaris their ability to infect murine macrophages was significantly decreased. B.vulgaris particularly berberine exhibited potent in vitro leishmanicidal effects against L. tropica and L.infantum. Further works are required to evaluate the antileishmanial effects of B.vulgaris on Leishmania species using clinical settings.

  6. Imported malaria in children: A national surveillance in the Netherlands and a review of European studies

    NARCIS (Netherlands)

    Driessen, G.J.; Pereira, R.R.; Brabin, B.J.; Hartwig, N.G.

    2008-01-01

    Background: Falciparum malaria or malaria tropica is one of the leading causes of childhood mortality worldwide. Malaria-related deaths occur mainly in sub-Saharan Africa, where an estimated 365 million clinical cases of Plasmodium falciparum malaria occur each year. In Europe, imported malaria

  7. Ongoing Epidemic of Cutaneous Leishmaniasis among Syrian Refugees, Lebanon1

    Science.gov (United States)

    Saroufim, Maya; Charafeddine, Khalil; Issa, Grace; Khalifeh, Haifaa; Habib, Robert H.; Berry, Atika; Ghosn, Nada; Rady, Alissar

    2014-01-01

    In September 2012, a cutaneous leishmaniasis outbreak began among Syrian refugees in Lebanon. For 948 patients in whom leishmaniasis was not confirmed, we obtained samples for microscopic confirmation and molecular speciation. We identified Leishmania tropica in 85% and L. major in 15% of patients. After 3 months of megulamine antimonite therapy, patients initial cure rate was 82%. PMID:25279543

  8. DNA repair characteristics of a hybrid cell clone between xeroderma pigmentosum and Potorous tridactilis

    International Nuclear Information System (INIS)

    Ida, Kenji

    1986-01-01

    A hybrid cell clone PX1 was isolated by fusing UV sensitive XP20S(SV)neo, an SV-40-transformed, neomycin-resistant xeroderma pigmentosum (XP) cell line, and Pt K2, a rat kangaroo (Potorous tridactilis) cell line. The UV-survival curve of PX1 cells fell midway between those of Pt K2 and XP20S(SV)neo cells, since mean lethal doses(D 0 ) were 2.5, 4.7 and 0.27 J/m 2 for PX1, Pt K2 and XP20S(SV)neo, respectively. Amounts of unscheduled DNA synthesis (UDS) after UV, relative to normal human cells, were 60.4 % for Pt K2, 37.7 % for PX1 and 0.1 % for XP20S(SV)neo. Such relative UDS capacities for excision repair of Pt K2, PX1 and XP20S(SV)neo were also consistent with the respective relative capacities of host cell reactivation (HCR) of UV-irradiated Herpes simplex virus. Apparently, there was no single Pt K2 chromosome in the PX1 cells. One possibility is that a gene which may account for the partial restoration of the UV resistance has been transferred from Pt K2 to PX1. (author)

  9. Porphyromonas loveana sp. nov., isolated from the oral cavity of Australian marsupials.

    Science.gov (United States)

    Bird, Philip S; Trott, Darren J; Mikkelsen, Deirdre; Milinovich, Gabriel J; Hillman, Kristine M; Burrell, Paul C; Blackall, Linda L

    2016-10-01

    An obligatory anaerobic, Gram-stain-negative coccobacillus with black-pigmented colonies was isolated from the oral cavity of selected Australian marsupial species. Phenotypic and molecular criteria showed that this bacterium was a distinct species within the genus Porphyromonas, and was closely related to Porphyromonas gingivalis and Porphyromonas gulae. This putative novel species and P. gulae could be differentiated from P. gingivalis by catalase activity. Further characterization by multi-locus enzyme electrophoresis of glutamate dehydrogenase and malate dehydrogenase enzyme mobility and matrix-assisted laser desorption ionization time-of-flight MS showed that this putative novel species could be differentiated phenotypically from P. gingivalis and P. gulae. Definitive identification by 16S rRNA gene sequencing showed that this bacterium belonged to a unique monophyletic lineage, phylogenetically distinct from P. gingivalis (94.9 % similarity) and P. gulae (95.5 %). This also was supported by 16S-23S rRNA intergenic spacer region and glutamate dehydrogenase gene sequencing. A new species epithet, Porphyromonas loveana sp. nov., is proposed for this bacterium, with DSM 28520T (=NCTC 13658T=UQD444T=MRK101T), isolated from a musky rat kangaroo, as the type strain.

  10. X-ray induction of mitotic and meiotic chromosome aberrations

    International Nuclear Information System (INIS)

    Yao, K.T.S.

    1980-01-01

    In 1964 six pairs of rat kangaroo (Potorous tridactylis) were obtained from Australia. The tissues of these animals were used to initiate cell lines. Since this species has a low chromosome number of six pairs, each pair with its own distinctive morphology, it is particularly favorable for cytogenetic research. In cell cultures derived from the corneal endothelial tissues of one animal there emerged a number of haploid cells. The number of haploid cells in the cultures reached as high as 20% of the total mitotic configurations. The in vitro diploid and haploid mixture cell cultures could be a resemblance or a coincidence to the mixture existence of the diploid primary spermatocytes and the haploid secondary spermatocytes (gametes) in the in vivo testicular tissues of the male animals. It would be interesting to compare reactions of the haploid and diploid cell mixture, either in the cultures or in the testes, to x-ray exposure. Two other studies involving x-ray effects on Chinese hamster oocyte maturation and meiotic chromosomes and the x-ray induction of Chinese hamster spermatocyte meiotic chromosome aberrations have been done in this laboratory. A review of these three studies involving diploid and haploid chromosomes may lead to further research in the x-ray induction of chromosome aberrations

  11. Induced DNA repair pathway in mammalian cells

    International Nuclear Information System (INIS)

    Overberg, R.

    1985-01-01

    The survival of cultured rat kangaroo cells (PtK-2) and human xeroderma pigmentosum cells incubated with 5 μM cycloheximide subsequent to ultraviolet irradiation is lower than that of cells incubated without cycloheximide. The drop in survival is considerably larger than that produced by incubation of unirradiated cells with cycloheximide. The phenomenon was also observed when PtK-2 cells were incubated with emetine, another protein synthesis inhibitor, or with 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole, a RNA synthesis inhibitor. PtK cells which received a preliminary UV treatment followed by an incubation period without cycloheximide and then a second irradiation and 24 hour incubation with cycloheximide, survived the effects of the second irradiation better than cells which were incubated in the presence of cycloheximide after the first and second UV irradiation. The application of cycloheximide for 24 hours after UV irradiation of PtK cells resulted in one-half as many 6-thioguanine resistant cells as compared to the number of 6-thioguanine resistant cells found when cycloheximide was not used. These experiments indicate that a UV-inducible cycloheximide-sensitive DNA repair pathway is present in PtK and xeroderma pigmentosum cells, which is error-prone in PtK cells

  12. RNA polymerase activity in PtK1 micronuclei containing individual chromosomes: an in vitro and in situ study

    International Nuclear Information System (INIS)

    Labidi, B.; Gregoire, M.; Frackowiak, S.; Hernandez-Verdun, D.; Bouteille, M.

    1987-01-01

    Micronuclei have been induced by colchicine in rat kangaroo (Potorous tridactylis) PtK1 cells. The synthesis of RNA was investigated both in isolated micronuclei by quantifying RNA polymerase activities at different ionic strengths with or without inhibitors, and in micronucleated cells by radioautography after [ 3 H]uridine pulse labeling. In vitro transcription shows that isolated micronuclei are able to take up [ 3 H]UTP. The rate curves of incorporation are close to those of isolated diploid nuclei, though the level of incorporation was relatively lower (65-70%) than control nuclei. This indicates that micronuclei react to the ionic environment and to inhibitors in the same manner as described for many species of isolated diploid nuclei. The labelling distributions plotted from radioautographs show that micronuclei were able to efficiently incorporate the hot precursor. Furthermore, for short pulses there is no homogeneity in the labelling density among the different micronuclei and there is no correlation between the labelling intensity and the size of micronuclei. After 60-min pulse time, there is an enhanced uptake of [ 3 H]uridine and all the micronuclei exhibit considerable labelling, although less than control cells. Thus, the micronuclei exhibit some characteristic RNA transcriptional activity in situ as well as after isolation. This material should be a particular interesting model with which to study the physiological activity and the role of each individual interphasic chromosome

  13. Epidemiological profile of cutaneous leishmaniasis in Morocco, 2004-2013.

    Science.gov (United States)

    Laboudi, M

    2017-02-01

    The main objective of our study is to provide a comprehensive overview of the situation of cutaneous leishmaniasis in Morocco. We therefore conducted a retrospective descriptive study covering the decade 2004-2013. Data were collected from the Moroccan Ministry of Health's annual reports about the national program against leishmaniasis. These data show a regression in the cases due to Leishmania major and the persistence of L. tropica transmission. Mapping cutaneous leishmaniasis cases reported between 2004 and 2013 showed geographical changes ; the number of cases due to L. tropica were concentrated in the regions of Marrakech-Tensift-Elhaouz, Souss Massa-Draa Tadla-Azila, and Taza-Al Hoceima-Taounate. Cutaneous disease due to L. major was most common in the Souss-Massa-Draa region. Continuous monitoring of the epidemiological situation is important to assess the actions taken to reduce its incidence.

  14. The United States Army Medical Department Journal, July-September 2009

    Science.gov (United States)

    2009-09-01

    including Escherichia coli O157:H7, Salmonella enteritidis, Yersinia pseudotuberculosis, Helicobactor pylori, Vibrio cholerae , and rotavirus,3-11 as shown...of houseflies (Musca domestica) in the transmission of Vibrio cholerae in India. Acta Tropica. 2001;78:31-34. 11. Tan SW, Yap KL, Lee HL. Mechanical...northeastern Mexico . J Am Mosq Control Assoc. 2001;17(2):93-97. 22. Cohnstaedt LW, Gillen JI, Munstermann LE. Light- emitting diode technology improves

  15. Molecular Characterization of Leishmania Parasites in Giemsa-Stained Slides from Cases of Human Cutaneous and Visceral Leishmaniasis, Eastern Algeria.

    Science.gov (United States)

    Beldi, Nadia; Mansouri, Roukaya; Bettaieb, Jihene; Yaacoub, Alia; Souguir Omrani, Hejer; Saadi Ben Aoun, Yusr; Saadni, Farida; Guizani, Ikram; Guerbouj, Souheila

    2017-06-01

    In Algeria, visceral leishmaniasis (VL) is due to Leishmania (L.) infantum, while three cutaneous forms (CL) are caused by Leishmania major, Leishmania tropica and Leishmania infantum. In this study, the use of Giemsa-stained slides was evaluated with two PCR techniques, in Eastern Algeria. A total of 136 samples corresponding to 100 CL smears (skin scrapings) and 36 VL slides (bone marrow aspirates) collected from 2008 to 2014 were tested. Upon DNA extraction, two PCRs were used to amplify the ribosomal Internal Transcribed Spacer 1 (ITS1) and mini-exon genes. Amplified products were digested (PCR-RFLP) and profiles analyzed for Leishmania species identification. A statistical analysis was also performed. ITS1-PCR was found significantly more sensitive than mini-exon-PCR (77.95% positives vs. 67.65%; p = 0.001). Comparison of PCR positivity showed statistically significant differences between old and recently prepared slides suggesting a better use of recent slides in PCR analyses. For species identification, PCR-restriction fragment length polymorphism (RFLP) results of ITS1 and mini-exon were concordant. L. infantum was identified from VL cases and L. infantum, L. major, and L. tropica from CL ones. According to geographical origin, L. infantum was found in North-Eastern provinces, while L. major was distributed from the North to the Center-East of Algeria. Interestingly, two L. tropica samples were identified in Annaba, located far North-East Algeria. Distribution of leishmaniasis in Eastern parts of Algeria, besides finding of L. tropica in the far North, is in this study described for the first time using molecular tools, thus confirming the usefulness of slides for PCR identification of Leishmania parasites in retrospective epidemiological investigations.

  16. Molecular identification and polymorphism determination of cutaneous and visceral leishmaniasis agents isolated from human and animal hosts in Iran.

    Science.gov (United States)

    Hajjaran, Homa; Mohebali, Mehdi; Mamishi, Setareh; Vasigheh, Farzaneh; Oshaghi, Mohammad Ali; Naddaf, Saied Reza; Teimouri, Aref; Edrissian, Gholam Hossein; Zarei, Zabiholah

    2013-01-01

    Amplification of internal transcript spacer 1 of ribosomal RNA (ITS1-RNA) gene followed by RFLP analysis and sequencing was used to identify the causing agents of cutaneous and visceral leishmaniasis (CL and VL) in humans and animal reservoir hosts from various geographical areas in Iran. We also used random amplified polymorphic DNA (RAPD-PCR) to obtain polymorphisms among isolates of Leishmania spp. Totally, 362 suspected human and animal cases including 173 CL, 49 VL, 60 rodents, and 80 domestic dogs were examined for Leishmania infection. From 112 culture-positive samples prepared from CL cases, 75 (67%) were infected with L. major and 37 (33%) with L. tropica. Of the 60 rodents examined, 25 (41.6%) harbored the Leishmania infection; 21 were infected with L. major and 4 with L. turanica. From 49 suspected VL, 29 were positive by direct agglutination test (DAT), whereas microscopy detected parasite in bone marrow of 25 and culture in 28 of the patients. Two VL patients were infected with L. tropica and 26 with L. infantum. Of the 80 domestic dogs, 56 showed anti-Leishmania antibodies with DAT. Of these, 55 were positive by both microscopy and culture. Molecular identity, obtained only for 47 samples, revealed L. infantum in 43 and L. tropica in 4 dogs. The polymorphisms among L. tropica and L. major isolates were 3.6% and 7.3%; the rate among human and canine VL isolates was 2.8% and 9.8%, respectively. Our results showed that at least four different Leishmania species with various polymorphisms circulate among humans and animal hosts in Iran.

  17. Molecular Identification and Polymorphism Determination of Cutaneous and Visceral Leishmaniasis Agents Isolated from Human and Animal Hosts in Iran

    Directory of Open Access Journals (Sweden)

    Homa Hajjaran

    2013-01-01

    Full Text Available Amplification of internal transcript spacer 1 of ribosomal RNA (ITS1-RNA gene followed by RFLP analysis and sequencing was used to identify the causing agents of cutaneous and visceral leishmaniasis (CL and VL in humans and animal reservoir hosts from various geographical areas in Iran. We also used random amplified polymorphic DNA (RAPD-PCR to obtain polymorphisms among isolates of Leishmania spp. Totally, 362 suspected human and animal cases including 173 CL, 49 VL, 60 rodents, and 80 domestic dogs were examined for Leishmania infection. From 112 culture-positive samples prepared from CL cases, 75 (67% were infected with L. major and 37 (33% with L. tropica. Of the 60 rodents examined, 25 (41.6% harbored the Leishmania infection; 21 were infected with L. major and 4 with L. turanica. From 49 suspected VL, 29 were positive by direct agglutination test (DAT, whereas microscopy detected parasite in bone marrow of 25 and culture in 28 of the patients. Two VL patients were infected with L. tropica and 26 with L. infantum. Of the 80 domestic dogs, 56 showed anti-Leishmania antibodies with DAT. Of these, 55 were positive by both microscopy and culture. Molecular identity, obtained only for 47 samples, revealed L. infantum in 43 and L. tropica in 4 dogs. The polymorphisms among L. tropica and L. major isolates were 3.6% and 7.3%; the rate among human and canine VL isolates was 2.8% and 9.8%, respectively. Our results showed that at least four different Leishmania species with various polymorphisms circulate among humans and animal hosts in Iran.

  18. In Vitro Inhibitory Effect of Berberis vulgaris (Berberidaceae and Its Main Component, Berberine against Different Leishmania Species.

    Directory of Open Access Journals (Sweden)

    Hossein Mahmoudvand

    2014-03-01

    Full Text Available Leishmaniasis has been identified as a major public health problem in tropical and sub-tropical countries. The present study was aimed to investigate antileishmanial effects of various extracts of Berberis vulgaris also its active compoenent, berberine against Leishmania tropica and L. infantum species on in vitro experiments.In this study in vitro antileishmanial activity of various extracts of B. vulgaris also its active compoenent, berberine against promastigote and amastigote stages of L. tropica and L. infantum was evaluated, using MTT assay and in a macrophage model, respectively. Furthermore, infectivity rate and cytotoxicity effects of B. vulgaris and berberine in murine macrophage cells were investigated.The findings of optical density (OD and IC50 indicated that B. vulgaris particulary berberine significantly (P<0.05 inhibited the growth rate of promastigote stage of L.tropica and L.infantum in comparison to meglumine antimoniate (MA. In addition, B. vulgaris and berberine significantly (P<0.05 decreased the mean number of amastigotes in each macrophage as compared with positive control. In the evaluation of cytotoxicity effects, it could be observed that berberine as compared with B. vulgaris exhibited more cytotoxicity against murine macrophages. Results also showed that when parasites were pre-incubated with B. vulgaris their ability to infect murine macrophages was significantly decreased.B.vulgaris particularly berberine exhibited potent in vitro leishmanicidal effects against L. tropica and L.infantum. Further works are required to evaluate the antileishmanial effects of B.vulgaris on Leishmania species using clinical settings.

  19. Impact of phlebotomine sand flies on U.S. military operations at Tallil Air Base, Iraq: 4. Detection and identification of leishmania parasites in sand flies.

    Science.gov (United States)

    Coleman, Russell E; Hochberg, Lisa P; Swanson, Katherine I; Lee, John S; McAvin, James C; Moulton, John K; Eddington, David O; Groebner, Jennifer L; O'Guinn, Monica L; Putnam, John L

    2009-05-01

    Sand flies collected between April 2003 and November 2004 at Tallil Air Base, Iraq, were evaluated for the presence of Leishmania parasites using a combination of a real-time Leishmania-generic polymerase chain reaction (PCR) assay and sequencing of a 360-bp fragment of the glucose-6-phosphate-isomerase (GPI) gene. A total of 2,505 pools containing 26,574 sand flies were tested using the real-time PCR assay. Leishmania DNA was initially detected in 536 pools; however, after extensive retesting with the real-time PCR assay, a total of 456 pools were considered positive and 80 were considered indeterminate. A total of 532 samples were evaluated for Leishmania GPI by sequencing, to include 439 PCR-positive samples, 80 PCR-indeterminate samples, and 13 PCR-negative samples. Leishmania GPI was detected in 284 samples that were sequenced, to include 281 (64%) of the PCR-positive samples and 3 (4%) of the PCR-indeterminate samples. Of the 284 sequences identified as Leishmania, 261 (91.9%) were L. tarentolae, 18 (6.3%) were L. donovani-complex parasites, 3 (1.1%) were L. tropica, and 2 were similar to both L. major and L. tropica. Minimum field infection rates were 0.09% for L. donovani-complex parasites, 0.02% for L. tropica, and 0.01% for the L. major/tropica-like parasite. Subsequent sequencing of a 600-bp region of the "Hyper" gene of 12 of the L. donovani-complex parasites showed that all 12 parasites were L. infantum. These data suggest that L. infantum was the primary leishmanial threat to U.S. military personnel deployed to Tallil Air Base. The implications of these findings are discussed.

  20. Development and Production of a Leishmania Skin Test

    Science.gov (United States)

    2009-03-01

    homologous species than to L.tropica. Phase II Study Report: USTA-06 Page 8 of 53 3. LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS: 21 AE CL CPM CRF...be recruited from the patient data base of the clinical site. If other recruiting measures are needed, such as placing an advertisement in the local...newspaper or by posting or distributing flyers in public areas, such advertisement or flyers will be sent to the IRB for review and approval. Refer to

  1. Activation of macrophages for microbicidal and tumoricidal effector functions by soluble factors from EL-4, a continuous T cell line.

    OpenAIRE

    Nacy, C A; James, S L; Benjamin, W R; Farrar, J J; Hockmeyer, W T; Meltzer, M S

    1983-01-01

    Macrophages treated with culture fluids from EL-4 cells, a continuous T cell line, were activated to kill mKSA-TU-5 fibrosarcoma cells, amastigotes of Leishmania tropica, and schistosomula of Schistosoma mansoni. Active EL-4 factors eluted from Sephadex G-100 in two distinct regions: molecular weight 45,000 (activities induced killing of unrelated intracellular and extracellular targets) and molecular weight 23,000 (activities induced killing of extracellular targets only). These results conf...

  2. Sensitive Molecular Diagnostics for Cutaneous Leishmaniasis.

    Science.gov (United States)

    Sagi, Orli; Berkowitz, Anat; Codish, Shlomi; Novack, Victor; Rashti, Aviv; Akad, Fouad; Shemer-Avni, Yonat

    2017-01-01

    Rapid diagnosis of cutaneous leishmaniasis (CL) and identification of Leishmania species is highly important for the disease management. In Israel, CL is caused mainly by Leishmania major and Leishmania tropica species. We established an easy to handle point of care lesion-swabbing, combined with a highly sensitive multiplex real time PCR (multiplex qPCR) for accurate and rapid diagnosis of Leishmania species. Using three probes: one general for: Leishmania species, and two specific for L major , and L tropica , we screened 1783 clinical samples collected during two years. Leishmania species was found in 1086 individuals, 1008 L major , and 70 L tropica . Eight samples positive for Leishmania species only, were further tested using a second set of multiplex qPCR developed, and were found positive for Leishmania braziliensis and Leishmania infantum/donovani (2 and 6 samples, concomitantly). Taken together, the test enabled diagnostics and better treatment of Leishmania infections from the Old World (1078 samples) and the New World (8 samples), and the subtyping of the dominant strains in the region, as well as in returning travelers'.

  3. Detection, molecular typing and phylogenetic analysis of Leishmania isolated from cases of leishmaniasis among Syrian refugees in Lebanon

    Directory of Open Access Journals (Sweden)

    Tamara Salloum

    2016-06-01

    Two molecular typing methods of 39 FFPE Leishmania isolates were used: the ITS1-PCR RFLP and the nested ITS1-5.8S rDNA gene amplification followed by sequencing and phylogenetic analysis. The efficiency of these two techniques in Leishmania identification was compared and the phylogenetic relationships among these isolates were illustrated based on the neighbor-joining (NJ method. The results were statistically correlated with the parasitic index (PI. The DNA storage in formalin-fixed paraffin embedded (FFPE tissues was assessed as well. The parasites identified were all L. tropica as determined by both techniques. ITS1-5.8S rDNA gene based typing proved to be more sensitive in the detection of parasites (positive in 69.2% of the isolates as opposed to the ITS1-PCR RFLP method that was successful in identifying L. tropica in only 43.6% of the isolates. Sequencing and phylogenetic analysis revealed high levels of heterogeneity. A statistically significant correlation was observed between PI and the results of the nested ITS1-5.8S rDNA gene PCR. Genotyping at the species level is essential for monitoring the relative frequency of CL in the Mediterranean area that is correlated to three different Leishmania species (Leishmania infantum, Leishmania major and L. tropica, each characterized by distinct epidemiological features. The obtained results highlight the need to find a universally accepted diagnostic tool for Leishmania typing.

  4. Receptors and cGMP signalling mechanism for E. coli enterotoxin in opossum kidney

    International Nuclear Information System (INIS)

    Forte, L.R.; Krause, W.J.; Freeman, R.H.

    1988-01-01

    Receptors for the heat-stable enterotoxin produced by Escherichia coli were found in the kidney and intestine of the North American opossum and in cultured renal cell lines. The enterotoxin markedly increased guanosine 3',5'-cyclic monophosphate (cGMP) production in slices of kidney cortex and medulla, in suspensions of intestinal mucosa, and in the opossum kidney (OK) and rat kangaroo kidney (PtK-2) cell lines. In contrast, atrial natriuretic factor elicited much smaller increases in cGMP levels of kidney, intestine, or cultured kidney cell lines. The enterotoxin receptors in OK cells had a molecular mass of approximately 120 kDa when measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of receptors crosslinked with 125 I-enterotoxin. The occurrence of receptors for the E. coli peptide in OK implies that these receptors may be involved in the regulation of renal tubular function in the opossum. E. coli enterotoxin caused a much larger increase in urine cGMP excretion than did atrial natriuretic factor when these peptides were injected intravenously into opossums. However, atrial natriuretic factor elicited a marked diuresis, natriuresis, and increased urinary excretion of calcium, phosphate, potassium, and magnesium. In contrast, the enterotoxin did not acutely influence OK fluid and electrolyte excretion. Thus the substantial increase in cGMP synthesis produced by the bacterial peptide in OK cortex and medulla in vitro and the increased renal excretion of cGMP in vivo were not associated with changes in electrolyte or water excretion. Whether cGMP represents a second messenger molecule in the kidney is an interesting question that was raised but not answered in this series of experiments

  5. Clinical Features of Anthroponotic Cutaneous Leishmaniasis in a Major Focus, Southeastern Iran, 1994-2014

    Directory of Open Access Journals (Sweden)

    Ahmad KHOSRAVI

    2017-12-01

    Full Text Available AbstractBackground: Cutaneous leishmaniasis (CL is associated with a broad and complex clinical spectrum of diseases. The objectives of this study were to assess the clinical features and identification of the causative agents of CL in a well-known focus of anthroponotic CL (ACL caused by Leishmania tropica, southeast Iran.Methods: This study was performed randomly as a descriptive cross-sectional survey to evaluate 2000 CL patients by active and passive case-detection approaches in Kerman Province from 1994 to 2014. The ACL patients were confirmed by direct smear and 600 cases by one or a combination of intrinsic methods.Results: Children aged <10 yr old were the most infected patients (P<0.001. The majority of the CL lesions were located in hands (46.3%, face (34.1%, legs (14.3%, and other parts of the body (5.3%. The mean number of lesions was 1.5 and most of the patients had single lesion (65%.Typical clinical lesions included papule (36.8%, followed by ulcerated nodule (20.7%, plaque (18.4%, and ulcerated plaque (18.5%. While among atypical clinical features, leishmaniasis recidivans (LR (4.7% and leishmanid (0.3% were the dominant forms, followed by diffuse, disseminated, sporotrichoid, and erysipeloid types, 0.1% each, and then lymphedematous, lymphadenic, hyperkeratotic, paronychial, and mutilating types, 0.05% each. Based on various intrinsic methods the parasites isolated from the lesions were characterized as L. tropica.Conclusion: ACL due to L. tropica presents numerous cases of localized form and diverse uncommon clinical presentations, which mimic other disease conditions. Therefore, physicians should be aware of such manifestations for selecting appropriate treatment modality.

