Effect of myrrh and thyme on Trichinella spiralis enteral and parenteral phases with inducible nitric oxide expression in mice

Trichinellosis is a serious disease with no satisfactory treatment. We aimed to assess the effect of myrrh (Commiphora molmol

Morphological and initial molecular characterization of Clogmia albipunctatus larvae (Diptera: Psychodidae) causing urinary myiasis in Egypt.

Myiasis is the infestation of human tissues by dipterous fly larvae of the class Insecta. Clogmia albipunctatus, family Psychodidae, is one of the most medically important insects that cause human myiasis. The aim of the present study is the morphological identification and the molecular characterization of moth flies causing many cases of urinary myiasis in Egypt, based on sequencing of the mitochondrial DNA of the larvae. Seven urinary samples of patients complaining of urinary symptoms and giving a history of low socioeconomic level were examined. Recovered larvae were identified using light microscopy and SEM. For molecular identification, the mitochondrial genes Cytochrome B (cytB), NADH1, NADH1, and 16S were sequenced and phylogenetically analyzed. The morphological and molecular characterization could accurately diagnose our patients to have C. albipunctatus infestation. Such results provided the initial set of data on the molecular identification and phylogenetic analysis of moth flies based on DNA barcoding in Egypt

Effect of myrrh and thyme on Trichinella spiralisenteral and parenteral phases with inducible nitric oxide expression in mice

Trichinellosis is a serious disease with no satisfactory treatment. We aimed to assess the effect of myrrh (Commiphora molmol) and, for the first time, thyme (Thymus vulgaris L.) against enteral and encysted (parenteral) phases of Trichinella spiralis in mice compared with albendazole, and detect their effect on inducible nitric oxide synthase (iNOS) expression. Oral administration of 500 mg/kg of myrrh and thyme led to adult reduction (90.9%, 79.4%), while 1,000 mg/kg led to larvae reduction (79.6%, 71.3%), respectively. Administration of 50 mg/kg of albendazole resulted in adult and larvae reduction (94.2%, 90.9%). Positive immunostaining of inflammatory cells infiltrating intestinal mucosa and submucosa of all treated groups was detected. Myrrh-treated mice showed the highest iNOS expression followed by albendazole, then thyme. On the other hand, both myrrh and thyme-treated groups showed stronger iNOS expression of inflammatory cells infiltrating and surrounding encapsulated T. spiralis larvae than albendazole treated group. In conclusion, myrrh and thyme extracts are highly effective against both phases of T. spiralis and showed strong iNOS expressions, especially myrrh which could be a promising alternative drug. This experiment provides a basis for further exploration of this plant by isolation and retesting the active principles of both extracts against different stages of T. spiralis

<i>Allovahlkampfia spelaea</i> Causing Keratitis in Humans

<div><p>Background</p><p>Free-living amoebae are present worldwide. They can survive in different environment causing human diseases in some instances. <i>Acanthamoeba</i> sp. is known for causing sight-threatening keratitis in humans. Free-living amoeba keratitis is more common in developing countries. Amoebae of family <i>Vahlkampfiidae</i> are rarely reported to cause such affections. A new genus, <i>Allovahlkampfia spelaea</i> was recently identified from caves with no data about pathogenicity in humans. We tried to identify the causative free-living amoeba in a case of keratitis in an Egyptian patient using morphological and molecular techniques.</p><p>Methods</p><p>Pathogenic amoebae were culture using monoxenic culture system. Identification through morphological features and 18S ribosomal RNA subunit DNA amplification and sequencing was done. Pathogenicity to laboratory rabbits and ability to produce keratitis were assessed experimentally.</p><p>Results</p><p><i>Allovahlkampfia spelaea</i> was identified as a cause of human keratitis. Whole sequence of 18S ribosomal subunit DNA was sequenced and assembled. The Egyptian strain was closely related to SK1 strain isolated in Slovenia. The ability to induce keratitis was confirmed using animal model.</p><p>Conclusions</p><p>This the first time to report <i>Allovahlkampfia spelaea</i> as a human pathogen. Combining both molecular and morphological identification is critical to correctly diagnose amoebae causing keratitis in humans. Use of different pairs of primers and sequencing amplified DNA is needed to prevent misdiagnosis.</p></div

Morphological characters of trophozoite and cyst.

<p>(A) Trophozoite showing unidirectional movement (arrows) and cyst aggregating to each other (arrow head) (inverted microscope using x20 objective lens). (B) Cysts in 10 days old culture (inverted microscope using x20 objective lens). (C) Trophozoite showing filopodia (arrow) (oil immersion x100 objective lens). (D) Cysts with perinuclear clear ring (x40 objective lens)</p

Primers designed to specifically amplify and sequence 18S ribosomal RNA gene (position is in relation to <i>Allovahlkampfia spelaea</i> SK1strain sequence).

<p>Primers designed to specifically amplify and sequence 18S ribosomal RNA gene (position is in relation to <i>Allovahlkampfia spelaea</i> SK1strain sequence).</p

PCR amplification of 18S ribosomal RNA gene.

<p>(A) Lane (1) shows faint band of JDP1 & JDP2 primers, lane (2) shows band amplified using short universal primers, while lane (3) shows band specific for 5.8S ITS with auxiliary band (arrow) indicating long variant. (B) Amplification of whole 18S gene using 3 pairs of primers, bands appear at expected molecular weight calculated during primer design.</p