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.

In Vitro Immunomodulatory Activity of Aqueous Quercus infectoria Gall Extract.

OBJECTIVES: Our study reports the immunomodulatory potency of Quercus infectoria gall extract in vitro. The aqueous extract was prepared and examined for its effects on cell proliferation, phagocytic activity, nitric oxide (NO) production, and cytokine synthesis by murine macrophages. METHODS: Proliferative, phagocytic activity, and NO production of extract-treated and control cells were studied using proliferative assay, flow cytometry, and Griess reaction, respectively. An enzyme-linked immunosorbent assay was performed to determine the levels of pro- and anti-inflammatory cytokines in the macrophage culture. RESULTS: Treated macrophages had a higher proliferative rate and phagocytic activity compared to untreated macrophages. The cell treatment with an extract concentration of 64 µg/mL demonstrated a significant decrease in NO production (p < 0.001). An increase in cytokine levels (IL-2, IL-5, IL-10, IL-17A, IL-23, TGF-ß1) was observed; however, this increase was not statistically significant. CONCLUSIONS: Our study suggests that gall extract possesses the potential for augmenting immunomodulatory activity by cellular mediated mechanism and could play a role in regulating the innate immune response.

In vitro anti-Candida activity of Quercus infectoria gall extract-based vaginal cream and its local tissue effects in vivo

Aims@#Aqueous extract of Quercus infectoria (QI) galls has been reported to possess anti-fungal and anti-inflammatory activities. Hence, this study aimed to determine in vitro antimicrobial activity of formulated QI gall extract-based vaginal cream against Candida albicans and to evaluate the possible side effects on the cervicovaginal epithelium of healthy rats. @*Methodology and results@#Three different cream formulations containing 10%, 20%, and 30% of QI gall extract respectively were tested for their antimicrobial activity against C. albicans (ATCC 10231) by using disc diffusion test. Microbroth serial dilution method was performed in determining the minimum inhibitory concentration (MIC) and fungicidal concentration (MFC). The 30% formulated extract cream (FEC) was applied topically on the cervicovaginal surface of healthy Sprague Dawley (SD) rats and examined for local tissue effects histologically. The mean scores of inhibition zone diameter were compared by one-way ANOVA and post-hoc test using PRISM software. All extract cream formulations displayed a relatively good anti-Candida activity. The MIC values exhibited by 10%, 20%, and 30% FEC against C. albicans were 1.094 mg/mL, 0.547 mg/mL, and 0.068 mg/mL, respectively. The 10% and 20% FECs showed a significant difference (P=0.0254) in the mean of inhibition zone diameter. The lowest MFC value (0.068 mg/mL) was shown by 30% FEC. There were no abnormal changes seen at the vagina and cervical mucosa after 2 weeks application of 30% FEC. @*Conclusion, significance and impact of study@#QI gall extract formulated in the cream base has an anti-Candida activity in vitro and the present finding suggests that this herbal cream formulation is potentially useful in preventing vaginal candidiasis without causing any unwanted local side effects.