PHARMACOLOGICAL EVALUATION OF ACETYL-11-α-KETO-β-BOSWELLIC ACID MEDIATED NANO SILVER IN EXPERIMENTAL MURINE MASTITIS
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Date
2013-03
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SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517502. (A.P.) INDIA
Abstract
ABSTRACT :
Acetyl-11-α-keto-β-Boswellic acid mediated nanosilver (AKBANS), was synthesized, characterized and evaluated in Staphylococcus aureus induced murine mastitis model. The characterization of AKBANS by UV visible absorption spectrum showed a maximum absorption around 200 nm and SEM images showed that AKBANS with irregular and spherical morphology of size 363 to 574 nm. The size of the particles measured by DLS technique was 262 nm. The results of FT-IR analysis for AKBANS showed the involvement of hydroxyl, carboxyl, amines, keto and nitrile groups in the synthesis of AKBANS. The MIC of the compound was found to be 3.6 ng/ml against Staphylococcus aureus showed an in vitro spleenocyte viability of more than 95% at the highest concentration of 87.5 ppm. No toxicity was found in the oral dose even at the limit dose. A total of 40 female mice of 10-15 days post partum were utilized for the study. The animals were divided into five groups of eight animals each. Group I served as lactating control, groups II to V were inoculated with 20 μl of 24h broth culture of S.aureus containing 4.0 x 105 cfu/quarter (Log10 5.60 cfu/quarter) under ketamine ananestheisa using 33G blunt hamilton needle. After 6h post inoculation, groups III and IV received 20 μl of AKBANS through intramammary and intra peritoneal routes respectively. Whereas, group V received Cefepime & tazobactum combination @ 1mg/kg body weight through intraperitoneal route. After 24 h post inoculation, 0.5ml of whole blood was collected from tail vein and serum was separated for the estimation of CRP. Later, the mice were euthanized and L4 mammary gland was collected under asceptic conditions into sterile eppendorf tubes. The mammary weights were recorded and tissue samples for histopathology and ultrastructural studies were collected in 10% neutral buffered formalin and 5% glutaraldehyde respectively. A 10% homogenate of the mammary glands was prepared in 0.5M phosphate buffer (pH 7.4) and was utilized for the evaluation of bacterial load and antioxidant parameters like TBARS, GSH, SOD and CAT.
All the groups inoculated with S.aureus showed significantly higher mammary weights, however, the bacterial load was found to be significantly higher in mastitis group II. Treatment with both AKBANS and antibiotic showed significantly reduced bacterial loads compared to mastitis control. Similarly, group II showed significantly elevated levels of CRP, which were significantly reduced in AKBANS treatment groups III and IV and antibiotic treated group V. However, AKBANS showed significant reduction compared to antibiotic treatment.
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