Plasmodium yoelii: axenic development of the parasite mosquito stages

Study of the parasite mosquito stages of Plasmodium and its use in the production of sporozoite vaccines against malaria has been hampered by the technical difficulties of in vitro development. Here, we show the complete axenic development of the parasite mosquito stages of Plasmodium yoelii. While we demonstrate that matrigel is not required for parasite development, soluble factors produced and secreted by Drosophila melanogaster S2 cells appear to be crucial for the ookinete to oocyst transition. Parasites cultured axenically are both morphologically and biologically similar to mosquito-derived ookinetes, oocysts, and sporozoites. Axenically derived sporozoites were capable of producing an infection in mice as determined by RT-PCR; however, the parasitemia was significantly much less than that produced by mosquito-derived sporozoites. Our cell free system for development of the mosquito stages of P. yoelii provides a simplified approach to generate sporozoites that may be for biological assays and genetic manipulations.     

Fine discrimination in the recognition of individual species of phosphatidyl-myo-inositol mannosides from Mycobacterium tuberculosis by C-type lectin pattern recognition receptors

The Mycobacterium tuberculosis (M.tb) envelope is highly mannosylated with phosphatidyl-myo-inositol mannosides (PIMs), lipomannan, and mannose-capped lipoarabinomannan (ManLAM). Little is known regarding the interaction between specific PIM types and host cell C-type lectin pattern recognition receptors. The macrophage mannose receptor (MR) and dendritic cell-specific ICAM-3-grabbing nonintegrin on dendritic cells engage ManLAM mannose caps and regulate several host responses. In this study, we analyzed the association of purified PIM families (f, separated by carbohydrate number) and individual PIM species (further separated by fatty acid number) from M.tb H(37)R(v) with human monocyte-derived macrophages (MDMs) and lectin-expressing cell lines using an established bead model. Higher-order PIMs preferentially associated with the MR as demonstrated by their reduced association with MDMs upon MR blockade and increased binding to COS-1-MR. In contrast, the lower-order PIM(2)f associated poorly with MDMs and did not bind to COS-1-MR. Triacylated PIM species were recognized by MDM lectins better than tetra-acylated species and the degree of acylation influenced higher-order PIM association with the MR. Moreover, only higher-order PIMs that bind the MR showed a significant increase in phagosome-lysosome fusion upon MR blockade. In contrast with the MR, the PIM(2)f and lipomannan were recognized by DC-SIGN comparable to higher-order PIMs and ManLAM, and the association was independent of their degree of acylation. Thus, recognition of M.tb PIMs by host cell C-type lectins is dependent on both the nature of the terminal carbohydrates and degree of acylation. Subtle structural differences among the PIMs impact host cell recognition and response and are predicted to influence the intracellular fate of M.tb.     

Decline in ascorbate peroxidase activity--a prerequisite factor for tepal senescence in gladiolus

Flower senescence was studied in Gladiolus cv. "Snow Princess" over five arbitrarily divided developmental stages (stage 1, half bloom; stage 2, full bloom; stage 3, beginning of wilting; stage 4, 50% wilting; stage 5, complete wilting) in terms of changes in fresh weight, antioxidant enzymes (superoxide dismutase, SOD; ascorbate peroxidase, APX; glutathione reductase, GR) activities and membrane integrity. A significant decrease in tepal fresh weight was observed over the senescence period (after stage 2). Membrane integrity was studied by measuring lipid peroxidation [in terms of thiobarbituric acid reactive substances (TBARS) content] and membrane stability index (MSI) percentage. Maximum TBARS content was recorded in stage 4 (50% wilting). This increase in lipid peroxidation over the senescence period was in close association with high degree of membrane deterioration expressed as decrease in membrane stability index percentage. A significant decrease (two and half-fold) in MSI% in stage 5 (as compared to stage 1) indicates complete membrane deterioration. Progressive increase in endogenous H2O2 level was recorded over senescence period. Maximum H2O2 content (19.7+/-1.4 micromol g(-1) DW) was recorded at stage 5 (complete wilting). Three different patterns were observed in antioxidant enzymes behavior over the senescence period. APX activity was declined significantly as, the flower entered stage 3 (beginning of wilting) from full bloom condition (stage 2). Progressive and significant increase in SOD activity was measured as a function of time. Maximum SOD activity (24.2+/-0.8 U mg(-1) DW) was recorded in stage 5 (three-fold increase over stage 1). GR activity initially increased up to stage 4 (50% wilting) and declined significantly thereafter (approximately seven-fold). An increase in endogenous H2O2 level during senescence may be the result of a programmed down-regulation of APX enzyme activity, which seems to be the prerequisite factor for initiating senescence process in gladiolus tepal.     

