Biogenic Synthesis of Silver Nanoparticles Using Pantoea stewartii and Priestia aryabhattai and Their Antimicrobial, Larvicidal, Histopathological, and Biotoxicity Potential.

In recent years, green nanotechnology has gained considerable importance for the synthesis of nanoparticles due to its economic viability and biosafety. In the current study, silver nanoparticles were synthesized using two bacterial isolates, H2 and H3, which were isolated from soil samples collected from the Western Ghats, Tamil Nadu, and identified at the species level as Pantoeastewartii (H2) and Priestiaaryabhattai (H3) by sequencing their 16s rRNA genes. Intracellularly synthesized silver nanoparticles were characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy, atomic force microscopy, and particle size analysis. AFM studies show that both of the bacterial synthesized Ag NPs were circular-shaped and disaggregated, with an average size distribution of 4 nm for Pantoeastewartii and 3.6 nm for Priestiaaryabhattai. Furthermore, their larvicidal activity, antimicrobial, histopathological, and biotoxicity effects were determined. The synthesized Ag NPs exhibited potent larvicidal activity against fourth instars of Ae.aegypti, An.stephensi, and Cx.quinquefasciatus exposed to a 50 µg/mL concentration for 24 h based on their LC50 and LC90 values. Histopathological studies of the affected mosquito larvae clearly show damage to the epithelial cells, food bolus, basement membrane, muscles, and midgut parts. The maximum antimicrobial activity of Priestiaaryabhattai-synthesized Ag NPs was observed for Streptomyces varsoviensis MTCC-1537, and that of Pantoea stewartii-synthesized Ag NPs was against Escherichia coli MTCC-43. The toxicity test on non-target organisms such as Artemia nauplii and zebrafish embryos indicates no visible abnormalities or mortality after their exposure for 48h. It is concluded that silver nanoparticles can easily be synthesized using Pantoea stewartii (H2) and Priestia aryabhattai (H3) as capping and reducing agents. Silver nanoparticles showed potent larvicidal activities and could potentially be used in integrated vector control programs because they are safe for other inhabitants of the same aquatic environment as mosquito larvae.

Histomorphology and Chemical Constituents of Interdigital Gland of Vembur Sheep, Ovis aries.

The interdigital gland is a specialized skin gland located between the digits of Artiodactyla (i.e., even-toed ungulates). Its secretion participates in semiochemical communication, and protects from ultraviolet radiation as well as fungal and bacterial infections of the feet. The present study aimed at finding if there are male-female differences in the anatomy, morphology, and volatile compounds of the interdigital gland of the South Indian breed of Vembur sheep. A total of 24 sheep (12 each of male and female) were spotted at the slaughterhouse and the interdigital gland was removed for examination. The anatomical examination revealed it to resemble a tobacco pipe and to consist of a body, flexure, and excretory duct with an external orifice located at the cleft of the digits. Morphometrically, the interdigital glands differed between males and females. The gland possesses a distinct fibrous capsule, epidermis, and dermis. The fibrous capsule contains several parallel bundles of collagen fibers, nerve fibers, and blood vessels, etc. The epidermis consists of keratinized squamous epithelium formed of stratum basale, stratum granulosum and stratum spinosum. The dermis consists of hair follicles, nerve plexuses, arrector pili muscles, and apocrine and sebaceous glandular lobules. The latter, lined by a simple cuboidal epithelium, are arranged in clusters of acini in the upper portion of the dermis. The apocrine secretory lobules, made up of parenchymal cells, are found in the lower portion of the dermis. The density and diameter of the apocrine and sebaceous secretory lobules were significantly higher in the males than females. Scanning electron microscopic (SEM) analysis confirmed the apocrine and sebaceous secretory components. Twenty-three major compounds were identified in the interdigital gland postings of male and female sheep, among which butanoic acid, 2-methylpropanoic acid, 1-heptanol and octadecanoic acid were present only in the male glandular post, whereas octane, 7-hexyl-tridecane, tetradecane, heptadecane and decanoic acid were present only in the female glandular post. Tetradecanol, tetradecanoic acid and hexadecanol peaks, reportedly antibacterial compounds in pronghorn antelopes, were highly prominent in both male and female sheep. Thus, the interdigital gland of Vembur sheep has two major secretory lobules, namely, sebaceous and apocrine, larger in males than females, which secrete a variety chemical compounds that may serve as chemical communication systems and protect the sheep from foot-borne diseases.

Synthesis of silver nanoparticles using Catharanthus roseus root extract and its larvicidal effects.

