Diversity and comparative studies on bulinus snails collected from two localities in Egypt

A comparative study of the shell structure, seasonal temperature and Calcium content of Bulinus snails from two areas [Damietta and Giza] in Egypt was done and compared with laboratory snails from Schistosome Biological Supply Center [SBSC]. The shells of collected snails identified as Bulinus truncatus, showed a wide variation in shape. The results showed a significant differences were detected between the populations from SBSC and Damietta [p<0.05] for mean of measured shell width, aperture length, length of spire and number of whorls. The populations from Giza and Damietta governorates showed significant differences [P<0.05] in mean of measured length of diagonal, length of body whorl above aperture, length of spire and number of whorls. There were no statically significant differences between the populations from SBSC and Giza. The seasonal temperature affected on susceptibility of snails to infection with Schistosoma haematobium. The mean prepatent period was short in summer and long in winter. The shells of S. haematobium - infected B. truncatus snails showed hypocalcification from all localities

Cytokines and mother sporocysts in susceptible and resistant bulinus truncatus snails infected with schistosoma haematobium

The presence of immunoreactive interleukin [IL-2] interferon gamma [FN-y] and tumor necrosis factor alpha [TNF-alpha] in addition to the citation of mother sporscytes in cephalopodal musculature in the susceptible and resistance Bulinus truncatus the specific intermediate host for the trematode Schistosoma haematobium were investigated. Using ELISA tests, Results indicated that the concentration of IL-2-like activity in the susceptible and resistant snails decreased significantly after infection then persisted at low levels until the 4[th] week post exposure [WPE] in susceptible snails, while in resistant snails elevated during the second WPE, and returned to initial level at 3 and 4 WPE. Susceptible snails had low detectable levels of TNF-alpha and INF-y like-activity after infection. However, the resistant snails had significant low levels of TNF-alpha and INF-y like-activity from 3 WPE until the 4th WPE without any sign of normalization. Histological sections in the head- foot region of susceptible and resistance B, truncatus infected with S. haematobium, mother sporocysts exists froml to 7 [day post exposure] DPE, in the susceptible snail the mother sporocysts were found as single, multiple and mature types. No mother sporocysts were appear in the lip and mantle of the snail on 2, 5, 7 DPE and on 1-3, 6 DPE respectively. In the resistant snails few mother sporocysts were found in the lip, mantle and tentacles. The results showed that schistosome-resistant Bulinus can be an alternative strategy for the control of schistosomiasis

Osteoclastogenesis and lymph node organogenesis in different species of Egyptian rats

The development of lymphoid organs depends on the correct expression of several molecules within a defined timeframe during ontogeny. Although this is an extremely complex process, with each secondary lymphoid tissue requiring subtly different signals, a common framework for lymphoid development is beginning to emerge. Bone remodeling is tightly regulated by a molecular trial composed of OPG/RANK/RANKL. The receptor activator of RANKL [localized on osteoblasts] enhances osteoclastogenesis via interaction with its receptor RANK [localized on osteoclasts], whereas osteoprotegerin [OPG] [produced by osteoblasts] inhibits this osteoclastogenesis by binding to RANKL. The RANK provides critical signals necessary for lymph node organogenesis and osteoclast differentiation. The TNF family molecule OPGL has been identified as a potential osteoclast differentiation factor and regulator of interactions between T cells and dendritic cells in vitro. Thus OPGL is a new regulator of lymph node organogenesis and lymphocyte development and is an essential osteoclast differentiation factor in vivo: So, the result of this study showed that lymph node organogenesis appears to require adequate quantity of RANKL, and this significant level can apparently persist despite marked overexpression of the soluble RANKL inhibitor OPG

Immunological and molecular detection of digenetic infections in different species of Egyptian freshwater snails

Due to the possibility of utilizing different snails in the combat of Schistosoma in Egypt; it is important to study the role it may play in transmitting other trematodes of medical and veterinary importance. Taking this background into consideration, polymerase chain reaction [PCR] assay was designed to identify trematode species at larval stages in intermediate hosts [cercariae in snails] using a combination of standard and molecular methods. This PCR assay was also applied to naturally infected molluscan in order to assess the use of the procedure for detection. The importance of the present study was to demonstrate the epidemiological situation and application in control

Immune response of mice infested by hyalomma dromedarii ticks infected with entomopathognic nematodes

In the present work, the total leucocytes count of infested mice with ticks Hyalomma dromedarii infected with nematodes showed significant increase than infested group with ticks only. The infested group of mice with ticks and infested mice with ticks and nematodes showed a markedly significant drop in the eryhrocytes count. The results showed a highly significant decrease in Hb concentration, and HCT percentage for mice infessted with ticks and a marked increase with ticks infested with nematodes. The results showed a significant decrease in level of IgG of mice infested with ticks, while in infected mice with ticks and nematodes, the IgG level increased to control level which might indicated that nematodes penetrate ticks that were feeding on mice through the thinner cuticle of engorging females. Differences in cytokines responses were observed between tick-infested and non-infested mice. Proinflammatory Th1 T lym-phocyte cytokines were suppressed and TH2 cytokines were enhanced for infestation with H. dromedarii. The date indicated a polarization of immune response towards Th2 lymphocytes.