  6. Assessment of nuclear and mitochondrial genes in precise identification and analysis of genetic polymorphisms for the evaluation of Leishmania parasites.

    Science.gov (United States)

    Fotouhi-Ardakani, Reza; Dabiri, Shahriar; Ajdari, Soheila; Alimohammadian, Mohammad Hossein; AlaeeNovin, Elnaz; Taleshi, Neda; Parvizi, Parviz

    2016-12-01

    The polymorphism and genetic diversity of Leishmania genus has status under discussion depending on many items such as nuclear and/or mitochondrial genes, molecular tools, Leishmania species, geographical origin, condition of micro-environment of Leishmania parasites and isolation of Leishmania from clinical samples, reservoir host and vectors. The genetic variation of Leishmania species (L. major, L. tropica, L. tarentolae, L. mexicana, L. infantum) were analyzed and compared using mitochondrial (COII and Cyt b) and nuclear (nagt, ITS-rDNA and HSP70) genes. The role of each enzymatic (COII, Cyt b and nagt) or housekeeping (ITS-rDNA, HSP70) gene was employed for accurate identification of Leishmania parasites. After DNA extractions and amplifying of native, natural and reference strains of Leishmania parasites, polymerase chain reaction (PCR) products were sequenced and evaluation of genetic proximity and phylogenetic analysis were performed using MEGA6 and DnaSP5 software. Among the 72 sequences of the five genes, the number of polymorphic sites was significantly lower as compared to the monomorphic sites. Of the 72 sequences, 54 new haplotypes (five genes) of Leishmania species were submitted in GenBank (Access number: KU680818 - KU680871). Four genes had a remarkable number of informative sites (P=0.00), except HSP70 maybe because of its microsatellite regions. The non-synonymous (dN) variants of nagt gene were more than that of other expression genes (47.4%). The synonymous (dS)/dN ratio in three expression genes showed a significant variation between five Leishmania species (P=0.001). The highest and lowest levels of haplotype diversity were observed in L. tropica (81.35%) and L. major (28.38%) populations, respectively. Tajima's D index analyses showed that Cyt b gene in L. tropica species was significantly negative (Tajima's D=-2.2, PLeishmania parasites. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis.

    Science.gov (United States)

    Nemati, Sara; Fazaeli, Asghar; Hajjaran, Homa; Khamesipour, Ali; Anbaran, Mohsen Falahati; Bozorgomid, Arezoo; Zarei, Fatah

    2017-08-01

    Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

  8. Antileishmanial effect of silver nanoparticles and their enhanced antiparasitic activity under ultraviolet light

    Directory of Open Access Journals (Sweden)

    Allahverdiyev AM

    2011-11-01

    Full Text Available Adil M Allahverdiyev1, Emrah Sefik Abamor1, Malahat Bagirova1, Cem B Ustundag2, Cengiz Kaya2, Figen Kaya2, Miriam Rafailovich3 1Department of Bioengineering; 2Department of Metallurgical and Materials Engineering, Yildiz Technical University, Esenler, Istanbul, Turkey; 3Department of Materials Science and Engineering, Stony Brook University, Stony Brook, NY, USA Abstract: Leishmaniasis is a protozoan vector-borne disease and is one of the biggest health problems of the world. Antileishmanial drugs have disadvantages such as toxicity and the recent development of resistance. One of the best-known mechanisms of the antibacterial effects of silver nanoparticles (Ag-NPs is the production of reactive oxygen species to which Leishmania parasites are very sensitive. So far no information about the effects of Ag-NPs on Leishmania tropica parasites, the causative agent of leishmaniasis, exists in the literature. The aim of this study was to investigate the effects of Ag-NPs on biological parameters of L. tropica such as morphology, metabolic activity, proliferation, infectivity, and survival in host cells, in vitro. Consequently, parasite morphology and infectivity were impaired in comparison with the control. Also, enhanced effects of Ag-NPs were demonstrated on the morphology and infectivity of parasites under ultraviolet (UV light. Ag-NPs demonstrated significant antileishmanial effects by inhibiting the proliferation and metabolic activity of promastigotes by 1.5- to threefold, respectively, in the dark, and 2- to 6.5-fold, respectively, under UV light. Of note, Ag-NPs inhibited the survival of amastigotes in host cells, and this effect was more significant in the presence of UV light. Thus, for the first time the antileishmanial effects of Ag-NPs on L. tropica parasites were demonstrated along with the enhanced antimicrobial activity of Ag-NPs under UV light. Determination of the antileishmanial effects of Ag-NPs is very important for the further

  9. Molecular Epidemiological Survey of Cutaneous Leishmaniasis in Two Highly Endemic Metropolises of Iran, Application of FTA Cards for DNA Extraction From Giemsa-Stained Slides.

    Science.gov (United States)

    Izadi, Shahrokh; Mirhendi, Hossein; Jalalizand, Niloufar; Khodadadi, Hossein; Mohebali, Mehdi; Nekoeian, Shahram; Jamshidi, Ali; Ghatee, Mohammad Amin

    2016-02-01

    PCR has been used for confirmation of leishmaniasis in epidemiological studies, but complexity of DNA extraction and PCR approach has confined its routine use in developing countries. In this study, recent epidemiological situation of cutaneous leishmaniasis (CL) in two hyper-endemic metropolises of Shiraz and Isfahan in Iran was studied using DNA extraction by commercial FTA cards and kinetoplastid DNA (kDNA)-PCR amplification for detection/identification of Leishmania directly from stained skin scraping imprints. Fifty four and 30 samples were collected from clinically diagnosed CL patients referred to clinical laboratories of leishmaniasis control centers in Isfahan and Shiraz cities, respectively. The samples were examined by direct microscopy and then scrapings of the stained smears were applied to FTA cards and used directly as DNA source in a nested-PCR to amplify kDNA to detect and identify Leishmania species. Fifty four of 84 (64.2%) slides obtained from patients had positive results microscopically, while 79/84 (94%) of slides had positive results by FTA card-nested-PCR. PCR and microscopy showed a sensitivity of 96.4% and 64.2% and specificity of 100% and 100%, respectively. Interestingly, Leishmania major as causative agent of zoonotic CL was identified in 100% and 90.7% of CL cases from Isfahan and Shiraz cities, respectively, but L. tropica was detected from only 9.3% of cases from Shiraz city. All cases from central regions of Shiraz were L. tropica and no CL case was found in Isfahan central areas. Filter paper-based DNA extraction can facilitate routine use of PCR for diagnosis of CL in research and diagnostic laboratories in Iran and countries with similar conditions. Epidemiologic changes including dominancy of L. major in suburbs of Shiraz and Isfahan metropolises where anthroponotic CL caused by L. tropica had been established, showed necessity of precise studies on CL epidemiology in old urban and newly added districts in the suburbs.

  10. Seed borne mycoflora of castor bean (ricinus communis l.) from pakistan

    International Nuclear Information System (INIS)

    Dawar, S.; Khalid, S.

    2014-01-01

    Castor bean seeds were analyzed by using ISTA (International seed testing association) for the detection of seed borne mycoflora. Thirty one fungal species belonging to 15 genera were isolated from 12 samples of castor bean seeds collected from different areas of Pakistan. Fusarium solani, Alternaria alternata, Cephaliophora tropica were most predominant fungal species isolated while the saprophytic fungi like A. niger, A. flavus were common in all samples of castor bean seed tested. Blotter method was considered to be better technique which gave maximum number of fungi followed by agar plate and deep freezing methods. (author)

  11. Brachionidae (Rotifera: Monogononta de la albufera El Paraíso y el reporte de Brachionus ibericusen el Perú

    Directory of Open Access Journals (Sweden)

    Emily Toscano

    2013-12-01

    Full Text Available En este trabajo se reportan las especies de Brachionidae reconocidos en las muestras recogidas en octubre de 2010 en la albufera Laguna El Paraiso, Huacho (11°13' −11° 10'S, 77° 35' −77°40'W. Cinco especies fueron determinadas: Keratella tropica, Brachionus quadridentatus, B. urceolaris, B. angularis y B. ibericusCiros-Pérez 2001. Este último es un nuevo reporte para el Perú y amplía su distribución a nivel mundial.

  12. Spatiotemporal and molecular epidemiology of cutaneous leishmaniasis in Libya.

    Directory of Open Access Journals (Sweden)

    Ahmad Amro

    2017-09-01

    Full Text Available Cutaneous leishmaniasis (CL is a major public health problem in Libya. In this paper, we describe the eco-epidemiological parameters of CL during the armed conflict period from January 2011 till December 2012. Current spatiotemporal distributions of CL cases were explored and projected to the future using a correlative modelling approach. In addition the present results were compared with our previous data obtained for the time period 1995-2008.We investigated 312 CL patients who presented to the Dermatology Department at the Tripoli Central Hospital and came from 81 endemic areas distributed in 10 districts. The patients presented with typical localized lesions which appeared commonly on the face, arms and legs. Molecular identification of parasites by a PCR-RFLP approach targeting the ITS1 region of the rDNA was successful for 81 patients with two causative species identified: L. major and L. tropica comprised 59 (72.8% and 22 (27.2% cases, respectively. Around 77.3% of L. tropica CL and 57.7% of L. major CL caused single lesions. Five CL patients among our data set were seropositive for HIV. L. tropica was found mainly in three districts, Murqub (27.3%, Jabal al Gharbi (27.3% and Misrata (13.7% while L. major was found in two districts, in Jabal al Gharbi (61% and Jafara (20.3%. Seasonal occurrence of CL cases showed that most cases (74.2% admitted to the hospital between November and March, L. major cases from November till January (69.4%, and L. tropica cases mainly in January and February (41%. Two risk factors were identified for the two species; the presence of previously infected household members, and the presence of rodents and sandflies in patient's neighborhoods. Spatiotemporal projections using correlative distribution models based on current case data and climatic conditions showed that coastal regions have a higher level of risk due to more favourable conditions for the transmitting vectors.Future projection of CL until 2060

  13. Arginase activity in pathogenic and non-pathogenic species of Leishmania parasites.

    Science.gov (United States)

    Badirzadeh, Alireza; Taheri, Tahereh; Taslimi, Yasaman; Abdossamadi, Zahra; Heidari-Kharaji, Maryam; Gholami, Elham; Sedaghat, Baharehsadat; Niyyati, Maryam; Rafati, Sima

    2017-07-01

    Proliferation of Leishmania (L.) parasites depends on polyamine availability, which can be generated by the L-arginine catabolism and the enzymatic activity of arginase (ARG) of the parasites and of the mammalian hosts. In the present study, we characterized and compared the arginase (arg) genes from pathogenic L. major and L. tropica and from non-pathogenic L. tarentolae. We quantified the level of the ARG activity in promastigotes and macrophages infected with pathogenic L. major and L. tropica and non-pathogenic L. tarentolae amastigotes. The ARG's amino acid sequences of the pathogenic and non-pathogenic Leishmania demonstrated virtually 98.6% and 88% identities with the reference L. major Friedlin ARG. Higher ARG activity was observed in all pathogenic promastigotes as compared to non-pathogenic L. tarentolae. In vitro infection of human macrophage cell line (THP1) with pathogenic and non-pathogenic Leishmania spp. resulted in increased ARG activities in the infected macrophages. The ARG activities present in vivo were assessed in susceptible BALB/c and resistant C57BL/6 mice infected with L. major, L. tropica and L. tarentolae. We demonstrated that during the development of the infection, ARG is induced in both strains of mice infected with pathogenic Leishmania. However, in L. major infected BALB/c mice, the induction of ARG and parasite load increased simultaneously according to the time course of infection, whereas in C57BL/6 mice, the enzyme is upregulated solely during the period of footpad swelling. In L. tropica infected mice, the footpads' swellings were slow to develop and demonstrated minimal cutaneous pathology and ARG activity. In contrast, ARG activity was undetectable in mice inoculated with the non-pathogenic L. tarentolae. Our data suggest that infection by Leishmania parasites can increase ARG activity of the host and provides essential polyamines for parasite salvage and its replication. Moreover, the ARG of Leishmania is vital for parasite

  14. Cutaneous leishmaniasis in the central provinces of Hama and Edlib in Syria: Vector identification and parasite typing.

    Science.gov (United States)

    Haddad, Nabil; Saliba, Hanadi; Altawil, Atef; Villinsky, Jeffrey; Al-Nahhas, Samar

    2015-10-12

    Cutaneous leishmaniasis is a disease transmitted by sand fly bites. This disease is highly prevalent in Syria where Leishmania major and Leishmania tropica are the known aetiological agents. In 2011, more than 58,000 cases were reported in the country by the Ministry of Health. The central region of the country harbors 20 % of the reported cases. However, the epidemiology of the disease in this area is not well understood. An epidemiological survey was conducted in 2010 to identity the circulating parasite and the sand fly vector in the central provinces of Edlib and Hama. Sand fly specimens were collected using CDC light traps and identified morphologically. Total DNA was extracted from the abdomens of female specimens and from Giemsa-stained skin lesion smears of 80 patients. Leishmania parasites were first identified by sequencing the ITS1 gene amplicons. Then polymorphism analysis was performed using the RFLP technique. A total of 2142 sand flies were collected. They belonged to eight species, among which Phlebotomus sergenti and Phlebotomus papatasi were the most predominant. L. tropica ITS1 gene was amplified from two pools of P. sergenti specimens and from skin smears of cutaneous leishmaniasis patients. This suggests that P. sergenti is the potential vector species in the study area. The digestion profiles of the obtained amplicons by TaqI restriction enzyme were identical for all analysed L. tropica parasites. Moreover, L. infantum ITS1 gene was amplified from two pools of Phlebotomus tobbi in the relatively humid zone of Edlib. L. tropica is confirmed to be the aetiological agent of cutaneous leishmaniasis cases in the central provinces. RFLP technique failed to show any genetic heterogeneity in the ITS1 gene among the tested parasites. The molecular detection of this parasite in human skin smears and in P. sergenti supports the vector status of this species in the study area. The detection of L. infantum in P. tobbi specimens indicates a potential

  15. Lysine and glutamate transport in the erythrocytes of common brushtail possum, Tammar Wallaby and eastern grey, kangaroo.

    Science.gov (United States)

    Ogawa, E; Kuchel, P W; Agar, N S

    1998-04-01

    It was recently coincidentally discovered, using 1H NMR spectroscopy, that the erythrocytes of two species of Australian marsupials, Tammar Wallaby (Macropus eugenii) and Bettong (Bettongia penicillata), contain relatively high concentrations of the essential amino acid lysine (Agar NS, Rae CD, Chapman BE, Kuchel PW. Comp Biochem Physiol 1991;99B:575-97). Hence, in the present work the rates of transport of lysine into the erythrocytes from the Common Brushtail Possum (Dactylopsilia trivirgata) and Eastern Grey Kangaroo (Macropus giganteus) (which both have low lysine concentrations), and Tammar Wallaby were studied, to explore the mechanistic basis of this finding. The concentration-dependence of the uptake was studied with lysine alone and in the presence of arginine, which may be a competitor of the transport in some species. In relation to GSH metabolism, glutamate uptake was determined in the presence and absence of Na+. The data was analysed to yield estimates of the maximal velocity (Vmax) and the Km in each of the species. Erythrocytes from Tammar Wallaby lacked saturable lysine transport in contrast to the other two species. The glutamate uptake was normal in all three animals for adequate GSH biosynthesis.

  16. Reversing functional extinction of mammals prompts a rethink of paradigms about seed fate in arid Australia.

    Science.gov (United States)

    Mills, Charlotte H; Letnic, Mike

    2018-01-01

    Functional extinction of once abundant species has frequently preceded understanding of their ecological roles. Consequently, our understanding of ecosystems is prone to shifting baselines because it often relies on observations made on depauperate species assemblages. In Australian deserts, current paradigms are that ants are the dominant granivores, mammals are unimportant seed predators and that myrmecochory in many Australian shrubs is an adaptation to increase dispersal distance and direct seeds to favourable germination sites. Here, we ask whether these paradigms could be artefacts of mammal extinction. We take advantage of a predator-proof reserve within which locally extinct native mammals have been reintroduced to compare seed removal by ants and mammals. Using foraging trays that selectively excluded mammals and ants we show that a reintroduced mammal, the woylie ( Bettongia penicillata ) was at least as important as ants in the removal of seeds of two shrub species ( Dodonaea viscosa and Acacia ligulata ). Our results provide evidence that the dominance of ants as granivores and current understanding of the adaptive benefit of myrmecochory in arid Australia may be artefacts of the functional extinction of mammals. Our study shows how reversing functional extinction can provide the opportunity to rethink contemporary understanding of ecological processes.

  17. A retrospective study of Babesia macropus associated with morbidity and mortality in eastern grey kangaroos (Macropus giganteus and agile wallabies (Macropus agilis

    Directory of Open Access Journals (Sweden)

    Shannon L. Donahoe

    2015-08-01

    Full Text Available This is a retrospective study of 38 cases of infection by Babesia macropus, associated with a syndrome of anaemia and debility in hand-reared or free-ranging juvenile eastern grey kangaroos (Macropus giganteus from coastal New South Wales and south-eastern Queensland between 1995 and 2013. Infection with B. macropus is recorded for the first time in agile wallabies (Macropus agilis from far north Queensland. Animals in which B. macropus infection was considered to be the primary cause of morbidity had marked anaemia, lethargy and neurological signs, and often died. In these cases, parasitised erythrocytes were few or undetectable in peripheral blood samples but were sequestered in large numbers within small vessels of visceral organs, particularly in the kidney and brain, associated with distinctive clusters of extraerythrocytic organisms. Initial identification of this piroplasm in peripheral blood smears and in tissue impression smears and histological sections was confirmed using transmission electron microscopy and molecular analysis. Samples of kidney, brain or blood were tested using PCR and DNA sequencing of the 18S ribosomal RNA and heat shock protein 70 gene using primers specific for piroplasms. The piroplasm detected in these samples had 100% sequence identity in the 18S rRNA region with the recently described Babesia macropus in two eastern grey kangaroos from New South Wales and Queensland, and a high degree of similarity to an unnamed Babesia sp. recently detected in three woylies (Bettongia penicillata ogilbyi in Western Australia.

  18. Genetic outcomes from the translocations of the critically endangered woylie

    Directory of Open Access Journals (Sweden)

    Carlo PACIONI, Adrian F.WAYNE, Peter B.S.SPENCER

    2013-06-01

    Full Text Available Translocations are an important conservation strategy for many species. However simply observing demographic growth of a translocated population is not sufficient to infer species recovery. Adequate genetic representation of the source population(s and their long-term viability should also be considered. The woylie Bettongia penicillata ogilbyi has been subject to more formal translocations for conservation than any other marsupial that, up until recently, has resulted in one of the most successful species recoveries in Australia. We used mitochondrial and nuclear DNA markers to assess the genetic outcomes of translocated woylie populations. These populations have lost genetic variability, differentiated from their source population and the supplementation program on two island populations appears to have failed. We discuss the conservation implications that our results have for managing threatened species, outline some general recommendations for the management of present and future translocations and discuss the appropriate sampling design for the establishment of new populations or captive breeding programs that may mitigate the genetic ‘erosion’ seen in our study species. This research provides some practical outcomes and a pragmatic understanding of translocation biology. The findings are directly applicable to other translocation programs [Current Zoology 59 (3: 294-310, 2013].

  19. Antileishmanial potential of medicinal plant extracts from the North-West of Morocco

    Directory of Open Access Journals (Sweden)

    Abdelhakim Bouyahya

    2018-03-01

    Full Text Available The aim of this study is to evaluate the antileishmanial activity of selected medicinal plants from the North-West of Morocco. Plant extracts were prepared by maceration using methanol, ethanol, and n-hexane. The antileishmanial activity was evaluated against Leishmania major, Leishmania tropica, and Leishmania infantum using MTT (3-(4.5-dimethylthiazol-2yl-2.5-diphenyltetrazolium bromide assay. All plant extracts showed a reducing in cell promastigotes viability with variability depending on tested strains and type of extracts. The n-hexane extract showed the highest antileishmanial activity and L. infantum was the most sensitive parasite. The best growth inhibition was observed with Cistus crispus n-hexane extract against L. major (IC50 = 47.29 ± 2.25 μg/mL, Arbutus unedo n-hexane extract against L. infantum (IC50 = 64.05 ± 1.44 μg/mL and Arbutus unedo n-hexane extract against L. tropica (IC50 = 79.57 ± 2.66 μg/mL. Considering these results, medicinal plants from the North-West of Morocco could constitute a promoter source for antileishmanial compounds.

  20. Antileishmanial, toxicity, and phytochemical evaluation of medicinal plants collected from Pakistan.

    Science.gov (United States)

    Shah, Naseer Ali; Khan, Muhammad Rashid; Nadhman, Akhtar

    2014-01-01

    Leishmaniasis is an important parasitic problem and is in focus for development of new drugs all over the world. Objective of the present study was to evaluate phytochemical, toxicity, and antileishmanial potential of Jurinea dolomiaea, Asparagus gracilis, Sida cordata, and Stellaria media collected from different areas of Pakistan. Dry powder of plants was extracted with crude methanol and fractionated with n-hexane, chloroform, ethyl acetate, n-butanol, and water solvents in escalating polarity order. Qualitative phytochemical analysis of different class of compounds, that is, alkaloids, saponins, terpenoids, anthraquinones, cardiac glycosides, coumarins, phlobatannins, flavonoids, phenolics, and tannins, was tested. Its appearance was observed varying with polarity of solvent used for fractionation. Antileishmanial activity was performed against Leishmania tropica KWH23 promastigote. Potent antileishmanial activity was observed for J. dolomiaea methanol extract (IC50 = 10.9 ± 1.1 μ g/mL) in comparison to other plant extracts. However, J. dolomiaea "ethyl acetate fraction" was more active (IC50 = 5.3 ± 0.2 μ g/mL) against Leishmania tropica KWH23 among all plant fractions as well as standard Glucantime drug (6.0 ± 0.1 μ g/mL). All the plants extract and its derived fraction exhibited toxicity in safety range (LC50 > 100) in brine shrimp toxicity evaluation assay.

  1. Epidemiology, pathology and treatment of cutaneous leishmaniasis in taif region of saudi arabia.

    Directory of Open Access Journals (Sweden)

    Wajihullah Khan

    2014-09-01

    Full Text Available Cutaneous leishmaniasis is an annoying and disfiguring disease affecting around 1,500,000 individuals globally. There are endemic pockets of this disease in Taif region. In some patients, lesion often weeps and leads to scar formation. The study was conducted to see the efficacy of fluconazole and itraconazole in the patients of cutaneous leishmaniasis and the effect of these drugs on liver enzymes and kidney markers.Positivity of Leishmania was recorded by microscopic examinations of smears. Specific diagnosis for Leishmania major and L. tropica was made with the help of nested polymerase chain reaction. Fluconazole was given at the rate of 200mg/day while itraconazole was given at 150mg/day for six weeks. AST, ALT, creatinine and urea were estimated during medication.Leishmania major and L. tropica were the species responsible for cutaneous leishmaniasis in Taif region. 81% patients had single lesions, mostly on face followed by hands and legs. 15% of the lesions had bacterial contamination. In terms of efficacy, fluconazole gave slightly better results compared to itraconazole. After 6 weeks of medications, slightly elevated values were recorded for liver enzymes and creatinine.Transmission of leishmaniasis in Taif region is probably because of poor coverage of residual insecticides spraying at hiding places in pile-ups of rocks and abandoned houses from where sand flies visit nearby houses and cattle sheds during night. Fluconazole and itraconazole may be used for the treatment of cutaneous leishmaniasis with good recovery rate and fewer side effects.

  2. A molecular and parasitological survey on cutaneous leishmaniasis patients from historical city of Kashan in Isfahan province, center of Iran

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Shiee

    2012-12-01

    Full Text Available Objective: To study cutaneous leishmaniasis (CL for identifying the dominant Leishmania species on CL patients referred to medical health centers of historical Kashan city and suburbs located in Isfahan province in central part of Iran during 2010 to 2011. Methods: From 137 CL cases, were microscopically positive, the skin lesion serosity materials of 103 cases were cultured in monophasic culture media (RPMI 1 640. We used the PCR-RFLP method for characterization the Leishmania isolates, by using specific internal transcribed spacer (ITS1 primers and HAEШ as the restriction fast enzyme. DNA was extracted from 63 samples. Results: L. tropica is main species in 58 (92.1% cases and L. major is identified in 5 (7.9% cases. Indeed randomly two isolates were the species characterized as L. major produced ulcer at the base tail of BALB/c mice after 3 weeks but from three L. tropica isolates none of them produced any lesion during 6 months post inoculation. Conclusions: The parasitological, epidemiological aspect and molecular methods of this study showed that, Kashan and suburb are anthroponetic CL area despite this city located in Isfahan province as an ancient focus of zoonotic CL in Iran.

