RESUMEN
Aedes aegypti and Ae. albopictus are among the most important vectors of arboviral diseases, worldwide. Recent studies indicate that diverse midgut microbiota of mosquitoes significantly affect development, digestion, metabolism, and immunity of their hosts. Midgut microbiota has also been suggested to modulate the competency of mosquitoes to transmit arboviruses, malaria parasites etc. Interestingly, the midgut microbial flora is dynamic and the diversity changes with the development of vectors, in addition to other factors such as species, sex, life-stage, feeding behavior and geographical origin. The aim of the present study was to investigate the midgut bacterial diversity among larva, adult male, sugar fed female and blood fed female Ae. albopictus collected from Tezpur, Northeastern India. Based on colony morphological characteristics, we selected 113 cultivable bacterial isolates for 16S rRNA gene sequence based molecular identification. Of the 113 isolates, we could identify 35 bacterial species belonging to 18 distinct genera under four major phyla, namely Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. Phyla Proteobacteria and Firmicutes accounted for majority (80%) of the species, while phylum Actinobacteria constituted 17% of the species. Bacteroidetes was the least represented phylum, characterized by a single species- Chryseobacterium rhizoplanae, isolated from blood fed individuals. Dissection of midgut microbiota diversity at different developmental stages of Ae. albopictus will be helpful in better understanding mosquito-borne diseases, and for designing effective strategies to manage mosquito-borne diseases.
Asunto(s)
Aedes , Microbioma Gastrointestinal , Estadios del Ciclo de Vida , Animales , Bacterias/clasificación , Bacterias/genética , Biodiversidad , Femenino , India , Masculino , Metagenoma , Metagenómica/métodos , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Thirty isolates of endophytic fungi were isolated from healthy asymptomatic leaves of tea plant (Camellia sinensis) and identified morphologically based on colony morphology, spore shape and size, growth and sporulation rate. Internal transcribed spacer r-DNA sequence analysis supported for molecular identification of all the isolates. Based on morphological and molecular characteristics the isolates were identified as Colletotrichum gloeosporioides. Variations on colony morphology which included the production of conidial masses, led to divide the isolates into different groups. Variations on spore size, growth rate and sporulation rate were exhibited by all the isolates. With RAPD molecular markers, genetic variations among the thirty isolates were observed. Genetic variations and relatedness among the thirty isolates were analyzed with UPGMA phylogram using NTSYS program. Two major groups were obtained among the thirty isolates. Group I comprised of 16 isolates which included three sub groups (Ia, Ib and Ic) and Group II constituted fourteen isolates and it also had three sub groups (IIa, IIb and IIc). A partial co-relationship among the isolates was established on the basis of morphological and molecular based clustering.
RESUMEN
OBJECTIVE: To study the morphology of the salivary gland of the female blackfly of the species Simulium indicum (S. indicum) along with protein profile and anticoagulant activity of the salivary gland extract. METHODS: Sodium dodecyl sulphate polyacrylamide gel electrophoresis was used to analyze the protein profile of the salivary gland extract (SGE) and anticoagulant activities against thrombin, and the extrinsic and intrinsic coagulation pathways were found in S. indicum SGE in the TT, PT and APTT assays, respectively. RESULTS: Results revealed that each gland consisted of a cylindrical U-shaped secretory lobe and a more or less spherical reservoir. The protein contents of whole salivary glands were also quantified and the amount of salivary gland proteins in the adult female S. indicum was found out to be approximately 1.12±0.13 µg/female. At least 16 major and several minor protein bands were detected in the female salivary glands. The molecular masses of these major protein bands were estimated at 69, 65, 61, 58, 44, 42, 39, 33, 30, 28, 27, 26, 23, 21, 18 and 16 kDa, consecutively. Anticoagulant activities were found in S. indicum SGE in all the assays. It was found that SGE prolonged human plasma clotting time in a dose-dependent manner. Factor Xa inhibition was shown by the SGE of S. indicum. Percent inhibition value was 93.8. A positive correlation (r=0.89) was observed between total protein and percent inhibition of factor Xa. CONCLUSIONS: The present study demonstrated that the mode of action of the anticoagulant(s) is mainly on the inhibition of thrombin and factor Xa along with other target factors of the coagulation cascade.
RESUMEN
An endophytic fungus isolated from Camellia sinensis, Assam, Northeastern India was identified as Colletotrichum gloeosporioides on the basis of morphological characteristics and rDNA ITS analysis. This endophytic fungus was evaluated for growth inhibition against tea pathogens Pestalotiopsis theae and Colletotrichum camelliae. One isolate of C. gloeosporioides showed strong antagonistic activity against Pestalotiopsis theae (64 %) and moderate activity against C. camelliae (37 %). Fifty percent cell-free culture filtrate from 5-day-old cultures showed highest antagonistic activity against both the pathogens although the inhibition percent was less as compared to dual culture. In the experiment of volatile compounds none of the isolates of C. gloeosporioides strains showed visible inhibition against P. theae and C. camelliae. The activity of extracellular hydrolytic enzymes chitinase and protease was also high in this culture fluid and measured 10 and 4.3 IU/µl, respectively.