Association of immune responses to mycobacterium tuberculosis peptide antigens with host genetic factors

Apolymerase chain reaction [PCR], based on insertion sequence IS6110,was developed to detect mycobacterium tuberculosis complex organisms in the blood samples of 56 tuberculosis patients and 34 healthy controls.the erly secreted antigenic target 6-KDa[ESAT-6] are used to stimulate T lymphocyte subsets from tuberculosis infected patients and the correltion of these immune responses to the genetic factoes[HLATYPE] which determined the host immune response is evaluated.ESAT-6 derived peptides:P1[1.05 +/- 0.084],P2[1.08 +/- 0.094],P3[1.02 +/- 0.086],P5[0.98 +/- 0.117] and P7[1.26 +/- 0.152] were significantly higer in the infected group than in non-infected one. Besides, 33ptients and 12 controls were tested for HL-DRB HLA DQB and HLA-DPB Only type HLA-DEB 15was significan tly associated with tuberculosis infection using the Chi-square test[X[2]=0.04311]. By using the relative risk some HL types were relatively more susceptible to be associated with tuberculosis infection. HLA-DR typing of patients showed that they covered a large spectrum of HLA-DR molecules encoded by HLA-DEB1,-DRB3,-DRB4,and DRB5genes.however,HLA-DQ typing showed that they HLA-DQB1 molecules, HLA-DP typing of patients showed that covered a large spectrum of HLA-DP molecules encoded by HLA-DPB1.

Biomphalaria Alexandrina: cellular responses of susceptible and resistant snails to Schistosma manson infection

The distribution pattern of Schistosoma mansoni miracidia among exposed Biomphalaria alexandrina determined by the level of host susceptibility/resistance and the basic cellular responses observed during the parasite development were studied. Several snail stocks showed a wide spectrum of host reaction to the parasite. From the present study, a vigorous "resistant-type" cellular response to invade miracidia was observed in the histological sections of the non- susceptible snails. In this respect, they were classified in this study as "resistant snails". B. alexandrina experimentally infected with S. Mansoni exhibited a wide range of histopathological and immunological changes. The rate of phagocytosis as well as the actual number of hemocytes were determined in different groups of snails during the infection cycle. The significant fluctuation [increase and/or decrease] in circulating hemocyte number was only correlated with a shift in the hemocyte populations in the first two days after the exposure to the infective stage. The in vitro phagocytic activity of the resistant snail hemocytes was found to be higher than that observed in the susceptible snails

possible rolefor an enkephalinergic system in the internal defense mechanism of Biomphalaria alexandrina exposed to Schistosoma mansoni

The present study revealed that in Biomphalaria alexandrina, the coordinated responses to Schistosoma mansoni infection are modulated by receptor-mediated opioid signals. The comprehensive tests in susceptible and resistant snails demonstrated the presence of endogenous opioids in the snail hemolymph [in particular, Leu- enkephalin-like material]. The in vitro treatment of snail hemocytes with synthetic Leu-enkephalin analogue [DADLE] resulted in the modulation of cellular adherence and phagocytic activity. The addition of naloxone, either alone or in combination with DADLE, generally reduced the hemocyte activity indicating opioid-receptor- mediated mechanism. The presence of DADLE or naloxone modulated the level of IL-2-, TNF-gamma- and FNF-alpha-like molecules in S. Mansoni resistant and susceptible snails. Specifically, DADLE and DADLE in combination with naloxone, generally, were found to be capable of modulating the resistant snail hemocytes at concentrations of 10-6 and 10-8 M. Similar actions after incubation with the same concentrations were not detected in the susceptible snails

Cellular response to Schistosoma mansoni infection in Biomphalaria alexandria strains selected for susceptibility and resistance

