Chitosan nanoparticles prepared from lucilia cuprina maggots as antibacterial agent

Chitosan were prepared from cuticle of Lucilia cuprina maggots with two steps; deproteinization and deacetylation. It was characterized with solubility and Fourier Transform Infrared spectroscopy [FT-IR]. Chitosan was ball-milled to obtain the chitosan nanoparticles which characterized with dynamic light scattering [DLS] and transmission electron microscope [TEM]. Chitosan nanoparticles with degree of deacetylation [DDA] 80.5% were showed antibacterial activities against Klebsiella pneumoniae and Bacillus subtilis. The mode of action of chitosan nanoparticles on the tested bacteria was studied by TEM. Leakage of some cell contents, cell deformation mi rupture of cell were observed, therefore, the chitosan nanoparticles were observed to be a powerful antibacterial agent

Antimicrobial activities of chitosan nanoparticles prepared from lucilia cuprina maggots [diptera: calliphoridae]

Chitosan nanoparticles were studied as antimicrobial agent. The antibacterial activity of chitosan nanoparticles were investigated against three Gram-negative bacteria; Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi, and three Gram-positive bacteria; Staphylococus aureus, Enterococcus faecalis and Streptococcus pyogenes. The antifungal activity were examined against three fungi; Geotrichum candidum, Candida krusei and Candida parapsilosis. The antiviral activities were tested against three viruses; Rift Valley Fever [RVFV], Herpes simplex-1 [HSV-1] and Coxsackie viruses. Chitosan nanoparticles were inhibited all bacteria and fungi except E.faecalis seemed to be resistant strain. Infectivity titers of all viruses were reduced by chitosan nanoparticles, which are a natural antimicrobial agent

Antimicrobial activity for excretion and secretion of the greenbottle fly larvae lucilia sericata [meigen] [diptera: calliphoridae]

Sterile larval excretion/secretion [ES] exhibited antibacterial activity against some species of bacteria. They were shown to inhibit the growth of Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis Gram-negative bacteria Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae and Fungi Geotricum candidum and Aspergillus fumigatus thus exhibited limited inhibitory effect towards Gram-positive bacteria Streptococcus pyogenes and Staphylococcus epidermidis and Gram-negative Proteous vulgaris and Fungi Syncephalastrum racemosum, Candida albicans, that effect was slowed down when challenged with secretion on a solid media but no zone of complete inhibition was detected. Growth inhibiting activity was determined in liquid growth media using the Gram-positive, Gram-negative bacterial and fungal strains as indicator organisms

using of lucilia cuprina maggots in the treatment of diabetic foot wounds

In the present study, Lucilia cuprina maggots were used for the treatment of diabetic foot wounds. An artificial wound was made in diabetic foot of rabbit. The maggots were sterilized arid put directly on the wound after dressing the wound without using any antibiotics. Several cycles of maggots were put on the wound. The present results showed that the treatment of the diabetic foot was observed after 13 days. After this period the wound was completely healed and become free of microbial contamination. The new tissues were observed to close the wound

Repellent effect of lagenaria siceraria extracts against culex pipiens

Ethanolic, acetone and petroleum ether extracts from leaves and stems of Lagenaria siceraria [Cucurbitaceae] were screened for their repellency effect against Culex pipiens L. mosquito. The repellent action of the present plant extracts were varied depending on the plant parts and the dose of extract. The petroleum ether extract of leaves showed the same repellency percent [100%] of commercial formulation, N. N.'diethyl toulamide [DEBT] at the higher dose [3.33 mg/cm[2]], while petroleum ether extract from stems exhibiting the repellent action [89.6%] at the same dose, respectively. Ethanolic extracts of leaves and stems exhibited the lowest repellent activity as it recorded [81.3% and 69.1%] at [6.67 mg/cm[2]], respectively. Results of this study may contribute to design an alternative way to control mosquitoes currently based on applications of synthetic insecticides. These extracts could be developed commercially as an effective personal protection measure against mosquito bites and thus to control diseases caused by mosquito-borne pathogens