  3. Molecular Characterization of Leishmania Species Isolated from Cutaneous Leishmaniasis in Yemen

    Science.gov (United States)

    Mahdy, Mohammed A. K.; Al-Mekhlafi, Hesham M.; Al-Mekhlafi, Abdulsalam M.; Lim, Yvonne A. L.; Bin Shuaib, Naemah O. M.; Azazy, Ahmed A.; Mahmud, Rohela

    2010-01-01

    Background Cutaneous leishmaniasis (CL) is a neglected tropical disease endemic in the tropics and subtropics with a global yearly incidence of 1.5 million. Although CL is the most common form of leishmaniasis, which is responsible for 60% of DALYs lost due to tropical-cluster diseases prevalent in Yemen, available information is very limited. Methodology/Principal Findings This study was conducted to determine the molecular characterization of Leishmania species isolated from human cutaneous lesions in Yemen. Dermal scrapes were collected and examined for Leishmania amastigotes using the Giemsa staining technique. Amplification of the ribosomal internal transcribed spacer 1(ITS-1) gene was carried out using nested PCR and subsequent sequencing. The sequences from Leishmania isolates were subjected to phylogenetic analysis using the neighbor-joining and maximum parsimony methods. The trees identified Leishmania tropica from 16 isolates which were represented by two sequence types. Conclusions/Significance The predominance of the anthroponotic species (i.e. L. tropica) indicates the probability of anthroponotic transmission of cutaneous leishmaniasis in Yemen. These findings will help public health authorities to build an effective control strategy taking into consideration person–to-person transmission as the main dynamic of transmission of CL. PMID:20862227

  4. Molecular identification of Leishmania species in Taybad district, Iran

    Directory of Open Access Journals (Sweden)

    Salehi Ghodratollah

    2014-09-01

    Full Text Available Objective: To identify Leishmania species in patients with cutaneous leishmaniasis in the city of Taybad in Razavi Khorasan Province from April 2012 to March 2013. Methods: Among 52 persons who referred to Health Center of Taybad with suspected skin lesions, stained slide smears of 35 patients showed positive result for Leishmania. Also polymerase chain reaction assay performed using specific kDNA primers. Data of patients were analyzed with SPSS. Results: Of 35 positive smears for Leishmania, 21 (60% belonged to males and 14 (40% belonged to females. Polymerase chain reaction bands were observed in all 35 samples of which 31 (88.6% samples showed Leishmania tropica and 4 (11.4% showed Leishmania major. The highest infected age group was 11-20 years old. Conclusions: Both anthroponotic cutaneous leishmaniasis and zoonotic cutaneous leishmaniasis are present in Taybad. Leishmania tropica is the dominant causative species for anthroponotic cutaneous leishmaniasis. Further study is recommended to discover probable reservoir and vector for Leishmania major in Taybad.

  5. Identification of galactose as the immunodominant sugar of leishmanial excreted factor and subsequent labeling with galactose oxidase and sodium boro[3H]hydride.

    Science.gov (United States)

    Slutzky, G M; Greenblatt, C L

    1982-01-01

    Inhibition by low-molecular-weight sugars of precipitin line formation between a polysaccharide (EF) excreted by Leishmania tropica subsp. major, Leishmania enriettii, and rabbit antileishmanial antibodies on double gel diffusion plates revealed that galactose residues, possibly as components of lactosyl groups, were the critical immunodominant sugars mediating antibody recognition of EF. The galactose residues of the EF of L. tropica subsp. major were specifically labeled with tritium via galactose oxidase and sodium boro[3H]hydride. The radioactive EF had an apparent molecular weight of about 85,000 on sodium dodecyl sulfate-polyacrylamide gels and was precipitated by antileishmanial antibodies as well as Ricinus communis lectins I and II (galactose specific). Lectins specific for glucose-mannose residues, fucose, N-acetylglucosamine, and N-acetylgalactosamine did not precipitate the labeled EF. Treatment of [3H]EF with proteolytic (trypsin, papain, protease) or glycosidic (alpha-amylase, beta-galactosidase) enzymes had no effect on either the electrophoretic pattern of the material or on its recognition by antileishmanial antibodies or R. communis lectin. This resistance to enzyme activity suggests that EF may be a useful marker for the presence of the parasite in vivo if it can be detected in minute quantities. PMID:6179874

  6. Infectious diseases of afghan immigrants in the united states: review of published reports

    International Nuclear Information System (INIS)

    Pilsczek, F.H.

    2011-01-01

    Infectious diseases of immigrants may differ from patients born and resident in the same country, especially if immigrants from Africa or Asia live in Europe or North America. Because the available information is limited published reports of infections of Afghan immigrants in the United States and other countries were analysed. Four reports from the US and 15 reports from other countries were identified [7, (46.7%) Pakistan, 5 (33.3%) Iran, 1 (6.7%) United Kingdom, 1 (6.7%) Germany, 1 (6.7%) Israel)]. Reports from the US were case reports or case series of infections with gastro-intestinal parasites and Mycobacterium tuberculosis (1, 25%), Echinococcus species (2, 50%), and Plasmodium vivax (1, 25%). Reports from other countries were case reports, case series, or surveys and investigated infections with Echinococcus species (2, 13%), Hepatitis B virus (HBV) (1, 6.7%), M. tuberculosis (6, 40%), P. falciparum (1, 6.7%), Leishmania tropica (3, 20%), Fasciola hepatica (1, 6.7%), and M. leprae (1, 6.7%). The reports suggest that Echinococcus species and L. tropica infections can be encountered in Afghan immigrants in the US, and the frequency of a positive PPD (purified protein derivative) response or HBsAg test was increased. An infectious diseases database specific for the country of residence readily available to clinicians treating Afghan patients outside of Afghanistan may be useful. (author)

  7. Antileishmanial, Toxicity, and Phytochemical Evaluation of Medicinal Plants Collected from Pakistan

    Directory of Open Access Journals (Sweden)

    Naseer Ali Shah

    2014-01-01

    Full Text Available Leishmaniasis is an important parasitic problem and is in focus for development of new drugs all over the world. Objective of the present study was to evaluate phytochemical, toxicity, and antileishmanial potential of Jurinea dolomiaea, Asparagus gracilis, Sida cordata, and Stellaria media collected from different areas of Pakistan. Dry powder of plants was extracted with crude methanol and fractionated with n-hexane, chloroform, ethyl acetate, n-butanol, and water solvents in escalating polarity order. Qualitative phytochemical analysis of different class of compounds, that is, alkaloids, saponins, terpenoids, anthraquinones, cardiac glycosides, coumarins, phlobatannins, flavonoids, phenolics, and tannins, was tested. Its appearance was observed varying with polarity of solvent used for fractionation. Antileishmanial activity was performed against Leishmania tropica KWH23 promastigote. Potent antileishmanial activity was observed for J. dolomiaea methanol extract (IC50=10.9±1.1 μg/mL in comparison to other plant extracts. However, J. dolomiaea “ethyl acetate fraction” was more active (IC50=5.3±0.2 μg/mL against Leishmania tropica KWH23 among all plant fractions as well as standard Glucantime drug (6.0±0.1 μg/mL. All the plants extract and its derived fraction exhibited toxicity in safety range (LC50 >100 in brine shrimp toxicity evaluation assay.

  8. Antifungal, Antileishmanial, and Cytotoxicity Activities of Various Extracts of Berberis vulgaris (Berberidaceae) and Its Active Principle Berberine.

    Science.gov (United States)

    Mahmoudvand, Hossein; Ayatollahi Mousavi, Seyyed Amin; Sepahvand, Asghar; Sharififar, Fariba; Ezatpour, Behrouz; Gorohi, Fatemeh; Saedi Dezaki, Ebrahim; Jahanbakhsh, Sareh

    2014-01-01

    In this study, in vitro antidermatophytic activity against Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, and Microsporum gypseum was studied by disk diffusion test and assessment of minimum inhibitory concentration (MIC) using CLSI broth macrodilution method (M38-A2). Moreover, antileishmanial and cytotoxicity activity of B. vulgaris and berberine against promastigotes of Leishmania major and Leishmania tropica were evaluated by colorimetric MTT assay. The findings indicated that the various extracts of B. vulgaris particularly berberine showed high potential antidermatophytic against pathogenic dermatophytes tested with MIC values varying from 0.125 to >4 mg/mL. The results revealed that B. vulgaris extracts as well as berberine were effective in inhibiting L. major and L. tropica promastigotes growth in a dose-dependent manner with IC50 (50% inhibitory concentration) values varying from 2.1 to 26.6  μ g/mL. Moreover, it could be observed that berberine as compared with B. vulgaris exhibited more cytotoxicity against murine macrophages with CC50 (cytotoxicity concentration for 50% of cells) values varying from 27.3 to 362.6  μ g/mL. Results of this investigation were the first step in the search for new antidermatophytic and antileishmanial drugs. However, further works are required to evaluate exact effect of these extracts in animal models as well as volunteer human subjects.

  9. Entomological and epidemiological study of a new focus of cutaneous leishmaniasis in Morocco.

    Science.gov (United States)

    Ramaoui, K; Guernaoui, S; Boumezzough, A

    2008-09-01

    Cutaneous leishmaniasis (CL) due to Leishmania tropica Wright has become an increasingly important problem in Al-Haouz province, Morocco. The objective of this paper was to undertake a retrospective analysis of CL between 2000 and 2006 and to study the sandfly fauna for the first time in this focus. The consultation of bulletins, registers and monthly and annual reports published by local and national medical services permitted a global synthesis of CL in Al-Haouz between 2000 and 2006. Sandflies were collected using sticky-paper traps in five selected stations in two communes, Ourika and Sti Fadma, the most affected by CL in Al-Haouz. Overall, 2,648 sandflies, belonging to five species of Phlebotomus and three of Sergentomyia, were collected. Phlebotomus sergenti Parrot (14.5%) was found in all studied stations where it was associated with domestic and peridomestic habitats. Based on this first report on ecology of sandflies and epidemiology of CL, Al-Haouz seems to be a typical focus of L. tropica and P. sergenti like the other Moroccan foci. Two communes, Sti Fadma and Ourika, constituted an important CL micro-focus in this region. Considering their economical importance (trade and tourism), they contribute probably to the dissemination of Leishmania parasite in bordering regions.

  10. Current knowledge of sand fly fauna (Diptera: Psychodidae) of northwestern Yemen and how it relates to leishmaniasis transmission.

    Science.gov (United States)

    El Sawaf, Bahira M; Kassem, Hala A; Mogalli, Nabil M; El Hossary, Shabaan S; Ramadan, Nadia F

    2016-10-01

    This report presents the results of the first entomological survey of the sand fly fauna in northwestern Yemen. Sand flies were collected using sticky paper traps and CDC light traps from Hajjah governorate, a cutaneous leishmaniasis focus due to Leishmania tropica. Six Phlebotomus species: P. alexandri, P. arabicus. P. bergeroti, P. orientalis, P. papatasi, P. sergenti and ten Sergentomyia species: S. africana, S. antennata, S. christophersi, S. dolichopa, S. dreyfussi, S. fallax, S. multidens, S. taizi, S. tiberiadis, S. yusafi were identified. P. alexandri was the most predominant Phlebotomus species and P. papatasi was a scarce species. S. fallax was the principal Sergentomyia species and S. dolichopa was the least species encountered. The diversity of the sand fly fauna within and among three altitudinal ranges using Simpson index and Jaccard's diversity coefficient respectively were measured. High species diversity was found in all altitude ranges. There seemed to be more association between sand fly fauna in higher altitudes with fauna from moderate altitudes. Sand fly seasonal activity showed a mono-modal trend in the lowland and a confluent bimodal trend in the highlands. Leishmania DNA could not be detected from 150 Phlebotomus females using PCR-RFLP. A possible zoonotic cutaneous transmission cycle due to Leishmania tropica in northwestern Yemen would involve P. arabicus as the sand fly vector and the rock hyrax as the reservoir host. The vector competence for P. alexandri as a vector of visceral leishmaniasis in Hajjah governorate is discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Phlebotomus (Paraphlebotomus) riouxi: a synonym of Phlebotomus chabaudi without any proven vectorial role in Tunisia and Algeria.

    Science.gov (United States)

    Tabbabi, A; Rhim, A; Ghrab, J; Martin, O; Aoun, K; Bouratbine, A; Ready, P D

    2014-08-01

    Phlebotomus (Paraphlebotomus) riouxi Depaquit, Léger & Killick-Kendrick (Diptera: Psychodidae) was described as a typological species based on a few morphological characters distinguishing it from Phlebotomus (Paraphlebotomus) chabaudi Croset, Abonnenc & Rioux. The naming of P. riouxi coincided with its incrimination as a rural vector of Leishmania tropica Wright (junior synonym: Leishmania killicki Rioux, Lanotte & Pratlong) in Tataouine governorate, an arid region of southern Tunisia. The current report finds insufficient evidence to incriminate either phlebotomine sandfly as a vector of L. tropica in North Africa. Phlebotomus riouxi was found not to have the characteristics of a phylogenetic or biological species, and therefore it is synonymized with P. chabaudi. Both taxa were recorded together for the first time in Tunisia, in Tataouine, where three of 12 males showed intermediate morphology and both sexes of each taxon were not characterized by specific lineages of the nuclear gene elongation factor-1α or the mitochondrial gene cytochrome b, for which a long 3' terminal fragment is recommended for phlebotomine phylogenetics. This case study indicates that the eco-epidemiology of leishmaniasis should focus more on identifying key components of vectorial transmission that are susceptible to interventions for disease control, rather than on defining sibling species of vectors. © 2014 The Royal Entomological Society.

  12. Reproductive fitness and dietary choice behavior of the genetic model organism Caenorhabditis elegans under semi-natural conditions.

    Science.gov (United States)

    Freyth, Katharina; Janowitz, Tim; Nunes, Frank; Voss, Melanie; Heinick, Alexander; Bertaux, Joanne; Scheu, Stefan; Paul, Rüdiger J

    2010-10-01

    Laboratory breeding conditions of the model organism C. elegans do not correspond with the conditions in its natural soil habitat. To assess the consequences of the differences in environmental conditions, the effects of air composition, medium and bacterial food on reproductive fitness and/or dietary-choice behavior of C. elegans were investigated. The reproductive fitness of C. elegans was maximal under oxygen deficiency and not influenced by a high fractional share of carbon dioxide. In media approximating natural soil structure, reproductive fitness was much lower than in standard laboratory media. In seminatural media, the reproductive fitness of C. elegans was low with the standard laboratory food bacterium E. coli (γ-Proteobacteria), but significantly higher with C. arvensicola (Bacteroidetes) and B. tropica (β-Proteobacteria) as food. Dietary-choice experiments in semi-natural media revealed a low preference of C. elegans for E. coli but significantly higher preferences for C. arvensicola and B. tropica (among other bacteria). Dietary-choice experiments under quasi-natural conditions, which were feasible by fluorescence in situ hybridization (FISH) of bacteria, showed a high preference of C. elegans for Cytophaga-Flexibacter-Bacteroides, Firmicutes, and β-Proteobacteria, but a low preference for γ-Proteobacteria. The results show that data on C. elegans under standard laboratory conditions have to be carefully interpreted with respect to their biological significance.

  13. New epidemiological pattern of cutaneous leishmaniasis in two pre-Saharan arid provinces, southern Morocco.

    Science.gov (United States)

    Ait Kbaich, Mouad; Mhaidi, Idriss; Ezzahidi, Abdelkacem; Dersi, Nouredine; El Hamouchi, Adil; Riyad, Myriam; Akarid, Khadija; Lemrani, Meryem

    2017-09-01

    Three Leishmania species are responsible of cutaneous leishmaniasis (CL) in Morocco. Zoonotic CL due to Leishmania major and Leishmania infantum, the first is known as established in the eastern arid regions, whereas the latter evolves sporadically, especially in the North. While Leishmania tropica, classically considered anthroponotic, is endemic in the semi-arid regions and is largely distributed throughout the country. The aim of this study was to identify the Leishmania species causing CL in two Provinces in arid pre-Saharan region known as zoonotic CL foci, and to contribute an update to the national data concerning the distribution of Leishmania species in both regions. The recruitment of patients was done in six localities in Ouarzazate and Zagoura provinces in 2015 and 2016. Out of 81 samples collected, 66 were positive (81%) by ITS1-PCR amplification of Leishmania DNA extracted from stained smears. The highest rate of Leishmania infection was registered in children aged 9 years or less (71,2%). The ITS1-PCR- RFLP analysis revealed the predominance of L. major infecting 52 patients (79%), followed by L. tropica in 12 patients (18%) and L. infantum in 2 patients who had no history of travel outside the studied area (3%). The sequencing of the ITS1 of both L. infantum, showed 100% similarities with L. infantum strains isolated from dogs and visceral leishmaniasis patients from the south and north of Morocco. The coexistence of the 3 Leishmania species in the same focus, and the difficult distinction of infections associated to the different Leishmania species based only on clinical lesions' aspects complicate the diagnosis and then the national control strategy, as well as the therapeutic management. The epidemiological pattern of CL in the studied areas appears to have changed during the last decades, from a predominant zoonotic CL caused by L. major to a polymorphic disease that can be due to any of the 3 Leishmania species. The expansion of L. infantum and

  14. Stage-specific adhesion of Leishmania promastigotes to sand fly midguts assessed using an improved comparative binding assay.

    Directory of Open Access Journals (Sweden)

    Raymond Wilson

    2010-09-01

    Full Text Available The binding of Leishmania promastigotes to the midgut epithelium is regarded as an essential part of the life-cycle in the sand fly vector, enabling the parasites to persist beyond the initial blood meal phase and establish the infection. However, the precise nature of the promastigote stage(s that mediate binding is not fully understood.To address this issue we have developed an in vitro gut binding assay in which two promastigote populations are labelled with different fluorescent dyes and compete for binding to dissected sand fly midguts. Binding of procyclic, nectomonad, leptomonad and metacyclic promastigotes of Leishmania infantum and L. mexicana to the midguts of blood-fed, female Lutzomyia longipalpis was investigated. The results show that procyclic and metacyclic promastigotes do not bind to the midgut epithelium in significant numbers, whereas nectomonad and leptomonad promastigotes both bind strongly and in similar numbers. The assay was then used to compare the binding of a range of different parasite species (L. infantum, L. mexicana, L. braziliensis, L. major, L. tropica to guts dissected from various sand flies (Lu. longipalpis, Phlebotomus papatasi, P. sergenti. The results of these comparisons were in many cases in line with expectations, the natural parasite binding most effectively to its natural vector, and no examples were found where a parasite was unable to bind to its natural vector. However, there were interesting exceptions: L. major and L. tropica being able to bind to Lu. longipalpis better than L. infantum; L. braziliensis was able to bind to P. papatasi as well as L. major; and significant binding of L. major to P. sergenti and L. tropica to P. papatasi was observed.The results demonstrate that Leishmania gut binding is strictly stage-dependent, is a property of those forms found in the middle phase of development (nectomonad and leptomonad forms, but is absent in the early blood meal and final stages (procyclic

  15. Analysis of kinetoplast cytochrome b gene of 16 Leishmania isolates from different foci of China: different species of Leishmania in China and their phylogenetic inference

    Science.gov (United States)

    2013-01-01

    Background Leishmania species belong to the family Trypanosomatidae and cause leishmaniasis, a geographically widespread disease that infects humans and other vertebrates. This disease remains endemic in China. Due to the large geographic area and complex ecological environment, the taxonomic position and phylogenetic relationship of Chinese Leishmania isolates remain uncertain. A recent internal transcribed spacer 1 and cytochrome oxidase II phylogeny of Chinese Leishmania isolates has challenged some aspects of their traditional taxonomy as well as cladistics hypotheses of their phylogeny. The current study was designed to provide further disease background and sequence analysis. Methods We systematically analyzed 50 cytochrome b (cyt b) gene sequences of 19 isolates (16 from China, 3 from other countries) sequenced after polymerase chain reaction (PCR) using a special primer for cyt b as well as 31 sequences downloaded from GenBank. After alignment, the data were analyzed using the maximum parsimony, Bayesian and netwok methods. Results Sequences of six haplotypes representing 10 Chinese isolates formed a monophyletic group and clustered with Leishmania tarentolae. The isolates GS1, GS7, XJ771 of this study from China clustered with other isolates of Leishmania donovani complex. The isolate JS1 was a sister to Leishmania tropica, which represented an L. tropica complex instead of clustering with L. donovani complex or with the other 10 Chinese isolates. The isolates KXG-2 and GS-GER20 formed a monophyletic group with Leishmania turanica from central Asia. In the different phylogenetic trees, all of the Chinese isolates occurred in at least four groups regardless of geographic distribution. Conclusions The undescribed Leishmania species of China, which are clearly causative agents of canine leishmaniasis and human visceral leishmaniasis and are related to Sauroleishmania, may have evolved from a common ancestral parasite that came from the Americas and may have

  16. Recent findings regarding non-native or poorly known diatom taxa in north-western Italian rivers

    Directory of Open Access Journals (Sweden)

    Elisa Falasco

    2013-02-01

    Full Text Available Diatoms of the major rivers of North-Western Italy were investigated to highlight the presence of species of particular ecological interest but not as yet recorded. The survey area included streams belonging to seven different hydroecoregions (HERs with a wide range of physical characteristics. Between 2008 and 2010, 200 samples were taken for the study of the diatom community composition, while a larger set of samples was examined to determine the presence or absence of the nuisance diatom species Didymosphenia geminata (Lyngbye Schmidt. A specific field study was performed in two rivers characterized by persistent blooms of this species to evaluate the effects of its proliferation on the benthic communities. D. geminata was present in almost 20% of the samples. From a comparison with published data, we can confirm that D. geminata has recently been expanding its ecological range, as it has been found also in mesotrophic lowlands water. In some instances the formation of massive proliferation has been recorded. The calculation of autecological values confirmed its preference for oligotrophic waters with low mineral content and organic loading, although with a wider ecological amplitude than recorded in the first studies on this species. Another four taxa of particular interest were detected: Achnanthidium subhudsonis (Hustedt Kobayasi (in 15 sites, Cymbella tropica Krammer (11 sites, Mayamaea cahabaensis Morales and Manoylov (2 sites and Reimeria uniseriata Sala, Guerrero and Ferrario (18 sites. The first three species must be considered new records for Northern Italy. A. subhudsonis and C. tropica reached up to 20% relative abundance. From the analysis of their distribution and autecological values, we can assert that A. subhudsonis and M. cahabaensis show a preference for high values of nitrogen, this latter preferring also quite high values of total phosphorus. C. tropica prefers intermediate values of nitrogen nutrients and R. uniseriata is

  17. Temporal Patterns in the Abundance of a Critically Endangered Marsupial Relates to Disturbance by Roads and Agriculture.

    Science.gov (United States)

    Yeatman, Georgina J; Wayne, Adrian F; Mills, Harriet R; Prince, Jane

    2016-01-01

    The aim of this study was to investigate how landscape disturbance associated with roads, agriculture and forestry influenced temporal patterns in woylie (Bettongia penicillata) abundance before, during and after periods of rapid population change. Data were collected from an area of approximately 140,000 ha of forest within the Upper Warren region in south-western Australia. Woylie abundance was measured using cage trapping at 22 grid and five transect locations with varying degrees of landscape disturbance between 1994 and 2012. We found evidence that the distribution and abundance of woylies over time appears to be related to the degree of fragmentation by roads and proximity to agriculture. Sites furthest from agriculture supported a greater abundance of woylies and had slower rates of population decline. Sites with fewer roads had a greater abundance of woylies generally and a greater rate of increase in abundance after the implementation of invasive predator control. The results of this study suggest that landscape disturbance is less important at peak population densities, but during times of environmental and population change, sites less dissected by roads and agriculture better support woylie populations. This may be due to the role these factors play in increasing the vulnerability of woylies to introduced predators, population fragmentation, weed species invasion, mortality from road collisions or a reduction in available habitat. Strategies that reduce the impact of disturbance on woylie populations could include the rationalisation of forest tracks and consolidation of contiguous habitat through the acquisition of private property. Reducing the impact of disturbance in the Upper Warren region could improve the resilience of this critically important woylie population during future environmental change.

