Atorvastatin: In-Vivo Synergy with Metronidazole as Anti-Blastocystis Therapy

Blastocystis is an enteric Straminopile in tropical, subtropical and developing countries. Metronidazole has been a chemotheraputic for blastocystosis. Failures in its regimens were reported and necessitate new studies searching for alternative therapeutic agents. Aim of current study is to investigate potential effects of Atorvastatin (AVA) compared to the conventional chemotherapeutic MTZ in experimentally Blastocystis-infected mice. Anti-Blastocystis efficacy of AVA was evaluated parasitologically, histopathologically and by transmission electron microscopy using MTZ (10 mg/kg) as a control. Therapeutic efficacy of AVA was apparently dose-dependent. Regimens of AVA (20 and 40 mg/kg) proved effective against Blastocystis infections with high reduction in Blastocystis shedding (93.4–97.9%) compared to MTZ (79.3%). The highest reductions (98.1% and 99.4%) were recorded in groups of combination treatments AVA 20–40 mg/kg and MTZ 10 mg/kg. Blastocystis was nearly eradicated by the 20th day post infection. Genotype analysis revealed that genotype I was most susceptible, genotype III was less. Histopathologic and ultrastructural studies revealed apoptotic changes in Blastocystis and significant improvement of intestinal histopathological changes more remarkable in combinational therapy groups. Thus, the present study offers AVA as a potential candidate for Blastocystis therapy combined with MTZ.

Genotypic Identification of Cystoisospora in Immunocompromised Patients Using T(m)-Variation Analysis

Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures (T(m)s) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified T(m)s at 85.8°C and 88.6°C, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using qPCR-T(m)s analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.

Potential use of Biomphalaria alexandrina snail antigens for serodiagnosis of Schistosomiasis Mansoni by immunoblot analysis

The aim of this study was to evaluate the possible use of Biomphalaria alexandrina snail antigens in diagnosis of schistosomiasis mansoni using enzyme linked immunolectrotransfere blot [EITB]. S. mansoni adult worm crude antigens [AWA], feet and visceral humps of B. alexandrina and Bulinus truncatus were used. Hyperimmune mice sera [HIS] versus each antigen were prepared for diagnosis of S. mansoni using western blot [WB]. Snail foot antigens were more specific in antibodies detection than visceral hump antigens. Three of five polypeptides of B. alexandrina foot antigen identified by S. mansoni HIS showed specific positive reactivity. These polypeptides were at MW of 31/32 and 43 kDa. While, only one of the six polypeptides of B. alexandrina hepatopancrease antigen identified by S. mansoni HIS, at a MW of 43 kDa was specific. Similarly, 2 polypeptides at MW of 44 and 55 kDa were specific in detection of anti-S. haematobium antibodies. However, the antigenically active polypeptide of B. truncatus hepatopancrease antigen had no specific reactivity towards anti-S. haematobium antibodies. B. alexandrina foot antigens were the most specific of the tested snail antigens in diagnosis of schistosomiasis mansoni

Therapeutic Potential of Myrrh and Ivermectin against Experimental Trichinella spiralis Infection in Mice

Trichinosis is a parasitic zoonosis caused by the nematode Trichinella spiralis. Anthelmintics are used to eliminate intestinal adults as well as tissue-migrating and encysted larvae. This study aimed to investigate the effects of ivermectin and myrrh obtained from the aloe-gum resin of Commiphora molmol on experimental trichinosis. Ninety albino mice were orally infected with 300 T. spiralis larvae. Drugs were tested against adult worms at day 0 and day 5 and against encysted larvae on day 15 and day 35 post-infection (PI). Mature worms and encysted larvae were counted in addition to histopathological examination of muscle specimens. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, globulin, urea, and creatinine values were estimated. Significant reductions in mean worm numbers were detected in ivermectin treated mice at day 0 and day 5 PI achieving efficacies of 98.5% and 80.0%, while efficacies of myrrh in treated mice were 80.7% and 51.5%, respectively. At days 15 and 35 post-infection, ivermectin induced significant reduction in encysted larval counts achieving efficacies of 76.5% and 54.0%, respectively, while myrrh efficacies were 76.6% and 35.0%, respectively. AST, ALT, urea, and creatinine levels were reduced, while total proteins were increased in response to both treatments compared to their values in the infected non-treated mice. Ivermectin use for controlling T. spiralis could be continued. Myrrh was effective and could be a promising drug against the Egyptian strains of T. spiralis with results nearly comparable to ivermectin.

