Nanotechnology: a contemporary therapeutic approach in combating infections from multidrug-resistant bacteria.

In the 20th century, the discovery of antibiotics played an essential role in the fight against infectious diseases, including meningitis, typhoid fever, pneumonia and Mycobacterium tuberculosis. The development of multidrug resistance in microflora due to improper antibiotic use created significant public health issues. Antibiotic resistance has increased at an alarming rate in the past few decades. Multidrug-resistant bacteria (superbugs) such as methicillin-resistant Staphylococcus aureus (MRSA) as well as drug-resistant tuberculosis pose serious health implications. Despite the continuous increase in resistant microbes, the discovery of novel antibiotics is constrained by the cost and complexities of discovery of drugs. The nanotechnology has given new hope in combating this problem. In the present review, recent developments in therapeutics utilizing nanotechnology for novel antimicrobial drug development are discussed. The nanoparticles of silver, gold and zinc oxide have proved to be efficient antimicrobial agents against multidrug-resistant Klebsiella, Pseudomonas, Escherichia Coli and MRSA. Using nanostructures as carriers for antimicrobial agents provides better bioavailability, less chances of sub-therapeutic drug accumulation and less drug-related toxicity. Nanophotothermal therapy using fullerene and antibody functionalized nanostructures are other strategies that can prove to be helpful.

Multiplex PCR for rapid differential diagnosis of co-prevalent species of Theileria (Theileria annulata and Theileria orientalis) in cattle.

Theileriosis is a tick-borne disease that causes enormous losses in the dairy industry. There are several species of Theileria that can infect bovines. Generally, more than one species are prevalent in any geographical area; thus, chances of co-infections are high. Differentiation of these species may not be possible by microscopic examination or serological tests. Therefore, in this study, a multiplex PCR assay was standardized and evaluated for rapid and simultaneous differential detection of two species of Theileria viz., Theileria annulata and Theileria orientalis. Species-specific primers were designed to target the merozoite piroplasm surface antigen gene (TAMS1) of T. annulata and the major piroplasm surface protein gene of T. orientalis, yielding specific amplicon of 229 bp and 466 bp, respectively. The sensitivity of multiplex PCR was 102 and 103 copies for T. annulata and T. orientalis, respectively. The simplex and multiplex PCRs were specific and showed no cross-reactivity with other hemoprotozoa for either primer. For comparative evaluation, blood samples from 216 cattle were tested by simplex and multiplex PCR for both species. Using multiplex PCR, 131 animals were found infected for theileriosis, of which 112 were infected with T. annulata, five were infected with T. orientalis, and 14 had mixed infections. This is the first report of T. orientalis from Haryana, India. Representative sequences of T. annulata (ON248941) and T. orientalis (ON248942) were submitted in GenBank. The standardized multiplex PCR assay used in this study was specific, sensitive, for the screening of field samples.

A Rapid and Novel Multiplex PCR Assay for Simultaneous Detection of Multiple Viruses Associated with Bovine Gastroenteritis.

Bovine viral diarrheal virus (BVDV) and bovine coronavirus (BCoV) are prevalent viral infections in buffalo calves that result in significant economic losses globally. However, Bovine picobirnavirus (BPBV) Group I and II has been an emerging causes of gastrointestinal infection as has been detected with mixed of BVDV as well as BCV. To combat economic losses and viral infection, a rapid and innovative multiplex-PCR assay (M-PCR) was developed to simultaneously identify BVDV, BCV, and BPBV. The assay employed three primer pairs, each specific to a particular virus. Notably, the primers for BCV and BVDV, targeting the transmembrane (M) Mebus gene and 5'UTR genes, respectively, were self-designed. To validate the assay, 300 samples of buffalo calf feces were subjected to the standardized multiplex PCR. The results demonstrated that 54 (18%) samples tested positive for multiple viruses, with 16.67% samples infected by BVDV, 0.9% by BCoV, and 0.13% by BPBV, as detected by the M-PCR assay. In summary, this developed assay is characterized by high specificity, sensitivity, throughput, and speed, enabling the simultaneous detection of the three viruses in a single reaction tube. Consequently, it holds potential for epidemiological investigations. It is worth noting that, to the best of our knowledge, this is the first reported multiplex assay for the worldwide detection of BVDV, BCoV, and BPBV. This novel assay promises to aid in the detection of mixed infections in the gastrointestinal tract.

