The course of infection with Toxoplasma gondii RH strain in mice pre-vaccinated with gamma irradiated tachyzoites.

Toxoplasma gondii is a ubiquitous protozoan of major medical and veterinary importance. Its treatment is difficult since the available drugs have severe side effects and reactivation may occur anytime. Vaccination with irradiated parasites exhibits ideal characteristics for vaccine development. In our experimental mice model, the protection against challenge with the virulent RH strain was assessed, using 255Gy irradiated tachyzoites. Eighty mice were allocated into 3 groups: naive control group, challenged with virulent RH tachyzoites group and a third group which is challenged with 1 × 106 irradiated tachyzoites, administered as two biweekly doses intraperitoneally. Protection was tested by challenging vaccinated mice with the virulent type RH tachyzoites 30 days after the 2nd vaccination dose. The assessment was built on qualitative clinical, quantitative parasitological, histopathological parameters and measurement of serum Nitric Oxide (NO). The results showed prolonged survival rate, absence of tachyzoites in the peritoneal aspirate by counting, absence of tachyzoites in all examined organs by impression smears, amelioration of histopathological changes in the liver, spleen, brain and lung specimens and increase of the serum NO level in the vaccinated group. Therefore, we propose that irradiated Toxoplasma tachyzoites confer protection for challenged mice and could be an alternative immunization schedule for vaccine development especially for who are at risk of severe immunosuppression.

Serological and molecular screening of umbilical cord blood for Toxoplasma gondii infection.

BACKGROUND: Human umbilical cord blood has proven to be a successful alternate source of hematopoietic stem cells for pediatric patients with major hematologic disorders. Toxoplasma gondii is a global opportunistic protozoan which cause fatal complications in immunocompromised individuals. AIM: Our goal is to study the prevalence of toxoplasmosis in umbilical cord blood (UCB) and to assess the sensitivity of ELISA and PCR for Toxoplasma infection screening. MATERIAL AND METHODS: One hundred cord blood samples were collected immediately after delivery. Anti-Toxoplasma IgG and IgM antibodies were determined using ELISA method; Toxoplasma DNA was detected using nested PCR technique. Total nucleated cells (TNC) and HB were also determined. Demographic data and risk factors data related to the transmission of toxoplasmosis, were collected from mothers. RESULTS: Among 100 cord blood samples, 36 (36%) were positive for anti-Toxoplasma IgG antibodies and 6 (6%) were positive for anti-Toxoplasma IgM antibodies. The nested PCR showed 11 (11%) samples containing Toxoplasma DNA from which, 6 (55%) samples were IgM positive. There was no significant association between the risk of Toxoplasma transmission and cord blood positivity for toxoplasmosis. CONCLUSION: Owing to the prevalence of toxoplasmosis, its rapid progression and its fatal outcome in immunocompromised patients, cord blood screening for toxoplasmosis with nested PCR should be incorporated into cord blood bank screening protocols.

Sex dichotomy in the course of experimental latent toxoplasmosis.

Toxoplasma gondii is an opportunistic zoonotic protozoan that exceeds neurological and congenital impact sequence to reactivating latent toxoplasmosis especially under immunosuppression. Sex-associated hormones influence the severity of Toxoplasma infection. Thus, our study aimed to compare toxoplasmosis associated morbidity in both male and female mice and to monitor the response to anti-Toxoplasma therapeutics fortified with sex hormones in comparison to presently used drugs. Twenty male and 20 female mice were infected with ME49 Toxoplasma strain. The morbidity was assessed in the chronic stage in both sexes by estimating brain cyst burden, brain histopathological examination and monitoring serum anti-Toxoplasma IL-12 using ELISA method. Another 40 male and 40 female mice were infected with ME49 Toxoplasma strain then after 6 weeks received different treatment regimens including Atovaquone, Spiramycin, Metronidazole, Estradiol benzoate and Testoserone propionate either as a monotherapy or as a combination. Treatment response was monitored by scoring mice activity and brain cyst burden. This study showed that female mice demonstrated higher cyst burden and manifested more pathological reactions than male mice. While, the IL-12 serum level was significantly higher in male than female mice. Also, it is proved that the Toxoplasma cyst number was reduced significantly when used testosterone/atovaquone, or testosterone/spiramycin/metronidazole combined regimen in female mice groups. While for male mice, the combined therapy of spiramycin/metronidazole was the superior one. Accordingly, combined therapy with sex hormones is a promising strategy for discovering new therapeutic regimens for treating latent toxoplasmosis especially in female.

Could Araucaria heterophylla resin extract be used as a new treatment for toxoplasmosis?