Effects of midgut bacteria and two protease inhibitors on the reproductive potential and midgut enzymes of culex pipiens infected with wuchereria bancrofti

Laboratory investigations were carried out to asses the effect of some proteases inhibitors on the reproductive potential of Culex pipiens females resulted from larvae treated with different protease inhibitors. The fecundity and engorgement of symbiotic and aposymbiotic C. pipiens females were significantly reduced. The blood meal digestion period increased significantly. On the other hand, enzyme band with molecular weight of 40 KDa which may be cysteine protease was detected in untreated symbiotic and aposymbiotic female midguts. The results may explain that the absence of this enzyme bands in treated female midguts may be due to the inhibition caused by [E-64] a cysteine protease inhibitor

Effects of midgut bacteria and two protease inhibitors on the transmission of wuchereria bancrofti by the mosquito vector, culex pipiens

Laboratory investigations were carried out to study the effect of two protease inhibitors on the transmission of W. bancrofti filarial by Culex pipiens and to study the susceptibility interaction between filaria and protease inhibitors. The results obtained revealed that, infection rates were variable among untreated and treated symbiotic and aposymbiotic Cx. pipiens females resulted from third instar larvae treated with E-64 and EDTA. The survival rate was variable among untreated and treated symbiotic and aposymbiotic females resulted from third instar larvae treated with E-64 and EDTA. Protease inhibitor [E-64] caused inhibition of the parasite development and transmission by means of ceasing catalytic activity- responsible for parasite migration-caused by parasitic larval stages inside the mosquito vector

Histological and histochemical study of testicular tissue used in cases of [ICSI] [intra cytoplasmic sperm injection]

The advances in the technology of in vitro fertilization [IVF] increases the incidence of success in treatment cases of infertile patients with non-obstructive azoospermia. Evaluation of the microscopic changes in the obtained testicular samples was one of the main aims in this study for better prognosis of the results. In this study one hundred human testicular biopsies were obtained from Islamic reproductive centre and Department of Urology Al-Hussein University Hospital. Cases in this study were classified into 4 groups: Group I includes 5 cases of normal fertile persons free of any medical problems and with normal seminal parameters. Patients consent was performed before sampling. Group II includes 35 cases of infertile obstructive azoospermic patients. All of the rest of cases [60] were considered infertile non-obstructive azoospermic cases and were included in both the third and the fourth groups. Group III: includes 20 cases of infertile non-obstructive positive patients. Group IV: includes 40 cases of infertile non-obstructive negative patients. Cases were either of normal sized testis or of hypoplastic testis and were subdivided into the following subgroups: I- Cases with normal sized testes [30] infertile patients: Group IV A: Normal sized testis with arrested spermatogenesis; Group IV B: Normal sized testis with SCOS; Group IVC: Normal sized testis with mixed atrophy. II- Cases with hypoplastic testes [10] infertile patients: Group IV D: Hypoplastic testis. The testicular samples were prepared for Histological and Histochemical examination. Different staining techniques were used: 1- Haematoxylin and Eosin stain; 2- Mallory's trichrome stain; and 3- Histochemical techniques: a- Periodic acid Schiff [PAS] technique. B- Methyl green pyronin. The obtained results showed no Morphological changes in group II except mild vascular dilation and congested peritubular capillaries. Cases of non-obstructive positive azoospermia showed reduced number of spermatogenic layers while cases of group IVa showed reduced size of the tubules and lined only by Sertoli cells and there was marked reduction in the number of leydig cells. In this study there were definite histochemical changes observed in both the PAS positive material and the nucleic acid in the different elements of the obtained testicular samples, Clinical and microscopic evaluations in this study could be of complementary importance and may increases of the incidence of success

preliminary pilot survey [culex pipiens], Sharkia governorate, Egypt

Sixteen different ecological areas in Sharkia Governorate were surveyed for adult Culex pipiens. The prevalence was investigated among 160 houses of 2800 individuals and their surrounding outdoors. The collected adults were 41488 from October 2003 to November 2004. The collection rates were higher in the rural areas than urban ones. Mosquitoes were common all the year round except January, February and March, with peaks in summer and autumn than in spring and winter. Larvae in descending order were Cx. pipiens the predominant larva followed by Ae. caspius, Culiseta sp. and then Cx. pusillus. Cx. pipiens was described as the most common larva

