Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit.

Contagious bovine pleuropneumonia (CBPP) is an infectious and contagious bacterial respiratory disease that affects cattle with significant economic losses to the African animal industry. The use of ELISA kits based on monoclonal antibodies (mAbs) will aid in quick and precise diagnosis of CBPP, contributing to disease control and prevention in cattle. Thus, this research aims to develop and evaluate monoclonal antibodies against CBPP (T1/44) antigen for use in ELISA kits for CBPP diagnosis. Hybridoma technology was used to develop monoclonal antibodies that recognize and bind to the CBPP (T1/44) antigen. The antibody-secreting hybridomas were produced after immunizing mice with purified CBPP antigens. The hybridomas were screened for high sensitivity, specificity, and liking to the antigen. The selected mAbs were assessed for sensitivity and specificity against CBPP antigen using different immunoassays, dot-blot, ELISA, and mouse mAb isotyping. The monoclonal antibodies were profoundly specific, with a higher hindrance to CBPP antigen (<0.50 OD) while lacking cross-reactivity to other antigens. The monoclonal antibodies could distinguish CBPP antigen at low concentrations, showing their high sensitivity (>80% PI). The isotyped mAbs of intrigued appeared to have a place in the IgG class. These identified monoclonal antibodies can be utilized to develop an ELISA kit for CBPP diagnosis, which would give a fast, precise, and cost-effective strategy for screening and checking CBPP in cattle herds.

In vitro fermentation characteristics and effective utilisable crude protein in leaves and green pods of Moringa stenopetala and Moringa oleifera cultivated at low and mid-altitudes.

This study was conducted to assess the in vitro nutrient digestibility and utilisation of leaves and green pods of two Moringa species in supplementing the feed of ruminant animals during the dry season. Samples were analysed for proximate nutrients using official methods. The metabolisable energy (ME), organic matter digestibility (OMD) and effective utilisable crude protein (uCP) were estimated using the Hohenheim in vitro gas test method. Gas volume in Moringa stenopetala leaves and green pods was generally higher than those of Moringa oleifera. Gas volume for leaves was similar between low and mid-altitudes but was higher for green pods at mid-altitude. M. stenopetala leaves contained significantly higher ME (9.8 MJ/kg DM) and OMD (75%) than those of M. oleifera. Similarly, M. stenopetala green pods had higher ME and OMD values than those of M. oleifera. For green pods, the ME and OMD values were significantly higher at mid-altitude than those at low altitude although these values for leaves were similar between both altitudes. Moringa oleifera leaves had higher effective uCP than those of M. stenopetala. Nevertheless, the effective uCP was higher for green pods of M. stenopetala than those of M. oleifera. The effective uCP for leaves cultivated at mid-altitude was slightly higher than those at low altitude. This study suggested that leaves and green pods could be used as alternative energy and protein supplements for tropical ruminants, particularly during dry periods. It was further concluded that leaves were generally better in nutrient compositions and in vitro nutrient digestibility characteristics than green pods.

Effect of long-term heat exposure on peripheral concentrations of heat shock protein 70 (Hsp70) and hormones in laying hens with different genotypes.

1. The effect of long-term, moderate heat stress (30 to 32 degrees C) on heat shock protein 70 (Hsp70) concentration in mononuclear blood cells and plasma concentrations of 3,5,3'-triiodothyronine (T3) and corticosterone in laying hens was investigated. 2. Three groups of 48 hens each (Ethopian line [Angete Melata, Na], New Hampshire [NH], F1 cross [Na x NH]) were divided into an experimental group (24 each) and a control group (24 each, ambient temperature 18 to 20 degrees C), respectively. All hens were kept in individual cages up to an age of 68 weeks and performance data were recorded. 3. Blood samples were taken from the wing vein of 12 hens from each group at weeks 22, 38, 51 and 65 (12 hens x 3 lines x 2 treatments). Mononuclear blood cells were isolated and Hsp70 concentrations were determined by Western Blot analysis with a monoclonal anti-Hsp70 antibody. T3 and corticosterone were measured with commercially available ELISA and RIA kits, respectively. 4. The moderate heat stress caused significantly increased Hsp70 levels compared with the control groups in weeks 51 and 65. However, the responses of the lines were not uniform at different ages. 5. In contrast, T3 levels were significantly decreased in stressed birds regardless of line and age. There was no effect of treatment and line on corticosterone levels during the experimental period. 6. Our results indicate that Hsp70 and T3 levels are affected by mild heat stress applied over a long period but are both involved in independent mechanisms of acquisition of thermotolerance. Further investigations are necessary to clarify whether the observed differences in Hsp70 response between the genotypes are indicators for differences in thermotolerance.