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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., Competing Interests: Conflict of Interest None., (© Penerbit Universiti Sains Malaysia, 2023.)

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.

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 (χ 2 ). 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., (© Penerbit Universiti Sains Malaysia, 2021.)

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-β1) 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., (The OMJ is Published Bimonthly and Copyrighted 2021 by the OMSB.)

Human fascioliasis is a public health problem particularly in areas where ruminants are raised. The aims of this study were to determine the seroprevalence of anti-Fasciola antibody and the associated risk factors among cattle farm workers and dwellers in Kelantan. A total of 90 blood samples were collected in this cross-sectional study. A set of validated questionnaire was used to obtain information on socio-demographic profiles and dietary habits of participants. The sera were subjected to enzyme linked immunosorbent assay (ELISA) for the detection of anti-Fasciola IgG antibody. The association between seropositivity and the significant risk factors were determined via logistic regression. From the result, serological screening revealed 60 (67%) participants positive for anti-Fasciola IgG antibody. The factors found to be significantly associated with seropositivity against anti-Fasciola IgG antibody were the age group of 18 years old and above with calculated odds ratio of 3.2 times (p=0.032) and the duration of farming activities of more than 5 years with calculated odds ratio of 2.6 times (p=0.036). In conclusion, Fasciola infection is prevalent among cattle farm workers and dwellers in Kelantan.

Rapid and inexpensive assays for drug susceptibility testing (DST) of Mycobacterium tuberculosis (MTB) are urgently required especially in developing countries where tuberculosis cases are prevalent. In response to this necessity, a direct microplate-based colorimetric assay which excludes the use of pre-testing culture isolate was evaluated. MTB susceptibility to the first line anti-tuberculosis drugs was tested directly on sputum specimens using tetrazolium microplate assay (TEMA) method and the sensitivity, specificity, accuracy as well as mean turn-around time of TEMA were compared to the standard absolute concentration method (ACM). TEMA was performed on 41 acid fast bacilli (AFB) positive sputum specimens by direct inoculation of the processed specimens into the microplate wells containing serialdiluted first line anti-tuberculosis drugs using tetrazolium dye as growth indicator. Indirect TEMA was performed on MTB isolates of the corresponding samples. The minimum inhibitory concentrations (MICs) of isoniazid (INH), rifampicin (RMP), ethambutol (EMB) and streptomycin (SM) were obtained for direct and indirect TEMA with reference to the absolute concentration method (ACM). After establishing the breakpoint MIC of each drug using receiver operating characteristics (ROC) curve, reliable results from direct TEMA were obtained for INH and SM, with excellent levels of sensitivity, specificity, and accuracy (more than 90%). The predictive values for susceptibility were 100% for INH, EMB and SM as well as 96% for RMP. A shorter mean turn-around time of 14 days was observed for direct TEMA (P < 0.05). Thus direct TEMA is potentially rapid, reliable and inexpensive DST screening method of MTB in countries with high prevalence rates of drug resistance tuberculosis.