New concept in histopathological grading and staging of chronic hepatitis C infection at Sharkia Governorate, Egypt

In this study, the Knodell histology activity index and the semi- quantitative reproducible description of the various morphological lesions of chronic hepatitis were applied on 109 liver biopsies taken from Egyptian patients infected with hepatitis C virus [HCV]. It was found that the presented histopathological features may be unusual for any of the known scoring systems. Therefore, a new system was suggested for grading and staging of liver diseases in Egyptian patients infected with HCV. Accordingly, the degrees of necro- inflammations are classified into 3 grades [1-3] and the progression of fibrosis is classified into 3 stages [1-3]. The reduced numbers of grades and stages proposed in this study may be attributed to the rapid course among Egyptians who differ in the environmental circumstances

Molecular cytogntic profiles of hepatitis C infection in patients at Sharkia Governorate, Egypt

In this study, 40 paraffin blocks liver tissues from HCV-PCR positive patients [HBV seronegative] were examined using DNA image cytometry to evaluate its role in diagnosing hepatocellular carcinoma [HCC] associated with hepatitis C virus [HCV] infection. Fluorescent in situ hybridization [FISH] technique using LSIZNF 217 chromosome 20q 13.2 probe was applied. The results showed a high percentage of S- phase fraction in cases of G2S2 and G3S3 with DNA diploidy. Only two cases of G3S3 showed DNA aneuploidy with a severe amplification of chromosome 20q 13.2. Consequently, DNA imaging cytometry is considered a good approach in differentiating dysplasia from well- differentiated HCC on the top of HCV infection. In conclusion, HCV has an acquired role in the development of HCC through the amplification of the aggressive tumor behavior oncogene LSIZNF 217 at chromosome 20q 13.2

Haematologial manifestations in HCV infected patients at Sharkia Governerate, Egypt

The aim of this study was to evaluate the hematological manifestations occurring in patients suffering from chronic hepatitis C virus [HCV] infection. Positive HCV-RNA cases [109] were subjected to complete blood count [CBC], prothrombin time [PT], partial thromboplastin time [PTT], bleeding time [BT], coagulation time [CT], detection of fibrinogen degradation products [FDPs], measurement of plasma alpha- antitrypsin [AAT], then bone marrow [BM] aspiration and examination for 20 cases. The patients were classified into three groups according to the histopathological staging and grading of liver biopsy. The comparison between groups according to histopathological grading and staging for hematological and chemical parameters revealed a significant statistical difference in platelets count, S albumin, ALT and AST levels. The comparison between groups according to histopathological grading and staging for coagulation profile, AAT level and FDPs revealed a significant statistical difference between all parameters. Bone marrow aspiration and examination revealed mild hypocellularity with an increased number of lymphocytes and a relevance of plasmacytoid-lymphocytes

Autoantibodies in HCV infected patients at Sharkia Governerate, Egypt

The aim of this study was to evaluate some immunological manifestations in chronic hepatitis C patients and to find out its relationship with liver pathology. The study included 109 positive HCV-RNA patients classified according to liver histopathology into three groups: Group I included 22 patients [G1S1], group II included 67 patients [G2S2] and group III included 20 patients [G3S3], where G = the degree of necro-inflammatory process and S = the stage of liver fibrosis. All patients were investigated for the presence of cryoglobulin, antineutrophil cytoplasmic [ANCA], anti-liver kidney microsomes [LKM], anti-double stranded DNA, [ds-DNA], anti-nuclear [ANA], anti-mitochondrial [AMA] and anti-smooth muscle [ASMA] autoantibodies. The high prevalence of autoantibodies in chronic HCV patients suggests that HCV may trigger an autoimmune reaction, but most probably do not indicate a distinct autoimmune mechanism. Cryoglobulins and ANCA may be considered as useful prognostic indicator for the increased risk of cirrhosis in chronic HCV patients. Follow up studies were recommended