Exposure of juvenile and adult Biomphalaria alexandrina to Schistosoma mansoni miracidia resulted, typically, in three susceptibility patterns: a]non-infected snails b] normal infections, and c] retarded infections. Under laboratory conditions, a vigorous resistant-type cellular response to invading miracidia was seen in the histological sections of non-susceptible snails. Accordingly, they were classified as resistant snails. Data pertaining to the influence of host size on suceptibility to S. mansoni indicates that adult snails [i.e.10-20 mm shell diameter] were significantly less likely to harbour sporocysts than juvenile ones [i.e. 5-10 mm shell diameter]. Cellular reaction to the infection varied with sporocysts location and length of infection. At 2 days post exposure [DPE], most sporocysts were viable. Approximately 8-12% of the sporocysts had elongated shaped transverse constriction and were categorized "normal", while those showing no elongation were categorized "retarded". All remaining sporocysts at 4 DPE were categorized "dead" while at 30 DPE most sporocysts were "amorphous" with eosinophilic masses. Although encapsulation of sporocysts never occurred in susceptible snails, haemocyte aggregations could sometimes be observed in the proximity of well developed sporocysts. In resistant snails, this cellular response continued to increase and resulted in the encapsulation of the sporocysts. In susceptible snails, the nucleus of secretory cells of the albumen cytoplasm were different in susceptible and resistant snails

Evaluation of serum total bile acids in the diagnosis of hepatocellular carcinoma

Fasting total serum bile acids [FSBA] concentrations were measured in 140 cases of chronic liver diseases including 50 patients with liver cirrhosis, 40 patients with chronic hepatitis and 50 patients with hepatocellular carcinoma [HCC]. FSBA concentrations were significantly higher in patients with HCC than those with chronic hepatitis and cirrhosis and both were significantly higher than the controls [p<0.0001]. FSBA concentrations were not correlated to the size of the tumor, the Child Pugh grades, the histopathologic grades and most of the liver function tests. Using the receiver operative characteristic [ROC] and the differential positive rate [DPR] analysis, 30 pmol/l was the optimal cut-off value that differentiates patients with HCC from those with cirrhosis. At this level, the sensitivity, the specificity and the diagnostic accuracy were 42%, 98% and 70% respectively. AFP was found to be significantly increased in patients with HCC than those with chronic hepatitis and cirrhosis. The best cut-off value of AFP was 100 ng/ml, at which the sensitivity, the speci-ficity and diagnostic accuracy were 52%, 96% and 74% respectively. The simultaneous determination of AFP and total bile acids raised the sensitivity of the test to 76%. The area under the ROC curve for AFP and total bile acids was 0.764 and 0.744 respectively, the difference is non-significant. In conclusion, both AFP and total bile acids are good markers for HCC and their simultaneous determination may improve the detection of HCC in cirrhotic patients negative for AFP

Possible role of schistosoma mansoni infection in male hypogonadism

Schistosomiasis has been suggested to decrease the reproductive potential or castrate both invertebrate and vertebrate hosts. Furthermore, schistosomiasis may cause anatomic anomalies of the reproductive organs responsible for permanent or reversible infertility. To specify the effect of schistosomiasis on gonadal functions, production of testosterone [TS], leutinizing hormone [LH] and estradiol [E2] in Egyptian men infected with Schistosomiasis were studied. All participants were tested for clinical examination, semen, liver function tests and blood level of IL-2. The mean TS levels were at the lowest limit of normal range for liver cirrhotic patients. Mean E2 levels were increased in all patients, but patients with liver cirrhosis-related schistosomiasis had higher E2 levels. Linear regression analysis showed that the sex hormone levels correlated best with the patient's liver function parameters. The present data suggested that a sex hormone imbalance played a role in patients with liver cirrhosis due to the inhibitory effects of schistosomiasis on gonadal functions

Immunoaffinity isolation of Schistosoma mansoni species specific molecules using monoclonal antibodies

Traditional diagnosis of schistosomiasis by stool and urine examination techniques is both insensitive and labor intensive; many efforts are directed towards the development of alternate sensitive and specific assays for diagnosis of schistosomiasis. With the development of hybridoma technology, monoclonal antibodies [MoAb] with high specificity to diagnostic antigens could be produced. In the present study, a species-specific fraction namely, Gp30 prepared from S. mansoni microsomal antigen [MAMA] was purified by a simple, easy and cheap apparatus [491-prep cell] [Bio-Rad]. Balb/c mice were immunized with the isolated glycoprotein [Gp] to produce a panel of MoAbs, which can be used to immunoaffinity purify target antigen. Among the produced panel, 6B3-1B432 MoAb of IgG2a isotope was able to detect 10-20 ng of Gp30 antigen in ELISA [O.D[650] = 1.985] and dot blot. In immunoblotting, 6B3-1B432 recognized only one band in the MAMA antigens, namely Gp30. This MoAb was purified with protein G Suprose using FPLC and PD-10 column and consequently used to immunoaffinity purify target antigen. The specificity and sensitivity of the immunoaffinity purified Gp30 [IP Gp30] was determined before using it in the dipstick assay. Our results indicated the absolute specificity of IP Gp30 and sensitivity results, 98%, was comparable to that obtained by the electroeluted GP30. The simplicity of the purification and the efficacy of the dipstick assay may render this diagnostic method for widespread use in the field of schistosomiasis