  18. Does bipedality predict the group-level manual laterality in mammals?

    Science.gov (United States)

    Giljov, Andrey; Karenina, Karina; Malashichev, Yegor

    2012-01-01

    Factors determining patterns of laterality manifestation in mammals remain unclear. In primates, the upright posture favours the expression of manual laterality across species, but may have little influence within a species. Whether the bipedalism acts the same in non-primate mammals is unknown. Our recent findings in bipedal and quadrupedal marsupials suggested that differences in laterality pattern, as well as emergence of manual specialization in evolution might depend on species-specific body posture. Here, we evaluated the hypothesis that the postural characteristics are the key variable shaping the manual laterality expression across mammalian species. We studied forelimb preferences in a most bipedal marsupial, brush-tailed bettong, Bettongia penicillata in four different types of unimanual behavior. The significant left-forelimb preference at the group level was found in all behaviours studied. In unimanual feeding on non-living food, catching live prey and nest-material collecting, all or most subjects were lateralized, and among lateralized bettongs a significant majority displayed left-forelimb bias. Only in unimanual supporting of the body in the tripedal stance the distribution of lateralized and non-lateralized individuals did not differ from chance. Individual preferences were consistent across all types of behaviour. The direction or the strength of forelimb preferences were not affected by the animals' sex. Our findings support the hypothesis that the expression of manual laterality depends on the species-typical postural habit. The interspecies comparison illustrates that in marsupials the increase of bipedality corresponds with the increase of the degree of group-level forelimb preference in a species. Thus, bipedalism can predict pronounced manual laterality at both intra- and interspecific levels in mammals. We also conclude that quadrupedal position in biped species can slightly hinder the expression of manual laterality, but the evoked biped

  19. A role for selective contraception of individuals in conservation.

    Science.gov (United States)

    Cope, Holly R; Hogg, Carolyn J; White, Peter J; Herbert, Catherine A

    2017-10-28

    Contraception has an established role in managing overabundant populations and preventing undesirable breeding in zoos. We propose that it can also be used strategically and selectively in conservation to increase the genetic and behavioral quality of the animals. In captive breeding programs, it is becoming increasingly important to maximize the retention of genetic diversity by managing the reproductive contribution of each individual and preventing genetically suboptimal breeding through the use of selective contraception. Reproductive suppression of selected individuals in conservation programs has further benefits of allowing animals to be housed as a group in extensive enclosures without interfering with breeding recommendations, which reduces adaptation to captivity and facilitates the expression of wild behaviors and social structures. Before selective contraception can be incorporated into a breeding program, the most suitable method of fertility control must be selected, and this can be influenced by factors such as species life history, age, ease of treatment, potential for reversibility, and desired management outcome for the individual or population. Contraception should then be implemented in the population following a step-by-step process. In this way, it can provide crucial, flexible control over breeding to promote the physical and genetic health and sustainability of a conservation dependent species held in captivity. For Tasmanian devils (Sarcophilus harrisii), black-flanked rock wallabies (Petrogale lateralis), and burrowing bettongs (Bettongia lesueur), contraception can benefit their conservation by maximizing genetic diversity and behavioral integrity in the captive breeding program, or, in the case of the wallabies and bettongs, by reducing populations to a sustainable size when they become locally overabundant. In these examples, contraceptive duration relative to reproductive life, reversibility, and predictability of the contraceptive

  20. Prevalence and Clinical Significance of Herpesvirus Infection in Populations of Australian Marsupials.

    Directory of Open Access Journals (Sweden)

    Kathryn Stalder

    Full Text Available Herpesviruses have been reported in several marsupial species, but molecular classification has been limited to four herpesviruses in macropodids, a gammaherpesvirus in two antechinus species (Antechinus flavipes and Antechinus agilis, a gammaherpesvirus in a potoroid, the eastern bettong (Bettongia gaimardi and two gammaherpesviruses in koalas (Phascolarctos cinereus. In this study we examined a range of Australian marsupials for the presence of herpesviruses using molecular and serological techniques, and also assessed risk factors associated with herpesvirus infection. Our study population included 99 koalas (Phascolarctos cinereus, 96 eastern grey kangaroos (Macropus giganteus, 50 Tasmanian devils (Sarcophilus harrisii and 33 common wombats (Vombatus ursinius. In total, six novel herpesviruses (one alphaherpesvirus and five gammaherpesviruses were identified in various host species. The overall prevalence of detection of herpesvirus DNA in our study population was 27.2% (95% confidence interval (CI of 22.6-32.2%, but this varied between species and reached as high as 45.4% (95% CI 28.1-63.7% in common wombats. Serum antibodies to two closely related macropodid herpesviruses (macropodid herpesvirus 1 and 2 were detected in 44.3% (95% CI 33.1-55.9% of animals tested. This also varied between species and was as high as 92% (95% CI 74.0-99.0% in eastern grey kangaroos. A number of epidemiological variables were identified as positive predictors for the presence of herpesvirus DNA in the marsupial samples evaluated. The most striking association was observed in koalas, where the presence of Chlamydia pecorum DNA was strongly associated with the presence of herpesvirus DNA (Odds Ratio = 60, 95% CI 12.1-297.8. Our results demonstrate the common presence of herpesviruses in Australian marsupials and provide directions for future research.

  1. Prevalence and Clinical Significance of Herpesvirus Infection in Populations of Australian Marsupials.

    Science.gov (United States)

    Stalder, Kathryn; Vaz, Paola K; Gilkerson, James R; Baker, Rupert; Whiteley, Pam; Ficorilli, Nino; Tatarczuch, Liliana; Portas, Timothy; Skogvold, Kim; Anderson, Garry A; Devlin, Joanne M

    2015-01-01

    Herpesviruses have been reported in several marsupial species, but molecular classification has been limited to four herpesviruses in macropodids, a gammaherpesvirus in two antechinus species (Antechinus flavipes and Antechinus agilis), a gammaherpesvirus in a potoroid, the eastern bettong (Bettongia gaimardi) and two gammaherpesviruses in koalas (Phascolarctos cinereus). In this study we examined a range of Australian marsupials for the presence of herpesviruses using molecular and serological techniques, and also assessed risk factors associated with herpesvirus infection. Our study population included 99 koalas (Phascolarctos cinereus), 96 eastern grey kangaroos (Macropus giganteus), 50 Tasmanian devils (Sarcophilus harrisii) and 33 common wombats (Vombatus ursinius). In total, six novel herpesviruses (one alphaherpesvirus and five gammaherpesviruses) were identified in various host species. The overall prevalence of detection of herpesvirus DNA in our study population was 27.2% (95% confidence interval (CI) of 22.6-32.2%), but this varied between species and reached as high as 45.4% (95% CI 28.1-63.7%) in common wombats. Serum antibodies to two closely related macropodid herpesviruses (macropodid herpesvirus 1 and 2) were detected in 44.3% (95% CI 33.1-55.9%) of animals tested. This also varied between species and was as high as 92% (95% CI 74.0-99.0%) in eastern grey kangaroos. A number of epidemiological variables were identified as positive predictors for the presence of herpesvirus DNA in the marsupial samples evaluated. The most striking association was observed in koalas, where the presence of Chlamydia pecorum DNA was strongly associated with the presence of herpesvirus DNA (Odds Ratio = 60, 95% CI 12.1-297.8). Our results demonstrate the common presence of herpesviruses in Australian marsupials and provide directions for future research.

  2. Prevalence and Clinical Significance of Herpesvirus Infection in Populations of Australian Marsupials

    Science.gov (United States)

    Stalder, Kathryn; Vaz, Paola K.; Gilkerson, James R.; Baker, Rupert; Whiteley, Pam; Ficorilli, Nino; Tatarczuch, Liliana; Portas, Timothy; Skogvold, Kim; Anderson, Garry A.; Devlin, Joanne M.

    2015-01-01

    Herpesviruses have been reported in several marsupial species, but molecular classification has been limited to four herpesviruses in macropodids, a gammaherpesvirus in two antechinus species (Antechinus flavipes and Antechinus agilis), a gammaherpesvirus in a potoroid, the eastern bettong (Bettongia gaimardi) and two gammaherpesviruses in koalas (Phascolarctos cinereus). In this study we examined a range of Australian marsupials for the presence of herpesviruses using molecular and serological techniques, and also assessed risk factors associated with herpesvirus infection. Our study population included 99 koalas (Phascolarctos cinereus), 96 eastern grey kangaroos (Macropus giganteus), 50 Tasmanian devils (Sarcophilus harrisii) and 33 common wombats (Vombatus ursinius). In total, six novel herpesviruses (one alphaherpesvirus and five gammaherpesviruses) were identified in various host species. The overall prevalence of detection of herpesvirus DNA in our study population was 27.2% (95% confidence interval (CI) of 22.6–32.2%), but this varied between species and reached as high as 45.4% (95% CI 28.1–63.7%) in common wombats. Serum antibodies to two closely related macropodid herpesviruses (macropodid herpesvirus 1 and 2) were detected in 44.3% (95% CI 33.1–55.9%) of animals tested. This also varied between species and was as high as 92% (95% CI 74.0–99.0%) in eastern grey kangaroos. A number of epidemiological variables were identified as positive predictors for the presence of herpesvirus DNA in the marsupial samples evaluated. The most striking association was observed in koalas, where the presence of Chlamydia pecorum DNA was strongly associated with the presence of herpesvirus DNA (Odds Ratio = 60, 95% CI 12.1–297.8). Our results demonstrate the common presence of herpesviruses in Australian marsupials and provide directions for future research. PMID:26222660

  3. Development and nitrate reductase activity of sugarcane inoculated with five diazotrophic strains.

    Science.gov (United States)

    Dos Santos, Silvana Gomes; da Silva Ribeiro, Flaviane; da Fonseca, Camila Sousa; Pereira, Willian; Santos, Leandro Azevedo; Reis, Veronica Massena

    2017-08-01

    Diazotrophs are able to stimulate plant growth. This study aimed at evaluating the effect of inoculation of five diazotrophic strains on growth promotion and nitrate reductase (NR, EC 1.7.1.1) activity in sugarcane. An experiment was carried out from three stages of cultivation: sprouting, tubes, and in hydroponics. On the first two stages, seven treatments were adopted: uninoculated control; mixed inoculation with five strains; and individual inoculation with Gluconacetobacter diazotrophicus (Gd), Herbaspirillum rubrisubalbicans (Hr), Herbaspirillum seropedicae (Hs), Nitrospirillum amazonense (Na), and Paraburkholderia tropica (Pt). The four treatments showing the best performance were transferred to the hydroponic system for analysis of NR activity. Hs, Pt, and the mixture of all strains led to the highest seedling biomass in tubes, followed by Hr. In hydroponics, the mixture and the strain Hr had the highest growth-promoting effect. NR activity was influenced by inoculation only under low N supply conditions, with positive effect of Hr, Pt, and the mixture.

  4. Adipose Tissue-Derived Mesenchymal Stem Cells as a New Host Cell in Latent Leishmaniasis

    Science.gov (United States)

    Allahverdiyev, Adil M.; Bagirova, Melahat; Elcicek, Serhat; Koc, Rabia Cakir; Baydar, Serap Yesilkir; Findikli, Necati; Oztel, Olga N.

    2011-01-01

    Some protozoan infections such as Toxoplasma, Cryptosporidium, and Plasmodium can be transmitted through stem cell transplantations. To our knowledge, so far, there is no study about transmission of Leishmania parasites in stem cell transplantation and interactions between parasites and stem cells in vitro. Therefore, the aim of this study was to investigate the interaction between different species of Leishmania parasites and adipose tissue-derived mesenchymal stem cells (ADMSCs). ADMSCs have been isolated, cultured, characterized, and infected with different species of Leishmania parasites (L. donovani, L. major, L. tropica, and L. infantum). Infectivity was examined by Giemsa staining, microculture, and polymerase chain reaction methods. As a result, infectivity of ADMSCs by Leishmania parasites has been determined for the first time in this study. According to our findings, it is very important that donors are screened for Leishmania parasites before stem cell transplantations in regions where leishmaniasis is endemic. PMID:21896818

  5. Unusual Presentation of Cutaneous Leishmaniasis: Ocular Leishmaniasis

    Directory of Open Access Journals (Sweden)

    Masoud Doroodgar

    2017-01-01

    Full Text Available The leishmaniases are parasitic diseases that are transmitted to humans by infected female sandflies. Cutaneous leishmaniasis (CL is one of 3 main forms of the disease. CL is the most common form of the disease and is endemic in many urban and rural parts of Iran and usually caused by two species of Leishmania: L. major and L. tropica. We report a case of unusual leishmaniasis with 25 lesions on exposed parts of the body and right eyelid involvement (ocular leishmaniasis. The patient was a 75-year-old male farmer referred to health care center in Aran va Bidgol city. The disease was diagnosed by direct smear, culture, and PCR from the lesions. PCR was positive for Leishmania major.

  6. Detection of Cutaneous Leishmaniasis by PCR in Fasa district in 2012

    Directory of Open Access Journals (Sweden)

    Mehdi Sharafi

    2013-09-01

    Material & Methods: Slit biopsies were collected from 138 suspected cases of cutaneous leishmaniasis who were presented, consecutively, in Fasa district from April 2011 to February 2012. After the microscopy, the entire smear was then scraped off the slide surface for DNA extraction and PCR assay. Results: All 138 investigated smears were reported positive by microscopy and nested PCR assay. In PCR, One of the smears had the 750-bp band, indicative of L. tropica and the rest had the 560-bp band indicative of L. major. Conclusion: In conclusion, the cutaneous leishmaniasis found in Fasa district is predominantly Rural Cutaneous Leishmaniasis. The PCR-based assay used in the present study appears to be a suitable and powerful tool for the characterization of Leishmanial species.

  7. Antileishmanial activities of dihydrochalcones from piper elongatum and synthetic related compounds. Structural requirements for activity.

    Science.gov (United States)

    Hermoso, Alicia; Jiménez, Ignacio A; Mamani, Zulma A; Bazzocchi, Isabel L; Piñero, José E; Ravelo, Angel G; Valladares, Basilio

    2003-09-01

    Two dihydrochalcones (1 and 2) were isolated from Piper elongatum Vahl by activity-guided fractionation against extracellular promastigotes of Leishmania braziliensis in vitro. Their structures were elucidated by spectral analysis, including homonuclear and heteronuclear correlation NMR experiments. Derivatives 3-7 and 20 synthetic related compounds (8-27) were also assayed to establish the structural requirements for antileishmanial activity. Compounds 1-11 that proved to be more active that ketoconazol, used as positive control, were further assayed against promastigotes of Leishmania tropica and Leishmania infantum. Compounds 7 and 11, with a C(6)-C(3)-C(6) system, proved to be the most promising compounds, with IC(50) values of 2.98 and 3.65 microg/mL, respectively, and exhibited no toxic effect on macrophages (around 90% viability). Correlation between the molecular structures and antileishmanial activity is discussed in detail.

  8. Dicty_cDB: Contig-U01510-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available .4 1 ( BE248608 ) NF021H01DT1F1013 Drought Medicago truncatula cDNA... 42 6.4 1 ( BE205651 ) AOB130 Onion seedling leaf cDNA library...-41B2_Sp6.1 CH216 Xenopus (Silurana) tropica... 42 6.4 1 ( CG770475 ) TcB41.2_F05_SP6 Tribolium BAC library ...ed ... 44 1.6 1 ( CK424003 ) AUF_IpSto_10_c04 Stomach cDNA library Ictalurus p........ 42 6.4 1 ( EI465423 ) PV_GBa0071A02.f PV_GBa Phaseolus vulgaris genomic... 42 6.4 1 ( ED568449 ) SBA034_G14.f Sugar beet BAC libra...ago trunca... 42 6.4 1 ( BF650883 ) NF097E12EC1F1097 Elicited cell culture Medicago t... 42 6.4 1 (

  9. Elephantiasis nostras verrucosa or "mossy foot lesions" in lymphedema praecox: report of a case.

    Science.gov (United States)

    Duckworth, Amy L; Husain, Jugnoo; Deheer, Patrick

    2008-01-01

    Elephantiasis nostras verrucosa is a rare disorder that results from chronic obstructive lymphedema. It is characterized clinically by deforming, nonpitting edema; malodorous hyperkeratosis with generalized lichenification; cobblestoned papules; and verrucous changes, that often result in extreme enlargement of the involved body part. Although elephantiasis nostras verrucosa is striking in clinical appearance, biopsy reveals only moderately abnormal findings: pseudoepitheliomatous hyperplasia with dilated lymphatic spaces in the dermis, accompanied by chronic inflammation and fibroblast proliferation. The term elephantiasis nostras (nostras means "from our region") has traditionally been used to differentiate temperate zone disease from the classic disease process, elephantiasis tropica, which is defined by chronic filarial lymphatic obstruction caused by Wuchereria bancrofti, Wuchereria malayi, or Wuchereria pacifica. We present a case report of elephantiasis nostras verrucosa arising as a result of lymphedema praecox.

  10. Detection and identification of cutaneous leishmaniasis isolates by culture, Polymerase chain reaction and sequence analyses in Syrian and Central Anatolia patients.

    Science.gov (United States)

    Beyhan, Yunus E; Karakus, Mehmet; Karagoz, Alper; Mungan, Mesut; Ozkan, Aysegul T; Hokelek, Murat

    2017-09-01

    To characterize the cutaneous leishmaniasis (CL) isolates of Syrian and Central Anatolia patients at species levels. Methods: Skin scrapings of 3 patients (2 Syrian, 1 Turkish) were taken and examined by direct examination, culture in Novy-MacNeal-Nicole (NNN) medium, internal transcribed spacer polymerase chain reaction and sequence analysis (PCR). Results:According to microscopic examination, culture and PCR methods, 3 samples were detected positive. The sequencing results of all isolates in the study were identified as Leishmania tropica. The same genotypes were detected in the 3 isolates and nucleotide sequence submitted into GenBank with the accession number: KP689599. Conclusion: This finding could give information about the transmission of CL between Turkey and Syria. Because of the Syrian civil war, most of the Syrian citizens circulating in Turkey and different part of Europe, this can be increase the risk of spreading the disease. So, prevention measurements must be taken urgently.

  11. Detection and identification of cutaneous leishmaniasis isolates by culture, Polymerase chain reaction and sequence analyses in Syrian and Central Anatolia patients

    Directory of Open Access Journals (Sweden)

    Yunus E. Beyhan

    2017-09-01

    Full Text Available Objectives: To characterize the cutaneous leishmaniasis (CL isolates of Syrian and Central Anatolia patients at species levels. Methods: Skin scrapings of 3 patients (2 Syrian, 1 Turkish were taken and examined by direct examination, culture in Novy-MacNeal-Nicole (NNN medium, internal transcribed spacer polymerase chain reaction and sequence analysis (PCR. Results:According to microscopic examination, culture and PCR methods, 3 samples were detected positive. The sequencing results of all isolates in the study were identified as Leishmania tropica. The same genotypes were detected in the 3 isolates and nucleotide sequence submitted into GenBank with the accession number: KP689599. Conclusion: This finding could give information about the transmission of CL between Turkey and Syria. Because of the Syrian civil war, most of the Syrian citizens circulating in Turkey and different part of Europe, this can be increase the risk of spreading the disease. So, prevention measurements must be taken urgently.

  12. Molecular detection and identification of Leishmania infection in naturally infected sand flies in a focus of cutaneous leishmaniasis in northern Morocco.

    Science.gov (United States)

    Es-Sette, Nargys; Ajaoud, Malika; Laamrani-Idrissi, Abderrahman; Mellouki, Fouad; Lemrani, Meryem

    2014-07-02

    Cutaneous leishmaniasis is an infectious disease caused by various species of the flagellate protozoan Leishmania. During the past 20 years, cutaneous leishmaniasis has emerged as a major public health threat in Morocco. The main objective of this study was to study the occurrence of Leishmania infection in vectors and to identify sand fly blood meal sources in an endemic locality of cutaneous leishmaniasis within Sefrou province, where the vectors of leishmaniasis were still unknown. 2650 sand flies were collected using CDC miniature light traps and identified morphologically. The identified sand flies were tested for Leishmania infection by nested PCR. The source of blood meal of 10 freshly engorged females: 6 Phlebotomus longicuspis and 4 Phlebotomus sergenti, was determined using the Cyt b sequence. The collected sand flies consisted of 10 species, seven of which belonged to the genus Phlebotomus and three to the genus Sergentomyia. The most abundant species was P. longicuspis, accounting for 72% of the total sand flies collected. In females of three P. longicuspis and four P. sergenti, Leishmania infantum and Leishmania tropica DNA was detected, respectively.The source of blood meal of engorged females showed that all sand flies tested fed on humans. We report for the first time the natural infection of P. longicuspis with L. infantum in Morocco. The high frequency of this species in this region, in addition to its anthropophilic character make P. longicuspis the putative vector of L. infantum in this cutaneous leishmaniasis focus where L. tropica is confirmed as the causative agent of the disease and P. sergenti as its vector. The presence of L. infantum, and its presumed vector in this area, makes this a site of high risk of visceral leishmaniasis, mostly because of the proximity of a focus of human and canine visceral leishmaniasis.

  13. In Vitro and In Vivo Antileishmanial Activities of Pistacia vera Essential Oil.

    Science.gov (United States)

    Mahmoudvand, Hossein; Saedi Dezaki, Ebrahim; Ezatpour, Behrouz; Sharifi, Iraj; Kheirandish, Farnaz; Rashidipour, Marzieh

    2016-03-01

    This study aims to evaluate the in vitro and in vivo antileishmanial activities of Pistacia vera essential oil and compare their efficacy with a reference drug, meglumine antimoniate (Glucantime®). This essential oil (0-100 µg/mL) was evaluated in vitro against the intracellular amastigote forms of Leishmania tropica (MHOM/IR/2002/Mash2) and then tested on cutaneous leishmaniasis of male BALB/c mice by Leishmania major (MRHO/IR/75/ER). In the in vitro assay, it could be observed that P. vera essential oil significantly (p essential oil had potent suppression effects on cutaneous leishmaniasis in BALB/c mice (87.5% recovery), while 10 and 20 mg/mL of the essential oil represented the suppression effects as weak to intermediate. The mean diameter of the lesions decreased about 0.11 and 0.27 cm after the treatment of the subgroups with the essential oil concentrations of 10 and 20 mg/mL, respectively. In contrast, in the subgroup treated with the essential oil concentration of 30 mg/mL, the mean diameter of the lesions decreased about 0.56 cm. In the control subgroups, the mean diameter of the lesions increased to 1.01 cm. The main components of P. vera essential oil were limonene (26.21%), α-pinene (18.07%), and α-thujene (9.31%). It was also found that P. vera essential oil had no significant cytotoxic effect on J774 cells. The present study found that P. vera essential oil showed considerable in vitro and in vivo effectiveness against L. tropica and L. major compared to the reference drug. These findings also provided the scientific evidence that natural plants could be used in traditional medicine for the prevention and treatment of cutaneous leishmaniasis. Georg Thieme Verlag KG Stuttgart · New York.

  14. A voltage-gated pore for translocation of tRNA

    Energy Technology Data Exchange (ETDEWEB)

    Koley, Sandip; Adhya, Samit, E-mail: nilugrandson@gmail.com

    2013-09-13

    Highlights: •A tRNA translocating complex was assembled from purified proteins. •The complex translocates tRNA at a membrane potential of ∼60 mV. •Translocation requires Cys and His residues in the Fe–S center of RIC6 subunit. -- Abstract: Very little is known about how nucleic acids are translocated across membranes. The multi-subunit RNA Import Complex (RIC) from mitochondria of the kinetoplastid protozoon Leishmania tropica induces translocation of tRNAs across artificial or natural membranes, but the nature of the translocation pore remains unknown. We show that subunits RIC6 and RIC9 assemble on the membrane in presence of subunit RIC4A to form complex R3. Atomic Force Microscopy of R3 revealed particles with an asymmetric surface groove of ∼20 nm rim diameter and ∼1 nm depth. R3 induced translocation of tRNA into liposomes when the pH of the medium was lowered to ∼6 in the absence of ATP. R3-mediated tRNA translocation could also be induced at neutral pH by a K{sup +} diffusion potential with an optimum of 60–70 mV. Point mutations in the Cys{sub 2}–His{sub 2} Fe-binding motif of RIC6, which is homologous to the respiratory Complex III Fe–S protein, abrogated import induced by low pH but not by K{sup +} diffusion potential. These results indicate that the R3 complex forms a pore that is gated by a proton-generated membrane potential and that the Fe–S binding region of RIC6 has a role in proton translocation. The tRNA import complex of L. tropica thus contains a novel macromolecular channel distinct from the mitochondrial protein import pore that is apparently involved in tRNA import in some species.

  15. Circulating species of Leishmania at microclimate area of Boulemane Province, Morocco: impact of environmental and human factors.