Modulatory role of immunization with secretory-excretory products of schistosoma haematobium eggs on morbidity in infected hamsters

A variety of secretory-excretory products [SEPs] from different stages of Schistosoma have been identified to induce a level of host-protective immune responses with amelioration of morbidity. Identification of SEPs complex components can be expected to facilitate discovery of new therapeutic drug targets and new diagnostic markers for schistosomiasis control. The present study was undertaken to evaluate the possible anti-morbidity effects of SEPs of S. haematobium eggs twelve weeks after exposure to infection. The liver and intestinal tissues of infected hamsters were selected for evaluation as in the murine models the urinary bladder exhibits minimal morbidity in response to S. haematobium infection. The experimental design included three groups of 10 hamsters each; SEPs immunized group, infected immunized group and infected control group. Multiple small doses of purified S. haematobium eggs SEPs were injected intra-peritoneally, followed 2 weeks later with 2 booster doses at weekly intervals. Animals were infected with S. haematobium cercariae 1 week following last booster immunization dose. All animals were sacrificed 12 weeks PI. Assessment of the modulatory role of SEPs immunization including worm burden, tissues egg loads, oogram pattern and histopathological examination of liver and intestinal tissues were carried out. Total and subclasses of anti SEPs of S. haematobium eggs IgG, IgM, IgG2 and polyvalent immunoglobulins [Igs] were measured using indirect ELISA at weeks 3, 6 and 9 post-infection [PI]. Immunization with SEPs of S. haematobium eggs produced significant reduction in worm load [61.37%], reduction in tissue egg loads [54.85% and 41.57% for hepatic and intestinal ova, respectively]; decreased percent of immature stages and increase in the percent of dead ova in oogram pattern. Pathological examination also revealed significant reduction in number of hepatic granuloma [46.06%]. At 3, 6 and 9 weeks PI, the level of Igs especially IgG was significantly higher in the infected immunized group, compared to both control groups, reached a peak value at 6 weeks PI [1.6] and remained elevated till the end of the experiment with slightly decreasing tendency at 9 weeks PI [1.3]. This study could represent an immunization model as a trial to decrease severe morbidity of schistosomiasis haematobium which may be aggravated by serious sequels

pathogenic role of different blastocystis hominis genotypes isolated from patients with irritable bowel syndrome

The pathogenic role of Blastocystis hominis is still regarded by some as controversial. Studies have been in progress for years to evaluate the role of blastocystosis in irritable bowel syndrome [IBS] and demonstrated that faecal carriage of B. hominis was frequent in these patients. This study attempted to distinguish different genotypes of B. hominis isolates obtained from patients with IBS and to evaluate their pathogenic potentials. One hundred subjects [51 patients with IBS and 49 asymptomatic infected subjects] harbouring B. hominis were investigated by a direct smear examination and in vitro culture of stool samples followed by genotyping of B. hominis by PCR using STS primers. Sigmoidoscopy was done in all subjects and biopsies were taken and subjected to histopathologic examination. Genotyping proved that only four genotypes of B. hominis were identified. In patients with IBS, genotypes III, I, and IV were detected [28, 15 and 14 isolates, respectively]. On the other hand, genotypes III, IV, and II were identified in asymptomatic infected individuals [21, 19 and 13 isolates, respectively]. The degrees of chronic inflammatory changes in sigmoidoscopic biopsies caused by B. hominis genotypes among IBS patients revealed that severe inflammation was present mainly in patients harboring genotype I isolates [4/15] [26.66%], while genotype III caused severe inflammation only in 9.09%. Genotype II isolates were not detected in IBS cases. Asymptomatic infected individuals harboring genotypes II, III and IV exhibited mild to moderate inflammatory changes. Genotype I isolates were not detected in asymptomatic infected group. The correlation between different B. hominis genotypes and degree of inflammation was statistically insignificant. Genotype I was the most pathogenic genotype of B. hominis isolates in patients with IBS while genotype II was not detected among those patients. Also, our results suggest the presence of pathogenic and non-pathogenic strains among genotypes III and IV