Phytosynthesis of silver nanoparticles has attracted considerable attention due to their biocompatibility, low toxicity, cost-effectiveness and being a novel method has an eco-friendly approach. Biological activity of root extracts as well as synthesized silver nanoparticles of Catharanthus roseus were evaluated against larvae of Aedes aegyptiand Culex quinquefasciatus. The structure and proportion of the synthesized nanoparticles was defined by exploitation ultraviolet spectrophotometry, X-ray diffraction, fourier transform infrared spectroscopy, energy dispersive X-ray spectroscopy and scanning electron microscopy methods. Reduction of silver ions occurred when silver nitrate solution was treated with aqueous root extract at 60°C. Synthesized silver nanoparticles (AgNPs) were confirmed by analyzing the excitation of surface plasmon resonance (SPR) using UV-vis spectrophotometer at 423 nm. FTIR showed aliphatic amines and alkanes corresponding peaks to be presence of responsible compounds to produced nanoparticles in the reaction mixture. Spherical shaped and crystalline nature of particles was recorded under XRD analysis. Presence of silver metal and 35-55nm sized particles were recorded using EDAX and SEM respectively. Larvicidal activitywas observed after24 hrs of exposure to root extracts and synthesized silver nanoparticles. The highest larval mortality was observed in synthesized silver nanopartiucles against Aedes aegypti (LC50= 2.01 ± 0.34; LC90= 5.29 ± 0.07 at 5.0 mg(-1) concentration) and Culex quinquefasciatus (LC50= 1.18 ± 0.15; LC90= 2.55 ± 0.76 at 3.5 to 5.0 mgl(-1) concentration) respectively. The present study provides evidence that synthesized silver nanoparticles of Catharanthus roseus offer potential source for larvicidal activity againstthe larvae of both dengue and filariasis vectors.

Preputial gland activates olfactory receptor neurons in rat: calcium imaging study using laser scanning confocal microscopy.

The rodent preputial gland is one of the major sources of odours and is reported to be involved in several behavioural activities. However, how the preputial gland initiates the olfactory response to manifest the effects is not known. Olfactory receptor neurons (ORNs) present in the olfactory epithelium are involved in the perception of odorant/pheromonal compounds. In the present study, the response of rat ORNs to preputial gland extract was evaluated by calcium imaging analysis. We found that some rat ORNs responded to the preputial gland extract by exhibiting an intracellular calcium response. By contrast, the ORNs did not respond at all to the foot pad extract (control). The results indicated that the substances contained in the preputial gland might interact with a type of receptor expressed in the female rat ORNs, suggested to manifest the behavioural responses, such as social and sexual interactions. This study provided the first evidence of activation of ORNs by the preputial gland extract.

1-Iodo-2 methylundecane [1I2MU]: an estrogen-dependent urinary sex pheromone of female mice.

In our previous investigations, urine of female mice contained specific compounds, namely isocroctylhydrazine, 4-methyl-2-heptanone, and azulene during proestrus, whereas during estrus it contained 1-H-cyclopop.e.azulene, caryophyllene, and copanene. Furthermore, 1-iodo-2 methyl undecane (1I2MU), present during both proestrus and estrus, was regarded as a putative estrus-specific chemo-signal. The primary objective of the present study was to determine the estrogen-dependency of the above-mentioned compounds, including 1I2MU. Furthermore, the effect of these compounds on pre-mating behavior, e.g., sniffing, licking, and grooming, were recorded to determine their role as sex pheromones. Based on gas chromatography linked mass spectrometry (GC-MS) of urine samples, profiles in oophorectomized female mice had 14 major peaks. Furthermore, neither 1I2MU (nor other estrus-specific compounds) were detected in the urine of these mice, although they were detected in urine of proestrus and estrus mice. In addition, 1I2MU was not detected in urine of prepubertal mice. It was noteworthy that both 1I2MU and 4-methyl-2-heptanone reappeared in estrogen-treated females. Based on pre-mating behavioral analysis, 1I2MU was the compound most preferred by males. In conclusion, production of 1I2MU was estrogen-dependent in females, and it enhanced reproductive activities in males.

Detection of estrus by male mice: synergistic role of olfactory-vomeronasal system.