    Science.gov (United States)

    Hmamouch, Asmae; El Alem, Mahmoud Mohamed; Hakkour, Maryam; Amarir, Fatima; Daghbach, Hassan; Habbari, Khalid; Fellah, Hajiba; Bekhti, Khadija; Sebti, Faiza

    2017-02-22

    Cutaneous leishmaniasis (CL) is widely distributed in Morocco where its geographical range and incidence are related to environmental factors. This study aimed to examine the impact of several factors on the distribution of CL in Boulemane Province, which is characterized by several microclimates, and to identify the Leishmania species circulating in these areas. Ordinary least squares regression (OLSR) analysis was performed to study the impact of poverty, vulnerability, population density, urbanization and bioclimatic factors on the distribution of CL in this province. Molecular characterization of parasites was performed using a previously described PCR-RFLP method targeting the ITS1 of ribosomal DNA of Leishmania. A total of 1009 cases were declared in Boulemane Province between the years 2000 and 2015 with incidences fluctuating over the years (P = 0.007). Analyzing geographical maps of the study region identified four unique microclimate areas; sub-humid, semi-arid, arid and Saharan. The geographical distribution and molecular identification of species shows that the Saharan microclimate, characterized by the presence of Leishmania major was the most affected (47.78%) followed by semi-arid area where Leishmania tropica was identified in three districts. Among several environmental factors included in the study, poverty had the greatest influence on the spatial extension of the disease in this province. The incidence of CL in Boulemane Province varies between microclimate areas, and environmental factors partly explain this variation. However, the existence of CL in the most affected districts is mainly related to poverty, population movement and human behavior. To our knowledge, this the first study utilizing molecular techniques to confirm L. tropica and L. major as the causative agents of CL in Boulemane Province. Our findings indicate that the spatial and temporal distribution of CL in Boulemane Province is strongly related to poverty and population

  16. Diagnosis of Cutaneous Leishmaniasis by Multiplex PCR

    Directory of Open Access Journals (Sweden)

    M Heiat

    2010-07-01

    Full Text Available Introduction: Annually, more than 14 million people are reported to be infected with Leishmaniasis all over the world. In Iran, this disease is seen in the form of cutaneous and visceral leishmaniasis, of which the cutaneous form is more wide spread. In recent years, cutaneous leishmaniaisis is diagnosed by PCR utilizing specific primers in order to amplify different parasite genes including ribosomal RNA genes, kinetoplast DNA or tandem repeating sequences. The aim of this research was to detect early stage cutaneous leishmaniasis using Multiplex-PCR technique. Methods: In this study, 67 samples were prepared from patients with cutaneous leishmaniasis. DNA was extracted with phenolchloroform. Each specimen was analyzed using two different pairs of PCR primers. The sensitivity of each PCR was optimized on pure Leishmania DNA prior to use for diagnosis. Two standard parasites L. major and L. tropica were used as positive control. Results: DNA amplification fragments were two 115 bp and 683 bp for AB and UL primers, respectively. The sensitivity of two primers was not equal for detection of L. major and L. tropica. The sensivity of PCR with AB primer was 35 cells, while that for UL primer was 40 cells. Conclusion: The results of this study indicate that PCR is a sensitive diagnostic assay for cutaneous leishmaniasis and could be employed as the new standard for routine diagnosis when species identification is not required. However, the ability to identify species is especially important in prognosis of the disease and in deciding appropriate therapy, especially in regions where more than one type of species and disease are seen by clinicians.

  17. Evaluation of the ecological characteristics in the vector of anthroponotic cutaneous leishmaniasis in a new focus of Mohammad Abad, Kerman, southeast of Iran

    Directory of Open Access Journals (Sweden)

    Abbas Aghaei Afshar

    2017-02-01

    Full Text Available Objective: To evaluate the fauna, seasonal activity and the host preference of sand flies and to determine the main vectors of anthroponotic cutaneous leishmaniasis in Mohammad Abad, Kerman Province, southeast of Iran. Methods: Sand flies were collected indoors and outdoors by sticky paper traps from May to November, 2012. Regarding the importance of host preference and its impact on leishmaniasis control, blood meal was analyzed by the restriction fragment length polymorphism method. The cytochrome b mitochondrial genomic regions (mitochondrial DNA and enzymatic digestion of Xho I and Hae III were used for the diagnosis of human blood feeding. In the detection of leptomonad (promastigote infection in sand flies, a nested PCR method and high resolution melt analysis were exploited. Results: A total of 919 sand flies were identified as belonging to 14 species in two genera, namely, Phlebotomus spp. (5 species and Sergentomyia spp. (9 species. The most frequently occurring species was Phlebotomus sergenti (P. sergenti (67.46%, followed by Phlebotomus papatasi (19.37%. The highest indoor collection of P. sergenti was realized in late July, and the highest outdoor collection of P. sergenti ensued in early July. A total of 250 sand flies were collected for host preference analysis, and blood meals of 120 sand flies belonged to the species of P. sergenti were subjected to restriction fragment length polymorphism confirmation. A total of 39 P. sergenti sand flies (32.5% were identified to have fed on human. NestedPCR and high resolution melt analyses confirmed that these sand flies had been infected with Leishmania tropica. Conclusions: The present study has confirmed P. sergenti as the main Phlebotomine sand fly vector for anthroponotic cutaneous leishmaniasis caused by Leishmania tropica in Southeastern Iran.

  18. Concept Mapping zur Unterstützung der differentialdiagnostischen Hypothesenbildung im fallbasierten Online-Lernsystem CASUS: Qualitative Verbesserung der Diagnosefindung durch ICD-10 Kodierung [Concept mapping for supporting the differential diagnostic generation of hypotheses in the case-based online learning system CASUS: Qualitative improvement of dagnostic performance through ICD-10 coding

    Directory of Open Access Journals (Sweden)

    Kernt, Marcus

    2008-08-01

    Full Text Available [english] Introduction: Concept mapping tools have long been established in medical education as an aid for visualizing learning processes in computer-based programs. The case-based learning system CASUS with its mapping tool for visualizing the differential diagnostic reasoning process is an example. It was shown that such tools are well accepted by users and lead to an increased number of diagnostic hypotheses being visualized as maps. However, there is scarce evidence on the quality of user-generated diagnostic hypotheses. This study examines the quality of diagnostic hypotheses obtained with CASUS and whether the quality can be improved through ICD-10 coding as compared with an expert’s solution. Methods: We randomized 192 third-year medical students at the University of Munich into two groups. The students worked in groups of two on one computer. Group A was asked to code their diagnostic hypotheses with an ICD-10 coding browser before entering them into the mapping tool. Group B generated their hypotheses without prior ICD-10 coding. The differential diagnostic reasoning visualizations were analyzed quantitatively and qualitatively. An expert solution was used as reference. Results: Eighty-seven differential diagnoses were evaluated. Group A, using ICD-10 coding, made the correct and precise diagnosis of malaria tropica significantly more often than Group B (p < 0.05. For additional alternative diagnostic hypotheses, no quantitative or qualitative differences were detected. Conclusions: ICD-10 coding in connection with a mapping tool supporting the diagnostic reasoning process improved the accuracy of diagnostic performance in third-year medical students in the case of malaria tropica. [german] Einleitung: Der Einsatz von Concept-Mapping-Tools in computergestützten Lernprogrammen ist in der medizinischen Ausbildung etabliert: Es konnte gezeigt werden, dass diese Werkzeuge zur Visualisierung von Differentialdiagnosen vom Anwender

  19. Rapid healing of cutaneous leishmaniasis by high-frequency electrocauterization and hydrogel wound care with or without DAC N-055: a randomized controlled phase IIa trial in Kabul.

    Directory of Open Access Journals (Sweden)

    Ahmad Fawad Jebran

    2014-02-01

    Full Text Available Anthroponotic cutaneous leishmaniasis (CL due to Leishmania (L. tropica infection is a chronic, frequently disfiguring skin disease with limited therapeutic options. In endemic countries healing of ulcerative lesions is often delayed by bacterial and/or fungal infections. Here, we studied a novel therapeutic concept to prevent superinfections, accelerate wound closure, and improve the cosmetic outcome of ACL.From 2004 to 2008 we performed a two-armed, randomized, double-blinded, phase IIa trial in Kabul, Afghanistan, with patients suffering from L. tropica CL. The skin lesions were treated with bipolar high-frequency electrocauterization (EC followed by daily moist-wound-treatment (MWT with polyacrylate hydrogel with (group I or without (group II pharmaceutical sodium chlorite (DAC N-055. Patients below age 5, with facial lesions, pregnancy, or serious comorbidities were excluded. The primary, photodocumented outcome was the time needed for complete lesion epithelialization. Biopsies for parasitological and (immunohistopathological analyses were taken prior to EC (1(st, after wound closure (2(nd and after 6 months (3(rd. The mean duration for complete wound closure was short and indifferent in group I (59 patients, 43.1 d and II (54 patients, 42 d; p = 0.83. In patients with Leishmania-positive 2(nd biopsies DAC N-055 caused a more rapid wound epithelialization (37.2 d vs. 58.3 d; p = 0.08. Superinfections occurred in both groups at the same rate (8.8%. Except for one patient, reulcerations (10.2% in group I, 18.5% in group II; p = 0.158 were confined to cases with persistent high parasite loads after healing. In vitro, DAC N-055 showed a leishmanicidal effect on pro- and amastigotes.Compared to previous results with intralesional antimony injections, the EC plus MWT protocol led to more rapid wound closure. The tentatively lower rate of relapses and the acceleration of wound closure in a subgroup of patients with parasite persistence warrant

  20. LOS HIDROZOOS (CNIDARIA, HYDROZOA DE LA CAYERÍA SUR DEL GOLFO DE BATABANÓ, CUBA

    Directory of Open Access Journals (Sweden)

    Susel Castellanos Iglesias

    2011-12-01

    Full Text Available Hidrozoos de aguas poco profundas fueron recolectados en 17 estaciones, en julio del 2004. Las estaciones se localizaron en manglares, pastos marinos y arrecifes coralinos. Se encontraron hidrozoos de dos subclases (Anthoathecata y Leptothecata, 11 familias, 27 géneros y 34 especies. Se indican la localidad de recolecta, la profundidad, el tipo de sustrato, la descripción y las figuras de 15 especies de hidrozoos  tecados. Se confirma la presencia de Hebellopsis communis Calder, 1991a y Antennella siliquosa (Hincks, 1877 para el Mar Caribe. Se expresa la abundancia relativa en tres categorías: abundante, común y rara. Las especies más abundantes fueron Dynamena crisioides Lamouroux, 1824 en el hábitat de manglar, Cnydoscyphus marginatus (Allman, 1877 y Sertularella diaphana (Allman, 1885 en los arrecifes. Monostaechas quadridens (McCrady, 1859 y Diphasia tropica Nutting, 1904 fueron de baja representación en las muestras. Los sustratos más colonizados fueron la raíz sumergida de mangle rojo y las esponjas en arrecife. El hidrozoo más colonizado fue C. marginatus. El material fue depositado en la colección de invertebrados marinos del Acuario Nacional de Cuba. ABSTRACT Hydrozoans from shallow waters were collected at 17 stations in July 2004. Stations were located in mangrove forests, seagrass meadows and coral reefs. Two subclases (Anthoathecata and Leptothecata of hydrozoans were found, including 11 families, 27 genera and 34 species. The collection site, depth, type of substrate, description and pictures are recorded for the fifteenth thecate hydrozoan species. The presence of Hebellopsis communis Calder, 1991a and Antennella siliquos (Hincks, 1877 is confirmed for the Caribbean Sea. The relative abundance is expressed in three categories: abundant, common and rare. Most abundant species were Dynamena crisioides Lamouroux, 1824 in mangrove habitat, Cnydoscyphus marginatus (Allman, 1877 and Sertularella diaphana (Allman, 1885 in

  1. Diversity and distribution of Brachionidae (Rotifera in Thailand, with a key to the species

    Directory of Open Access Journals (Sweden)

    Sujeephon Athibai

    2013-08-01

    Full Text Available We investigated the diversity of brachionid rotifers in Thailand. Rotifers were collected from a wide range of habitat types (294 sites, 508 samples including canals, lakes, peat swamps, ponds, reservoirs, rice fields, rivers, swamps and temporary ponds from April 1998 to January 2004. Twenty-eight species including 11 infrasubspecific forms belonging to five genera were identified. The most frequently encountered species was Brachionus falcatus (50.7% of the sampling sites, followed by Keratella cochlearis (50%, B. angularis (49.7%, K. tropica (46.3% and B. forficula (45.9%. Less frequently encountered species were B. caudatus, B. durgae and B. urceolaris. These species were found in low abundances and only in a single site. Two sites in the Northeast – Nong Changpeuak and Bueng Srithat – recorded the greatest richness with 15 taxa, whereas no brachionid rotifers were found in 31 sites. On average, the species richness was five taxa per sampling site. A key to the genera and species of Thai brachionid rotifers is provided.

  2. Cutaneous leishmaniasis in children: A case series

    Directory of Open Access Journals (Sweden)

    Mrinal Gupta

    2018-01-01

    Full Text Available Background: Cutaneous leishmaniasis (CL is a protozoal disease usually caused by Leishmania major and Leishmania tropica and transmitted by the bite of a sandfly. It is usually characterized by a single, polymorphous lesion located in an uncovered area. Aims: This study aimed to evaluate the clinico-epidemiological characteristics of CL among children. Materials and Methods: It was a retro-prospective study carried out over a period of 18 months in our center, in which the clinico-epidemiological features of children presenting with CL were assessed. Results: A total of 10 children (male:female - 7:3 were included in the study with the age range of 1–15 years, with a mean age of 8.8 years. Face was the most commonly affected site (n = 8, followed by neck and hands (1 each, and nodulo-ulcerative was the most common clinical type seen in nine patients. Skin smears for Leishman–Donovan (LD bodies were positive in five patients while the skin biopsy, which was done in four cases, showed the presence of LD bodies in only one patient. Conclusion: CL is a common presentation among children, especially in the endemic areas, and the clinical features are similar to the adult onset disease.

  3. Richness of endophytic fungi isolated from Opuntia ficus-indica Mill. (Cactaceae) and preliminary screening for enzyme production.

    Science.gov (United States)

    Bezerra, J D P; Santos, M G S; Svedese, V M; Lima, D M M; Fernandes, M J S; Paiva, L M; Souza-Motta, C M

    2012-05-01

    Opuntia ficus-indica Mill. (forage cactus) is farmed with relative success in the semi-arid region of the Brazilian northeast for commercial purposes, particularly as forage and food. Endophytic microorganisms are those that can be isolated inside plant tissues and can be a new source to production of enzymes with different potentialities. The objective of this study was to describe the richness of endophytic fungi from O. ficus-indica and to detect the capacity of these species to produce extracellular hydrolytic enzymes. Forty-four endophytic fungi species were isolated. Among them, the most commonly found were Cladosporium cladosporioides (20.43%) and C. sphaerospermum (15.99%). Acremonium terricola, Monodictys castaneae, Penicillium glandicola, Phoma tropica and Tetraploa aristata are being reported for the first time as endophytic fungi for Brazil. The majority of isolated fungi exhibited enzymatic potential. Aspergillus japonicus and P. glandicola presented pectinolytic activity. Xylaria sp. was the most important among the other 14 species with positive cellulase activity. All 24 isolates analysed were xylanase-positive. Protease was best produced by isolate PF103. The results indicate that there is a significant richness of endophytic fungi in O. ficus-indica, and that these isolates indicate promising potential for deployment in biotechnological processes involving production of pectinases, cellulases, xylanases and proteases.

  4. History on the biological nitrogen fixation research in graminaceous plants: special emphasis on the Brazilian experience.

    Science.gov (United States)

    Baldani, José I; Baldani, Vera L D

    2005-09-01

    This review covers the history on Biological Nitrogen Fixation (BNF) in Graminaceous plants grown in Brazil, and describes research progress made over the last 40 years, most of which was coordinated by Johanna Döbereiner. One notable accomplishment during this period was the discovery of several nitrogen-fixing bacteria such as the rhizospheric (Beijerinckia fluminensis and Azotobacter paspali), associative (Azospirillum lipoferum, A. brasilense, A. amazonense) and the endophytic (Herbaspirillum seropedicae, H. rubrisubalbicans, Gluconacetobacter diazotrophicus, Burkholderia brasilensis and B. tropica). The role of these diazotrophs in association with grasses, mainly with cereal plants, has been studied and a lot of progress has been achieved in the ecological, physiological, biochemical, and genetic aspects. The mechanisms of colonization and infection of the plant tissues are better understood, and the BNF contribution to the soil/plant system has been determined. Inoculation studies with diazotrophs showed that endophytic bacteria have a much higher BNF contribution potential than associative diazotrophs. In addition, it was found that the plant genotype influences the plant/bacteria association. Recent data suggest that more studies should be conducted on the endophytic association to strengthen the BNF potential. The ongoing genome sequencing programs: RIOGENE (Gluconacetobacter diazotrophicus) and GENOPAR (Herbaspirillum seropedicae) reflect the commitment to the BNF study in Brazil and should allow the country to continue in the forefront of research related to the BNF process in Graminaceous plants.

  5. The rare fluorinated natural products and biotechnological prospects for fluorine enzymology.

    Science.gov (United States)

    Chan, K K Jason; O'Hagan, David

    2012-01-01

    Nature has hardly evolved a biochemistry of fluorine although there is a low-level occurrence of fluoroacetate found in selected tropical and subtropical plants. This compound, which is generally produced in low concentrations, has been identified in the plants due to its high toxicity, although to date the biosynthesis of fluoroacetate in plants remains unknown. After that, fluorinated entities in nature are extremely rare, and despite increasingly sophisticated screening and analytical methods applied to natural product extraction, it has been 25 years since the last bona fide fluorinated natural product was identified from an organism. This was the reported isolation of the antibiotic 4-fluorothreonine and the toxin fluoroacetate in 1986 from Streptomyces cattleya. This bacterium has proven amenable to biochemical investigation, the fluorination enzyme (fluorinase) has been isolated and characterized, and the biosynthetic pathway to these bacterial metabolites has been elucidated. Also the fluorinase gene has been cloned into a host bacterium (Salinispora tropica), and this has enabled the de novo production of a bioactive fluorinated metabolite from fluoride ion, by genetic engineering. Biotechnological manipulation of the fluorinase offers the prospects for the assembly of novel fluorinated metabolites by fermentation technology. This is particularly attractive, given the backdrop that about 15-20% of pharmaceuticals licensed each year (new chemical entities) contain a fluorine atom. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Cryopreservation of Leishmania Species in Manisa Province.

    Science.gov (United States)

    Çavuş, İbrahim; Ocak, Fulya; Kaya, Tuğba; Özbilgin, Ahmet

    2017-09-01

    It was aimed to assess the success of the cryopreservation process which is carried out in order to preserve the genetic material and the virulence of the Leishmania species that are an important health problem in our region. Leishmania tropica, L. infantum, L. major, and L. donovani strains in Novy-MacNeal-Nicolle (NNN) medium in MCBU were used. Promastigotes cultured in the NNN medium were transferred to RPMI 1640 medium; promastigotes in the logarithmic phase were washed three times with PBS, and 15% dimethylsulfoxide (DMSO) was added. Leishmania species were transferred to 12 separate tubes. The tubes were stored at -86°C for one night by placing them in Coolcell boxes. The tubes were transferred into a liquid nitrogen tank. One cryotube per Leishmania strain is thawed monthly and cultured in NNN medium. For the duration of study it was observed that each Leishmania isolate preserved 60-65% of their viability and entered the logarithmic phase on the 7th day following the inoculation in the NNN medium. Abnormalities in the structures and movements of the promastigotes were not observed in microscopic examinations. The following conclusions were made: cryopreservation is important for studies planned related to leishmaniasis and cryopreservation with DMSO is successful.

  7. The proliferation potential of promastigotes of the main Leishmania species of the old world in NNN culture medium prepared using blood of four different mammals.

    Science.gov (United States)

    Ladopoulos, Theodoros; Ntais, Pantelis; Tsirigotakis, Nikolaos; Dokianakis, Emmanouil; Antoniou, Maria

    2015-10-01

    The efficacy of the in vitro cultivation of promastigotes of four Leishmania spp. was tested in the biphasic Novy-MacNeal-Nicolle (NNN) medium prepared using blood from different animals (horse, donkey, goat and sheep). The aim was to test which NNN preparation gave the best yield in the shortest time for different parasite species, in order to obtain a large crop of promastigotes for experimental work and for antigen preparation. Promastigotes of Leishmania infantum, Leishmania donovani, Leishmania tropica and Leishmania major, the four main parasite species occurring in the old world, were defrosted from -80 °C and placed, at equal numbers, in the 4 different NNN preparations. At the end of the 7th day, the NNN medium using horse blood produced the greatest number of promastigotes for all Leishmania spp. tested, whilst goat blood proved the poorest medium, providing culture results only for L. infantum. This finding may be explained by the fact that Leishmania is a nicotinamide adenine dinucleotide (NAD) auxotroph and horse erythrocytes support NAD-dependent microorganisms. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species.

    Science.gov (United States)

    Zampieri, Ricardo Andrade; Laranjeira-Silva, Maria Fernanda; Muxel, Sandra Marcia; Stocco de Lima, Ana Carolina; Shaw, Jeffrey Jon; Floeter-Winter, Lucile Maria

    2016-02-01

    Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol. Exploring the High Resolution Melting (HRM) dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR) targeting heat-shock protein 70 coding gene (hsp70) revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania) infantum chagasi, L. (L.) amazonensis, L. (L.) mexicana, L. (Viannia) lainsoni, L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi and L. (V.) shawi, and three species found in Eurasia and Africa, including L. (L.) tropica, L. (L.) donovani and L. (L.) major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol. HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA.

  9. High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species.

    Directory of Open Access Journals (Sweden)

    Ricardo Andrade Zampieri

    2016-02-01

    Full Text Available Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol.Exploring the High Resolution Melting (HRM dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR targeting heat-shock protein 70 coding gene (hsp70 revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania infantum chagasi, L. (L. amazonensis, L. (L. mexicana, L. (Viannia lainsoni, L. (V. braziliensis, L. (V. guyanensis, L. (V. naiffi and L. (V. shawi, and three species found in Eurasia and Africa, including L. (L. tropica, L. (L. donovani and L. (L. major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol.HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA.

  10. Detection and molecular identification of leishmania RNA virus (LRV) in Iranian Leishmania species.

    Science.gov (United States)

    Hajjaran, Homa; Mahdi, Maryam; Mohebali, Mehdi; Samimi-Rad, Katayoun; Ataei-Pirkooh, Angila; Kazemi-Rad, Elham; Naddaf, Saied Reza; Raoofian, Reza

    2016-12-01

    Leishmania RNA virus (LRV) was first detected in members of the subgenus Leishmania (Viannia), and later, the virulence and metastasis of the New World species were attributed to this virus. The data on the presence of LRV in Old World species are confined to Leishmania major and a few Leishmania aethiopica isolates. The aim of this study was to survey the presence of LRV in various Iranian Leishmania species originating from patients and animal reservoir hosts. Genomic nucleic acids were extracted from 50 cultured isolates belonging to the species Leishmania major, Leishmania tropica, and Leishmania infantum. A partial sequence of the viral RNA-dependent RNA polymerase (RdRp) gene was amplified, sequenced and compared with appropriate sequences from the GenBank database. We detected the virus in two parasite specimens: an isolate of L. infantum derived from a visceral leishmaniasis (VL) patient who was unresponsive to meglumine antimoniate treatment, and an L. major isolate originating from a great gerbil, Rhombomys opimus. The Iranian LRV sequences showed the highest similarities to an Old World L. major LRV2 and were genetically distant from LRV1 isolates detected in New World Leishmania parasites. We could not attribute treatment failure in VL patient to the presence of LRV due to the limited number of specimens analyzed. Further studies with inclusion of more clinical samples are required to elucidate the potential role of LRVs in pathogenesis or treatment failure of Old World leishmaniasis.

  11. Fractional ablative carbon dioxide laser followed by topical sodium stibogluconate application: A treatment option for pediatric cutaneous leishmaniasis.

    Science.gov (United States)

    Hilerowicz, Yuval; Koren, Amir; Mashiah, Jacob; Katz, Oren; Sprecher, Eli; Artzi, Ofir

    2018-05-01

    Leishmaniasis is a protozoan zoonotic parasitic infection with cutaneous, mucocutaneous, and visceral manifestations. Israel is endemic for cutaneous leishmaniasis, which is a self-limited disease but is associated with scarring, which is often a source of psychological and social burden for patients. Scars can be especially devastating for children and teenagers. A wide range of physical and medical approaches is used to treat cutaneous leishmaniasis, among which intralesional injections of sodium stibogluconate rank among the most frequently used. Unfortunately, despite being effective, this therapeutic modality can be very painful. Fractional ablative laser creates a controlled mesh-like pattern of tissue ablation in the skin that promotes dermal remodeling and collagen production while at the same time facilitating enhanced delivery of topically applied medications. Patients were treated with fractional ablative carbon dioxide laser followed by immediate topical application of sodium stibogluconate. All children were diagnosed with cutaneous leishmaniasis prior to treatment initiation.. Ten children were treated. One leishmania tropica-positive girl failed to respond. The other nine patients achieved clinical cure and demonstrated good to excellent final cosmesis. Self-rated patient satisfaction and tolerance were high No adverse effects were observed or reported during treatment. Fractional ablative carbon dioxide laser followed by topical sodium stibogluconate application appears to be a safe and promising treatment for cutaneous leishmaniasis infection in children. Future controlled studies are required to validate these findings and compare this technique with traditional approaches. © 2018 Wiley Periodicals, Inc.

  12. The Emergence of Anthroponotic Cutaneous Leishmaniasis Following the Earthquake in Southern Villages of Bam District, Southeastern Iran, 2010

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Aflatoonian

    2013-03-01

    Full Text Available Background: The objective of this study was to assess the epidemiological characteristics of a new emerging focus of cutaneous leishmaniasis (CL in southern villages of Bam District, southeastern Iran, 2010.Methods: A house-to- house census survey of 5544 individuals were interviewed and physically examined for the presence of active lesions or scars. Diagnosis was confirmed by direct smears, cultures and identification by PCR. The data were entered into a computer and SPSS ver. 15.Results: Overall, 1.2% of the inhabitants were infected, 0.5% active and 0.7% scars and females were more signifi­cantly infected (1.7% than males (0.8%, (P= 0.003. All age groups were equally affected. Most of the lesions were on the face and majority had single lesion. Most of the cases appeared from 2006 to 2008 during the CL epidemic in the city of Bam. PCR indicated L. tropica as the causative agent.Conclusion: The presence of non-immune individuals along with suitable ecological conditions could induce a new emerging focus of ACL in villages.

  13. Identification of Tunisian Leishmania spp. by PCR amplification of cysteine proteinase B (cpb) genes and phylogenetic analysis.