HCV and associated concomitant infections at Sharkia Governorate Egypt

This study was performed on 109 cases divided into 6 groups according to the concomitant infection with hepatitis C virus [HCV]. The results proved that groups 1, 3 and 5 had a higher level of viremia than the other groups and a higher risk was found in these groups, as 56.4% and 34.6% were in G2S2 and G3S3, respectively. All cases of liver cell dysplasia and hepatocellular carcinoma in this study were seen in these groups. The study concluded that these factors play an important role in the progression of HCV infection. The death of the patients of this progressive condition occurs in younger age and due to liver failure more than to HCC

Non-invasive markers and predictors of severity of hepatic fibrosis in HCV patients at Sharkia Governorate, Egypt

This study included 109 patients with detectable hepatitis C virus [HCV] by real time PCR. The patients were classified into three different pathological stages and grades according to the new concept of histopathological staging and grading. The different clinical, biochemical, virological and ultrasonographic parameters were assessed and analyzed and the variables that showed a significant association with the histopathological staging and grading were included in the multivariate logistic regression analysis. The regression model revealed that platelet count, matrix metalloproteinase-9 [MMP-9], portal vein diameter, splenic longitudinal axis, alanine transaminase, aspartate transaminase and viral load added a significance to the model in a decreasing order of significance. From these findings, a new score ranged from 0-9 was generated. The score model was applied to the patients to assess its validity, where it proved to be accurate in discriminating patients with mild inflammation and fibrosis [sensitivity 81.8%, specificity 80.5% and accuracy 80.7%] and more accurate in detecting patients with cirrhosis [specificity 96.6%, sensitivity 80% and accuracy 93.6%], but less accurate in detecting patients with moderate to severe fibrosis [specificity 66.7%, sensitivity 68.7% and accuracy 67.9%]. Also, the results revealed that co-infection with schistosomiasis, old age >/45 years and positive history of blood transfusion as a source of infection were significantly associated with severe hepatic pathology

role of aflatoxin-contaminated food materials and HCV in developing hepatocellular carcinoma in Al-Sharkia Governorate, Egypt

In this study, the role of aflatoxin contamination in the onset of liver cancer in HCV-infected patients was studied. The quantitative identification of the possible aflatoxins contamination in six urban and eleven rural areas using high performance liquid chromatography technique revealed that corn, wheat, peanut, lupine "tennis", white rice, cowpea "lobiya", fava bean and brown rice showed a prevalence of AFB1 [64.7%, 53%, 53%, 47%, 47%, 41%, 29.4% and 29.4%, respectively]. A positive correlation was found between aflatoxin and positive HCV- PCR together with liver disease progression to G3S3, the indicative of hepatocellular carcinoma. Such correlation was not fully understood, but the oncogene amplification caused by HCV- infection may be aggravated by the consumption of aflatoxin contaminated raw food materials or their products

Experimental demonstration of hepatitis C virus [HCV] in an Egyptian strain of Culex pipiens complex

Reverse transcriptase [RT] polymerase chain reaction [PCR] was used to detect hepatitis C virus [HCV] RNA among heads, guts, larvae and eggs of Culex pipiens complex. The mosquitos were trapped from homes of hepatitis C patients or among the same organs of symbiotic [normal gut bacteria] and aposymbiotic [without gut bacteria] mosquitos fed HCV positive blood by an artificial membrane feeder. The eggs and larvae resulted from symbiotic females fed HCV positive blood were tested for HCV-RNA. Hepatitis C virus RNA was detected only in the heads of symbiotic mosquitos collected from homes of HCV positive patients at 3 and 6 hours after feeding. The virus was detected at 3 and 8 days after being fed on HCV-RNA positive blood in guts of the same group. The virus was not detected in the eggs or larvae resulted from female mosquitos fed on HCV-RNA positive blood