RESUMEN
The chemical treatment of gastrointestinal parasitic diseases has been undermined by increasing resistance and high toxicity. There is an urgent need to search for alternative natural sources for the treatment of such parasites. In this respect, the present study aims to quantify phenolic compounds of chamomile (Matricaria recutita L.) and to study their in vitro anti-oxidant and anthelmintic activities in solvents with increasing polarity. In vitro determination of anti-oxidant capacity was carried out using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical cation methods. In vitro anthelmintic activity was investigated on egg-hatching inhibition and loss of motility of adult worms of Haemonchus contortus from sheep. The results showed that methanolic and aqueous extracts contain more total polyphenols, total flavonoids and condensed tannins than chloroformic and hexanic extracts. ABTS and DPPH assays showed that methanolic extracts had the highest anti-oxidant potency (IC50 = 1.19 µg/ml and 1.18 µg/ml, respectively). In vitro anthelmintic activity showed that both methanolic (IC50 = 1.559 mg/ml) and aqueous (IC50 = 2.559 mg/ml) extracts had the greatest effect on egg hatching and motility of worms (100% after 8 h post exposure at 8 mg/ml). A significant and positive correlation between DPPH and ABTS tests was observed for all tested extracts. Therefore, total phenolic, total flavonoid and condensed tannin values were correlated with IC50 from both ABTS and DPPH, and with inhibition of egg hatching. To our knowledge, this report is the first of its kind to deal with in vitro anthelmintic activities of chamomile extracts.
Asunto(s)
Manzanilla/química , Haemonchus/efectos de los fármacos , Extractos Vegetales/farmacología , Polifenoles/farmacología , Animales , Antihelmínticos/farmacología , Benzotiazoles/farmacología , Compuestos de Bifenilo/farmacología , Hemoncosis/parasitología , Aceites Volátiles/farmacología , Picratos/farmacología , Ovinos/parasitología , Enfermedades de las Ovejas/parasitología , Ácidos Sulfónicos/farmacología , TúnezRESUMEN
Two peptides, HP6-2 and Ts45S-10, were used as antigens for the detection of antibodies against Taenia saginata cysticercosis in serum and meat juice samples using enzyme-linked immunosorbent assay (ELISA). Positive control samples were obtained from animals experimentally infected (serum) and from animals naturally infected (meat juice). The two peptides and a pooled preparation of both peptides were evaluated, and their cut-off points with both sample categories were calculated. ELISA results from these different peptides were compared. Sensitivity and specificity of HP6-2 using serum were calculated as being 100 and 98%, respectively, showing to be higher than the values for the other antigens used. The average optical density (OD) value for negative samples was 0.646, whereas it was 1.702 for the positive control samples. This peptide was used to examine serum samples from animals with cysts and random field serum samples. For meat juice samples the pooled peptides showed the highest sensitivity and specificity, as they were 100 and 95%, respectively. The average OD values for the negative and the positive reference meat juice samples were 0.379 and 1.291, respectively. The optimal dilution of the meat juice samples for the ELISA was very low, as it was 1:20 using the pooled peptides, compared with 1:800 serum dilution using HP6-2. To the authors' knowledge, this is the first report of a successful testing for T. saginata cysticercosis using meat juice.
Asunto(s)
Enfermedades de los Bovinos/diagnóstico , Cisticercosis/veterinaria , Pruebas Serológicas/veterinaria , Taenia saginata/aislamiento & purificación , Animales , Bovinos , Enfermedades de los Bovinos/sangre , Cisticercosis/sangre , Cisticercosis/diagnóstico , Carne/parasitología , Sensibilidad y EspecificidadRESUMEN
A total of 267 cysts were collected from March to December 2004 from two main abattoirs in northern Germany. The cysts were classified by the usual organoleptic methods during meat inspection as Cysticercus bovis. The reported prevalence of cysticercosis in the abattoirs was 0.48 and 1.08%, respectively. The cysts were examined macroscopically for description of their morphology and constituents and classified as viable or degenerating (dead). The DNA was extracted from these cysts and subjected to polymerase chain reaction (PCR) for evaluation of the detection methods used and to make certain that the cysts did indeed belong to C. bovis, as indicated at the slaughterhouses. Two sets of primers were used with different sensitivity levels. The first, HDP1, was able to detect 200 fg of Taenia saginata DNA and 100 pg of C. bovis DNA. The other primer set, HDP2, was able to detect 1 pg of T. saginata DNA and 1 ng of C. bovis DNA. No more than 52.4% of the samples tested positive for C. bovis in the PCR using both primers, while 20% of the viable cysts and 49.2% of the degenerating cysts tested negative with both primers.