Deep intraspecific divergences in the medically relevant fat-tailed scorpions (Androctonus, Scorpiones)

The genus Androctonus, commonly known as fat-tailed scorpions, contains 22 species distributed from Togo and Mauritania in the west, North Africa, through the Middle East and to as far east as India. With 13 species, a substantial amount of this genus’ diversity occurs in North Africa, which is a major hotspot of scorpion sting incidents. Androctonus are among the most medically relevant animals in North Africa. Since venom composition within species is known to vary regionally, the improvement of therapeutic management depends on a correct assessment of the existing regional specific and sub-specific variation. In this study, we assessed the phylogeographical patterns in six species of Androctonus scorpions from North Africa using mitochondrial DNA markers. We sequenced COX1, 12S, 16S and ND1 genes from 110 individuals. Despite lacking basal resolution in the tree, we found taxonomical and geographically coherent clades. We discovered deep intraspecific variation in the widespread Androctonus amoreuxi and Androctonus australis, which consisted of several well-supported clades. Genetic distances between some of these clades are as high as those found between species. North African A. australis have a deep split in Tunisia around the Chott el-Djerid salt-lake. A novel split between A. amoreuxi scorpions was found in Morocco. We also found deep divergences in Androctonus mauritanicus, corresponding to areas attributed to invalidated subspecies. In addition we uncovered a clade of specimens from coastal south Morocco, which could not be ascribed to any know species using morphological characters. Based on these findings we recommend a reassessment of venom potency and anti-venom efficacy between these deep intraspecific divergent clades.     

Inter- and intrapopulational genetic variability of Tityus serrulatus (Scorpiones, Buthidae)

In Brazil, there are near 20 genera and almost 120 species of scorpions of which 95% reproduce sexually. Parthenogenetic reproduction, however, may also take place. To gain insight into useful molecular markers in parthenogenetic scorpion species, we studied DNA polymorphism using two molecular approaches: simple sequence repeat anchored polymerase chain reaction (SSR-PCR) and sequencing of the cytochrome C oxidase subunit I of the mitochondrial genome, mtDNA (COXI), of Tityus serrulatus. Three different groups were used: group 1, composed of 1 female and 14 descendants; group 2 with 1 female and 17 descendants, both from the city of Uberlândia, State of Minas Gerais (MG), Brazil, and the third group that consisted of three adult scorpions from the city of Belo Horizonte, MG.The profiles generated by SSR-PCR were identical for all specimens, while partial sequencing of COXI showed the presence of SNPs. After aligning COXI contigs, one of the groups presented 18 SNPs and the second 8 SNPs. The two groups were differentiated by two diagnostic SNPs. We did not find evidence of mitochondrial recombination.The results are in agreement with the parthenogenetic mode of reproduction of this species and sequencing of the COXI gene enabled the separation of scorpions groups.     

Genetic analysis of ticks belonging to the Rhipicephalus sanguineus group in Latin America

Phylogenetic analyses based on mitochondrial 16S rDNA sequences were generated from Rhipicephalus sanguineus group specimens collected in 29 localities among 9 Latin-American countries, plus ticks collected in South Africa, Spain, and Italy. Sequences from Latin America generated six different haplotypes (A, B, C, D, E, and F). Phylogenetic analyses generated trees that segregated our tick sequences into two distinct clades: one is represented by haplotypes A–C, and South African R. sanguineus and Rhipicephalus turanicus ticks; the second clade is represented by haplotypes D–F, and European R. sanguineus and R. turanicus ticks. When haplotypes A–F are plotted in the Latin America map according to their geographical coordinates, it is clearly seen that haplotypes D–F are restricted to the southern portion of this continent, whereas haplotypes A–C are distributed in areas between northern Mexico and Brazil (except for the extreme south of this last country, where haplotype E was present). Hence, our phylogenetic analyses separated New World specimens of R. sanguineus into two distinct clades, one represented by tropical and subtropical populations (haplotypes A–C), here designated as the ‘tropical’ species. On the other hand, haplotypes D–F are here designated as the ‘temperate’ species because of their distribution in the southern portion of South America. Until recently, it was assumed that the R. sanguineus group was represented by a single species in the New World, namely R. sanguineus. While the present results coupled with recent studies support the presence of at least two species under the taxon R. sanguineus in the New World, they also show that even in the Old World, the taxon R. sanguineus might be represented by more than one species, since our phylogenetic analysis segregated European and South African R. sanguineus ticks into two distinct clades. The same can be applied for Spanish and South African R. turanicus.     

