Antimicrobial activities of certain seaweeds from muddy shore places of kerala

A study was carried out to reveal the growth inhibitory effect of methanol crude extract (MCE) and methanol supernatant extract (MSE) of sea weeds: 1) Gracillaria corticata, 2) Hypnea musciforms, 3) Gelidium micropterum and 4) Hypnea valentiae against six bacterial pathogens, 1) Pseudomonas aeruginosa, 2) Bacillus licheniformis, 3) Serratia marcescens, 4) Aeromonas hydrophila, 5) Acinetobacter baumanii, 6) Escherichia coli and two fungal strains, 1) Aspergillus niger and 2) Candida albicans respectively. Well diffusion method using zone of inhibition as indicator for growth inhibition was adopted. The results showed that methanol extracts of seaweeds viz., Gracillaria corticata, Hypnea musciforms, and Hypnea valentiae prevented the growth of pathogenic bacteria and fungi. The effect on growth was observed as zone of inhibition, the diameter of which was indicated in the units of a millimeter. The growth of the bacterium, Serratia marcescens was affected by methanol supernatant extract of the three types of seaweeds, Gracillaria corticata, Hypnea musciforms, and Hypnea valentiae, and by the methanolic crude extract of Hypnea musciforms and Hypnea valentiae. However, the growths of other species of bacteria were not controlled by either of the extracts of the seaweeds except, Bacillus licheniformis which was controlled by only Gracillaria corticata. The growth of fungi: Aspergillus niger and Candida albicans were inhibited by the methanol extracts of Gracillaria corticata. Between the two forms of methanolic extracts i.e., supernatant and crude, the efficiency of the supernatant extract was greater than that of crude one. Further, among the three types of seaweeds which showed an effect on the growth of microbes, the level of the zone of inhibition caused by Gracillaria corticata was statistically higher than that of the other two, Hypnea musciforms, and Hypnea valentiae.

Macrophage supernatant infected with Leishmania major mediates the cytology of fibroblast cells in skin wounds

Objective: Healing of Cutaneous Leishmaniasis relies on the effective and modulates protective immune responses. Although the immune system is necessary to eliminate the parasite, it could be considered as the main cause of ulcers. Therefore, main aim of this study was to explore the possible regulatory functions of macrophage supernatant infected with Leishmania major on the fibroblast cells. Materials and Methods: In this experimental study, different concentrations of infected macrophage supernatant extract (50, 100, 150, 200, and 250μg/mL) were tested at different times (6, 24, 48, and 72h) and the effect of the leishmanicidal extract on fibroblast cells was determined by MTS assay. Also, the flow-cytometry technique was used for the investigation of apoptosis induction percentage. Results: MTS assay showed that the leishmanicidal effect of infected macrophage supernatant extract was dependent on the concentration and the time of treatment. So, the best efficacy was observed in 200 μg/mL with 72 hours exposure time. Flow cytometry analysis showed that the infected macrophage supernatant extract could induce apoptosis in cultured fibroblasts. Conclusions: We have demonstrated that reduction of survival rate and induction of apoptosis in fibroblasts displayed a similar manner to keratinocytes when exposed to infected macrophages with L. major. Our data suggest that such a phenomenon can be the underlying cause of lesions with scarring, and future, the mechanism remains to be elucidated.

THE ANTIBACTERIAL EVALUATION OF HALICLONA ASSOCIATED BACTERIA AND THE RELATING COMPOUNDS DERIVED FROM THE HOST

Objective: The objective of this study was to investigate the active compounds derived from Haliclona sp. associated bacteria collected from Untung Jawa Island, Jakarta.Methods: The bacterial isolation, screening of antibacterial activity, purification, and prediction of potential compounds using liquid chromatography– mass spectroscopy/mass spectroscopy (LC–MS/MS) as well as 16S rDNA bacterial characterization were conducted.Results: The potential extract of bacterial strain UJ.17.10-4 showed the highest anti-Gram-positive bacteria. The 16S rDNA gene of this strain had 99% similarity with the actinobacteria Agromyces tropicus. Chemical separation of supernatant extract yielded 13 potent fractions. Identification of antibacterial compounds contained in active fractions using LC-MS/MS were halistanol sulphate C([M+H]+:703), cyclic bis-1,3- dialkylpyridinium([M+H]+: 213.72). In previous research reported that these compounds were isolated from Haliclona sponge and showed anticancer activity.Conclusion: This result supported the ideas that A. tropicus plays an important role in synthesizing, halistanol sulfate C, haliclorensin, and cyclic bis- 1,3-dialkylpyridinium metabolite derived from the host.