Toxoplasmosis is a worldwide parasitic disease responsible for serious health problems to human. The currently available drugs used for toxoplasmosis treatment showed a limited efficacy and cause serious host toxicity. The in vitro screening for toxoplasmicidal activity of Araucaria heterophylla resin (AHR) extract and its major component 13-epi-cupressic acid (CUP) showed that both AHR (EC50 = 3.90) and CUP (EC50 = 3.69) have high toxoplasmicidal activity in comparison with standard cotrimoxazole (EC50 = 4.28). The antiprotozoal effects of AHR and CUP were investigated against acute and chronic toxoplasmosis using mice models. Two groups of Swiss albino mice were infected by RH Toxoplasma strain intraperitoneally and by Me49 strain orally. Both groups were treated with AHR and CUP in different doses. Their effects were evaluated by survival rate, peritoneal, spleen and liver parasite burdens, brain cyst burden, NO serum level and histopathological lesions. The ultrastructural changes of tachyzoites of acutely infected mice were studied using scanning electron microscopy (SEM). There is an evidence of toxoplasmicidal activity of AHR and CUP in acute and chronic experimental toxoplasmosis. In the acute model, mice treated with AHR and CUP showed prolonged survival rates, a significant decrease in the parasite density in peritoneal lavage and pathological insult in both liver and spleen compared with that of untreated ones. SEM results denote evident morphological alterations of treated tachyzoites. In chronic experimental toxoplasmosis, AHR and CUP treated groups could significantly reduce brain cyst burden by 96.05% and 98.02% respectively. This study indicates that AHR and CUP showed potent toxoplasmicidal activities experimentally and could be used as a potential natural nontoxic agent for treatment of toxoplasmosis.

Seroprevalence of Toxoplasma gondii infection in children with central nervous system disorders in Mansoura, Egypt: a case-control study.

Background: Toxoplasma gondii is a global infection with a crucial role in the development of neurological diseases. Data concerning the association between T. gondii and neurological illnesses in Egyptian children is scarce. Methods: A case-control study was conducted on 60 patients divided into children suffering from central nervous system manifestations without apparent chromosomal anomalies (n=30) and children with Down syndrome (n=30) recruited from Mansoura University Children's Hospital, Mansoura, Egypt. A total of 30 healthy children were included as controls. Demographics and clinical data were collected from all cases and Toxoplasma immunoglobulin (Ig) M and G antibodies were assessed by enzyme-linked immunosorbent assay. Results: Anti-T. gondii IgG was the most frequent antibody detected and the highest seropositivity rates were ranked for the neurologically disabled non-syndromic children, followed by Down syndrome, compared with controls (p≤0.001). Statistically significant (p=0.05) associations were found between Toxoplasma IgG seropositivity and hydrocephalus and between Toxoplasma IgM and a history of contact with farm animals, soil and cats in children with Down syndrome. Conclusions: The association between Toxoplasma infection and neurological disorders in children should be kept in mind by paediatricians and assessment of T. gondii antibodies in early childhood is needed for timely management of afflicted patients.

Epidemiological, clinical and laboratory findings of infectious keratitis at Mansoura Ophthalmic Center, Egypt.

AIM: To analyze the epidemiological, clinical and laboratory findings of infectious keratitis. METHODS: A retrospective study on cases of infective keratitis, attended our institution from Mar. 2013 to Feb. 2015, was done at Mansoura Ophthalmic Center, Egypt. Corneal scrapings were performed and processed for direct microscopy and culture in appropriate media using standard laboratory protocols. RESULTS: Out of 245 patients enrolled for study, 247 corneal scrapings were obtained. Ocular trauma was the most common predisposing factor (51.4%), followed by diabetes mellitus (15.1%). Cultures were positive in 110 scraping samples (44.5%): 45.5% samples had pure fungal infection, 40% had pure bacterial infections and 10% had mixed fungal and bacterial growths. Acanthamoeba was detected in 5 (4.5%) samples. The most common fungal pathogen was Aspergillus spp. (41%). The most common bacterial isolates were Staphylococcus aureus (38.2%) and Pseudomonas aeruginosa (21.8%). CONCLUSION: Incidence of fungal keratitis is high in our region. Therapeutic approach can initially be based on clinical features and sensitivity/resistance patterns. Microbiological research should direct the antimicrobial treatment. Antibiotic resistance to fluoroquinolones and aminoglycosides is an important consideration.

Helminthes and insects: maladies or therapies.