Resistance to pyrethroid insecticides in Culex pipiens pipiens (Diptera: Culicidae) from Tunisia

Resistance toward 2 pyrethro?d insecticides (permethrin and deltamethrin), and an organochloric one (DDT) was analyzed in 18 samples of Culex pipiens pipiens common mosquitoes collected from different Tunisian areas between March 2002 and November 2005. Bioassays were performed over different larvae samples. The recorded mortalities, after 24h exposure to increased doses of insecticides, were compared to those obtained on a susceptible reference strain (S-LAB). All samples were resistant to permethrin. However, a large variation in the tolerance to this insecticide was observed between samples. Resistance ratio levels at LC50 (RR50) for the analyzed samples were ranged from 1.5 to 9092 folds. For deltamethrin, 14 among 17 studied samples were resistant. RR50, in the resistant samples, were ranged from 3 to 453 folds. Resistance to the two tested pyrethroids and DDT were correlated. The use of synergists showed that the cytochrome P450-dependent monooxygenases had a major contribution to the permethrin and deltamethrin resistance and that the esterases (and/or glutathione-S-transferases) had only a minor contribution. Results were discussed in relation to resistance mechanisms and mosquito's control.     

Species B of Anopheles culicifacies (Diptera: Culicidae) is reproductively less fit than species A and C of the complex

Anopheles culicifacies, the most important malaria vector of peninsular India exist as a complex of five sibling species. The member species of the complex have various biological differences including their susceptibility to malaria parasites. The present attempt is made to study and compare the fecundity of the differentially susceptible members of the An. culicifacies complex. Gravid female mosquitoes of species A, B and C were allowed to lay their eggs individually during first and second gonotrophic cycle. The eggs were counted after hatching and categorized as ‘hatched eggs’, ‘unhatched eggs’, ‘embryonated eggs’, ‘unembryonated eggs’ and ‘non/partially melanized eggs’. The data was analyzed using Student's t test, ANOVA, Chi-square and Pearson's correlation analysis. All females that were visually categorized as ‘gravid’ did not lay eggs. Species C laid maximum number of eggs per female. The eggs laid per female mosquito of each species were found to be significantly higher during second gonotrophic cycle as compared with the first gonotrophic cycle. The eggs hatched per female in species C were found to be significantly higher than that of species A and B. The poor-vector species B mosquitoes were found to be the least fecund among the members of the species complex. The unembryonated eggs constitute the biggest proportion of the unhatched eggs in species A, B and C of the species complex.     

Endoparasites of red fox (Vulpes vulpes) in the Slovak Republic with the emphasis on zoonotic species Echinococcus multilocularis and Trichinella spp.

Summary  Due to specific geographical localization, climatic and geomorphologic conditions, several serious parasitic diseases circulate in the territory of the Slovak Republic that makes this area an ideal model territory of the central European red fox system. The red fox is an important reservoir host of parasites, which can be spread to another animals and humans. Our study was aimed at determining the current prevalence of certain parasites in red foxes from the entire territory of the Slovak Republic and identifies some ecological factors influencing their epidemiology. Within the first systematic investigation of red foxes carried out between the years 2000 and 2006 in total 4026 foxes were examined for Echinococcus multilocularis (prevalence 31.1 %) and 4699 foxes were investigated for the presence of Trichinella spp. larvae (10.4 % infected). The results of the next separate study revealed that 83.3 % of 1198 red foxes in the Slovak Republic had coccidian oocysts and helminth eggs in their faeces. Fifteen helminth species including two trematode, four cestode and nine nematode species were detected by coprological examination. Nine of these parasite taxa have zoonotic potential: Capillaria spp. (prevalence 22.4 %), Ancylostoma caninum (18.1 %), Toxocara canis (12.5 %), Taenia spp. (12.2 %), Mesocestoides spp. (5.8 %), Strongyloides stercoralis (1.6 %), Hymenolepis diminuta (0.6 %), Dipylidium caninum (0.4 %) and Opisthorchis felineus (0.3 %). Toxascaris leonina was the most common helminth species found in this survey (42.9 %).     