The anthelmintic efficacy of natural plant cysteine proteinases against the rat tapeworm Hymenolepis diminutain vivo

Hymenolepis diminuta is a natural parasite of the common brown rat Rattus norvegicus, and provides a convenient model system for the assessment of the anthelmintic activity of novel drugs against cestodes. The experiments described in this paper indicate that treatment of rats infected with H. diminuta with a supernatant extract of papaya latex, containing a mixture of four cysteine proteinases, was moderately efficacious, resulting in a significant, but relatively small, reduction in worm burden and biomass. However, faecal egg output was not affected by treatment. In our experiments these effects were only partially dose-dependent, although specific inhibition by E-64 confirmed the role of cysteine proteinases as the active principles in papaya latex affecting worm growth but not statistically reducing worm burden. Data collected for a further 7 days after treatment indicated that the effects of papaya latex supernatant on worm loss and on worm growth were not enhanced. Our findings provide a starting point for further refinement in formulation and delivery, or assessment of alternative natural plant-derived cysteine proteinases in efforts to develop these naturally occurring enzymes into broad-spectrum anthelmintics, with efficacy against cestodes as well as nematodes.

Identification of a NewAlcaligenes faecalisStrain MOR02 and Assessment of Its Toxicity and Pathogenicity to Insects

We report the isolation of a bacterium fromGalleria mellonellalarva and its identification using genome sequencing and phylogenomic analysis. This bacterium was namedAlcaligenes faecalisstrain MOR02. Microscopic analyses revealed that the bacteria are located in the esophagus and intestine of the nematodesSteinernema feltiae, S. carpocapsae, andH. bacteriophora. UsingG. mellonellalarvae as a model, when the larvae were injected with 24,000 CFU in their hemocoel, more than 96% mortality was achieved after 24 h. Additionally, toxicity assays determined that 1 μg of supernatant extract fromA. faecalisMOR02 killed more than 70%G. mellonellalarvae 96 h after injection. A correlation of experimental data with sequence genome analyses was also performed. We discovered genes that encode proteins and enzymes that are related to pathogenicity, toxicity, and host/environment interactions that may be responsible for the observed phenotypic characteristics. Our data demonstrates that the bacteria are able to use different strategies to colonize nematodes and kill insects to their own benefit. However, there remains an extensive group of unidentified microorganisms that could be participating in the infection process. Additionally, a nematode-bacterium association could be established probably as a strategy of dispersion and colonization.

Sorption and desorption of picloram in soils under pastures in Brazil

The objective of this work was to determine the coefficients of sorption and desorption of picloram in Ultisol (PVA) and Oxisol (LVA), displaying different physical and chemical characteristics. Samples of soil were collected at the 0 20 cm depth in degraded pasture areas in Viçosa-MG. Firstly, the equilibrium time between the herbicide in solution and the herbicide which was sorbed in the soil was determined by the Batch Equilibrium method. The time required was 24 hours. Sorption and desorption studies were carried out under controlled laboratory conditions; the sorption evaluation consisted in adding 10.0 mL of herbicide solutions at different concentrations to tubes containing 2.00 g of soil, with vertical rotary agitation being maintained during the pre-determined equilibrium time. After centrifugation, supernatant extract cleaning and filtration, herbicide concentration was determined by high performance liquid chromatography (HPLC) with UV detection at 254 nm. Desorption was evaluated using the samples in the tubes after the sorption tests. The Freundlich model was used for interpretation of the sorption process. Ultisol showed higher adsorption coefficient (Kf a) compared with Oxisol, which may be attributed to the lower pH of the soil and its higher organic matter content. Desorption process occurred in both soils; the LVA allowed greater release of the previously sorbed molecules.

The anthelmintic efficacy of papaya latex in a rodent–nematode model is not dependent on fasting before treatment

In earlier studies of the anthelmintic activity of plant cysteine proteinases (CPs), a period of food deprivation was routinely employed before administration of CPs, but there has been no systematic evaluation as to whether this does actually benefit the anthelmintic efficacy. Therefore, we assessed the effect of fasting on the efficacy of CPs from papaya latex (PL) against Heligmosomoides bakeri in C3H mice. We used a refined, supernatant extract of papaya latex (PLS) with known active enzyme content. The animals were divided into three groups (fasted prior to treatment with PLS, not fasted but treated with PLS and fasted but given only water). The study demonstrated clearly that although food deprivation had been routinely employed in much of the earlier work on CPs in mice infected with nematodes, fasting has no beneficial effect on the efficacy of PLS against H. bakeri infections. Administration of CPs to fed animals will also reduce the stress associated with fasting.