Immunodiagnosis of Fascioliasis in Ruminants by ELISA Method: A Mini-Review.

Fascioliasis is an important zoonotic disease prevalent in domestic animals and it leads to socioeconomic impact in rural farming communities of the developing world. The gold standard diagnosis of ruminant fascioliasis involves coprological detection of Fasciola spp. eggs or recovery of flukes in infected livers. Coprological analysis is unreliable in the patent period of chronic infection, and even then, its sensitivity is relatively low. Robust diagnostic tools that can promptly and accurately detect an active infection are crucial to avoid complications and further losses in ruminant livestock productivity, as well as to preserve the livelihood of communities at risk. Immunodiagnosis determined by antibody and antigen detection in the sera and faeces of infected ruminants provides a valuable alternative to the parasitological diagnostic approach. This review discusses current developments in immunological techniques by enzyme-linked immunosorbent assay (ELISA) in the detection of ruminant fascioliasis and summarises the performance of various ELISAs in studies conducted to date. Indirect ELISAs demonstrated effective immunodiagnostic performance with high sensitivities and specificities. Cathepsin L ELISA is the most favourable antigen in serodiagnosis, among other recombinant and native proteins evaluated. Sandwich ELISA provides excellent sensitivity and specificity, which correlates well with the fluke burden. Utilising monoclonal antibodies in sandwich ELISA reduces the detection time and performance variations that commonly occur in polyclonal antibody ELISA.

Seasonal Occurrence of Cattle Fascioliasis in Kelantan, Malaysia.

A longitudinal study was conducted in five randomly selected farms in Kelantan, Malaysia to determine the seasonal occurrence of cattle fascioliasis and its association with climatic factors. A total of 480 faecal samples were collected by a random purposive sampling method from July 2018 to June 2019. The faecal samples were examined for the presence of Fasciola eggs using a formalin ether sedimentation technique. Meteorological data including temperature, humidity, rainfall, and pan evaporation were obtained from a local meteorological station. The overall prevalence of cattle fascioliasis in Kelantan was 45.8%. The prevalence was observed to be slightly higher during the wet season from August to December (50-58%) than during the dry season from January to June (30-45%). Meanwhile, the mean eggs per gram (EPG) were highest in June (191.1 ± 0.48) and lowest in October (77.62 ± 95.5). However, there were no significant differences in the mean of EPG between the monthly prevalence, tested using one-way ANOVA (p = 0.1828). A statistically significant association (p = 0.014) was observed between the disease and cattle breeds, with Charolais and Brahman showing lower odds of having the disease. There were significant moderate-to-strong positive correlations between cattle fascioliasis and rainfall (r = 0.666; p = 0.018) and humidity (r = 0.808; p = 0.001), as well as strong negative correlations with evaporation (r = -0.829; p = 0.001). The results indicated that the higher prevalence of cattle fascioliasis in Kelantan was correlated with the climatic factors, which include higher rainfall and humidity and lower evaporation.

Prevalence and Risk Factors of Bovine Fascioliasis in Kelantan, Malaysia: A Cross-Sectional Study.

Bovine fascioliasis is an important zoonotic parasitic disease that causes significant economic losses to the livestock industry. The aim of this study was to determine the prevalence and risk factors of bovine fascioliasis in Kelantan. In this cross-sectional study, a total of 308 stool and blood samples of farmed cattle were collected from December 2017 to June 2018. The stool samples were examined microscopically for the presence of Fasciola spp. eggs following a formalin-ether sedimentation process. The blood samples were subjected to a commercial ELISA kit (Bio-X-Diagnostic, Rochefort, Belgium) for the detection of anti-Fasciola IgG antibody. The association between coprological findings and risk factors was determined using Pearson's chi-square (χ2). The coproprevalence and seroprevalence of bovine fascioliasis was 14.6% and 37.3%, respectively. There were significant (P < 0.05) associations between the risk of infections and the sex, type of feedings, anthelmintic treatment and farm hygiene. Female cattle (OR: 3.104; 95% CI: 1.265, 7.615), feeding by grazing (OR: 4.458; 95% CI: 1.823, 10.90), untreated cattle (OR: 3.833; 95% CI: 1.620, 9.071), non-schedule anthelminthic treatment (OR: 3.927; 95% CI: 1.685, 9.152) and farm that have never been cleaned (OR: 2.829; 95% CI: 1.428, 5.608) showed higher odds of Fasciola spp. infection. These findings suggested bovine fascioliasis is a serious veterinary disease in Kelantan. Thus, appropriate control, prevention and monitoring strategies of this parasitic infection are urgently needed to reduce the burden of the disease.