    Science.gov (United States)

    Chaouch, Melek; Fathallah-Mili, Akila; Driss, Mehdi; Lahmadi, Ramzi; Ayari, Chiraz; Guizani, Ikram; Ben Said, Moncef; Benabderrazak, Souha

    2013-03-01

    Discrimination of the Old World Leishmania parasites is important for diagnosis and epidemiological studies of leishmaniasis. We have developed PCR assays that allow the discrimination between Leishmania major, Leishmania tropica and Leishmania infantum Tunisian species. The identification was performed by a simple PCR targeting cysteine protease B (cpb) gene copies. These PCR can be a routine molecular biology tools for discrimination of Leishmania spp. from different geographical origins and different clinical forms. Our assays can be an informative source for cpb gene studying concerning drug, diagnostics and vaccine research. The PCR products of the cpb gene and the N-acetylglucosamine-1-phosphate transferase (nagt) Leishmania gene were sequenced and aligned. Phylogenetic trees of Leishmania based cpb and nagt sequences are close in topology and present the classic distribution of Leishmania in the Old World. The phylogenetic analysis has enabled the characterization and identification of different strains, using both multicopy (cpb) and single copy (nagt) genes. Indeed, the cpb phylogenetic analysis allowed us to identify the Tunisian Leishmania killicki species, and a group which gathers the least evolved isolates of the Leishmania donovani complex, that was originated from East Africa. This clustering confirms the African origin for the visceralizing species of the L. donovani complex. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Elephantiasis nostras verrucosa in a patient with systemic sclerosis.

    Science.gov (United States)

    Chatterjee, S; Karai, L J

    2009-12-01

    Elephantiasis nostras verrucosa (ENV) is an unusual skin condition characterized by dermal fibrosis and hyperkeratotic verrucous lesions resulting from chronic nonfilarial lymphoedema. The condition is similar to 'elephantiasis tropica', in which elephantiasis develops secondary to filariasis. Lymphatic obstruction can be primary or due to various causes such as surgery, tumour, radiation, congestive heart failure or obesity. Recurrent attacks of cellulitis lead to further impairment of lymphatic drainage, causing permanent swelling, dermal fibrosis and epidermal thickening. We report a case of a 56-year-old man with systemic sclerosis (SS), who presented with painful lesions on both legs, consistent with ENV. He developed extensive, fungating, papillomatous lesions on the skin of the legs, toes and dorsa of the feet over a period of 3 years. Histology revealed dense dermal fibrosis, oedema of the papillary dermis and extensive pseudo-epitheliomatous changes. To our knowledge, this is the first report of ENV in which SS was considered to be the primary cause for the impairment of lymphatic flow.

  15. Characterization of Endotrypanum Parasites Using Specific Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Ramos Franco Antonia Maria

    1997-01-01

    Full Text Available A large number of Endotrypanum stocks (representing an heterogeneous population of strains have been screened against a panel of monoclonal antibodies (MAbs derived for selected species of Endotrypanum or Leishmania, to see whether this approach could be used to group/differentiate further among these parasites. Using different immunological assay systems, MAbs considered specific for the genus Endotrypanum (E-24, CXXX-3G5-F12 or strain M6159 of E. schaudinni (E-2, CXIV-3C7-F5 reacted variably according to the test used but in the ELISA or immunofluorescence assay both reacted with all the strains tested. Analyses using these MAbs showed antigenic diversity occurring among the Endotrypanum strains, but no qualitative or quantitative reactivity pattern could be consistently related to parasite origin (i.e., host species involved or geographic area of isolation. Western blot analyses of the parasites showed that these MAbs recognized multiple components. Differences existed either in the epitope density or molecular forms associated with the antigenic determinants and therefore allowed the assignment of the strains to specific antigenic groups. Using immunofluorescence or ELISA assay, clone E-24 produced reaction with L. equatorensis (which is a parasite of sloth and rodent, but not with other trypanosomatids examined. Interestingly, the latter parasite and the Endotrypanum strains cross-reacted with a number of MAbs that were produced against members of the L. major-L. tropica complex

  16. Clinicoepidemiologic pattern of cutaneous leishmaniasis and molecular characterization of its causative agent in Hajjah governorate, northwest of Yemen.

    Science.gov (United States)

    Mogalli, Nabil M; El Hossary, Shabaan S; Khatri, Mishri Lal; Mukred, Abdualdaim M; Kassem, Hala A; El Sawaf, Bahira M; Ramadan, Nadia F

    2016-11-01

    The clinicoepidemiologic profile of 143 cases (93 males and 50 females) with cutaneous leishmaniasis from 18 villages of Hajjah governorate, Yemen was studied. Dry-type lesions were seen in 98.6% and wet-type lesions in 1.4% of patients. Lesions were localized in all cases with different morphological patterns. Microscopic examination of Giemsa-stained slit smears revealed amastigotes in 74.1% of patients with dry-type lesions and 0% in patients with wet-type lesions. The burden of the parasites in the lesions was high indicating active transmission of the disease. Most cases were from villages with moderate altitude range (8001-1600m). All age groups were affected, but most cases were seen in ages from 5 to 15 years. Leishmania species identification was done for all cases by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The biopsic material was scraped from both Giemsa-stained and methanol-fixed smears. The molecular characterization of Leishmania species revealed Leishmania tropica as the causative agent of cutaneous leishmaniasis in Hajjah, Yemen. The risk factors associated with the transmission of the disease and recommendations for improving case detection were discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Cutaneous Leishmaniasis – Dermoscopic Findings And Cryotherapy

    Directory of Open Access Journals (Sweden)

    Dobrev Hristo P.

    2015-03-01

    Full Text Available We present a 60-year-old male patient who, three months after a holiday in Southern Greece, found a small ‘pimple’ on his back, which gradually got as big as a small walnut, the central part becoming ulcerated and scabby. Dermatological examination found an erythematous-to-livid nodular lesion on the right shoulder; it was 16 mm in diameter with central ulceration, covered with brownish crust which discharged pus-like secretion upon pressure. Microscope examination of Romanowsky-Giemsa stained lesion material detected amastigote forms of Leishmania tropica. The culture investigation and serological tests for leishmaniasis were negative. Dermoscopy of the lesion found the following features: erythema, hyperkeratosis, central ulceration covered with brownish crust, “yellow tears-like” structures and “white starburst-like” patterns, and various vascular structures (including dotted vessels, comma-shaped vessels, hairpin- and glomerular-like vessels. The patient was diagnosed with cutaneous leishmaniasis and underwent four cryotherapy sessions every other week with excellent therapeutic results - complete resolution of infiltrate with subsequent gentle hypopigmented scarring. In conclusion, dermoscopy is an easily accessible non-invasive method which can be useful for the diagnosis of cutaneous leishmaniasis. Cryotherapy is the treatment of choice for single skin lesions.

  18. Canine visceral leishmaniasis in kerman, southeast of iran: a seroepidemiological, histopathological and molecular study.

    Directory of Open Access Journals (Sweden)

    Mehdi Bamorovat

    2014-09-01

    Full Text Available Canine visceral leishmaniasis (CVL is a systemic disease with a high mortality rate, caused by a diphasic protozoan parasite, Leishmania infantum/chagasi in the world. The objective of the present study was to determine the presence of CVL in the city and suburbs of Kerman, using a range of serological, histopathological and molecular methods.Blood samples were taken from 80 clinically symptomatic stray dogs All the collected blood samples were tested by direct agglutination test (DAT to detect the anti-Leishmania antibodies in dogs, using a cut-off value of ≥1:320. Pathological specimens including spleen, liver and lymph nodes were prepared for paraffin blocks, sectioning, staining and final microscopic examination in the pathology laboratory. PCR amplification of kDNA from 9 samples of DAT positive stray dogs was studied.The anti-Leishmania antibody was detected in 9 dogs (11.25 % of the total 80 studied dogs. No significant difference was found between VL infection and gender. In contrast, there was a significant difference between seropositivity and age (P<0.05. Pathological samples showed changes including hyperplasia of infected macrophages and inflammatory cells that occupied sinusoids and splenic cords. Among the samples which was characterized by PCR, only one specimen revealed to be mixed infection between L. infantum and L. tropica.The results revealed a high prevalence of L. infantum infection in stray dogs in Kerman. This kind of information is needed for implementation of future control programs.

  19. Design and Validation of Real-Time PCR: Quantitative Diagnosis of Common Leishmania Species in Iran.

    Science.gov (United States)

    Fekri Soofi Abadi, Maryam; Dabiri, Shahriar; Fotouhi Ardakani, Reza; Fani Malaki, Lina; Amirpoor Rostami, Sahar; Ziasistani, Mahsa; Dabiri, Donya

    2016-07-01

    Design and validation of Real-time PCR on the protected gene region ITS2 to quantify the parasite load in common leishmania (L) species. Probe and primer were designed from the ITS2 region between the rRNA genes with minimum gene variation in three common leishmania species followed by a Real-time PCR using the Taq man probe method in the form of absolute quantification. A series of different concentrations of leishmania were analyzed. After the purified PCR product was successfully placed in a PTG19-T plasmid vector, specialized ITS2 region was cloned in this plasmid. In the last phase, the cloned gene was transferred to the Ecoli.Top10F bacteria. The standard plasmid was provided in 10(7) to 10(1) copies/rxn concentrations. The specification and clinical sensitivity of the data was analyzed using inter and intra scales. The probe and primer were designed using three species, including L. infantum, L. major, and L.tropica. Seven concentrations of purified parasite in culture media showed that the selected region for quantifying the parasite is suitable. Clinical and analytical specificity and sensitivity were both 100%, respectively. The Taq man method for the ITS2 region in leishmania is one the most sensitive diagnostic test for identifying the parasite load and is suggested as a tool for fast identification and quantification of species.

  20. Quality Evaluation of Ayurvedic Crude Drug Daruharidra, Its Allied Species, and Commercial Samples from Herbal Drug Markets of India

    Directory of Open Access Journals (Sweden)

    Sharad Srivastava

    2013-01-01

    Full Text Available Berberis aristata known as “Daruharidra” in Ayurveda is a versatile medicinal plant used singly or in combination with other medicinal plants for treating a variety of ailments like jaundice, enlargement of spleen, leprosy, rheumatism, fever, morning/evening sickness, snakebite, and so forth. A major bioactive marker of this genus is an alkaloid berberine, which is known for its activity against cholera, acute diarrhea, amoebiasis, and latent malaria and for the treatment of oriental sore caused by Leishmania tropica. Although the roots of B. aristata are considered as the official drug (Ayurvedic Pharmacopoeia of India, the study revealed that different species of Berberis, namely. B. asiatica, B. chitria, and B. lycium are also used under the name of Daruharidra in different parts of the country. Detailed physicochemical and phytochemical studies of subjects like total ash, acid insoluble ash, tannins, and total alkaloids were calculated from the shade dried powdered material according to the recommended procedures. Further, heavy metal studies and quantitative estimation of berberine through HPTLC have also been performed as per ICH guidelines. A detailed study of four Berberis species, namely B. aristata, B. asiatica, B. chitria, and B. lycium, which are implicated as Daruharidra and collected from wild and ten commercial samples procured from various important drug markets in India has been carried out, which may be useful to pharmaceutical industries for the authentication of the commercial samples and exploring the possibilities of using other species as a substitute of B. aristata.

  1. Endemic infectious diseases and biological warfare during the Gulf War: a decade of analysis and final concerns.

    Science.gov (United States)

    Hyams, K C; Riddle, J; Trump, D H; Graham, J T

    2001-11-01

    Infectious diseases were one of the first health threats confronted by Coalition troops deployed to the Arabian desert in August 1990. On the basis of experiences in World War II, the major endemic infectious disease risks were thought to be sandfly fever, cutaneous leishmaniasis, diarrheal disease, and malaria. Although there was active surveillance, no case of sandfly fever and few other endemic infectious diseases were identified among over 500,000 U.S., British, and Canadian ground troops. In addition, there was no diagnosis of biological warfare (BW) exposure, and BW agents were not detected in clinical, environmental, or veterinary samples. The most common infectious disease problems were those associated with crowding (acute upper respiratory infections) and reduced levels of sanitation (travelers-type diarrhea). Only one endemic infectious disease has been confirmed as causing chronic health problems: visceral Leishmania tropica infection (viscerotropic leishmaniasis). However, this protozoan infection was diagnosed in only 12 U.S. veterans, and no new cases have been identified during the last 8 years. Infectious diseases were not a serious problem for Gulf War troops because of extensive preventive medicine efforts and favorable weather and geographic factors. Moreover, it is unlikely that an endemic infectious disease or a BW agent could cause chronic health problems and remain undetected over a 10-year period.

  2. Burkholderia monticola sp. nov., isolated from mountain soil.

    Science.gov (United States)

    Baek, Inwoo; Seo, Boram; Lee, Imchang; Yi, Hana; Chun, Jongsik

    2015-02-01

    An ivory/yellow, Gram-stain-negative, short-rod-shaped, aerobic bacterial strain, designated JC2948(T), was isolated from a soil sample taken from Gwanak Mountain, Republic of Korea. 16S rRNA gene sequence analysis indicated that strain JC2948(T) belongs to the genus Burkholderia. The test strain showed highest sequence similarities to Burkholderia tropica LMG 22274(T) (97.6 %), Burkholderia acidipaludis NBRC 101816(T) (97.5 %), Burkholderia tuberum LMG 21444(T) (97.5 %), Burkholderia sprentiae LMG 27175(T) (97.4 %), Burkholderia terricola LMG 20594(T) (97.3 %) and Burkholderia diazotrophica LMG 26031(T) (97.1 %). Based on average nucleotide identity (ANI) values, the new isolate represents a novel genomic species as it shows less than 90 % ANI values with other closely related species. Also, other phylosiological and biochemical comparisons allowed the phenotypic differentiation of strain JC2948(T) from other members of the genus Burkholderia. Therefore, we suggest that this strain should be classified as the type strain of a novel species of the genus Burkholderia. The name Burkholderia monticola sp. nov. (type strain, JC2948(T) = JCM 19904(T) = KACC 17924(T)) is proposed. © 2015 IUMS.

  3. Molecular characterization of leishmania infection from naturally infected sand flies caught in a focus of cutaneous leishmaniasis (eastern iran.

    Directory of Open Access Journals (Sweden)

    Mohammad Akhoundi

    2013-12-01

    Full Text Available Cutaneous leishmaniasis due to Leishmania major is a serious and increasing problem affecting many rural areas of 17 out of 31 provinces in Iran. Little is known about sand fly fauna and leishmaniases in Eastern Iran and no study has been carried out in Sarbisheh County. The aim of this study was to determine sand flies composition and probable Leishmania infection to find the probable vectors of leishmaniasis in Sarbisheh district.Sand flies were caught using both sticky papers and CDC light traps in August 2010. They were identified morphologically and analyzed for Leishmania infection by amplification of ITS-rDNA.Totally, 842 specimens were caught and 8 species recorded. They belonged to the genera Phlebotomus and Sergentomyia: P. (Phlebotomus papatasi, P. (Paraphlebotomus sergenti, P. (Pa. caucasicus, P. (Pa. mongolensis, P. (Pa. jacusieli, S. (Sergentomyia dentata, S. (Se. sintoni and S. (Sintonius clydei. All collected females were processed for Leishmania DNA detection by PCR amplifying of Internal Transcribed Spacer1 (partial sequence, 5.8S (complete sequence and ITS2 (partial sequence fragments. Thirteen females were positive for Leishmania DNA. The sequencing of the 430 bp amplicons indicated that 9 P. papatasi and 3 females belonging to the Caucasicus group carried L. major DNA whereas one P. sergenti carried L. tropica DNA.Phlebotomus papatasi and P. sergenti are, like in several places, the probable vectors of cutaneous leishmaniases in this emerging or unknown focus of cutaneous leishmaniases.

  4. Compilation of a global N{sub 2}O emission inventory for tropical rainforest soils using a detailed biogeochemical model

    Energy Technology Data Exchange (ETDEWEB)

    Werner, C.

    2007-09-15

    Nitrous oxide (N{sub 2}O) is a potent trace gas contributing to approximately 6% to the observed anthropogenic global warming. Soils have been identified to be the major source of atmospheric N{sub 2}O and tropical rainforest soils are thought to account for the largest part. Furthermore, various studies have shown that the magnitude of N{sub 2}O emissions from tropical rainforest soil is highly variable on spatial and temporal scales. Detailed, process-based models coupled to Geographic Information Systems (GIS) are considered promising tools for the calculation of N{sub 2}O emission inventories. This methodology explicitly accounts for the governing microbial processes as well as the environmental controls. Moreover, mechanistic biogeochemical models operating in daily time-steps (e.g. ForestDNDC-tropica) have been shown to capture the observed intra- and inter-annual variations of N{sub 2}O emissions. However, detailed N{sub 2}O emission datasets are required for model calibration and testing, but are currently few in numbers. In this study an automated measurement system was used to derive detailed datasets of N{sub 2}O, methane (CH{sub 4}) and carbon dioxide (CO{sub 2}) soil-atmosphere exchange and important environmental parameters from tropical rainforest soils in Kenya and Southwest China. Distinct differences were identified in the magnitude of the C and N soil-atmosphere exchange at the investigated sites and forest types. However, common features such as N{sub 2}O pulse emissions after dry season or the pronounced soil moisture dependency of N{sub 2}O emissions were observed at both sites. The derived datasets are unique for these tropical regions as so far no information about the source strength of these regions was available and, for the first time, the N{sub 2}O, CH{sub 4} and CO{sub 2} soil-atmosphere exchange was recorded in sub-daily resolution. The datasets were utilized in conjunction with available high-resolution datasets from Australian

  5. History on the biological nitrogen fixation research in graminaceous plants: special emphasis on the Brazilian experience

    Directory of Open Access Journals (Sweden)

    José I. Baldani

    2005-09-01

    Full Text Available This review covers the history on Biological Nitrogen Fixation (BNF in Graminaceous plants grown in Brazil, and describes research progress made over the last 40 years, most of whichwas coordinated by Johanna Döbereiner. One notable accomplishment during this period was the discovery of several nitrogen-fixing bacteria such as the rhizospheric (Beijerinckia fluminensis and Azotobacter paspali, associative (Azospirillum lipoferum, A. brasilense, A. amazonense and the endophytic (Herbaspirillum seropedicae, H. rubrisubalbicans, Gluconacetobacter diazotrophicus, Burkholderia brasilensis and B. tropica. The role of these diazotrophs in association with grasses, mainly with cereal plants, has been studied and a lot of progress has been achieved in the ecological, physiological, biochemical, and genetic aspects. The mechanisms of colonization and infection of the plant tissues are better understood, and the BNF contribution to the soil/plant system has been determined. Inoculation studies with diazotrophs showed that endophytic bacteria have a much higher BNF contribution potential than associative diazotrophs. In addition, it was found that the plant genotype influences the plant/bacteria association. Recent data suggest that more studies should be conducted on the endophytic association to strengthen the BNF potential. The ongoing genome sequencing programs: RIOGENE (Gluconacetobacter diazotrophicus and GENOPAR (Herbaspirillum seropedicae reflect the commitment to the BNF study in Brazil and should allow the country to continue in the forefront of research related to the BNF process in Graminaceous plants.A presente revisão aborda a história da Fixação Biológica de Nitrogênio (FBN em Gramíneas no Brasil, procurando mostrar a evolução da pesquisa na área iniciada a mais de 40 anos sob a liderança da pesquisadora Johanna Döbereiner. Um aspecto marcante deste período foi a descoberta de diversas bactérias fixadoras de nitrogênio atmosf

  6. Fearful foragers: honey bees tune colony and individual foraging to multi-predator presence and food quality.

    Directory of Open Access Journals (Sweden)

    Ken Tan

    Full Text Available Fear can have strong ecosystem effects by giving predators a role disproportionate to their actual kill rates. In bees, fear is shown through foragers avoiding dangerous food sites, thereby reducing the fitness of pollinated plants. However, it remains unclear how fear affects pollinators in a complex natural scenario involving multiple predator species and different patch qualities. We studied hornets, Vespa velutina (smaller and V. tropica (bigger preying upon the Asian honey bee, Apis cerana in China. Hornets hunted bees on flowers and were attacked by bee colonies. Bees treated the bigger hornet species (which is 4 fold more massive as more dangerous. It received 4.5 fold more attackers than the smaller hornet species. We tested bee responses to a three-feeder array with different hornet species and varying resource qualities. When all feeders offered 30% sucrose solution (w/w, colony foraging allocation, individual visits, and individual patch residence times were reduced according to the degree of danger. Predator presence reduced foraging visits by 55-79% and residence times by 17-33%. When feeders offered different reward levels (15%, 30%, or 45% sucrose, colony and individual foraging favored higher sugar concentrations. However, when balancing food quality against multiple threats (sweeter food corresponding to higher danger, colonies exhibited greater fear than individuals. Colonies decreased foraging at low and high danger patches. Individuals exhibited less fear and only decreased visits to the high danger patch. Contrasting individual with emergent colony-level effects of fear can thus illuminate how predators shape pollination by social bees.

  7. Investigation of the cytotoxic and antileishmanal effects of fagonia indica l. extract and extract mediated silver nanoparticles (AgNPs)

    International Nuclear Information System (INIS)

    Ullah, I.; Shinwari, Z. K.; Khalil, A. T.

    2017-01-01

    Leishmaniasis is one of the globally neglected tropical disease cause by protozoan parasite of the genus Leishmania. In Pakistan, cutaneous leishmaniasis is more sporadic in the Afghan refugee camps, which is concern for the local villager and Pakistani population. In the current study an approache was made to synthesise biogenic silver nanoparticles using Fagonia indica leaf extract. Furthermore, the antileishmanial activity of the nanoparticles was evaluated compared to the crude extracts against Leishmania tropica which is the causative agent of cutenious leishmaniasis. MTT cell viability assay was used to determine the non toxicological concentration of the extract and nanoparticle in macrophage cell lines (J774), and the antileishmanial activity. We found that silver nanoparticles are not toxic to macropage cell above 30 μg/ml. where as the IC/sub 50/ against leishmania parsites was calculated as 8.16+-0.63 μg/ml and 4.8+-0.819 μg/ml for extract and AgNPs respectively. We also determine the infection index of the parasite in the macrophage cell. The infectivity of parasites also decreases as compared to control group after activation of macrophages. We further, evaluate the mechanism of growth inhibition using Griess reagent for the estimation of nitrogen oxide. We found that both the extract and AgNPs produce an elevated level of nitrogen oxide free radical. These radical produce oxidative stress in the cell that lead to the reduced metabolic activities of the parasites and ultimate death. Overall, the results indicate that Fagonia indica leves extract and AgNPs are potent antileishmanial agents. (author)

  8. Review: Improving our knowledge of male mosquito biology in relation to genetic control programmes.

    Science.gov (United States)

    Lees, Rosemary Susan; Knols, Bart; Bellini, Romeo; Benedict, Mark Q; Bheecarry, Ambicadutt; Bossin, Hervé Christophe; Chadee, Dave D; Charlwood, Jacques; Dabiré, Roch K; Djogbenou, Luc; Egyir-Yawson, Alexander; Gato, René; Gouagna, Louis Clément; Hassan, Mo'awia Mukhtar; Khan, Shakil Ahmed; Koekemoer, Lizette L; Lemperiere, Guy; Manoukis, Nicholas C; Mozuraitis, Raimondas; Pitts, R Jason; Simard, Frederic; Gilles, Jeremie R L

    2014-04-01

    The enormous burden placed on populations worldwide by mosquito-borne diseases, most notably malaria and dengue, is currently being tackled by the use of insecticides sprayed in residences or applied to bednets, and in the case of dengue vectors through reduction of larval breeding sites or larviciding with insecticides thereof. However, these methods are under threat from, amongst other issues, the development of insecticide resistance and the practical difficulty of maintaining long-term community-wide efforts. The sterile insect technique (SIT), whose success hinges on having a good understanding of the biology and behaviour of the male mosquito, is an additional weapon in the limited arsenal against mosquito vectors. The successful production and release of sterile males, which is the mechanism of population suppression by SIT, relies on the release of mass-reared sterile males able to confer sterility in the target population by mating with wild females. A five year Joint FAO/IAEA Coordinated Research Project brought together researchers from around the world to investigate the pre-mating conditions of male mosquitoes (physiology and behaviour, resource acquisition and allocation, and dispersal), the mosquito mating systems and the contribution of molecular or chemical approaches to the understanding of male mosquito mating behaviour. A summary of the existing knowledge and the main novel findings of this group is reviewed here, and further presented in the reviews and research articles that form this Acta Tropica special issue. Copyright © 2013 International Atomic Energy Agency 2013. Published by Elsevier B.V. All rights reserved.

