Scrub typhus is a mite-borne, acute febrile illness caused by the bacterium Orientia tsutsugamushi. It is a re-emerging infectious disease of the tsutsugamushi triangle. Scrub typhus is transmitted through bites of contaminated chiggers (larval stage). Diagnosis of scrub typhus is challenging as its symptoms mimic with other acute febrile illnesses. Several methods are effectual for diagnosis of scrub typhus that includes enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay (IFA), immunochromatographic test (ICT), Weil-Felix, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP). Weil-Felix test was initially used for the diagnosis of scrub typhus in underdeveloped countries but not preferred due to a lack of both specificity and sensitivity. Other immuno-based methods like IFA and ELISA are most outrank for detection of scrub typhus due to their higher sensitivity and specificity, but not vigorous to lay bare the infection at early stages and need the convalescent sampling for verification of positive samples. On another deed, PCR based methods becoming acceptable over era due to its dexterity of early-stage diagnosis with higher specificity and sensitivity but lack its applicability in circumstances of scrub typhus due to the variegated genetic makeup of Orientia tsutsugamushi among its serotypes. The present review focused on various detection methods along with their advantages and disadvantages used in the diagnosis of scrub typhus. A comparison between available methods of diagnosis with challenges in the detection of scrub typhus is also summarized.

Coronaviruses silently circulate in human and animal populations, causing mild to severe diseases. Therefore, livestock are important components of a "One Health" perspective aimed to control these viral infections. However, at present there is no example that considers pig genetic resources in this context. In this study, we investigated the variability of four genes (ACE2, ANPEP and DPP4 encoding for host receptors of the viral spike proteins and TMPRSS2 encoding for a host proteinase) in 23 European (19 autochthonous and three commercial breeds and one wild boar population) and two Asian Sus scrofa populations. A total of 2229 variants were identi ed in the four candidate genes: 26% of them were not previously described; 29 variants affected the protein sequence and might potentially interact with the infection mechanisms. The results coming from this work are a rst step towards a "One Health" perspective that should consider conservation programmes of pig genetic resources with twofold objectives: i) genetic resources could be reservoirs of host gene variability useful to design selection programmes to increase resistance to coronaviruses; ii) the described variability in genes involved in coronavirus infections across many different pig populations might be part of a risk assessment including pig genetic resources.

Progress in the medical profession is determined by the achievements and effectiveness of new antibiotics in the treatment of microbial infections. However, the development of multiple-drug resistance in numerous bacteria, especially Gram-negative bacteria, has limited the treatment options. Due to this resistance, the resurgence of cyclic polypeptide drugs like colistin remains the only option. The drug, colistin, is a well-known growth inhibitor of Gram-negative bacteria like Acinetobacter baumanni, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Technological advancements have uncovered the role of the mcr-1(mobilized colistin resistance) gene, which is responsible for the development of resistance in Gram-negative bacteria, which make them distinct from other bacteria without this gene. Additionally, food animals have been determined to be the reservoir for colistin resistance microbes, from which they spread to other hosts. Due to the adverse effects of colistin, many developed countries have prohibited its usage in animal foods, but developing countries are still using colistin in animal food production, thereby imposing a major risk to the public health. Therefore, there is a need for implementation of sustainable measures in livestock farms to prevent microbial infection. This review highlights the negative effects (increased resistance) of colistin consumption and emphasizes the different approaches used for detecting colistin in animal-based foods as well as the challenges associated with its detection.

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A novel approach has been developed for the detection of 56 kDa tissue-specific antigen (TSA) gene of Orientia tsutsugamushi a causative agent of scrub typhus disease. The approach was developed by immobilization of 5′ NH2 labeled ssDNA probe selective to 56 kDa TSA gene, to the surface of AuNPs/CNF modified screen-printed electrode. An electrochemical response was recorded with single stranded genomic DNA (ssDNA) of O. tsutsugamushi isolated from patient sample, using cyclic voltammetry and electrochemical impedance spectroscopy. The electrode surface was characterized by Field-Emission Scanning electron microscope (FE-SEM), Fourier Transform Infrared Spectroscopy (FTIR) and Raman Spectroscopy at each step of fabrication. The DNA biosensor shows optimum response within 50-60 s at room temperature (25 ± 3 °C). The sensor shows higher sensitivity [7849 (µA/cm 2 )/ng DNA], fast response time (60 s), wider linear range (0.04-2.6 ng) with limit of detection of 0.02 ng/µl of ssDNA sample.

An optimized DNA-based bioassay for Leptospira interrogans detection has been developed. Electrochemical studies of the developed biosensor were done using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Surface characterization of the biosensor was done using scanning electron microscopy (SEM). The biosensor showed specificity to L. interrogans as determined by specificity studies. The sensitivity of the biosensor was 264.5 µA/cm 2 /ng and lower limit of detection (LOD) was 0.015 ng/6 µl using CV. The biosensor was also validated with serum samples spiked with singlestranded leptospiral DNA. The developed biosensor also showed good stability for a period of 6 months at 4 °C as shown by the DPV analysis. Research highlightsThe developed DNA-based bioassay is the first report on biosensor-based leptospirosis diagnosis. The developed bioassay is a DNA-based amperometric biosensor specific to L. interrogans causing leptospirosis. The biosensor has a sensitivity of 264.5 µA/cm 2 /ng and a lower limit of detection (LOD) of 0.015 ng/6µl. The biosensor was stable for 6 months with only 13% loss in DPV peak current.