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Because of resistance and side effects of common anti-parasite and antifungal drugs, there have been many studies on the use of the herbal antifungal Herbal Extract. In this study, the anti-Candida and anti-Fasiola activities of Sage Herb (Salvia Officinalis) extract in comparison with anti-fungal and anti -parasite drugs were examined. The research disc diffusion method was used to study the inhibitory effects of the extract of Sage Herb (Salvia officinalis) and Miconazole, Triclabendazole and fluconazole on species of Candida Albicans and Fasciola Hpatitca, which are isolated from patients who suffered from vulvovaginal candidiasis and Fasciolasis at 25°C and 37°C. The transparent area around the manuscript discs showed that thyme extract has the most effect against Candida Albicans. The inhibitory effects of Sage Herb (Salvia Officinalis) extract on the growth of Candida and Fasciolaat 37°C were better than 25°C. The inhibitory effect of Miconazole, Triclabendazole was better than fluconazole. The results of the present study showed that Triclabendazole and Sage Herb (Salvia Officinalis) extract had inhibitory effects on the growth of Candida Albicans and Fasciola Hpatitca.
This review summarises the findings of a series of studies in which the histological changes, induced in the reproductive system of Fasciola hepatica following treatment of the ovine host with the anthelmintic triclabendazole (TCBZ), were examined. A detailed description of the normal macroscopic arrangement and histological features of the testes, ovary, vitelline tissue, Mehlis' gland and uterus is provided to aid recognition of the drug-induced lesions, and to provide a basic model to inform similar toxicological studies on F. hepatica in the future. The production of spermatozoa and egg components represents the main energy consuming activity of the adult fluke. Thus the reproductive organs, with their high turnover of cells and secretory products, are uniquely sensitive to metabolic inhibition and sub-cellular disorganisation induced by extraneous toxic compounds. The flukes chosen for study were derived from TCBZ-sensitive (TCBZ-S) and TCBZ-resistant (TCBZ-R) isolates, the status of which had previously been proven in controlled clinical trials. For comparison, flukes collected from flocks where TCBZ resistance had been diagnosed by coprological methods, and from a dairy farm with no history of TCBZ use, were also examined. The macroscopic arrangement of the reproductive system in flukes was studied using catechol/carmine stained whole mounts, and the histology of the main organs was examined using conventional haematoxylin-eosin stained sections. Validation of apoptosis in the fluke sections was carried out using an in situ hybridisation method designed to label endonuclease-induced DNA strand breaks. In TCBZ-S flukes exposed to TCBZ metabolites for 24-96 h in vivo, but not in TCBZ-R flukes, those tissues where active meiosis and/or mitosis occurred (testis, ovary, and vitelline follicles), were found to display progressive loss of cell content. This was due to apparent failure of cell division to keep pace with expulsion of the mature or effete products. Further, actively dividing cell types tended to become individualised, rounded and condensed, characteristic of apoptotic cell death. In the treated TCBZ-S flukes, strong positive labelling indicating apoptosis was associated with the morphologically abnormal cells undergoing mitosis or meiosis in the testis, ovary and vitelline follicles. In treated flukes from field outbreaks of suspected TCBZ-R fasciolosis, no significant histological changes were observed, nor was there any positive labelling for apotosis. On the other hand, sections of TCBZ treated flukes derived from a field case of fasciolosis where TCBZ resistance was not suspected displayed severe histological lesions, and heavy positive labelling for apoptosis. The triggering of apoptosis is considered to be related to failure of spindle formation at cell division, supporting the contention that TCBZ inhibits microtubule formation. In treated TCBZ-S flukes, protein synthesis and transport was apparently inhibited in the Mehlis' secretory cells, perhaps due to energy uncoupling or to microtubule defects. In the uterus, successful formation of shelled eggs represents the culmination of a complex sequence of cytokinetic, cytological and synthetic activity involving the vitelline follicles, the ovary and the Mehlis' gland. Histological evidence indicating failure of ovigenesis in TCBZ-S flukes was evident from as early as 24 h post-treatment onwards. Light labelling for apoptosis was associated with the testis of untreated Cullompton (TCBZ-S) and Sligo type 2 (TCBZ-R) flukes, which exhibit abnormal spermatogenesis and spermiogenesis, respectively. This was attributed to apoptosis and to heterophagy of effete germ line cells by the sustentacular tissue. The studies summarised in this review illustrate the potential utility of histological techniques for conveniently screening representative samples of flukes in field trials designed to validate instances of drug resistance. Histology can also be used to test the efficacy of new products against known drug-resistant and drug-susceptible fluke isolates. The account also provides reference criteria for drug-induced histopathological changes in fluke reproductive structures, examination of which may supplement and augment conventional coprological testing, and aid interpretation of TEM findings.