  9. Progress in research, control and elimination of helminth infections in Asia.

    Science.gov (United States)

    Utzinger, Jürg; Brattig, Norbert W; Leonardo, Lydia; Zhou, Xiao-Nong; Bergquist, Robert

    2015-01-01

    Global health has substantially improved over the past 20 years. In low- and middle-income countries, in particular, great strives have been made in the control of communicable diseases, including helminth infections. Nevertheless, the most marginalised communities still suffer from infectious diseases that are intimately connected with poverty and lack of access to essential commodities and services, such as clean water, improved sanitation and sufficient food. A two-pronged approach is thus necessary: (i) intensifying control in remaining high-endemicity areas and pockets of high transmission; and (ii) moving from morbidity control to interruption of disease transmission in low-endemicity areas with the goal of local elimination. The latter will require new tools and strategies, going hand-in-hand with strong partnerships and new strategic alliances. In this special issue of Acta Tropica, 35 articles are featured that, together, provide an up-to-date overview of the latest progress made in research, control and elimination of helminth infections in East and Southeast Asia. The first 12 articles expound tools and approaches for improved detection, surveillance and monitoring of helminth infections. Control and elimination approaches for the most important helminth infections are revisited in the next 20 articles. The three remaining articles are cross-cutting pieces examining the interface of agriculture, environment and helminth infections and providing a rationale for integrated, multi-sectorial control approaches that are necessary for sustaining helminthiasis control and progressively moving towards elimination. An interesting aspect revealed through an in-depth analysis of the provenance of the 35 contributions is that the People's Republic of China emerges as a key player in global health, which is documented through its prominent role in research and control of helminth infection and networking throughout Asia. Policy implications are discussed and will

  10. The origin and dispersion of human parasitic diseases in the old world (Africa, Europe and Madagascar).

    Science.gov (United States)

    Nozais, Jean-Pierre

    2003-01-01

    The ancestors of present-day man (Homo sapiens sapiens) appeared in East Africa some three and a half million years ago (Australopithecs), and then migrated to Europe, Asia, and later to the Americas, thus beginning the differentiation process. The passage from nomadic to sedentary life took place in the Middle East in around 8000 BC. Wars, spontaneous migrations and forced migrations (slave trade) led to enormous mixtures of populations in Europe and Africa and favoured the spread of numerous parasitic diseases with specific strains according to geographic area. The three human plasmodia (Plasmodium falciparum, P. vivax, and P. malariae) were imported from Africa into the Mediterranean region with the first human migrations, but it was the Neolithic revolution (sedentarisation, irrigation, population increase) which brought about actual foci for malaria. The reservoir for Leishmania infantum and L. donovani--the dog--has been domesticated for thousands of years. Wild rodents as reservoirs of L. major have also long been in contact with man and probably were imported from tropical Africa across the Sahara. L. tropica, by contrast, followed the migrations of man, its only reservoir. L. infantum and L. donovani spread with man and his dogs from West Africa. Likewise, for thousands of years, the dog has played an important role in the spread and the endemic character of hydatidosis through sheep (in Europe and North Africa) and dromadary (in the Sahara and North Africa). Schistosoma haematobium and S. mansoni have existed since prehistoric times in populations living in or passing through the Sahara. These populations then transported them to countries of Northern Africa where the specific, intermediary hosts were already present. Madagascar was inhabited by populations of Indonesian origin who imported lymphatic filariosis across the Indian Ocean (possibly of African origin since the Indonesian sailors had spent time on the African coast before reaching Madagascar

  11. Genomic islands predict functional adaptation in marine actinobacteria

    Energy Technology Data Exchange (ETDEWEB)

    Penn, Kevin; Jenkins, Caroline; Nett, Markus; Udwary, Daniel; Gontang, Erin; McGlinchey, Ryan; Foster, Brian; Lapidus, Alla; Podell, Sheila; Allen, Eric; Moore, Bradley; Jensen, Paul

    2009-04-01

    Linking functional traits to bacterial phylogeny remains a fundamental but elusive goal of microbial ecology 1. Without this information, it becomes impossible to resolve meaningful units of diversity and the mechanisms by which bacteria interact with each other and adapt to environmental change. Ecological adaptations among bacterial populations have been linked to genomic islands, strain-specific regions of DNA that house functionally adaptive traits 2. In the case of environmental bacteria, these traits are largely inferred from bioinformatic or gene expression analyses 2, thus leaving few examples in which the functions of island genes have been experimentally characterized. Here we report the complete genome sequences of Salinispora tropica and S. arenicola, the first cultured, obligate marine Actinobacteria 3. These two species inhabit benthic marine environments and dedicate 8-10percent of their genomes to the biosynthesis of secondary metabolites. Despite a close phylogenetic relationship, 25 of 37 secondary metabolic pathways are species-specific and located within 21 genomic islands, thus providing new evidence linking secondary metabolism to ecological adaptation. Species-specific differences are also observed in CRISPR sequences, suggesting that variations in phage immunity provide fitness advantages that contribute to the cosmopolitan distribution of S. arenicola 4. The two Salinispora genomes have evolved by complex processes that include the duplication and acquisition of secondary metabolite genes, the products of which provide immediate opportunities for molecular diversification and ecological adaptation. Evidence that secondary metabolic pathways are exchanged by Horizontal Gene Transfer (HGT) yet are fixed among globally distributed populations 5 supports a functional role for their products and suggests that pathway acquisition represents a previously unrecognized force driving bacterial diversification

  12. Phylogenetic analysis of HSP70 and cyt b gene sequences for Chinese Leishmania isolates and ultrastructural characteristics of Chinese Leishmania sp.

    Science.gov (United States)

    Yuan, Dongmei; Qin, Hanxiao; Zhang, Jianguo; Liao, Lin; Chen, Qiwei; Chen, Dali; Chen, Jianping

    2017-02-01

    Leishmaniasis is a worldwide epidemic disease caused by the genus Leishmania, which is still endemic in the west and northwest areas of China. Some viewpoints of the traditional taxonomy of Chinese Leishmania have been challenged by recent phylogenetic researches based on different molecular markers. However, the taxonomic positions and phylogenetic relationships of Chinese Leishmania isolates remain controversial, which need for more data and further analysis. In this study, the heat shock protein 70 (HSP70) gene and cytochrome b (cyt b) gene were used for phylogenetic analysis of Chinese Leishmania isolates from patients, dogs, gerbils, and sand flies in different geographic origins. Besides, for the interesting Leishmania sp. in China, the ultrastructure of three Chinese Leishmania sp. strains (MHOM/CN/90/SC10H2, SD, GL) were observed by transmission electron microscopy. Bayesian trees from HSP70 and cyt b congruently indicated that the 14 Chinese Leishmania isolates belong to three Leishmania species including L. donovani complex, L. gerbilli, and L. (Sauroleishmania) sp. Their identity further confirmed that the undescribed Leishmania species causing visceral Leishmaniasis (VL) in China is closely related to L. tarentolae. The phylogenetic results from HSP70 also suggested the classification of subspecies within L. donovani complex: KXG-918, KXG-927, KXG-Liu, KXG-Xu, 9044, SC6, and KXG-65 belong to L. donovani; Cy, WenChuan, and 801 were proposed to be L. infantum. Through transmission electron microscopy, unexpectedly, the Golgi apparatus were not observed in SC10H2, SD, and GL, which was similar to previous reports of reptilian Leishmania. The statistical analysis of microtubule counts separated SC10H2, SD, and GL as one group from any other reference strain (L. donovani MHOM/IN/80/DD8; L. tropica MHOM/SU/74/K27; L. gerbilli MRHO/CN/60/GERBILLI). The ultrastructural characteristics of Leishmania sp. partly lend support to the phylogenetic inference that

  13. Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay.

    Science.gov (United States)

    de Almeida, Marcos E; Koru, Ozgur; Steurer, Francis; Herwaldt, Barbara L; da Silva, Alexandre J

    2017-01-01

    Leishmaniasis in humans is caused by Leishmania spp. in the subgenera Leishmania and Viannia Species identification often has clinical relevance. Until recently, our laboratory relied on conventional PCR amplification of the internal transcribed spacer 2 (ITS2) region (ITS2-PCR) followed by sequencing analysis of the PCR product to differentiate Leishmania spp. Here we describe a novel real-time quantitative PCR (qPCR) approach based on the SYBR green technology (LSG-qPCR), which uses genus-specific primers that target the ITS1 region and amplify DNA from at least 10 Leishmania spp., followed by analysis of the melting temperature (T m ) of the amplicons on qPCR platforms (the Mx3000P qPCR system [Stratagene-Agilent] and the 7500 real-time PCR system [ABI Life Technologies]). We initially evaluated the assay by testing reference Leishmania isolates and comparing the results with those from the conventional ITS2-PCR approach. Then we compared the results from the real-time and conventional molecular approaches for clinical specimens from 1,051 patients submitted to the reference laboratory of the Centers for Disease Control and Prevention for Leishmania diagnostic testing. Specimens from 477 patients tested positive for Leishmania spp. with the LSG-qPCR assay, specimens from 465 of these 477 patients also tested positive with the conventional ITS2-PCR approach, and specimens from 10 of these 465 patients had positive results because of retesting prompted by LSG-qPCR positivity. On the basis of the T m values of the LSG-qPCR amplicons from reference and clinical specimens, we were able to differentiate four groups of Leishmania parasites: the Viannia subgenus in aggregate; the Leishmania (Leishmania) donovani complex in aggregate; the species L (L) tropica; and the species L (L) mexicana, L (L) amazonensis, L (L) major, and L (L) aethiopica in aggregate. Copyright © 2016 American Society for Microbiology.

  14. Natural infection of bats with Leishmania in Ethiopia.

    Science.gov (United States)

    Kassahun, Aysheshm; Sadlova, Jovana; Benda, Petr; Kostalova, Tatiana; Warburg, Alon; Hailu, Asrat; Baneth, Gad; Volf, Petr; Votypka, Jan

    2015-10-01

    The leishmaniases, a group of diseases with a worldwide-distribution, are caused by different species of Leishmania parasites. Both cutaneous and visceral leishmaniasis remain important public health problems in Ethiopia. Epidemiological cycles of these protozoans involve various sand fly (Diptera: Psychodidae) vectors and mammalian hosts, including humans. In recent years, Leishmania infections in bats have been reported in the New World countries endemic to leishmaniasis. The aim of this study was to survey natural Leishmania infection in bats collected from various regions of Ethiopia. Total DNA was isolated from spleens of 163 bats belonging to 23 species and 18 genera. Leishmania infection was detected by real-time (RT) PCR targeting a kinetoplast (k) DNA and internal transcribed spacer one (ITS1) gene of the parasite. Detection was confirmed by sequencing of the PCR products. Leishmania kDNA was detected in eight (4.9%) bats; four of them had been captured in the Aba-Roba and Awash-Methara regions that are endemic for leishmaniasis, while the other four specimens originated from non-endemic localities of Metu, Bedele and Masha. Leishmania isolates from two bats were confirmed by ITS1 PCR to be Leishmania tropica and Leishmania major, isolated from two individual bats, Cardioderma cor and Nycteris hispida, respectively. These results represent the first confirmed observation of natural infection of bats with the Old World Leishmania. Hence, bats should be considered putative hosts of Leishmania spp. affecting humans with a significant role in the transmission. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  15. Isatis tinctoria mediated synthesis of amphotericin B-bound silver nanoparticles with enhanced photoinduced antileishmanial activity: A novel green approach.

    Science.gov (United States)

    Ahmad, Aftab; Wei, Yun; Syed, Fatima; Khan, Shafiullah; Khan, Gul Majid; Tahir, Kamran; Khan, Arif Ullah; Raza, Muslim; Khan, Faheem Ullah; Yuan, Qiping

    2016-08-01

    After malaria, Leishmaniasis is the most prevalent infectious disease in terms of fatality and geographical distribution. The availability of a limited number of antileishmanial agents, emerging resistance to the available drugs, and the high cost of treatment complicate the treatment of leishmaniasis. To overcome these issues, critical research for new therapeutic agents with enhanced antileishmanial potential and low treatment cost is needed. In this contribution, we developed a green protocol to prepare biogenic silver nanoparticles (AgNPs) and amphotericin B-bound biogenic silver nanoparticles (AmB-AgNPs). Phytochemicals from the aqueous extract of Isatis tinctoria were used as reducing and capping agents to prepare silver nanoparticles. Amphotericin B was successfully adsorbed on the surface of biogenic silver nanoparticles. The prepared nanoparticles were characterized by various analytical techniques. UV-Visible spectroscopy was employed to detect the characteristic localized surface plasmon resonance peaks (LSPR) for the prepared nanoparticles. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) studies revealed the formation of spherical silver nanoparticles with an average particle size of 10-20nm. The cubic crystalline structure of the prepared nanoparticles was confirmed by X-ray diffraction (XRD) study. FTIR spectroscopic analysis revealed that plant polyphenolic compounds are mainly involved in metal reduction and capping. Under visible light irradiation, biogenic silver nanoparticles exhibited significant activity against Leishmania tropica with an IC50 value of 4.2μg/mL. The leishmanicidal activity of these nanoparticles was considerably enhanced by conjugation with amphotericin B (IC50=2.43μg/mL). In conclusion, the findings of this study reveal that adsorption of amphotericin B, an antileishmanial drug, to biogenic silver nanoparticles, could be a safe, more effective and economic alternative to the available

  16. Histological grading patterns in patients of cutaneous leishmaniasis

    International Nuclear Information System (INIS)

    Saleem, K.; Ayaz, B.; Shaikh, A.

    2007-01-01

    To determine the histological grading patterns in a cohort of hospitalized patients of cutaneous leishmaniasis. One hundred patients of Cutaneous Leishmaniasis (CL), admitted in dermatology wards at PNS Shifa Hospital, Karachi, were examined. Only admitted patients of all ages and both sexes were included in the study. Patients of CL, who had received or were receiving systemic treatment were excluded. The lesions having marked secondary bacterial infection were also excluded. Initial diagnosis was clinical. History of being to an endemic area supported the diagnosis. The lesions were divided in two groups. Early, with duration less than 03 months and late, with duration between 3 and 12 months. The clinical lesions were noted as nodules, plaques, ulcers, crusted ulcers, lupoid lesions and plaques with scarring. Three types of skin smears (slit skin smear, saline aspirate smear and dab smear) were taken and examined with Giemsa stain. Cultures were performed on Nicolle-Novy-MacNeal (NNN) culture medium from Defense Scientific and Technology Organization (DESTO) Lab., Pakistan. Incisional skin biopsies were done. The biopsy specimens were examined by hemotoxylin and eosin stain (H and E stain). The number of Leishmania Tropica (LT) bodies was graded according to modified Ridley's parasitic index 1983. Clinical features were correlated with the histological patterns. Five histological patterns were identified in current study: 1) diffuse dermal infiltration without necrosis, 2) patchy dermal infiltration, 3) diffuse dermal infiltration with necrosis, 4) early reactive granuloma formation and 5) established epithelioid granuloma formation. LT bodies were identified in 75% of cases. Epidermal features were non-specific. The early lesions presented with diffuse infiltrate and late lesions showed granuloma formation. Five distinct types of histological patterns of CL have been recognized in this study. The early lesions presented with diffuse infiltrate and late lesions

  17. Detection of Leishmania parasites in the blood of patients with isolated cutaneous leishmaniasis.

    Science.gov (United States)

    Nakkash-Chmaisse, Hania; Makki, Raja; Nahhas, Georges; Knio, Khouzama; Nuwayri-Salti, Nuha

    2011-07-01

    The consequences of the spread of Leishmania parasites to the blood from lesions in patients with cutaneous leishmaniasis are numerous. To assess the magnitude of this invasion we conducted the present study on patients referred to the American University of Beirut Medical Center for cutaneous leishmaniasis. Patients referred for the management of cutaneous leishmaniasis were included in the study. Skin and blood cultures for Leishmania were taken from these patients. One hundred sixty-two patients were proven to have cutaneous leishmaniasis by pathology; 52% were males and 44% females (gender information was missing for 4%). Patient age ranged from 5 months to 70 years. None of the patients had received treatment for Leishmania. We obtained parasite isolates from 85 patients (52.5%), proven by cultures from skin and blood/blood components. Interestingly, the parasite was isolated in the blood and blood components of 50 patients (30.9%). Isoenzyme analysis confirmed the fact that the organisms in blood and skin were the same; from the 28 isolates that were positive in both skin and blood, eight isolates were Leishmania major and two were Leishmania tropica. The remaining isolates, whether positive in both blood and skin or in either of these tissues, skin or blood and its products, were Leishmania infantum sensu lato. In the current study, the detection rate of parasites in the blood of patients with cutaneous leishmaniasis was high. This illustrates the invasive characteristic of the parasite that has escaped the skin. Testing should be considered in areas other than Lebanon, especially around the Mediterranean basin. Whether these findings support the administration of systemic treatment for cutaneous leishmaniasis or not needs to be confirmed in larger prospective studies. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  18. Epidemiology of elephantiasis with special emphasis on podoconiosis in Ethiopia: A literature review.

    Science.gov (United States)

    Yimer, Mulat; Hailu, Tadesse; Mulu, Wondemagegn; Abera, Bayeh

    2015-06-01

    Elephantiasis is a symptom of a variety of diseases that is characterized by the thickening of the skin and underlying tissues, especially in the legs, male genitals and female breasts. Some conditions having this symptom include: Elephantiasis nostras, due to longstanding chronic lymphangitis; Elephantiasis tropica or lymphatic filariasis, caused by a number of parasitic worms, particularly Wuchereria bancrofti; non-filarial elephantiasis or podoconiosis, an immune disease caused by heavy metals affecting the lymph vessels; proteus syndrome, the genetic disorder of the so-called Elephant Man, etc. Podoconiosis is a type of lower limb tropical elephantiasis distinct from lymphatic filariasis. Lymphatic filariasis affects all population at risk, whereas podoconiosis predominantly affects barefoot subsistence farmers in areas with red volcanic soil. Ethiopia is one of the countries with the highest number of podoconiosis patients since many people are at risk to red-clay soil exposure in many parts of the country. The aim of this review was to know the current status and impact of podoconiosis and its relevance to elephantiasis in Ethiopia. To know the epidemiology and disease burden, the literatures published by different scholars were systematically reviewed. The distribution of the disease and knowledge about filarial elephantiasis and podoconiosis are not well known in Ethiopia. It is relatively well studied in southern Ethiopia but data from other parts of the country are limited. Moreover, programmes that focus on diagnosis, treatment, prevention and control of filarial elephantiasis and podoconiosis are also non-existent even in endemic areas. Furthermore, the disease mapping has not been carried out country-wide. Therefore, in order to address these gaps, Ethiopian Ministry of Health needs to take initiative for undertaking concrete research and mapping of the disease in collaboration with stakeholders.

  19. The origin and dispersion of human parasitic diseases in the Old World (Africa, Europe and Madagascar

    Directory of Open Access Journals (Sweden)

    Jean-Pierre Nozais

    2003-01-01

    Full Text Available The ancestors of present-day man (Homo sapiens sapiens appeared in East Africa some three and a half million years ago (Australopithecs, and then migrated to Europe, Asia, and later to the Americas, thus beginning the differentiation process. The passage from nomadic to sedentary life took place in the Middle East in around 8000 BC. Wars, spontaneous migrations and forced migrations (slave trade led to enormous mixtures of populations in Europe and Africa and favoured the spread of numerous parasitic diseases with specific strains according to geographic area. The three human plasmodia (Plasmodium falciparum, P. vivax, and P. malariae were imported from Africa into the Mediterranean region with the first human migrations, but it was the Neolithic revolution (sedentarisation, irrigation, population increase which brought about actual foci for malaria. The reservoir for Leishmania infantum and L. donovani - the dog - has been domesticated for thousands of years. Wild rodents as reservoirs of L. major have also long been in contact with man and probably were imported from tropical Africa across the Sahara. L. tropica, by contrast, followed the migrations of man, its only reservoir. L. infantum and L. donovani spread with man and his dogs from West Africa. Likewise, for thousands of years, the dog has played an important role in the spread and the endemic character of hydatidosis through sheep (in Europe and North Africa and dromadary (in the Sahara and North Africa. Schistosoma haematobium and S. mansoni have existed since prehistoric times in populations living in or passing through the Sahara. These populations then transported them to countries of Northern Africa where the specific, intermediary hosts were already present. Madagascar was inhabited by populations of Indonesian origin who imported lymphatic filariosis across the Indian Ocean (possibly of African origin since the Indonesian sailors had spent time on the African coast before

  20. Miltefosina oral para el tratamiento de la leishmaniasis

    Directory of Open Access Journals (Sweden)

    Jaime Soto

    2006-10-01

    Full Text Available La pérdida de la eficacia, las dificultades para su administración y el incremento en la frecuenciay la severidad de los efectos secundarios de las sales de antimonio pentavalente han forzadola búsqueda de un medicamento que pueda remplazarlas en el tratamiento de la leishmaniasis.Los estudios adelantados con medicamentos inyectables, orales y tópicos han arrojadoresultados inconsistentes y de poca utilidad en el Nuevo Mundo. Desde 1998 lahexadecilfosfocolina (miltefosina viene siendo utilizada con éxito en India en enfermos conleishmaniasis visceral y en 1999 se iniciaron los estudios clínicos en Colombia en pacientescon leishmaniasis cutánea. A la fecha se han tratado mas de 2.500 pacientes en India(leishmaniasis visceral por L. donovani y Colombia (leishmaniasis cutánea por L. panamensisy se han logrado tasas de curación superiores a 91% a la dosis recomendada de 2,5 mg / kgpeso / día durante 28 días continuos, sin que haya diferencia en la respuesta entre pacientesnuevos y los que presentan fallas o recaídas después de recibir antimoniales. En Guatemalala respuesta fue de 53% ( 33% para L. braziliensis, 60% para L. mexicana mientras que la deL. tropica en Afganistán fue de 63%. Pacientes con leishmaniasis cutánea difusa, leishmaniasismucosa y con co-infección por HIV+, han tenido una buena respuesta inicial pero deberecordarse la tendencia a recaer que tienen estos pacientes. Entre el 35% al 60% de lospacientes presentan reacciones adversas gastrointestinales leves y 10% al 20% hacenelevación leve de transaminasas y creatinina.

  1. Neglected tropical diseases of the Middle East and North Africa: review of their prevalence, distribution, and opportunities for control.

    Directory of Open Access Journals (Sweden)

    Peter J Hotez

    Full Text Available The neglected tropical diseases (NTDs are highly endemic but patchily distributed among the 20 countries and almost 400 million people of the Middle East and North Africa (MENA region, and disproportionately affect an estimated 65 million people living on less than US$2 per day. Egypt has the largest number of people living in poverty of any MENA nation, while Yemen has the highest prevalence of people living in poverty. These two nations stand out for having suffered the highest rates of many NTDs, including the soil-transmitted nematode infections, filarial infections, schistosomiasis, fascioliasis, leprosy, and trachoma, although they should be recognized for recent measures aimed at NTD control. Leishmaniasis, especially cutaneous leishmaniasis, is endemic in Syria, Iran, Iraq, Libya, Morocco, and elsewhere in the region. Both zoonotic (Leishmania major and anthroponotic (Leishmania tropica forms are endemic in MENA in rural arid regions and urban regions, respectively. Other endemic zoonotic NTDs include cystic echinococcosis, fascioliasis, and brucellosis. Dengue is endemic in Saudi Arabia, where Rift Valley fever and Alkhurma hemorrhagic fever have also emerged. Great strides have been made towards elimination of several endemic NTDs, including lymphatic filariasis in Egypt and Yemen; schistosomiasis in Iran, Morocco, and Oman; and trachoma in Morocco, Algeria, Iran, Libya, Oman, Saudi Arabia, Tunisia, and the United Arab Emirates. A particularly noteworthy achievement is the long battle waged against schistosomiasis in Egypt, where prevalence has been brought down by regular praziquantel treatment. Conflict and human and animal migrations are key social determinants in preventing the control or elimination of NTDs in the MENA, while local political will, strengthened international and intersectoral cooperative efforts for surveillance, mass drug administration, and vaccination are essential for elimination.

  2. Role of phosphate solubilizing Burkholderia spp. for successful colonization and growth promotion of Lycopodium cernuum L. (Lycopodiaceae) in lateritic belt of Birbhum district of West Bengal, India.

    Science.gov (United States)

    Ghosh, Ranjan; Barman, Soma; Mukherjee, Rajib; Mandal, Narayan C

    2016-02-01

    Profuse growth of Lycpodium cernuum L. was found in phosphate deficient red lateritic soil of West Bengal, India. Interaction of vesicular-arbuscular mycorrhiza (VAM) with Lycopodium rhizoids were described earlier but association of PGPR with their rhizoids were not studied. Three potent phosphate solubilizing bacterial strains (P4, P9 and P10) associated with L. cernuum rhizoids were isolated and identified by 16S rDNA homologies on Ez-Taxon database as Burkholderia tropica, Burkholderia unamae and Burkholderia cepacia respectively. Day wise kinetics of phosphate solubilization against Ca3(PO4)2 suggested P4 (580.56±13.38 μg ml(-1)) as maximum mineral phosphate solubilizer followed by P9 (517.12±17.15 μg ml(-1)) and P10 (485.18±14.23 μg ml(-1)) at 28 °C. Release of bound phosphates by isolated strains from ferric phosphate (FePO4), aluminum phosphate (AlPO4) and four different complex rock phosphates indicated their very good phosphate solubilizng efficacy. Nitrogen independent solubilizition also supports their nitrogen fixing capabilities. Inhibition of P solubilization by calcium salts and induction by EDTA suggested pH dependent chelation of metal cations by all of the isolates. Rhizoidal colonization potentials of Burkholderia spp. were confirmed by in planta experiment and also using scanning electron microscope (SEM). Increases of total phosphate content in Lycopodium plants upon soil treatment with these isolates were also recorded. In addition siderophore production on CAS agar medium, tryptophan dependent IAA production and antifungal activities against pathogenic fungi by rhizospheric isolates deep-rooted that they have definite role in nutrient mobilization for successful colonization of L. cernuum in nutrient deficient lateritic soil. Copyright © 2015 Elsevier GmbH. All rights reserved.

