On-farm colorimetric detection of Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in crude bovine nasal samples

Abstract

This work modifies a loop-mediated isothermal amplification (LAMP) assay to detect the bovine respiratory disease (BRD) bacterial pathogens Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in a colorimetric format on a farm. BRD causes a significant health and economic burden worldwide that partially stems from the challenges involved in determining the pathogens causing the disease. Methods such as polymerase chain reaction (PCR) have the potential to identify the causative pathogens but require lab equipment and extensive sample processing making the process lengthy and expensive. To combat this limitation, LAMP allows accurate pathogen detection in unprocessed samples by the naked eye allowing for potentially faster and more precise diagnostics on the farm. The assay developed here offers 66.7–100% analytical sensitivity, and 100% analytical specificity (using contrived samples) while providing 60–100% concordance with PCR results when tested on five steers in a feedlot. The use of a consumer-grade water bath enabled on-farm execution by collecting a nasal swab from cattle and provided a colorimetric result within 60 min. Such an assay holds the potential to provide rapid pen-side diagnostics to cattle producers and veterinarians.

A Comprehensive Review on Therapeutic Perspectives of Phytosterols in Insulin Resistance: A Mechanistic Approach

“…Phytosterol has been studied extensively for the treatment and management of diabetes, a variety of malignancies, cardiovascular disorders, atherosclerosis, and skin problems [ 9 , 10 ]. These phytosterols have been shown to have multiple therapeutic implications, such as hypolipidemic, anti-inflammatory, antioxidant, antiproliferative, and hypoglycemic potential through experimental evidence ( Table 1 ).…”

“…The significance of phytosterols in the treatment of T2DM and its implications, as well as the theoretical potential for long-term pharmacological research and clinical diabetic treatment, are the focus of this review. Phytosterol has been studied extensively for the treatment and management of diabetes, a variety of malignancies, cardiovascular disorders, atherosclerosis, and skin problems [9,10]. These phytosterols have been shown to have multiple therapeutic implications, such as hypolipidemic, anti-inflammatory, antioxidant, antiproliferative, and hypoglycemic potential through experimental evidence (Table 1).…”

A low-cost, portable, and practical LAMP device for point-of-diagnosis in the field

“…LAMP technique has overcome majority of these limitations [2]. Improved and more sample-specific LAMP protocols have also been developed and published in recent decade [9][10][11][12][13][14][15][16][17][18][19]. In contrast to LAMP methods, technology development for LAMP-based detection has not been completely accelerated yet.…”

“…Hence, LAMP methods decrease cost, time and the need of expensive instruments or trained personnel. It has a great potential for breaking through the walls of laboratories to fields [9,10,11], clinics [12,13,14], farms [15,16,17], crime scenes [18,19] or even excavation sites.…”

Microfluidic Point-of-Care (POC) Devices in Early Diagnosis: A Review of Opportunities and Challenges

“…Pascual-Garrigos et al, improved the loop mediated isothermal amplification (LAMP) assay to realize the early POC detection of bovine respiratory diseases (BRD), so as to reduce the risk of morbidity and mortality of dairy cows on the farm and reduce the economic burden of farmers. It can be seen that POCT technologies can be widely used in different occasions [18].…”