Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis

A recombinant antigen-based single serum dilution enzyme-linked immunosorbent assay (ELISA) was developed to measure the specific antibody activity in sera of dogs with leptospirosis. The recombinant antigen developed and used in the assay was specific for the pathogenic serovars of Leptospira. A linear relationship was found to exist between the predicted antibody titres at a single working dilution of 1:1000 and the corresponding observed serum titres as determined by the standard serial-dilution method. Regression analysis was used to determine a standard curve from which an equation can be derived that allows demonstration of the mentioned correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay was proved to be sensitive, specific and accurate as compared to the standard microscopic agglutination test (MAT).     

Leaching of dissolved organic carbon and nitrogen under cotton farming systems in a Vertisol

Leaching with deep drainage is one of the loss pathways of carbon (C) and nitrogen (N) in cropping fields. However, field studies in irrigated row cropping systems are sparse. A 3‐year investigation on C and N leaching associated with deep drainage was overlaid on a long‐term experiment on tillage practices and crop rotations in Australia. The treatments included cotton (Gossypium hirsutum L.) monoculture and cotton–wheat (Triticum aestivum L.) or maize (Zea maize L.) rotations with maximum or minimum tillage. The deep drainage C and N concentrations at 0.6 and 1.2 m depth were measured after furrow irrigation with ceramic cup samplers during the 2014–15, 2015–16 and 2016–17 cotton seasons. Pre‐planting dissolved organic carbon (DOC) concentration in soil at 0.6–1.2 m depth during 2016–17 was 64 mg kg^?1 for maximum tilled cotton monoculture, 36 mg kg^?1 for minimum tilled cotton monoculture and 39 mg kg^?1 for cotton–wheat, and in maize and cotton subplots 51 and 41 mg kg^?1, respectively. Post‐harvest DOC values in soil were similar in all treatments (average of 32 mg DOC kg^?1). Total organic carbon (TOC) losses in deep drainage were equal to 2%–30% of TOC gained in irrigation water. Oxidized N losses in deep drainage ranged from 0.7% to 12% of applied N (260 kg ha^?1). NOx‐N concentrations in leachate under maize systems (20 mg L^?1) were up to 73% lower than those in cotton systems (75 mg L^?1). Maize sown in rotation with cotton can improve cotton yield, reduce N leaching and improve N use efficiency of subsequent cotton.     

A rapid latex agglutination test for detection of leptospiral antibodies

A rapid semi-quantitative latex agglutination test (LAT) has been standardized for the detection of leptospiral antibodies in serum samples of man and animals. The efficacy of the LAT was compared with the plate enzyme linked immunosorbent assay (ELISA). A total of 276 human serum samples were analyzed by both LAT and ELISA and percentage positives were 84.8 and 85.9%, respectively. Similarly, of 65 animal samples tested, 63.1 and 69.2% positivity were observed in LAT and ELISA, respectively. Even though the ELISA test was slightly more sensitive than LAT, the rapidity, simplicity and economics of the LAT were found to fulfill the requirements of a screening test for leptospiral antibodies.     

Demonstration of Alternative and Classical Complement Pathway Activity in Colostrum from Buffalo (Bubalus bubalis)

Buffalo colostrum caused lysis of unsensitized red blood cells (RBC) from sheep, goats, rabbits and chickens. RBC from cattle and buffalo were resistant to lysis. That lysis was due to the presence of natural antibodies to these RBC was ruled out since there was no reduction in haemolytic titres even after adsorption with the respective RBC. The addition of EGTA to the diluent had no effect on the haemolytic activity. These findings indicate the presence of alternative complement pathway (ACP) activity in buffalo colostrum. The haemolytic activity of buffalo complement for unsensitized rabbit RBC was reduced to very low levels by heating at 50°C for 45 min. Treatment with zymosan also inhibited the haemolytic activity, while inulin had no effect. The maximum activity of ACP occurred in the presence of 4 mmol/L Mg²⁺ in the diluent. The range of ACP activities in colostrum from buffaloes varied from 4.06 to 8.48 CH₅₀ units/ml. Using a standard system for titrating the classical complement pathway and rabbit red blood cells sensitized with goat haemolysin, the range of complement activity in buffalo colostrum was 4.81–6.77 CH₅₀/ml.     

Development of two cell culture systems from Asian seabass Lates calcarifer (Bloch)

Two new cell culture systems namely epitheloid cells of Lates (LCE) and fibroblastic cells of Lates (LCF) have been developed from fry and fingerling of the economically important brackishwater fish Lates calcarifer. Primary cultures were initiated by explant technique using caudal fin of fingerling and whole body tissue of the fry. The nutritional requirements and the growth pattern in response to different culture environment were similar for the two cell cultures. The culture medium used was Leibovitz‐15 supplemented with 20% fetal bovine serum (FBS) and 1% fish serum. The LCE comprised of epithelioid cells and LCF cells were fibroblastic. With a split ratio of 1:2, the confluency of cells was attained in 8–10 days at an incubation temperature of 28°C. The cells were found to grow well in a wide range of temperature (24–32°C) and stable at 20 and 36°C. The growth rate of LCF and LCE cells increased proportionately with the concentration of FBS from 5% to 20%. A decrease of serum level to 10% after eight subcultures produced no apparent change in cell morphology and growth rate. The viability of cells was found to be 70% when revived after a month of storage in liquid nitrogen (?196°C).     

A dot enzyme linked immunosorbent assay (dot ELISA): Comparison with standard fluorescent antibody test (FAT) for the diagnosis of rabies in animals

A modified enzyme linked immunosorbent assay (Dot ELISA) is described for visual detection of rabies antigen in animals. The test materials were dotted onto the nitrocellulose paper and allowed to react with rabies antiserum. The bound antigen—anti-body were reacted with a peroxidase conjugated antirabbit immunoglobulin. Positive reactions were easily visualized as brown dots after enzyme degradation of the substrate. A total of 400 specimens from various geographical locations were tested with the dot ELISA technique, and also with the fluorescent antibody test (FAT), which was used as a reference method. The concordance between the two tests was 95.25%. The dot ELISA may have potential applications as a rapid, simple and economical field test in the diagnosis of rabies.