Fascioliasis pada lembu adalah penyakit parasit zoonotik penting yang menyebabkan kerugian ekonomi yang signifikan pada industri ternakan. Tujuan kajian ini adalah untuk menentukan prevalens dan faktor risiko fascioliasis pada lembu ternak di Kelantan. Dalam kajian keratan rentas ini, sejumlah 308 sampel tinja dan sampel darah daripada lembu ternak telah diambil dari Disember 2017 hingga Jun 2018. Sampel tinja telah diperiksa secara mikroskopik bagi mengesan kehadiran telur Fasciola spp. melalui proses sedimentasi formalin-eter. Sampel darah telah disaring menggunakan kit ELISA komersial (Bio-X-diagnostics) untuk pengesanan antibodi anti-Fasciola IgG. Hubungan antara penemuan koprologi dan faktor risiko telah ditentukan dengan menggunakan ujian Chi-ganda dua (χ2). Koproprevalens dan seroprevalens fascioliasis pada lembu masingmasing adalah 14.6% dan 37.3%. Terdapat perbezaan signifikan (P < 0.05) antara risiko jangkitan dan jantina, kaedah pemakanan, penggunaan ubat cacing dan kebersihan ladang. Lembu betina (OR: 3.104; 95% CI: 1.265, 7.615), kaedah pemakanan melalui teknik ragut (OR: 4.458; 95% CI: 1.823, 10.90), lembu yang tidak dirawat (OR: 3.833; 95% CI: 1.620, 9.071), rawatan secara tidak berkala (OR: 3.927; 95% CI: 1.685, 9.152) dan ladang yang tidak pernah dibersih (OR: 2.829; 95% CI: 1.428, 5.608) merupakan antara risiko lebih tinggi untuk dijangkiti Fasciola spp.. Penemuan ini mencadangkan bahawa fascioliasis pada lembu adalah penyakit veterinar yang serius di Kelantan. Oleh itu, strategi kawalan, pencegahan dan pengawasan yang sesuai bagi jangkitan parasit ini amat diperlukan untuk mengurangkan beban penyakit ini.

Proapoptotic activities of Oroxylum indicum leave extract in HeLa cells

Objective: To examine the proapoptotic properties of Oroxylum indicum methanol extract on cervical cancer cells. Methods: Methylene blue assay was used to determine the IC

Proapoptotic activities of Oroxylum indicum leave extract in HeLa cells

Objective: To examine the proapoptotic properties of Oroxylum indicum methanol extract on cervical cancer cells. Methods: Methylene blue assay was used to determine the IC50 value of the extract. Western blotting assays were done to analyze the expression of HPV oncoproteins (HPV18 E6 and E7) and apoptotic molecules (caspase-3 and caspase-8). Reverse transcriptase PCR assays were performed to determine genetic alteration of tumor suppressors p53 and pRb and apoptosis markers Fas and FasL. Enzyme-linked immunosorbent assay (ELISA) was done to determine the expression of cytokine levels (IL-6 and IL-12). Results: The determination of IC50 value indicated a higher anti-proliferative activity of the extract compared to cisplatin. After 24 hours of treatment, Western blot analysis showed that treated HeLa cells exhibited a significant down-regulation of HPV18 oncoproteins E6 and E7, and a significant induction of caspase-8 and caspase-3 activation level. Meanwhile, the mRNA expressions of p53, pRb, Fas and FasL were significantly upregulated in treated cells. Moreover, ELISA showed an increased IL-12 and decreased IL-6 production after Oroxylum indicum treatment. Conclusions: The methanol extract of Oroxylum indicum has an anti-proliferative activity and proapoptotic potential. It induces localized-immunity improvements by altering cytokine production in HPV-positive cervical cancer cells.