Effects and mechanisms of action of polyene macrolide antibiotic nystatin on Babesia gibsoni in vitro

Nystatin is a membrane-active polyene macrolide antibiotic and a channel-forming ionophore. Nystatin exhibits in vitro activity against Babesia gibsoni infecting normal canine erythrocytes containing low potassium (LK) and high sodium concentrations, i.e., LK erythrocytes. The calculated IC(50) value of nystatin against B. gibsoni infecting LK erythrocytes was 31.96 μg/ml. The anti-babesial activity of nystatin disappeared when B. gibsoni in LK erythrocytes were incubated in culture media containing high potassium concentrations (HK). Moreover, when the parasites were harbored in canine HK erythrocytes, which contained high potassium and low sodium concentrations as a result of high Na-K-ATPase activity, the in vitro anti-babesial activities of nystatin also disappeared, apparently due to protection by HK erythrocytes. This suggested that nystatin could show in vitro anti-babesial activity against B. gibsoni by its ionophorous activity, the same as other ionophores such as valinomycin. Subsequently, the effects of nystatin on the host cells were observed. Nystatin could not modify the intracellular concentrations of potassium, sodium, adenosine triphosphate, or glucose in either LK or HK erythrocytes, although it caused weak hemolysis in HK erythrocytes. In addition, nystatin did not affect the survival of canine peripheral polymorphonuclear leukocytes. In conclusion, nystatin destroyed B. gibsoni by ionophorous activity but did not affect either canine erythrocytes or leukocytes in vitro.     

Use of polypropylene bags for mass rearing Neoseiulus baraki (Acari: Phytoseiidae), a predatory mite of Aceria guerreronis Keifer (Acari: Eriophyidae)

An efficient, low cost and practicable mass rearing method for the predatory mite, Neoseiulus baraki Athias-Henriot (Acari: Phytoseiidae) was developed using a bag made of two-ply polypropylene (gauge 150, 24 cm × 36 cm) sheets. Introducing 20 N. baraki females into the bag produced a mean number 5218 ± 212.10 offspring in 6 weeks with a 260-fold increase of the initial population.     

Structure/activity studies of anti-inflammatory peptides based on a conserved peptide region of the lectin domain of E-, L- and P-selectin

Previously, it was established that the peptide YYWIGIRK-NH₂inhibits both myeloid cell adhesion to selectins in vitro andneutrophil influx into inflammatory sites in vivo (Briggs etal., 1995). Initial structure/activity studies revealed thatat least one Y residue at the N-terminus of the peptide wasessential for these bioactivities but that the C-terminal Kresidue was unnecessary for inhibitory activity. We have nowsynthesized a new series of peptides which contain single residuesubstitutions at each position of the reference peptide, YYWIGIR-NH₂and have tested these peptides for inhibitory activity in aselectin cell binding assay. In addition, peptides containingsingle D-amino acids at selected positions, or an all D-configuredreference peptide sequence, or the retro-inverso version (rigiwyy-NH₂of the reference peptide sequence have also been analyzed forinhibitory activity in the same assays. Finally, the abilityof the reference peptide and a specifically designed controlsequence (YY(AIB)IGIR-NH₂ to discriminate between potentialsynthetic saccharide ligands, including sialyl-Lewis x, Lewisx, and sialyl-N-acetyl-lactosamine, was investigated using isothermaltitration calorimetry. The results of these studies demonstratethat whereas many single amino acid substitutions are toleratedin the peptide without complete loss of inhibitory activity,substitution at some positions (e.g., the W residue) resultsin relatively inactive compounds, clearly pointing to the importanceof these residues in making critical contacts with the appropriatesaccharide ligand. Titration calorimetry revealed that the referencepeptide does not discriminate between Lewis x or sialyl-Lewisx in vitro, but binds these saccharides with nearly 40-foldhigher affinity (K_D 25 µM) than the nonfucosylated trisaccharide,sialyl-N-acetyl-lactosamine. We can infer from these studiesthat the presence of a sialyl group, per Se, is not a requisitefor complex formation between the reference peptide and itssaccharide ligand. Substitution of single D-amino acid residuesat various po sitions in the reference peptide sequence reducesor elimi nates all inhibitory properties. However, the all Dconfigured peptide or the retro-inverso peptide sequence havegreater activity than the all L-configured reference peptidein the in vitro biological assays, and each was an effectiveinhibitor of neutrophil infiltration in a thioglycol late-inducedmouse peritonitis model. These results, combined with the resultsof titration, allow us to conclude that binding between thereference peptide and its saccharide ligand, which affords itsinhibitory properties, is mediated by the presence of a contiguous,nonpolar surface, or face, presented at the N-terminus of thereference peptide, likely encompassing the sequence YYWI. Furthermore,the W plays a critical role in binding, probably through formationof an essential hydrogen bond with a suitably juxtaposed groupcarried on the saccharide ligand. selectin peptides inhibition saccharide ligand     