Molecular epidemiology of anaplasmosis in Indian dromedary camels.

The present study aimed to investigate the molecular epidemiology, risk factors, and haemato-biochemical changes in anaplasmosis in Indian dromedary camels. Blood samples collected from 103 camels were analysed using blood smear examination and semi-nested PCR based on the 16s rRNA gene to diagnose anaplasmosis. The prevalence of anaplasmosis was estimated to be 42.72% (95% CI: 33.59-52.37) by PCR assay and 16.5% (95% CI: 10.47-24.95%) by blood smear examination. Phylogenetic analysis of six partial sequences of 16s rRNA gene obtained in the present study indicated the involvement of multiple Anaplasma species, including A. marginale and A. platys, showing genetic similarity with cattle strains. A novel genotype related to A. camelii/A. cinensis/A. platys group was also identified. Hemato-biochemical examination revealed mild anaemia, increased serum urea nitrogen and creatinine levels, and decreased total protein and albumin levels in Anaplasma-positive animals. The infections were largely subclinical in nature, except in one camel that revealed fever, inappetence, and pale mucous membrane and responded well to treatment with oxytetracycline. To our knowledge, this is the first molecular study on camel anaplasmosis in India, indicating a high prevalence of infection and involvement of multiple Anaplasma species with potential risk for interspecies transmission.

Molecular investigations of camelpox virus circulating in dromedary camel population in Rajasthan, India.

Camelpox is an important viral disease of dromedary camel in Rajasthan, India. In the present study, partial C18L gene sequences (n = 6) of camelpox virus (CMLV) obtained in an outbreak in Bikaner, Rajasthan, India in year 2022 were compared with other similar sequences obtained in the past in similar geographical location. Clinical and epidemiological features of the disease were also compared. Genomic study suggested variations in C18L gene sequences obtained in the present outbreak from those obtained during the past outbreaks. CMLV were genetically different from cowpox viruses, but appeared identical to CMLV causing disease in Israel, Egypt and Kazakhstan. Genomes of CMLV virus circulating in dromedary camel population of Rajasthan, India appeared diverse and changing, hence complete genome sequencing and identification of genomic changes altering infectivity and pathogenicity is warranted for designing control strategies.

Molecular and Physiological Mechanisms to Mitigate Abiotic Stress Conditions in Plants.

Agriculture production faces many abiotic stresses, mainly drought, salinity, low and high temperature. These abiotic stresses inhibit plants' genetic potential, which is the cause of huge reduction in crop productivity, decrease potent yields for important crop plants by more than 50% and imbalance agriculture's sustainability. They lead to changes in the physio-morphological, molecular, and biochemical nature of the plants and change plants' regular metabolism, which makes them a leading cause of losses in crop productivity. These changes in plant systems also help to mitigate abiotic stress conditions. To initiate the signal during stress conditions, sensor molecules of the plant perceive the stress signal from the outside and commence a signaling cascade to send a message and stimulate nuclear transcription factors to provoke specific gene expression. To mitigate the abiotic stress, plants contain several methods of avoidance, adaption, and acclimation. In addition to these, to manage stress conditions, plants possess several tolerance mechanisms which involve ion transporters, osmoprotectants, proteins, and other factors associated with transcriptional control, and signaling cascades are stimulated to offset abiotic stress-associated biochemical and molecular changes. Plant growth and survival depends on the ability to respond to the stress stimulus, produce the signal, and start suitable biochemical and physiological changes. Various important factors, such as the biochemical, physiological, and molecular mechanisms of plants, including the use of microbiomes and nanotechnology to combat abiotic stresses, are highlighted in this article.