Bovine mastitis is an inflammation of the mammary gland caused by a multitude of pathogens with devastating consequences for the dairy industry. Global annual losses are estimated to be around €30 bn and are caused by significant milk losses, poor milk quality, culling of chronically infected animals, and occasional deaths. Moreover, mastitis management routinely implies the administration of antibiotics to treat and prevent the disease which poses serious risks regarding the emergence of antibiotic resistance. Conventional diagnostic methods based on somatic cell counts (SCC) and plate-culture techniques are accurate in identifying the disease, the respective infectious agents and antibiotic resistant phenotypes. However, pressure exists to develop less lengthy approaches, capable of providing on-site information concerning the infection, and in this way, guide, and hasten the most adequate treatment. Biosensors are analytical tools that convert the presence of biological compounds into an electric signal. Benefitting from high signal-to-noise ratios and fast response times, when properly tuned, they can detect the presence of specific cells and cell markers with high sensitivity. In combination with microfluidics, they provide the means for development of automated and portable diagnostic devices. Still, while biosensors are growing at a fast pace in human diagnostics, applications for the veterinary market, and specifically, for the diagnosis of mastitis remain limited. This review highlights current approaches for mastitis diagnosis and describes the latest outcomes in biosensors and lab-on-chip devices with the potential to become real alternatives to standard practices. Focus is given to those technologies that, in a near future, will enable for an on-farm diagnosis of mastitis.

The accurate diagnosis of bacterial infections is of critical importance for effective treatment decisions. Due to the multietiologic nature of most infectious diseases, multiplex assays are essential for diagnostics. However, multiplexability in nucleic acid amplification-based methods commonly resorts to multiple primers and/or multiple reaction chambers, which increases analysis cost and complexity. Herein, a polymerase chain reaction (PCR) offer method based on a universal pair of primers and an array of specific oligonucleotide probes was developed through the analysis of the bacterial 16S ribosomal RNA gene. The detection system consisted of DNA hybridization over an array of magnetoresistive sensors in a microfabricated biochip coupled to an electronic reader. Immobilized probes interrogated single-stranded biotinylated amplicons and were obtained using asymmetric PCR. Moreover, they were magnetically labelled with streptavidin-coated superparamagnetic nanoparticles. The benchmarking of the system was demonstrated to detect five major bovine mastitis-causing pathogens: Escherichia coli, Klebsiella sp., Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. All selected probes proved to specifically detect their respective amplicon without significant cross reactivity. A calibration curve was performed for S. agalactiae, which demonstrates demonstrating a limit of detection below 30 fg/µL. Thus, a sensitive and specific multiplex detection assay was established, demonstrating its potential as a bioanalytical device for point-of-care applications.

The negative effect of high levels of heavy metals on the activity of soil microorganisms is well-known. However, some microorganisms survive even in high levels of heavy metals, and the microbial activity can therefore, help to recover these polluted soils. Microbial metal uptake in contaminated soils has to be tightly regulated to avoid toxic effects for the cells. These mechanisms of metal resistance are frequently associated to transport-related membrane proteins that mediate bacterium's direct metabolic interactions with the complex soil and aquatic harsh environments. This study reports the identification of gene clusters in rhizobial strains that are regulated by heavy metals, particularly chromium. A DNA fragment was amplified from R. leguminosarum, and in silico analysis of the sequence obtained revealed a putative protein homologue to a cation/multidrug efflux pump component (GenBank DQ398937). Another amplified DNA fragment, with 960 bp, has strong homology with anion ABC transporters (GenBank ZP_002212691) and a peptide ABC transporter (GenBank NP_766950), was identified in Mesorhizobium loti (GenBank DQ398941) and Sinorhizobium meliloti. Using Chromosome Walking technique, a single product from Sinorhizobium meliloti was cloned and sequenced. This new fragment enlarged more 302 bp to the initial sequence corresponding to the ABC transporter, confirming homology with an ATPase from PP superfamily (GenBank ZP_00197146.1).