By definition, parasites cause harm to their hosts. But, considerable evidence from ancient traditional medicine has supported the theory of using parasites and their products in treating many diseases. Maggots have been used successfully to treat chronic, long-standing, infected wounds which failed to respond to conventional treatment by many beneficial effects on the wound including debridement, disinfection, and healing enhancement. Maggots are also applied in forensic medicine to estimate time between the death and discovery of a corpse and in entomotoxicology involving the potential use of insects as alternative samples for detecting drugs and toxins in death investigations. Leeches are segmented invertebrates, famous by their blood-feeding habits and used in phlebotomy to treat various ailments since ancient times. Leech therapy is experiencing resurgence nowadays in health care principally in plastic and reconstructive surgery. Earthworms provide a source of medicinally useful products with potential antimicrobial, antiviral, and anticancer properties. Lumbrokinases are a group of fibrinolytic enzymes isolated and purified from earthworms capable of degrading plasminogen-rich and plasminogen-free fibrin and so can be used to treat various conditions associated with thrombotic diseases. Helminth infection has been proved to have therapeutic effects in both animal and human clinical trials with promising evidence in treating many allergic diseases and can block the induction of or reduce the severity of some autoimmune disorders as Crohn's disease or ulcerative colitis. What is more, venomous arthropods such as scorpions, bees, wasps, spiders, ants, centipedes, snail, beetles, and caterpillars. The venoms and toxins from these arthropods provide a promising source of natural bioactive compounds which can be employed in the development of new drugs to treat diseases as cancer. The possibility of using these active molecules in biotechnological processes can make these venoms and toxins a valuable and promising source of natural bioactive compounds. The therapeutic use of helminthes and insects will be of great value in biomedicine and further studies on insect toxins will contribute extensively to the development of Biomedical Sciences.

Toxoplasma gondii infection among chronic hepatitis C patients: a case-control study.

OBJECTIVE: To determine the detection rate of anti-Toxoplasma gondii (T. gondii) IgG and IgM in chronic HCV patients attending the Department of Tropical Medicine Mansoura University hospital in Egypt. METHODS: This study included 120 adult chronic HCV patients, 81 decompensate cirrhosis (late-stage) and 39 chronic HCV non cirrhotic patients (early-stage) and 40 healthy blood donors as controls. Serum samples were examined for anti-Toxoplasma IgM and anti-Toxoplasma IgG antibodies by ELISA. Real-time RT-polymerase chain reaction assay was done for quantitation of hepatitis C virus. RESULTS: Anti-T. gondii IgG antibodies were detected in 75 (92.6%) of 81 late-stage cirrhotic patients, 30 (76.9%) of the 39 chronic HCV non cirrhotic patients (early-stage) and in 6 (15%) of 40 controls with statistically significant difference (P<0.001). Anti-T. gondii IgM antibodies were found in 11 (13.6%) in late stage patients, 5 (12.8%) in early stage and in 3 (7.5%) of controls with no statistical significant difference (P=0.610). There was no correlation between stage of fibrosis and IgM or IgG antibodies positivity in our studied groups (P=0.526). High IgG levels significantly correlated with high viral load (P=0.026). CONCLUSIONS: Our findings suggest that the serious opportunistic T. gondii infection represent a potential significant risk for chronic HCV patients. So, toxoplasmosis should be considered in their investigations and follow-up.

Performance of rapid immunochromatographic assay in the diagnosis of Trichomoniasis vaginalis.

Trichomonas vaginalis accounts for nearly half of all curable sexually transmitted diseases worldwide with serious health consequences. Effort to increase the sensitivity of its diagnosis is critical to both control measures and epidemiologic studies. This study was conducted to evaluate the OSOM® Trichomonas Rapid Test (Sekisui Diagnostics, Framingham, MA, USA), a qualitative antigen-detection immunochromatographic (IC) assay in the diagnosis of vaginal trichomoniasis comparable to the conventional methods. The study enrolled 258 females aged 18-50 years classified into symptomatic (185) and asymptomatic (73) groups. Vaginal swab specimens were obtained for wet mount, stained preparation (Giemsa, acridine-orange), culture (InPouch TV™, modified Diamond's), and for rapid OSOM testing. Trichomonas vaginalis was detected in 67, 66, 71, 99, 96, and 97 using wet mount, acridine-orange stain, Giemsa stain, modified Diamond's, InPouch media, and OSOM test, respectively. In comparison to a composite reference standard (CRS) of wet mount microscopy and culture, OSOM test reported 97.98%, 99.37%, 98.98%, 98.75%, and 98.84% for sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy, respectively. The OSOM test proved to be a simple and objective test. This rapid point-of-care-test will contribute significantly in the diagnosis of vaginal trichomoniasis and will increase the understanding of its still vague epidemiology.