Sagerinic acid, a novel cyclobutane and salvianolic acid K, derived from rosmarinic acid,were isolated together with the parent compound from polar solvent extracts of Salvia officinalis.Their chemical structures were elucidated by NMR and, for sagerinic acid, the stereochemistry ofthe substituents on the cyclobutane moiety was established as 3β,4a-diaryl-1a,2β-dicarboxylicacid diester (m-truxinate form).
The antioxidant activities of the sage polyphenols, consisting of flavone glycosides and a range of rosmarinic acid derivatives, were evaluated for their capacity to scavenge DPPH and superoxide anion radicals and also to reduce Mo(VI) to Mo(V). The rosmarinic acid derivatives all showed potent antioxidant activity in three test systems and their capacity to reduce Mo(VI) to Mo(V) and their superoxide radical scavenging activities, with values ranging from 220 to 300 SOD units/mg, in particular, were 4–6 and 15–20 times greater than trolox, respectively. The high SOD activity of rosmarinic acids could be attributed to the radical-scavenging catechols and the xanthine oxidase-inhibiting caffeic acid moieties contained in them. The antioxidant activity of the flavonoids was variable and those with a catechol B-ring (luteolin glycosides) were more active than those without (apigenin glycosides).
FhSAP-2 is a novel member of the Fasciola hepatica saposin-like protein family that induces protection in rabbits against a challenge infection. We investigated the presence
of lineal B-cell epitopes within this protein using a set of overlapping synthetic peptides. Peptides were tested in enzyme-linked
immunosorbent assays against sera from rabbits infected with F. hepatica. Two dominant epitopes were identified, which were also highly reactive with sera from mice infected with Schistosoma mansoni. These peptides may be suitable to incorporate into a polyepitope-based vaccine formulation against F. hepatica.
Effects of particle size, temperature, contact time, solvent-to-sage ratio and the ethanol–water ratio on the extraction of the active compounds rosmarinic acid, carnosic compounds and essential oil from dried sage (Salvia officinalis) were studied. Optimal extraction conditions giving highest yield of all three active compounds were particle diameter 1 mm, extraction temperature 40 °C, solvent-to-sage ratio of 6:1 and 55–75 wt% ethanol for up to 3 h. This gave an extract equivalent to 14.9% of dry sage, containing 6.9% rosmarinic acid (55% recovery), 10.6% carnosic compounds (75% recovery) and 7.3% essential oil (42% recovery). Scale up of the process by a factor of 100 demonstrated that the optimised laboratory scale process can be carried out without any loss of efficiency at an industrial scale.
The factors affecting oil yield and quality of essential oils from Dalmatian sage (Salvia officinalis L.) are analyzed. Distillations of oils from individual plants and GC analyses revealed the presence of three chemotypes with different proportions of alpha- and beta-thujone (alpha/beta 10:1, 1.5:1, and 1:10). Different accessions could also be classified as having high (39-44%), medium (22-28%), or low (9%) total thujone contents. Flowering parts of S. officinalis had higher oil contents (1.6 versus 1.1%) and beta-pinene levels (27 versus 10%) than leaves and lower thujone levels (16 versus 31%). Major seasonal changes were found in the composition of oil distilled from a flowering type of Dalmatian sage, but oil yields from healthy, established plants did not vary greatly. Total thujone levels were lowest (25%) around flowering in spring and summer, so autumn or winter was the best harvest time to obtain oils with high thujone levels.
The cytosolic superoxide dismutase (SOD) of Fasciola hepatica, a causative agent of fascioliasis, was purified and characterized. The enzyme consists of two identical subunits, each with an apparent molecular mass of 17.5 kDa. An analysis of the enzyme's primary structure and inhibition studies revealed that the enzyme is a copper/zinc-containing SOD (Cu/Zn-SOD). The enzyme activity was relatively stable in a broad pH range, from pH 7.0 to 10.0, and the enzyme showed maximum activity at pH 7.5. This enzyme also displayed strong antigenicity against sera of bovine and human subjects with fascioliasis. The SOD gene fragment was amplified by PCR with degenerate oligonucleotide primers derived from amino acid sequences conserved in the Cu/Zn-SODs of other organisms. An F. hepatica cDNA library was screened with the SOD gene fragment as a probe. As a result, a complete gene encoding the Cu/Zn-SOD was identified, and its nucleotide sequence was determined. The gene had an open reading frame of 438 bp and 146 deduced amino acids. Comparison of the deduced amino acid sequence of the enzyme with previously reported Cu/Zn-SOD amino acid sequences revealed considerably high homologies. The coding region of the F. hepatica Cu/Zn-SOD was cloned and expressed in Escherichia coli. Staining of native polyacrylamide gel for SOD activity of the expressed protein revealed SOD activity that was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide. This means that the presence of the recombinant fusion protein is indicative of Cu/Zn-SOD. The expressed protein also reacted with sera of bovine and human subjects with fascioliasis, but it did not react with sera of uninfected bovine and human subjects.