Veterinary Research Forum

Abstract The penicillin allergy is being increasingly recognized as a significant public health problem. Immunological responses to penicillin and other beta-lactam antibiotics are classified as immediate and non-immediate responses. This research aimed to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of the reactive antibody value against penicillin in various species of animals. The serum samples were collected from nine species (forty mature animals in each species) including horse, dog, goat, sheep, buffalo, cattle, donkey, chicken, and fish. The concentrations of total antibody and immunoglobulin M (IgM) against penicillin were detected using an in-house ELISA test. The total anti-penicillin antibodies concentration from high to low in animals was as chicken, horse, fish, donkey, dog, goat, sheep, buffalo, and cattle, respectively. In cattle and sheepthe level of anti-penicillin IgM (APM) was significantly higher than non-IgM antibodies (APNM); moreover, levels of APNM were very low in chicken and fish serums; no difference was seen regarding these values in buffalo and goat. The other species had significantly lower APM than the APNM. The ani-penicillin antibody levels in the noted animals were successfully detected using the developed ELISA. Most of the species have anti-penicillin antibodies; however, they have reactive antibodies with differences in levels and isotypes.

Abstract Neospora caninum is an obligate intracellular parasite causing abortion and reproductive failure in ruminants. Here, the seroprevalence of Neospora DNA and anti-Neospora antibodies and the correlation between the DNA and the antibody using polymerase chain reaction (PCR) and a new developed whole cell-based enzyme-linked immunosorbent assay (ELISA) in water buffalo (Bubalus bubalis) were investigated. To determine the level of anti-Neospora antibody, 83 serum samples were collected from buffaloes in the northwest of Iran. Plates were coated with 2 × 10⁶ whole Neospora tachyzoites and the anti-Neospora antibody level was determined by calculating the ratio of sample/positive control (S/P) optical densities (ODs) in the ELISA. All samples with the ration of 0.50 or above were accounted as positive. To confirm the presence of Neospora DNA, the serum samples were directly subjected to PCR and nested PCR for detection of Neospora NC5 gene without the DNA isolation process. A total number of 83 buffalo serum samples were examined for the presence of anti-N. caninum immunoglobulin G and Neospora DNA. All samples with the S/P ratio of 0.50 or above (16 samples, 19.27%) were also positive for Neospora DNA. All samples with OD less than 0.50 (34 samples, 40.96%) were negative for Neospora DNA. However, 33 samples with the S/P ratio of bellow 0.50 (39.75%) showed a significant level of antibody. A 100% correlation was observed between high levels of the anti-Neospora antibody and Neospora DNA in the serum of water buffalo, and the whole N. caninum tachyzoites have the potency to be used as antigens for detection of the parasite in ELISA.

Abstract Excretory/Secretory antigen was prepared by culturing live adult worms of Haemonchus contortus in RPMI 1640 medium at a concentration of 50 worms per mL in a culture-flask at 37 ˚C for 24 hr and the culture supernatant was used as antigen. The E/S antigen was purified by thiol-sepharose affinity chromatography. On western blot analysis, it was demonstrated that thiol-purified antigen showed a single reactive band at 66 kDa. In immunization trial, sheep were administered intramuscularly with 500 μg of thiol-purified excretory/secretory antigen along with montanide as adjuvant on day 0, 30 and 60. On ELISA, it was observed that the mean absorbance values were significantly (p ≤ 0.01) higher up to 20 weeks post immunization in Group-I (purified antigen) compared to Group- II (unimmunized control). Further, the mean EPG values was lower in Group I (200.00 ± 40.82 to 400.00 ± 91.29) than Group II (2200.00 ± 108.01 to 5100.00 ± 169.56) and the percentage reduction in mean fecal egg counts was 88.50%. Similarly, the mean abomasal worm counts was lower in Group I (808.33 ± 78.29) than Group II (3280.00 ± 147.19) and the percentage reduction in mean abomasal worm count was 75.40%.