New molecular marker for Trypanosoma (Duttonella) vivax identification

Trypanosoma vivax is a widespread hemoparasite in tropical areas and is pathogenic to ruminant domestic livestock as well as wild ruminants. The accurate identification of parasites in both hosts and vectors is crucial for epidemiological studies and disease control programs. We describe here the development of molecular markers specific for T. vivax identification. These markers were used to identify mouthpart infections in field-collected tsetse flies from Cameroon. The markers target the genomic sequence of a species-specific antigen from the bloodstream stages. No cross amplification with other trypanosome species was observed, which makes the markers a reliable tool to detect T. vivax infections, both in hosts and vectors. The PCR-amplified sequence contains a (CA)_n microsatellite repeat for which 11 different alleles were identified. This microsatellite, which showed high polymorphism, provides a suitable marker for population genetic studies.     

Human cutaneous leishmaniasis due to Leishmania infantum in the Sidi Bourouis focus (Northern Tunisia): epidemiological study and isoenzymatic characterization of the parasites

The authors report an increase of the number of case of cutaneous leishmaniasis in the Sidi Bourouis region community of Siliana Governorate (Tunisia), with 38 cases diagnosed in 6 months (1st November 2000-30th April 2001), contrary to its usual sporadic character. The isoenzymatic identification of 15 isolated strains emphasizes the role of the Leishmania infantum zymodeme MON-1 as an important factor in the genesis of the sporadic cutaneous leishmaniasis of Northern Tunisia. In fact it was isolated in 6 cases while L. infantum MON-24, the usual agent was isolated in 9 cases.     

Molecular characterization of the invasive Asian tiger mosquito, Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Corsica

The Asian tiger mosquito Aedes albopictus, vector of various human viruses and parasites, has recently spread and established in many temperate regions including European countries. In the present study, we developed a simple PCR-based assay (the amplification of the internal transcribed spacer ITS2 within nuclear ribosomal rDNA) for molecular identification of A. albopictus and confirmed its presence in Corsica island. This assay may (i) facilitate future large scale studies and avoid misidentifications, especially because of the presence of co-occurring close species in this island and (ii) contribute to the monitoring of A. albopictus populations required for targeted control programs.     

Fuhrmanolepis beskydensis n.sp. (Cestoda: Dilepididae) from woodcock Scolopax rusticola L.(Aves, Charadriformes) in Slovakia with comments on sytematics and nomenclature of the genus Fuhrmanolepis Spassky et Spasskaya, 1965

Summary   Fuhrma n olepis beskydensis n.sp. from woodcock, Scolopax rusticola L. in Slovakia is described based upon light microscope observation. The medium sized species possess a single crown of 21‘diorchid’ (wrench-shaped) hooks, 28–30 μm long. Irregularly alternating genital pores were combined with abnormal multiple shifting of pores within the lateral margins of strobila. Number of testes 18–25. Cirrus-sac and evaginated cirrus are 115–135 × 7–12 and 26 × 6–11 μm, respectively. The species is differentiated from closely related congeneric taxons and some other morphologically similar dilepidids. An attention is being paid to taxonomy and nomenclature of Fuhrma n olepis Spassky et Spaskaja, 1965 regarding an emendation of mentioned genus to Fuhrma nn olepis by Bona (1994a) and modification of its diagnosis.     