The evolutionary origin of Xanthomonadales genomes and the nature of the horizontal gene transfer process

Determining the influence of horizontal gene transfer (HGT) on phylogenomic analyses and the retrieval of a tree of life is relevant for our understanding of microbial genome evolution. It is particularly difficult to differentiate between phylogenetic incongruence due to noise and that resulting from HGT. We have performed a large-scale, detailed evolutionary analysis of the different phylogenetic signals present in the genomes of Xanthomonadales, a group of Proteobacteria. We show that the presence of phylogenetic noise is not an obstacle to infer past and present HGTs during their evolution. The scenario derived from this analysis and other recently published reports reflect the confounding effects on bacterial phylogenomics of past and present HGT. Although transfers between closely related species are difficult to detect in genome-scale phylogenetic analyses, past transfers to the ancestor of extant groups appear as conflicting signals that occasionally might make impossible to determine the evolutionary origin of the whole genome.     

The endangered cyprinodont <Emphasis Type="Italic">Aphanius ginaonis</Emphasis> (Holly, 1929) from southern Iran is a valid species: evidence from otolith morphology

Aphanius Nardo, 1827 (Actinopterygii, Cyprinodontidae) is a widely distributed genus in the Mediterranean and Persian Gulf area and includes several endangered species. The otolith morphology in Aphanius is known to represent a valuable tool for the taxonomy, and is also indicative for the genetic diversity of a particular population. The present study focuses on the otoliths of the endangered A. ginaonis (Holly, 1929), which is endemic to the Geno hot spring in southern Iran. The taxonomic status of A. ginaonis has repeatedly been questioned, and some scholars have argued that it merely represents a morphological variation of the widespread A. dispar. We present a comparison of the otolith morphology of A. ginaonis (52 specimens) with that of A. dispar (Rüppell, 1828) from the Mehran River Basin (southern Iran) (17 specimens) and an A. dispar population from the Persian Gulf coast of the United Arab Emirates (32 specimens). Our data obtained from SEM pictures, otolith morphometry and statistical analyses suggest that A. ginaonis represents a valid species. In A. ginaonis individuals with a standard length exceeding 23 mm, the otolith variables length–height and rostrum length represent useful complementary diagnostic characters discriminating this species from other Aphanius species. Besides ontogenetic variation, we found extremely high otolith form variability in A. ginaonis, including some otoliths with a morphology distinctly deviating from the basic morphology type. We hypothesize that these variations may be a result of the artificial introduction of A. dispar into the Geno hot spring during the last years and subsequent hybridisation.     

Variability in Symbiotic Effectiveness of Native Rhizobia in Acid Stress

Two strains of gram-negative bacteria isolated because of their abilities to decompose xenobiotic compounds were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA gene sequence analysis, the two strains were found to belong to the genus Pseudomonas. Benzene degrading strain DSM 8628^T was moderately related to P. flavescens NCPP 3063^T (98.3% similarity), P. monteilii CIP 104883^T, and P. plecoglossicida FPC 951^T (98.1%). Strain DSM 9751^T capable to grow with cetyltrimethylammonium chloride as the sole carbon source showed the highest similarity values with P. tremae CFBP 2341^T and P. meliae MAFF 301463^T (98.0%), both related to Pseudomonas syringae. The fatty acid pattern of strain DSM 8628^T was distinct from patterns of other members of the genus Pseudomonas in combining a high ratio of 3OH-C_12:1 (5.1%), a low ratio of 2OH-C_12:0 (0.2%) and a relatively low ratio of C_18:1ω7c (23.8%). On the basis of phylogenetic analysis, physiological properties and the composition of whole cell fatty acids, two novel species, Pseudomonas benzenivorans sp. nov. with the type strain DSM 8628^T (=CIP 109857^T) and Pseudomonas saponiphila sp. nov. with the type strain DSM 9751^T (=CIP 109856^T), are proposed.     

Detection, identification and molecular typing of Leishmania major in Phlebotomus papatasi from a focus of zoonotic cutaneous leishmaniasis in central of Iran

Leishmania major is the causative agent and Phlebotomus papatasi is the main vector of rural zoonotic cutaneous leishmaniasis (ZCL) in Iran and elsewhere. Nested PCR protocols were used to amplify a region of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene) in female P. papatasi. In the current investigation, L. major was found in Natanz, Isfahan province in centre of Iran, in a focus of rural ZCL. Ten (1.8%) out of 549 female P. papatasi was found to be infected with L. major based on the PCR detection and sequencing of parasite ITS-rDNA.Nine (1.8%) out of 498 female P. papatasi infected with L. major came from animal shelters, inside houses and yards. And one (1.9%) out of 51 female P. papatasi infected with L. major came from gerbil borrows. Infection rates were higher for females containing red blood meals, large eggs (semi-mature and mature) than for those without either blood meals or eggs. From the 10 infections detected three different haplotypes of L. major were identified. Two haplotypes were found to be novel. The other haplotypes of L. major was found to be identical to that of isolates of L. major from Iran and in elsewhere using GenBank data. Comparisons of infection rates between habitats will be inaccurate when the proportions of blood-fed and gravid flies differ among sandfly samples.     

