Optimizing Enzyme-Linked Immunosorbent Assay Parameters for Chlamydial Abortion Diagnosis in Sheep

11 June, 2026

Background and Objective: Chlamydial abortion is one of the leading causes of reproductive loss in sheep, yet diagnostic accuracy remains limited due to suboptimal assay conditions. To address this gap, the present study aimed to optimize the conditions of an Enzyme-Linked Immunosorbent Assay (ELISA) for the detection of chlamydial abortion in sheep, using TT and MM strains of Chlamydia previously isolated from affected animals. Materials and Methods: Yolk sac suspensions from 6-7-day-old chicken embryos infected with the causative agent of sheep enzootic abortion were used as starting material for antigen preparation. The MM strain was employed to develop an optimal method for the purification and concentration of antigens. Hyperimmunization schemes were designed using purified proteins in combination with biostimulants to obtain positive serum. Conjugates were prepared from isolated immunoglobulins, achieving an activity of 1:800. Immunoglobulins were purified by triple precipitation with saturated ammonium sulfate solution, followed by gel chromatography on Sephadex G-200 and identification by immunoelectrophoresis. Statistical analyses were performed using Student’s t-test (p≤0.05) for continuous data and a one-sided Fisher’s exact test (α<0.05) for comparing group efficacy. Results: The study established optimal contact parameters for antigen antibody interactions in the ELISA system. Results showed that effective binding occurred within 3 hrs at 37±1°C or 18 hrs at 4±2°C, while specific conjugate-antigen interaction was optimal for 1 hr at 37±1°C. These optimized parameters significantly enhanced assay sensitivity and reliability, providing reproducible detection of antibodies against the chlamydial abortion pathogen. Conclusion: This study developed an optimized ELISA protocol for the diagnosis of sheep enzootic abortion by refining antigen preparation, immunoglobulin purification and incubation conditions. The findings contribute to improving diagnostic accuracy and may serve as a foundation for developing more rapid and cost-effective diagnostic kits for field application. Future research should validate these optimized conditions across broader sheep populations and different chlamydial strains.

Functional Evaluation of Moringa Seed Extracts as Plant-Based Coagulants for Camel Milk

11 June, 2026

Background and Objective: Cheese production from camel milk is limited by poor coagulation, leading to weak curd structures. Plant-based coagulants, such as Moringa oleifera seeds, have shown potential, but little is known about Moringa peregrina, a related species native to the Middle East and Northeast Africa. This study evaluated crude seed extracts of M. peregrina as a novel plant-based coagulant for camel milk and compared their performance with M. oleifera. Materials and Methods: Extracts were biochemically characterized, and their antimicrobial and antioxidant activities were assessed. Milk-clotting performance was tested under varying pH and temperature conditions. Trisodium citrate was evaluated as a milk-standardizing agent. Statistical analysis was performed using SPSS v19 with ANOVA and mean comparisons at p<0.05. Results: Both extracts exhibited strong antimicrobial activity against Gram-positive bacteria, notably Staphylococcus aureus and Bacillus cereus. Antioxidant activity was higher in M. oleifera (63.32%) than M. peregrina (49.93%). Temperature had a greater impact on clotting than pH, with firm curd formation at 60°C and weaker coagulation at 45°C, especially for M. peregrina. Trisodium citrate (≥30 mmol/L) inhibited curd formation. High milk-clotting to proteolytic activity ratios indicated strong enzyme specificity. Conclusion: M. peregrina seed extract is a promising plant-based coagulant for camel milk, offering a viable alternative to animal rennet. Future studies should focus on enzyme purification, molecular characterization, and detailed evaluation of cheese curd quality.

International Journal of Dairy Science - Current Issue

Optimizing Enzyme-Linked Immunosorbent Assay Parameters for Chlamydial Abortion Diagnosis in Sheep

Functional Evaluation of Moringa Seed Extracts as Plant-Based Coagulants for Camel Milk