  3. Burkholderia metalliresistens sp. nov., a multiple metal-resistant and phosphate-solubilising species isolated from heavy metal-polluted soil in Southeast China.

    Science.gov (United States)

    Guo, Jun Kang; Ding, Yong Zhen; Feng, Ren Wei; Wang, Rui Gang; Xu, Ying Ming; Chen, Chun; Wei, Xiu Li; Chen, Wei Min

    2015-06-01

    A metal-resistant and phosphate-solubilising bacterium, designated as strain D414(T), was isolated from heavy metal (Pb, Cd, Cu and Zn)-polluted paddy soils at the surrounding area of Dabao Mountain Mine in Southeast China. The minimum inhibitory concentrations of heavy metals for strain D414(T) were 2000 mg L(-1) (Cd), 800 mg L(-1) (Pb), 150 mg L(-1) (Cu) and 2500 mg L(-1) (Zn). The strain possessed plant growth-promoting properties, such as 1-aminocyclopropane-1-carboxylate assimilation, indole production and phosphate solubilisation. Analysis of 16S rRNA gene sequence indicated that the isolate is a member of the genus Burkholderia where strain D414(T) formed a distinct phyletic line with validly described Burkholderia species. Strain D414(T) is closely related to Burkholderia tropica DSM 15359(T), B. bannensis NBRC E25(T) and B. unamae DSM 17197(T), with 98.5, 98.3 and 98.3 % sequence similarities, respectively. Furthermore, less than 34 % DNA-DNA relatedness was detected between strain D414(T) and the type strains of the phylogenetically closest species of Burkholderia. The dominant fatty acids of strain D414(T) were C14:0, C16:0, C17:0 cyclo and C18:1 ω7c. The DNA G+C content was 62.3 ± 0.5 mol%. On the basis of genotypic, phenotypic and phylogenetic data, strain D414(T) represents a novel species, for which the name Burkholderia metalliresistens sp. nov. is proposed, with D414(T) (=CICC 10561(T) = DSM 26823(T)) as the type strain.

  4. Species composition of sand flies and bionomics of Phlebotomus papatasi and P. sergenti (Diptera: Psychodidae) in cutaneous leishmaniasis endemic foci, Morocco.

    Science.gov (United States)

    Boussaa, Samia; Kahime, Kholoud; Samy, Abdallah M; Salem, Abdelkrim Ben; Boumezzough, Ali

    2016-02-02

    Cutaneous Leishmaniasis (CL) is one of the most neglected tropical diseases in Morocco. Leishmania major and L. tropica are the main culprits identified in all endemic foci across the country. These two etiological agents are transmitted by Phlebotomus papatasi and P. sergenti, the two most prevalent sand fly species in Morocco. Previous studies reflected gaps of knowledge regarding the environmental fingerprints that affect the distribution of these two potential vectors across Morocco. The sand flies were collected from 48 districts across Morocco using sticky paper traps. Collected specimens were preserved in 70% ethanol for further processing and identification. Male and female densities were calculated in each site to examine their relations to the environmental conditions across these sites. The study used 19 environmental variables including precipitation, aridity, elevation, soil variables and a composite representing maximum, minimum and mean of day- and night-time Land Surface Temperature (LST), and Normalized Difference Vegetation Index (NDVI). A total of 11,717 specimens were collected during this entomological survey. These specimens represented 11 species of two genera; Phlebotomus and Sergentomyia. Correlations of the sand fly densities with the environmental variables were estimated to identify the variables which influence the distribution of the two potential vectors, Phlebotomus papatasi and P. sergenti, associated with all CL endemic foci across the country. The density of P. papatasi was most affected by temperature changes. The study showed a significant positive correlation between the densities of both sexes of P. papatasi and night-time temperatures. Both P. papatasi and P. sergenti showed a negative correlation with aridity, but, such correlation was only significant in case of P. papatasi. NDVI showed a positive correlation only with densities of P. sergenti, while, soil PH and soil water stress were negatively correlated with the

  5. Antimony susceptibility of Leishmania isolates collected over a 30-year period in Algeria.

    Science.gov (United States)

    Eddaikra, Naouel; Ait-Oudhia, Khatima; Kherrachi, Ihcen; Oury, Bruno; Moulti-Mati, Farida; Benikhlef, Razika; Harrat, Zoubir; Sereno, Denis

    2018-03-01

    In Algeria, the treatment of visceral and cutaneous leishmanioses (VL and CL) has been and continues to be based on antimony-containing drugs. It is suspected that high drug selective pressure might favor the emergence of chemoresistant parasites. Although treatment failure is frequently reported during antimonial therapy of both CL and VL, antimonial resistance has never been thoroughly investigated in Algeria. Determining the level of antimonial susceptibility, amongst Leishmania transmitted in Algeria, is of great importance for the development of public health policies. Within the framework of the knowledge about the epidemiology of VL and CL amassed during the last 30 years, we sampled Leishmania isolates to determine their susceptibility to antimony. We analyzed a total of 106 isolates including 88 isolates collected between 1976 and 2013 in Algeria from humans, dogs, rodents, and phlebotomines and 18 collected from dogs in France. All the Algerian isolates were collected in 14 localities where leishmaniasis is endemic. The 50% inhibitory concentrations (IC50) of potassium antimony tartrate (the trivalent form of antimony, Sb(III)) and sodium stibogluconate (the pentavalent form of antimony, Sb(V)) were determined in promastigotes and intramacrophage amastigotes, respectively. The epidemiological cutoff (ECOFF) that allowed us to differentiate between Leishmania species causing cutaneous or visceral leishmaniases that were susceptible (S+) or insusceptible (S-) to the trivalent form of antimony was determined. The computed IC50 cutoff values were 23.83 μg/mL and 15.91 μg/mL for VL and CL, respectively. We report a trend of increasing antimony susceptibility in VL isolates during the 30-year period. In contrast, an increase in the frequency of S- phenotypes in isolates causing CL was observed during the same period. In our study, the emergence of S- phenotypes correlates with the inclusion of L. killicki (syn: L. tropica) isolates that cause cutaneous

  6. Development of a rapid loop-mediated isothermal amplification assay for diagnosis and assessment of cure of Leishmania infection.

    Science.gov (United States)

    Verma, Sandeep; Singh, Ruchi; Sharma, Vanila; Bumb, Ram Avtar; Negi, Narendra Singh; Ramesh, V; Salotra, Poonam

    2017-03-23

    Leishmaniasis is a spectrum of diseases with great relevance to public health. Conventional diagnostic methods are time consuming, needing trained personnel. A robust, rapid and cost effective diagnostic test is warranted for on-time diagnosis and field application. We have developed a loop mediated isothermal amplification (LAMP) assay with primers (n = 6) based on Leishmania donovani kDNA for detection of Leishmania infection, using a closed tube to prevent cross-contamination. The assay was used to detect Leishmania infection in biological samples obtained from patients of visceral leishmaniasis (VL), post kala-azar dermal leishmaniasis (PKDL) and cutaneous leishmaniasis (CL). The assay was positive for L. donovani, L. tropica and L. major parasites, with the highest sensitivity towards L. donovani (1 fg DNA). The high sensitivity of the assay for detection of L. donovani was reflected in its ability to detect parasite DNA within 30 min of amplification time with a threshold detection limit of ≥25 copies per reaction. The assay detected parasite in 64 of 66 VL blood samples (sensitivity, 96.9%; 95% CI: 89.6-99.2%), 15 of 15 VL bone marrow aspirate samples (sensitivity, 100%; 95% CI:79.6-100%), 65 of 67 PKDL tissue biopsy samples (sensitivity, 97%; 95% CI:89.7-99.2%). The assay was evaluated in a few cases of CL wherein it was found positive in 8 of 10 tissue biopsies (sensitivity, 80%; 95% CI: 49-94.3%). The assay was negative in all control blood (n = 76) and tissue biopsy (n = 24) samples (specificity, 100%; 95% CI: 96.3-100%). Further, the assay was evaluated for its utility in assessment of cure in treated VL and PKDL patients. The assay detected parasite DNA in 2 of 20VL blood samples and 2 of 21 PKDL tissue samples. Out of 4 cases that were positive for parasite DNA at post treatment stage, 2 patients (1VL and 1 PKDL) returned with relapse. The study demonstrated a Leishmania genus specific closed tube LAMP assay for reliable and rapid

  7. Anti-leishmanial evaluation of fraxinus xanthoxyloides (G. don) DC. collected from district Islamabad

    International Nuclear Information System (INIS)

    Younis, T.; Khan, M.R.; Zai, J.A.; Khan, H.; Shah, N.A.

    2016-01-01

    In this study, we have investigated phytochemical classes, antileishmanial and cytotoxic activity of Fraxinus xanthoxyloides (Oleaceae) leaves. Powder of F. xanthoxyloides leaves was extracted with methanol to obtain the crude extract (FXM) and the resultant was fractionated with solvents in escalating polarity; n-hexane (FXH), chloroform (FXC), ethyl acetate (FXE), n-butanol (FXB) and the residual aqueous (FXA) fraction. Quantitative estimation of terpenoids, coumarins, flavonoids, phenolics and tannins was conducted. Anti-leishmanial activity was performed against Leishmania tropica promastigote stage parasite while insecticidal activity was assessed through brine shrimps lethality assay. Our results showed the maximum concentration of terpenoids in FXC while the highest quantity of coumarins, flavonoids, phenolics and tannins was recorded in FXE. Presence of terpenoids was not detected in FXB and in FXA. Among the extract/fractions, FXC exhibited the highest anti-leishmanial activity with LD/sub 50/ of 15.239+-0.9 mu g/ml to that of glucantime (LD/sub 50/ = 5.6+-2.4 mu g/ml) a reference drug. FXH exhibited the anti-leishmanial activity of LD/sub 50/ = 40.68+-1.9 mu g/ml followed by FXE (LD/sub 50/ = 102.9+-3.1 mu g/ml). Similarly potent insecticidal activity was recorded (LD/sub 50/ = 28.15+-1.8 mu g/ml) for FXC followed by FXH (LD/sub 50/ = 67.59+-2.3 mu g/ml). However, other fractions exhibited low level of anti-leishmanial and insecticidal activity. Correlation analysis exhibited a strong association (p 0.05) with the insecticidal activity. The coumarins established a medium association with the insecticidal activity. Other chemical classes exhibited a moderate to low level of association with the anti-leishmanial and the insecticidal activity. On the basis of these results we can conclude that chloroform fraction of F. xanthoxyloides is a potential source for anti-leishmanial and insecticidal activities and further studies are required to isolate the active

  8. The new situation of cutaneous leishmaniasis after Syrian civil war in Gaziantep city, Southeastern region of Turkey.

    Science.gov (United States)

    Özkeklikçi, Ahmet; Karakuş, Mehmet; Özbel, Yusuf; Töz, Seray

    2017-02-01

    Cutaneous leishmaniasis (CL) is an important public health problem with around 2.000 autochthonous reported cases each year in Turkey. Due to the civil war in Syria, Turkey received around three million refugees and they are mainly located at either camps or homes in south/southeastern part of Turkey. In the present study, we aimed to collect samples from CL suspected patients admitting to State Hospital in Gaziantep City and perform parasitological and DNA-based techniques for diagnosis as well as species identification of the parasite for better understanding the prevalence of each species among Turkish and Syrian patients in the region. The collection of samples was carried out between January 2009 and July 2015. The lesion aspiration samples were taken and stained with Giemsa stain followed by microscopical examination for parasitological diagnosis. After the DNA extraction from Giemsa stained slides, real time and semi-nested PCRs both targeting ITS1 region were performed for molecular diagnosis and species identification. A total of 567 people were admitted to the hospital with the suspicion of CL and 263 (46.4%) of them were found to be positive by parasitological examination. One hundred seventy-four (66.15%), 88 (33.46%) and 1 (0.38%) of them were Turkish, Syrians and Afghan, respectively. Slide samples obtained from 34 CL suspected patients were analyzed by PCR and 20 of them were found positive. Eighteen (13 Turkish and 13 Syrians) of the positive samples were identified as L. tropica, while two (1 Turkish and 1 Syrian) of them were L. infantum. In conclusion, the effects of Syrian civil war on the epidemiology of CL in Gaziantep city is demonstrated in the present study. The use of molecular tool in the diagnosis of leishmaniasis is effective, sensitive and time saving which will enable the species typing. Species typing of the causative agent in endemic areas will bring valuable data to epidemiological knowledge. Copyright © 2016 Elsevier B.V. All

  9. Diagnosis of clinical samples spotted on FTA cards using PCR-based methods.

    Science.gov (United States)

    Jamjoom, Manal; Sultan, Amal H

    2009-04-01

    The broad clinical presentation of Leishmaniasis makes the diagnosis of current and past cases of this disease rather difficult. Differential diagnosis is important because diseases caused by other aetiologies and a clinical spectrum similar to that of leishmaniasis (e.g. leprosy, skin cancers and tuberculosis for CL; malaria and schistosomiasis for VL) are often present in endemic areas of endemicity. Presently, a variety of methods have been developed and tested to aid the identification and diagnosis of Leishmania. The advent of the PCR technology has opened new channels for the diagnosis of leishmaniasis in a variety of clinical materials. PCR is a simple, rapid procedure that has been adapted for diagnosis of leishmaniasis. A range of tools is currently available for the diagnosis and identification of leishmaniasis and Leishmania species, respectively. However, none of these diagnostic tools are examined and tested using samples spotted on FTA cards. Three different PCR-based approaches were examined including: kDNA minicircle, Leishmania 18S rRNA gene and PCR-RFLP of Intergenic region of ribosomal protein. PCR primers were designed that sit within the coding sequences of genes (relatively well conserved) but which amplify across the intervening intergenic sequence (relatively variable). These were used in PCR-RFLP on reference isolates of 10 of the most important Leishmania species: L. donovani, L. infantum, L. major & L. tropica. Digestion of PCR products with restriction enzymes produced species-specific restriction patterns allowed discrimination of reference isolates. The kDNA minicircle primers are highly sensitive in diagnosis of both bone marrow and skin smears from FTA cards. Leishmania 18S rRNA gene conserved region is sensitive in identification of bone marrow smear but less sensitive in diagnosing skin smears. The intergenic nested PCR-RFLP using P5 & P6 as well as P1 & P2 newly designed primers showed high level of reproducibility and sensitivity

  10. The development of Leishmania turanica in sand flies and competition with L. major.

    Science.gov (United States)

    Chajbullinova, Alsu; Votypka, Jan; Sadlova, Jovana; Kvapilova, Katerina; Seblova, Veronika; Kreisinger, Jakub; Jirku, Milan; Sanjoba, Chizu; Gantuya, Sambuu; Matsumoto, Yoshitsugu; Volf, Petr

    2012-10-02

    In Central Asian foci of zoonotic cutaneous leishmaniases, mixed infections of Leishmania turanica and L. major have been found in a reservoir host (the great gerbil, Rhombomys opimus) as well as in the sand fly vector Phlebotomus papatasi, but hybrids between these two Leishmania species have never been reported. In addition, the role of sand fly species other than P. papatasi in L. turanica circulation is not clear. In this work we compared the development of L. turanica in three sand fly species belonging to different subgenera. In addition, we studied experimental co-infections of sand flies by both Leishmania species using GFP transfected L. turanica (MRHO/MN/08/BZ18(GFP+)) and RFP transfected L. major (WHOM/IR/-/173-DsRED(RFP+)). The possibility of Leishmania genetic exchange during the vectorial part of the life cycle was studied using flow cytometry combined with immunofluorescent microscopy. Late-stage infections of L. turanica with frequent colonization of the stomodeal valve were observed in the specific vector P. (Phlebotomus) papatasi and in the permissive vector P. (Adlerius) arabicus. On the other hand, in P. sergenti (the specific vector of L. tropica), L. turanica promatigotes were present only until the defecation of bloodmeal remnants. In their natural vector P. papatasi, L. turanica and L. major developed similarly, and the spatiotemporal dynamics of localization in the sand fly gut was the same for both leishmania species. Fluorescence microscopy in combination with FACS analyses did not detect any L. major / L. turanica hybrids in the experimental co-infection of P. papatasi and P. duboscqi. Our data provide new insight into the development of different leishmania parasite species during a mixed infection in the sand fly gut. Despite the fact that both Leishmania species developed well in P. papatasi and P. duboscqi and did not outcompete each other, no genetic exchange was found. However, the ability of L. turanica to establish late

  11. Desenvolvimento Inicial de Duas Variedades de Cana-de-açúcar Inoculadas com Bactérias Diazotróficas

    Directory of Open Access Journals (Sweden)

    Valfredo Almeida Chaves

    2015-12-01

    Full Text Available RESUMO A cana-de-açúcar é uma cultura de grande destaque na economia, em razão da produção de açúcar, etanol e energia. Tecnologias que possam contribuir para o aumento da produtividade e qualidade da cultura com mínimos danos ao meio ambiente são necessárias. Objetivou-se avaliar a produção de ácido indol acético de cinco estirpes de bactérias diazotróficas e o efeito da inoculação delas na brotação de duas variedades de cana-de-açúcar, RB867515 e IACSP95-5000. A produção de auxina foi determinada pelo teste colorimétrico, usando o reagente de Salkowski. Para avaliar a germinação, foi conduzido um experimento em casa de vegetação, utilizando-se delineamento experimental em blocos ao acaso com quatro repetições e sete tratamentos: controle não inoculado; inoculação mista com as cinco estirpes e inoculação individual com Gluconacetobacter diazotrophicus (Gd estirpe BR11281T(PAL-5T, Herbaspirillum seropedicae (Hs - BR11335 = HRC54, Herbaspirillum rubrisubalbicans (Hr - BR11504 = HCC103, Burkholderia tropica (Bt - BR11366T = PPe 8 T e Azospirillum amazonense (Aa - BR11145 = CBAMc. As bactérias mais eficientes na produção de auxina foram Hs e Hr, declinando 48 h após o crescimento. Hr, Aa e Bt aumentaram o índice de velocidade de germinação e o número de brotações nas duas variedades. Na var. RB867515, a velocidade de germinação ainda foi positivamente influenciada pela inoculação mista, sendo o mesmo observado pela inoculação de Gd na var. IACSP95-5000.

  12. Chemical composition of Mentha pulegium and Rosmarinus officinalis essential oils and their antileishmanial, antibacterial and antioxidant activities.

    Science.gov (United States)

    Bouyahya, Abdelhakim; Et-Touys, Abdeslam; Bakri, Youssef; Talbaui, Ahmed; Fellah, Hajiba; Abrini, Jamal; Dakka, Nadia

    2017-10-01

    The aim of the study was the determination of the chemical composition of Mentha pulegium L. and Rosmarinus officinalis L. essential oils and the evaluation of their antileishmanial, antibacterial and antioxidant activities. Essential oils (EOs) were isolated using steam distillation and the chemical composition was determined using GC-MS analysis. The antibacterial activity was tested against ten pathogenic strains using the diffusion method, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) by microtitration assay. The antioxidant activity was estimated by DPPH free radical scavenging ability and ferric-reducing power. The antileishmanial activity was tested against Leishmania major, Leishmania tropica and Leishmania infantum using MTT (3-(4.5-dimethylthiazol-2yl)-2.5-diphenyltetrazolium bromide) assay. The yield of essential oils (v/w %) M. puleguim and R. officinalis based on dry weight were 5.4 and 2.7% respectively. GC/MS analysis of R. officinalis essential oil (ROEO) revealed the presence of 29 components, mainly represented by oxygenated monoterpenes (63.743%) and hydrocarbons monoterpenes (21.231%). Mentha pulegium essential oil (MPEO) revealed 21 components, mainly represented by oxygenated monoterpenes (83.865%). The major components of ROEO were α-pinene (14.076), 1,8-Cineole (23.673) and camphor (18.743), while menthone (21.164) and pulegone (40.98) were the main major components of MPEO. M. pulegium and R. officinalis EOs showed a significant antioxidant activity compared with ascorbic acid and Trolox to the IC 50 values of 58.27 ± 2.72 and 85.74 ± 7.57 μg/mL respectively revealed by reducing power assay. As for the antibacterial effect, the highest zone diameters were shown by the MPEO against Bacillus subtilis (30 ± 1.43 mm) and Proteus mirabilis (28 ± 1.32 mm). These values are significantly important compared with those of the commercialized antibiotic (Erythromycin and

  13. The spider family Micropholcommatidae (Arachnida: Araneae: Araneoidea: a relimitation and revision at the generic level

    Directory of Open Access Journals (Sweden)

    Michael Rix

    2010-02-01

    .; the 20 previously described species of Textricella are thus transferred into Eterosonycha or other newly described genera. The Textricellini includes 10 genera from Australasia and Chile: Eterosonycha has four species, including the type E. alpina Butler (=Textricella parva Hickman syn. n., E. complexa (Forster, E. aquilina sp. n. and E. ocellata sp. n.; Epigastrina gen. n. has three species, including the type E. fulva (Hickman, E. loongana sp. n. and E. typhlops sp. n.; Guiniella gen. n. is monotypic, with G. tropica (Forster; Raveniella gen. n. has three species, including the type R. luteola (Hickman, R. hickmani (Forster and R. peckorum sp. n.; Rayforstia gen. n. has 12 species, including the type R. vulgaris (Forster, the two new species R. lordhowensis sp. n. and R. raveni sp. n., and the nine additional species R. antipoda (Forster, R. insula (Forster, R. mcfarlanei (Forster, R. plebeia (Forster, R. propinqua (Forster, R. salmoni (Forster, R. scuta (Forster, R. signata (Forster and R. wisei (Forster; Normplatnicka gen. n. has three species, including the type N. lamingtonensis (Forster, N. chilensis sp. n. and N. barrettae sp. n.; Eperiella gen. n. has two species, including the type E. alsophila sp. n., and E. hastings sp. n.; Algidiella gen. n. is monotypic, with A. aucklandica (Forster; Taliniella gen. n. has two species, including the type T. nigra (Forster, and T. vinki sp. n.; and Tinytrella gen. n. is monotypic, with T. pusilla (Forster. The micropholcommatine tribe Patelliellini trib. n. includes only one monotypic genus, Patelliella gen. n., represented by the enigmatic species Patelliella adusta sp. n. from Lord Howe Island. The subfamily Taphiassinae subfam. n. includes two genera of distinctive, heavily punctate Micropholcommatidae from Australasia: Taphiassa Simon has six species, including the type T. impressa Simon, T. punctata (Forster, T. castanea sp. n., T. globosa sp. n., T. magna sp. n. and T. robertsi sp. n.; the genus Parapua Forster

  14. Alterações cutâneas do cão no Kala-Azar sul-americano

    Directory of Open Access Journals (Sweden)

    C. Magarinos Torres

    1941-01-01

    YAKIMOFF & KOHL-YAKIMOFF (1911 in Tunis, by YAKIMOFF (1915 in Turkestan. Eczematous areas or a condition described as "eczema furfurace" is sometimes noted in the areas of depilation (DONATIEN & LESTOCQUARD. The skin changes noticed by ADLER & THEODOR (1932 in dogs naturally infected with Mediterranean Kala Azar can be briefly summarized as a selective infiltration of macrophages around hair follicles including the sebaceous glands and the presence of infected macrophages in normal dermis. The latter phenomenon in the complete absence of secondary infiltration of round cells and plasma cells is the most striking characteristic of canine Kala Azar and differentiates it from L. tropica. In the more advanced stages the dermis is more cellular than that of normal dogs and may even contain a few small dense areas of infiltration with macrophages and some round cells and polymorphs. The external changes, i. e., seborrhea and depilation are roughly proportional to the number of affected hair follicles. In dogs experimentally infected with South-American Kala Azar the parasites were regularly found in blocks of skin removed from the living animal every fortnight (CUNHA, 1938. The changes noticed by CUNHA, besides the presence of Leishmania, were perivascular and diffuse infiltration of the cutis with mononuclears sometimes more marked near hair follicles, as well as depilation, seborrhea and ulceration. The parasites were first discovered and very numerous in the paws. Our material was obtained from dogs experimentally infected by Dr. A. MARQUES DA CUNHA< and they were the subject of a previous paper by CUNHA (1938. In this study, however, several animals were discarded as it was found that they did develop a superimposed infection by Demodex canis. This paper deals with the changes found in 88 blocks of skin removed from five dogs, two infected with two different canine strains, and three with two distinct human strains of South-American Kala Azar. CUNHA'S valuable material

  15. Catalogue of Tenebrionidae (Coleoptera of North America

    Directory of Open Access Journals (Sweden)

    Yves Bousquet

    2018-01-01

    Conte, 1858]; Metablapylis Blaisdell, 1909 [Eleodes nigrina LeConte, 1858]; Steneleodes Blaisdell, 1909 [Eleodes longicollis LeConte, 1851]; Scaptes Champion, 1886 [Scaptes squamulatus Champion, 1886 (= Asida tropica Kirsch, 1866]; Aspidius Mulsant and Rey, 1859 [Blaps punctata Fabricius, 1792]; Cryptozoon Schaufuss, 1882 [Cryptozoon civile Schaufuss, 1882]; Halophalerus Crotch, 1874 [Phaleria rotundata LeConte, 1851]; Dignamptus LeConte, 1878 [Dignamptus stenochinus LeConte, 1878]. Two species previously known from South America [Nilio lebasi J. Thomson and Platydema erotyloides Chevrolat] are reported for the first time from North America.