Characterization of Novel Lactobacillus fermentum from Curd Samples of Indigenous Cows from Malnad Region,Karnataka, for their Aflatoxin B1 Binding and Probiotic Properties

Thirty-four isolates of Lactobacillus spp. (LAB) from 34 curd samples were evaluated for their aflatoxin B₁ (AFB₁) binding and probiotic properties. Upon characterization, four LAB isolates (LC3/a, LC4/c, LC/5a, and LM13/b) were found to be effective in removing AFB₁ from culture media with a capacity of above 75%. Staining reaction, biochemical tests, pattern of sugar utilization, and 16s rRNA gene sequence analysis revealed the identity of all the four isolates as L. fermentum. All of them could tolerate acidic pH, salt, and bile, which promise the use of these probiotic bacterial isolates for human applications. These isolates showed poor hydrophobicity and higher auto-aggregation properties. All L. fermentum isolates were found susceptible to gentamycin, chloramphenicol, cefoperazone, ampicillin, and resistant to ciprofloxacin and vancomycin. Results of hemolytic and DNase activity indicated their nonpathogenic nature. Though all L. fermentum isolates found inhibiting the growth of Salmonella ebony, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, maximum inhibition was obtained with isolate LC5/a. Kinetic studies revealed that all four bacteria required a minimum of 2 h to reach stationary phase of AFB₁ binding. AFB₁ binding ability varied from 66 to 85.2% among these four isolates. Bile (0.4%) was significant (P ≤ 0.05) in reducing the AFB₁ binding property of isolates LC3/a, LC4/c, and LM13/b, while increased AFB₁ binding ability was recorded at acidic pH (2.0). AFB₁ binding properties of isolate LC5/a were found least affected by acidic pH and bile. The findings of our study revealed the higher efficiency of L. fermentum isolate LC5/a in reducing the bioavailability of AFB1 in gut, and additionally, it improves the consumers’ health by its various probiotic characters. These beneficial characters, L. fermentum isolates, promise them to use as probiotic formulations alone or in combinations with other beneficial probiotic-bacterial isolates.

     

Prioritizing Remnant Forests for the Conservation of Mysore Slender Lorises (<Emphasis Type="Italic">Loris lyddekerianus lyddekerianus</Emphasis>) in Karnataka,India Through Estimation of Population Density

Information on a species’ distribution, abundance, and habitat requirements is important for formalizing a comprehensive management and conservation strategy. This becomes an even higher priority when the distribution of a species lies largely outside demarcated protected areas for wildlife. We used line transect distance sampling to estimate the density of an arboreal, nocturnal, and threatened primate, the slender loris (Loris lyddekerianus lyddekerianus) across diverse habitat types including both reserve forests and production landscapes in the southern part of the state of Karnataka, India. Abundance estimates varied from 0.16 individuals/ha in Devrayandurga State Forest to 2.57 individulas/ha in Ippadi State Forest, with the mean density estimate being highest in the Forest Division of Tumkur, at 165 individuals/km². Based on density estimates for the survey regions, their current habitat status, and perceived threats, we identify Ippadi, Ujjani, Devrayandurga, and Savanadurga state forests as priority areas for conservation of the slender loris. We propose that these areas be declared protected. The required management intervention is preventing further degradation of the habitat and enhancing canopy contiguity to facilitate loris movement.     

Isolation and characterization of immunoglobulin of the Indian major carp, rohu [Labeo rohita (Ham.)

Information on the structure and character of immunoglobulin of fishes is essential in health management. A study was carried out to characterize the serum immunoglobulin (IgM) of the Indian major carp, rohu Labeo rohita (Ham.). Rohu (500g) were immunised with bovine serum albumin (BSA) and the anti-BSA antibody was purified employing BSA-CL agarose affinity column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified Ig in a 3% gel under non-reduced conditions revealed a single protein having a molecular weight of 850kDa. Analysis of the purified serum in 10% SDS-PAGE under reduced conditions revealed that the immunoglobulin contained heavy and light chains with molecular weights of 85 and 23kDa, respectively. A polyclonal mouse anti-rohu IgM was prepared and used in an immunodot test which showed a specific reaction of the crude rohu anti-BSA antiserum and the purified anti-BSA IgM with BSA. Results indicate that the immunoglobulin of L. rohita is tetrameric IgM, similar to that of other fishes.     