In rodents, olfactory pathway comprises two distinct systems viz, the main olfactory and vomeronasal systems, both differing in anatomy, physiology and function. The precise role of the main olfactory/vomeronasal system in estrus detection is yet to be explored. Therefore, the present investigation was planned to elucidate the role of main olfactory and vomeronasal system in the estrus discriminating ability of male mice. Female urine samples of proestrus, estrus, metestrus, diestrus, ovarectomized, ovarectomized plus estrogen treated and prepubertal mice were used for the present study. In addition, the urine from intact, castrated and castrated with testosterone treated mice was also tested for odour preference by male mice. The male responders were categorized into three groups namely (a) normal, (b) ZnSO(4)-irrigated and (c) vomeronasal organ (VNO)-ablated. The behavioural responses such as frequency and duration of visits to urine samples were carried out in a Y-maze apparatus to assess odour preference. The normal mice displayed more frequent visits to estrus urine samples than to non-estrus samples. In contrast, ZnSO(4)-irrigated mice showed significant reduction in the frequency of visits towards estrus urine, whereas, the vomeronasal (VNO)-ablated mice did not show any noticeable preference. With regard to the duration of visits the VNO-ablated mice showed significant reduction in visiting time when compared to ZnSO(4)-irrigated mice. This finding indicated that the main olfactory system (MOS) was involved primarily in the attraction from a distance, while the VNO played a major role in close proximity (pre-copulatory behaviour). The males spent less time with the urine of same-sex; however, the response was higher with castrated male urine which was reduced on testosterone treatment indicating that a specific odour in intact male causes aversive behaviour in male. This study provides support to the fact that volatile compounds could also be perceived by VNO, probably when the main olfactory system is in functional state. The study implies that the olfactory-vomeronasal system plays a synergistic role in the detection of estrus.

Identification of testosterone-dependent volatile compounds and proteins in the preputial gland of rat Rattus norvegicus.

Preputial gland is one of the best known and most odour-producing organs in many non-primate mammals. It is generally believed that the development of this gland and functions are regulated by testosterone. To substantiate this point, the present study was aimed to evaluate the testosterone-dependent volatile compounds and proteins in the preputial gland of rat adopting castration and testosterone supplementation. The results revealed that four compounds, geranyl linalool isomer, oxirane, farnesol and lanosterol, are testosterone-dependent. Similarly, a low molecular mass protein with molecular weight 18kDa, supposed to be a pheromone carrier, also is shown to be testosterone-dependent. This study leads to the conclusion that testosterone-dependent compounds and sex-associated protein are present in the preputial gland of rat which may act as a sex pheromone and pheromone carrier, respectively.

Protective effect of Gymnema montanum against renal damage in experimental diabetic rats.

Gymnema montanum Hook (Asclepiadaceae), is an endemic plant species of India, traditionally used for diabetes and its management. In this experiment, the ethanol extract of G. montanum (GLEt) at a dose of 200mg/kg body weight was tested to evaluate its effect on renal damage in alloxan-induced diabetic rats and the efficacy was compared with standard hypoglycemic drug, glibenclamide (600 microg/kg body weight). The GLEt and glibenclamide were administered orally for 3 weeks and the effects on glucose, insulin, renal markers including urea, creatinine and uric acid, lipid peroxidation markers including thiobarbituric reactive substances (TBARS) and hydroperoxides and antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities in kidney were studied. In addition, the urinary protein profile was studied using SDS-PAGE. The results indicated that the GLEt significantly normalized the elevated blood glucose, renal markers and lipid peroxidation markers and increased antioxidant levels in diabetic kidney. The diabetic rats excreted large amount of proteins than untreated rats which was normalized during the treatment with GLEt. In conclusion, the GLEt was found to be more effective in reducing oxidative stress, thus confirming the ethnopharmacological use of G. montanum in protecting diabetes and its complications.

Antihyperlipidaemic effect of Gymnema montanum: a study on lipid profile and fatty acid composition in experimental diabetes.

In the present study, the antihyperlipidaemic efficacy of ethanol extract of Gymnema montanum leaves was investigated in alloxan-induced diabetic rats and the effect was compared to standard hypoglycaemic drug, glibenclamide. Male adult albino Wistar rats were injected with freshly prepared solution of alloxan monohydrate (150 mg/kg body weight) to induce diabetes. After 2 weeks, the rats with moderate diabetes were administered G. montanum leaves (200 mg/kg body weight) for 21 days by gastric lavage, after which serum, liver and kidney samples were analysed for lipid profile, lipoprotein changes and fatty acid composition. While the alloxan-induced diabetic rats showed a significant increase in the levels of cholesterol, triglycerides and free fatty acids, the levels in the animals treated with G. montanum leaves were considerably reduced and restored to near normal values. Antihyperlipidaemic effects of G. montanum leaves were found to be comparable with that of glibenclamide. Similarly, G. montanum leaves treatment resulted in reversal of alterations observed in the plasma lipoproteins (high-density lipoprotein, low-density lipoprotein and very high-density lipoprotein-cholesterol) and fatty acid composition in serum, liver and kidney of alloxan-induced rats. Our study suggests that phytochemicals present in G. montanum may play an important role in suppressing the elevated lipid profile in diabetes and may be useful for the prevention and/or early treatment of diabetes-associated hyperlipidaemia.