Molecular phylogeography of Culex quinquefasciatus mosquitoes in central Bangladesh

Mosquitoes in the Culex pipiens complex are a major vector of numerous parasitic and arboviral diseases. Here we report the phylogeography of a prevalent Culex mosquito, Cx. quinquefasciatus, from three locations in Bangladesh: Dhaka, Savar and Mymensingh. Sequence analysis of the genes encoding mitochondrial cytochrome oxidase subunit II, nuclear elongation factor-1 alpha, and acetylcholinesterase-2 revealed the lack of a population genetic structure among the three locations. Moreover, the highly divergent ribosomal internal transcribed spacer 2 suggests that this locus has not evolved in concert. The results further show evidence of historical introgression of internal transcribed spacer 2 from Cx. pipiens to Cx. quinquefasciatus of Bangladesh, and that the introgression occurred before Cx. quinquefasciatus had dispersed within this region. The study also reveals historical population expansion in this region, followed by a post-expansion Wolbachia sweep.     

Stichorchis subtriquetrus in two beavers (Castor fiber) from Slovenia

Summary  Intestines of two beavers (Castor fiber), one killed by a car accident and one drowned were examined for parasites. Examination revealed the presence of 1 and 14 adult trematodes, Stichorchis subtriquetrus in the caecum and colon, respectively. The flotation method revealed no ova from faecal material. The European beaver was present in the territory of Slovenia until the beginning of the 18^th century. Recently, in 1998, the species reappeared in southeast of Slovenia as a result of the reintroduction of European beaver in Croatia. This is the first report of veterinary examination of beaver and the first evidence of S. subtriquetrus in Slovenia.     

Stenotrophomonas (Xanthomonas) maltophilia infection in necrotizing pancreatitis

Summary Conclusion: Although the therapy of infected pancreatic collections or organized pancreatic necrosis remains surgical, we have demonstrated that infected organized pancreatic necrosis can be treated endoscopically. Background: Stenotrophomonas (Xanthomonas) maltophilia has been increasingly recognized as a nosocomial pathogen associated with meningitis, pneumonia, conjunctivitis, soft tissue infections, endocarditis, and urinary tract infections. This organism is consistently resistant to imipenem, a drug commonly employed in patients with necrotizing pancreatitis to prevent local and systemic infections. Methods and Results: We report the first case of infected pancreatic necrosis by S. (X.) maltophilia. Our patient was treated successfully with endoscopic drainage of the pancreatic fluid collection and appropriate antibiogram-based antibiotic therapy. Endoscopic drainage has emerged as one of the treatment modalities for pancreatic fluid collections.     

Redescription of Desportesius brevicaudatus (Spirurida, Acuariidae) based on nematodes from Ixobrychus minutus (Aves, Ciconiiformes) from Bulgaria

Summary   Desportesius brevicaudatus (Dujardin, 1845) is redescribed on the basis of light-microscopy and SEM observations on specimens collected from the oesophagus and under the lining of the gizzard of Ixobrychus minutus (Ardeidae) from Bulgaria. New metrical data expand the known ranges of variation of the measurements of the body, tail, cordons, oesophagus and spicules. New information is provided on the variation of the shape of the deirids, the structure of the vagina, the complexity of the cordons, the pattern of the cuticular striation and the shape of the postdeirids. Cordons are described as consisting of a single row of serrate cuticular plates and a longitudinal cuticular ridge along the outer rims of the cuticular plates. The cuticular ridge is interpreted as homologous to the outer row of plates in the cordons of the genera Acuaria, Cheilospirura and Echinuria.     

Inhibition of the growth of Plasmodium falciparum and Plasmodium berghei in vitro by an extract of Cochlospermum angolense (Welw.)

An extract of Cochlospermum angolense (Welw.) is used in the traditional medicine of Angola for the therapy of icterus and for the prophylaxis of malaria. From the roots of this plant red crystalline substances have been isolated and tested for their effect on Plasmodium falciparum in vitro and on the DNA and protein synthesis of Plasmodium berghei. The multiplication of P. falciparum was decreased to 50% of the control in the presence of 10 μg/ml extracted material and there was a total inhibition at a concentration of 50 μ/ml. If mice erythrocytes infected by P. berghei were incubated for 6 h with 25 μg/ml of the extract DNA synthesis was depressed to nearly background level. And, even more important, this effect could be demonstrated immediately. On the contrary, protein synthesis continued for at least 90 min at a reduced rate and stopped then. The results obtained show the direct antiparasitic effect of the substances extracted from C. angolense. The activity seems to be directed against DNA synthesis.     