Design and Synthesis of Heterocyclic Conjugated Peptides as Novel Antimicrobial Agents

Antimicrobial peptides have been recognized as a novel class of antibiotics and several candidates are currently in clinical trials. In the present study, new antimicrobial compounds were synthesized by coupling quinazolinone moiety with the fragments of elastin sequences VP, GVP, VGVP and GVGVP. They were evaluated for their antibacterial activity against both gram positive and gram negative bacterial strains. We are here reporting that heterocyclic conjugated tetra peptide and penta peptide showed enhanced antibacterial activity compare to the conventional antimicrobial drugs.     

High mangrove density enhances surface accretion, surface elevation change, and tree survival in coastal areas susceptible to sea-level rise

Survival, growth, aboveground biomass accumulation, sediment surface elevation dynamics and nitrogen accumulation in sediments were studied in experimental treatments planted with four different densities (6.96, 3.26, 1.93 and 0.95 seedlings m⁻²) of the mangrove Rhizophora mucronata in Puttalam Lagoon, Sri Lanka. Measurements were taken over a period of 1,171 days and were compared with those from unplanted controls. Trees at the lowest density showed significantly reduced survival, whilst measures of individual tree growth did not differ among treatments. Rates of surface sediment accretion (means ± SE) were 13.0 (±1.3), 10.5 (±0.9), 8.4 (±0.3), 6.9 (±0.5) and 5.7 (±0.3) mm year⁻¹ at planting densities of 6.96, 3.26, 1.93, 0.95, and 0 (unplanted control) seedlings m⁻², respectively, showing highly significant differences among treatments. Mean (±SE) rates of surface elevation change were much lower than rates of accretion at 2.8 (±0.2), 1.6 (±0.1), 1.1 (±0.2), 0.6 (±0.2) and −0.3 (±0.1) mm year⁻¹ for 6.96, 3.26, 1.93, 0.95, and 0 seedlings m⁻², respectively. All planted treatments accumulated greater nitrogen concentrations in the sediment compared to the unplanted control. Sediment %N was significantly different among densities which suggests one potential causal mechanism for the facilitatory effects observed: high densities of plants potentially contribute to the accretion of greater amounts of nutrient rich sediment. While this potential process needs further research, this study demonstrated how higher densities of mangroves enhance rates of sediment accretion and surface elevation processes that may be crucial in mangrove ecosystem adaptation to sea-level rise. There was no evidence that increasing plant density evoked a trade-off with growth and survival of the planted trees. Rather, facilitatory effects enhanced survival at high densities, suggesting that managers may be able to take advantage of high plantation densities to help mitigate sea-level rise effects by encouraging positive sediment surface elevation.     

Birth Patterns in Wild and Captive Lion-Tailed Macaques (Macaca silenus)

During a 5-yr study of lion-tailed macaques in their natural habitat, we found that: 1) most births occurred from January to April (70%) and from September to December (19%), showing a bimodal pattern with a major and a minor birth peak; 2) the period of peaks remained the same over 5 yr; and 3) a similar pattern of birth peaks occurred both in groups in large forest complexes that had overlapping home ranges with other groups and in single groups that were isolated in forest fragments in the same region but with similar ecological conditions. The results suggest more of a birth seasonality than mere breeding synchronization in the wild lion-tailed macaques. We also analyzed data on births in captivity in European Zoos for 10 yr. We observed no seasonality or peaks in births, and the pattern was the same over the years. Data on rainfall suggest that resource availability in the wild habitat may not be uniform throughout the year; hence, ecological factors may play an important role in determination of birth patterns in the natural habitats of lion-tailed macaques.     

Preparation and characterization of a thermostable and biodegradable biopolymers using natural cross-linker

The present study describes preparation and characterization of a thermally stable and biodegradable biopolymer using collagen and a natural polymer, alginic acid (AA). Required concentration of alginic acid and collagen was optimized and the resulting biopolymer was characterized for, degree of cross-linking, mechanical strength, thermal stability, biocompatibility (toxicity) and biodegradability. Results reveal, the degree of cross-linking of alginic acid (at 1.5% concentration) with collagen was calculated as 75%, whereas it was 83% with standard cross-linking agent, glutaraldehyde (at 1.5% concentration). The AA cross-linked biopolymer was stable up to 245°C and Exhibits 5-6-fold increase in mechanical (tensile) strength compared to plain collagen (native) materials. However, glutaraldehyde cross-linked material exhibits comparatively less thermal stability and brittle in nature (low tensile strength). With regard to cell toxicity, no cytotoxicity was observed for AA cross-linked material when tested with mesenchymal cells and found degradable when treated with collagenase enzyme. The nature of bonding pattern and the reason for thermal stability of AA cross-linked collagen biopolymer was discussed in detail with the help of bioinformatics. A supplementary file on efficacy of AACC as a wound dressing material is demonstrated in detail with animal model studies.