Distribution of arbuscular mycorrhizal fungi in marshy and shoreline vegetation of Deepar Beel Ramsar Site of Assam,India

Arbuscular mycorrhizal fungal (AMF) symbiosis was thought to be rare in wetland plant roots, although several recent studies suggested that this association might be important in wetland ecosystems. In this research work we have studied the distribution of AMF in the marshy and shoreline vegetation of Deepar Beel Ramsar site of Assam, India. The study reveals the percentage of mycorrhizal colonization in the roots of different plant species which were observed from 20.89 to 86.47% and particularly found larger among the members of the family Poaceae. The Vetiveria zizanioides L. from the family Cyperaceae showed the highest (86.47%) percentage of root colonization, however, only one plant species viz. Scirpus lateriflorus Gmel. from the same family was found to be nonmycorrhizal. The rhizospheric soil samples of most of the plant species were found to be dominated by Glomus morphotypes. All total 18 AMF morphotypes were recorded which comprises four genera viz. Glomus (66.67%), Acaulospora (16.66%), Gigaspora (11.11%) and Scutellospora (5.56%). The observation of diversity of AMF in 25 different plant species among the wetland plants gives a glimpse of AMF diversity and their host selectivity in the said ecosystem.     

Envelope Proteins Pertain with Evolution and Adaptive Mechanism of the Novel Influenza A/H1N1 in Humans

The novel swine-origin influenza A/H1N1 virus (S-OIV) first detected in April 2009 has been identified to transmit from human to human directly and is the cause of currently emerged pandemic. In this study, nucleotide and deduced amino acid sequences of hemagglutinin (HA) and neuraminidase (NA) of the S-OIV and other influenza A viruses were analyzed through bioinformatic tools for phylogenetic analysis, genetic recombination and point mutation to investigate the emergence and adaptation of the S-OIV in human. The phylogenetic analysis showed that the HA comes from triple reassortant influenza A/H1N2 and the NA from Eurasian swine influenza A/H1N1 indicating HA and NA to descend from different lineages during the genesis of the S-OIV. Recombination analysis nullified the possibility of occurrence of recombination in HA and NA denoting the role of reassortment in the outbreak. Several conservative mutations are observed in the amino acid sequences of the HA and NA and this mutated residues are identical in the S-OIV. The results reported herein suggested the notion that the recent pandemic is the result of reassortment of different genes from different lineages of two envelope proteins, HA and NA which are responsible for antigenic activity of virus. This study further suggests that the adaptive capability of the S-OIV in human is acquired by the unique mutations generated during emergence.     

Metabolic characteristics and oxidative damage to skeletal muscle in broiler chickens exposed to chronic heat stress

Emerging evidence has shown that acute heat exposure affects metabolic characteristics and causes oxidative damage to skeletal muscle in birds. Little is known, however, about such phenomena under chronic heat stress conditions. To address this, we designed the present study to determine the influence of cyclic (32 to 24 to 32 °C: 32 °C for 8 h/d, 32–24–32HS ), and constant (32 and 34 °C, 32HS and 34HS, respectively) heat exposure on the metabolic and peroxide status in skeletal muscle of 4-wk-old male broiler chickens. Heat stress, particularly in the 32HS and 34HS groups, depressed feed intake and growth, while cyclic high temperature gave rise to a less severe stress response in performance terms. Malondialdehyde (MDA) levels in skeletal muscle were enhanced (P < 0.05) by constant heat treatment; the degree of enhancement was not as large as the changes observed in our previous ‘acute’ heat stress model. The 3HADH (3-hydroxyacyl CoA dehydrogenase related to fatty acid oxidation) and CS (citrate synthase) enzyme activities were lowered (P < 0.05) by both the cyclic and constant 34HS treatments, and constant 34HS group, respectively. These results suggest that chronic heat exposure decreases metabolic oxidation capacity in skeletal muscle of broiler chickens. On exposure to chronic heat stress, GPx activity remained relatively constant, though a temperature-dependent elevation in Cu/Zn-SOD activity was observed, implying that anti-oxidation ability was disturbed by the chronic stress condition. From these results it can be concluded that chronic heat stress did not induce oxidative damage to a major extent. This may probably be due to a decrease in metabolic oxidation capacity or due to a self-propagating scavenging system, though the system was not fully activated.