First record of Steinernema feltiae (Filipjev) (Rhabditida: Steinernematidae) in Slovenia

Summary  In October 2007 we examined 80 soil samples from 16 different locations in the central part of Slovenia (the Notranjska region) and confirmed the presence of entomopathogenic nematodes only in two soil samples. This represents the first recorded instance of an entomopathogenic nematode in Slovenia. In sample B30 we confirmed the presence of Steinernema feltiae (Rhabditida: Steinernematidae) by means of a molecular technique. In Slovenia the application of entomopathogenic nematodes was hitherto possible only in laboratory experiments, while the Rules on Biological Plant Protection made the practical application of exotic organisms in the domestic environment entirely impossible. After the first record of the entomopathogenic nematode S. feltiae we expect the aformentioned agent to become an important alternative to insecticides in plant protection against pest insects.     

Helminthofauna of Pelophylax kl. esculentus (Linne, 1758) from Petrovaradinski Rit Marsh (Serbia)

Summary  The helminth fauna of lungs, small intestine, and rectum of 83 female specimens of Pelophylax kl. esculentus was investigated. Three helminth groups, Trematoda, Nematoda, and Acanthocephala were recorded. The sample was dominated by the class Trematoda. This class was represented by 8 adult and 2 larval species. Opisthioglyphe ranae and Pleurogenoides medians were the dominant fluke species. The data show the occurrence of Acanthocephalus ranae in frogs inhabiting Vojvodina Province. The most frequent Nematoda was Oswaldocruzia filiformis. The greatest number of parasitic species and individuals was isolated from the small intestine. Infestation of individual parasitic groups was greater than that of mixed groups. The most frequent combination was T/N while analysed organs were most frequently invaded by a parasitic species only. Antagonism between genus Rhabdias and Haematoloechus and between species Aplectana acuminata and Opisthioglyphe ranae was ascertained.     

Distribution of the cestode Taenia parva (Taeniidae) along the digestive tract of the common genet (Genetta genetta)

Summary  The small intestine of the common genet is the habitat of Taenia parva (Taeniidae), a common parasite whose intermediate host is Apodemus spp. (Rodentia). Here we divided the small intestine of 51 common genets into sections and analysed them separately. Cestodes were classified into four categories: larval, pre-adult, adult and gravid specimens. A total of 4,443 individuals were recovered and classified: 543 larval stages, 2,326 pre-adults, 872 adults and 702 gravid specimens. A double analysis was performed by checking the distribution of the parasite along the digestive tract and comparing this distribution with reproductive fitness. This is the first study to test the border effect on the distribution of T. parva in its habitat. We also tested and demonstrated the Ideal Free Distribution Model (IFM model). A non-random distribution was found, with the lowest parasite frequencies in the top and the bottom portions of the small intestine. Maturity classes of cestodes vs. intestine portion were tested and no differences between portions were detected. We also analysed seasonal changes in parasite frequency. Log-linear analysis showed that the mean frequency of cestode occurrence per individual host was higher during winter, followed by spring, autumn and summer.     

A low concentration of azithromycin inhibits the mRNA expression of N-acyl homoserine lactone synthesis enzymes, upstream of lasI or rhlI, in Pseudomonas aeruginosa

Low-dose macrolides are effective therapy in patients with chronic lung infections, but the mechanisms of action are unclear. In this study, we performed DNA microarray analysis of Pseudomonas aeruginosa after treatment with a low concentration of azithromycin. We found that a sub-MIC of azithromycin didn't change mRNA expression of quorum-sensing related genes (lasI, lasR, rhlI, rhlR, vft, rsaL), but lowered expression of most N-acyl homoserine lactone (AHL) synthesis enzymes upstream of lasI and rhlI. We propose that small down regulation of these enzymes cumulatively resulted in a larger decrease of AHL production and inhibition of quorum-sensing in P. aeruginosa.