Keywords : Lipopolysaccharide

The effect of lipopolysaccharide extracted from Escherichia coli on total WBCs, granulocytes and on phagocytic activity in female rats

Aziz T. Koro; Adeba Y. Sharif

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 285-289
DOI: 10.33899/ijvs.2021.130030.1721

The present study was conducted to compare the total white blood cells count, granulocytes and phagocytic activity in female rats immunized with lipopolysaccharide (LPS) extracted from two different isolates of Escherichia coli, one normal isolate from the gut of healthy adult and the other was pathogenic strain isolated from patients suffering from urinary tract infection. The study was included 35 female albino rats divided randomly into seven groups, three groups injected with different concentrations of LPS of normal flora of E.coli 90, 120 and 150 microgram/kilogram body weight respectively and the other three groups were injected with the same concentrations of LPS extracted from pathogenic strain of E.coli, the last group (7th) represented the control group which given normal saline only. The results showed that the LPS from normal isolate causes a significant increase in the total number of leukocytes, granulocytes and phagocytic activity in animals immunized with the concentration of 120 µg/kg compared with the control group, and the concentration of 150 µg/kg of the same extract resulted in a high significant increase in phagocytic activity and the concentration of 90 µg/kg did not stimulate any significant differences in all the variables, while immunization with the extract of pathogenic strain increases the total leukocytes at a concentration of 90 µg/kg only with a significant increases in phagocytic activity at all concentration, and it also did not cause any significant differences in the granulocytes count.

Extraction and purification of lipopolysaccharide from Escherichia coli (local Isolate) and study its pyrogenic activity

Ammar M. Al-AAlim; Ali A. Al-Iedani; Mohammad A. Hamad

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 45-51
DOI: 10.33899/ijvs.2021.128963.1614

In this study, we tried to extract and purify the LPS from E. coli local isolate and determine the molecular weight, purity, and pyrogenic effect of the product and compare it with standard E. coli O55:B5 LPS, the E. coli LPS was extracted by using hot phenol method then SDS- PAGE was used with both Coomassie blue and silver nitrate stain to determine its molecular weight and protein contamination also we used HPLC to the estimation of E. coli LPS purity and finally the pyrogenicity of extracted E. coli LPS was tested by using rabbit pyrogen test. The result showed that the hot phenol method with enzymatic treatment gave highly pure LPS with a high yield reach up to 242.4 mg, staining the SDS page gel with Coomassie blue and silver nitrate uncover the high purification of the extracted LPS (ELPS) with no protein contamination, with a molecular weight range between 15-23 kDa, HPLC test reveals that purity of ELPS was 100 % compared with standard LPS. The rabbits' pyrogen test confirmed that the biological activity of ELPS. In conclusion, the LPS was extracted with high purity compare with standard LPS and without any protein or DNA contamination by using the hot phenol method also the extracted rough LPS was slightly lighter than the standard LPS used but this did not affect its biological activity which remained intact 

Development of experimentally-induced periodontitis in a Sprague Dawley rat model

Hana H. Mustafa; Ahmed Kh. Ali; Chen Cheng; Rozanaliza Radzi; Lau Fong; Noordin Mustapha; Hewa O. Dyary

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 4, Pages 765-769
DOI: 10.33899/ijvs.2021.128422.1573

Periodontitis is a common inflammatory disease that leads to the degradation of periodontium and results in alveolar bone loss. The development of a suitable animal model of periodontitis is a prerequisite to understanding better the mechanisms that underly this disease. This study evaluated periodontal disease induction via retentive ligature, intragingival injection of lipopolysaccharide (LPS), and their combination in a rat model.Seventy-two Sprague Dawley rats were divided into four groups. The first group (control) did not receive any treatment. The second group underwent the application of 4/0 nylon ligature around the second maxillary molars. The third group was treated with an intragingival injection of Porphyromonas gingivalis LPS into the palatal mucosa of the second maxilla molars, and the fourth group was treated with a combination of ligature and LPS injection (ligature-LPS). Morphological changes in the gingival tissues were evaluated after 7, 14, and 30 days of treatment.Significant degenerative changes were observed in the periodontal tissues and alveolar bone in the third and fourth groups, which were evident as early as seven days. The lesions remained until 14 days and declined with time in the third and fourth groups. The changes induced by ligature and ligature-LPS were not different. Injection with LPS alone resulted in minimal increases in the Gingival and Plaque Indices.The ligature technique induced periodontal disease successfully, more effective than the injection of LPS. The combination of ligature with LPS injection added no significant effect compared to ligature alone.