Identification of circulatory microRNA based biomarkers for early pregnancy diagnosis in buffalo.

Introduction: The most crucial factor in improving animal reproduction efficiency is early pregnancy diagnosis. Early diagnosis not only reduces the time interval between two calvings but also aids farmers in identifying open animals, thereby preventing significant milk production losses. Therefore, the objective of this study was to discover circulatory miRNAs that would be useful for early pregnancy diagnosis in buffalo. Material and methods: Blood samples were taken on 0, 6th, 12th, and 18th day after artificial insemination from pregnant animals (n = 30) and non-pregnant animals (n = 20). During these stages of pregnancy, total RNA was extracted, and a small RNA library was subsequently generated and sequenced on the Illumina platform. Subsequently, Real-time PCR was used to validate the findings. Results and discussion: There were 4,022 miRNAs found during the pregnancy, with 15 of those lacking sequences and 4,007 having sequences already in the database. From the beginning of pregnancy until the 18th day, 25 of these miRNAs showed a substantial shift in expression levels in the maternal blood, with a change more than two logs. Furthermore, based on qPCR results, 19 miRNAs were found to be more abundant in pregnant animals than in non-pregnant animals. We used target prediction analysis to learn how maternally expressed miRNAs relate to fetal-maternal communication. In conclusion, miRNA based biomarkers that could be associated with the diagnosis of pregnancy were identified including miR-181a and miR-486 highly upregulated on the 18th day of pregnancy. This study also provides a comprehensive profile of the entire miRNA population in maternal buffalo blood during the early stages of pregnancy.

Temporal analysis of circulating nucleic acid in early days of pregnancy in buffalo.

Improving reproductive efficiency in livestock relies mainly on the ability to detect pregnancy quickly and accurately. Recently, circulating nucleic acids (CNAs) have been exploited for prenatal diagnosis in humans and animals. In the current investigation, serum samples were collected from pregnant animals (n = 30) and non-pregnant animals (n = 20) on 0th, 6th, 12th, and 18th day post artificial insemination. Total DNA was isolated from these serum samples. Two CNA tags (Bov-B and ART2A) derived from repetitive sections of the bovine genome were amplified using DNA extracted from serum samples. The expression analysis of these CNAs was done using real-time polymerase chain reaction assay, and copy number of each tag was calculated in pregnant and non-pregnant animals. The average number of copies of Art2A increased approximately threefold (P < 0.01) from day zero of pregnancy (7,000 copies) to the day 18 of pregnancy (> 21,000). Similarly, BovB levels in the pregnant group increased significantly (approximately 2.9-fold) from day zero (93,900 copies) till day 18 (> 2, 72,310 copies) (P < 0.01). There was no significant change observed on the 6th and 12th day of pregnancy and on the 18th day in the non-pregnant animals. In conclusion, based on these findings, the defined cut-off value can distinguish between pregnant and non-pregnant animals with a sensitivity of nearly 80% and specificity of nearly 70%. It is possible to employ these two CNA tags as biomarkers for early detection of pregnancy in buffaloes.

The involvement of organic acids in soil fertility, plant health and environment sustainability.

Increasing demand for safe food by an ever-growing human population emphasizes the urgency for increasing crop yields and reducing the losses caused by abiotic and biotic stresses; a partial solution to this problem is to develop a better understanding of plant-microbe interactions. Plant roots continuously release a wide range of compounds including organic acids in root exudates. These root exudates stimulate growth of specific microbial communities in the rhizosphere, which affect complex biological and physico-chemical interactions occurring between plant roots and the surrounding soil environment. In addition, organic acids are also released by different microbes and during decomposition of organic matter and plant residues in the soil. Interestingly, the available organic acids in the rhizosphere play crucial roles in various physio-chemical processes including the chemoattraction of microbes (both beneficial and pathogenic), mineralization and solubilization of complex minerals (P, K and Zn), biocontrol of phytopathogens, induction of systemic resistance, biogas formation, mitigation of abiotic stresses and, detoxification of metals and residual pesticides. Thus, organic acids play a significant role in the sustainable management of the soil ecosystem and in environmental sustainability. This review discusses the role of organic acids in the stimulation or enrichment of specific root-associated microbial communities and their effect on plant-microbe interactions at the root surface. In addition, the potential for root microbiome modification to enhance nutrient cycling and nutrient acquisition, and in amelioration of environmental stresses for increasing food production is discussed.

An Insight on RNA Based Therapeutics and Vaccines: Challenges and Opportunities.

In this era, RNA molecules have provided a unique opportunity to researchers all over the world for expanding their range of targets in the development of drugs. Due to the unique pharmacological as well as physicochemical characteristics of different RNA molecules such as aptamers, small interfering RNAs (siRNA), antisense oligonucleotides (ASO) and guide RNAs (gRNA), they have emerged recently as a new class of drugs. They are used for selective action on proteins and genes that were not possible to target by conventional drug molecules. These RNA molecules like guide RNAs are also components of novel gene editing mechanisms which can modify the genome nearly in all cells. Vaccines based on RNA molecules have also provided a promising alternative to conventional live attenuated vaccines. RNA based vaccines have high potency, can be rapidly developed, and have potential for manufacturing at a cheaper rate and safe administration. However, the application of these RNAs has been restricted by the high instability and inefficient in vivo delivery. Technological advancement needs to overcome these issues so that RNA based drugs targeting several diseases can be developed. This article emphasizes the potential of RNA based drugs and the major barriers associated with the development of RNA therapeutics. Additionally, the role of RNA based vaccines and their challenges in advancing this promising vaccine platform for the prevention of infectious diseases have been discussed.

Bovine brucellosis - a comprehensive review.

Brucellosis is a zoonotic disease of great animal welfare and economic implications worldwide known since ancient times. The emergence of brucellosis in new areas as well as transmission of brucellosis from wild and domestic animals is of great significance in terms of new epidemiological dimensions. Brucellosis poses a major public health threat by the consumption of non-pasteurized milk and milk products produced by unhygienic dairy farms in endemic areas. Regular and meticulous surveillance is essentially required to determine the true picture of brucellosis especially in areas with continuous high prevalence. Additionally, international migration of humans, animals and trade of animal products has created a challenge for disease spread and diagnosis in non-endemic areas. Isolation and identification remain the gold standard test, which requires expertise. The advancement in diagnostic strategies coupled with screening of newly introduced animals is warranted to control the disease. Of note, the diagnostic value of miRNAs for appropriate detection of B. abortus infection has been shown. The most widely used vaccine strains to protect against Brucella infection and related abortions in cattle are strain 19 and RB51. Moreover, it is very important to note that no vaccine, which is highly protective, safe and effective is available either for bovines or human beings. Research results encourage the use of bacteriophage lysates in treatment of bovine brucellosis. One Health approach can aid in control of this disease, both in animals and man.

Interkingdom signaling in plant-rhizomicrobiome interactions for sustainable agriculture.

The rhizosphere is a complex ecosystem around plant roots that comprises an integrated network of plant roots, the microbiome and soil. Wide spread communication between prokaryotes and eukaryotes occurs within this integrated network via a complex set of signal molecules secreted during both beneficial and harmful interactions. Intra- and inter-species communication among microbes occurs through various signal molecules that coordinate and control the behaviours of microorganisms in mixed communities. In addition, interkingdom signal exchange between plants and microbes occurs through the release of root exudates from the host plants. The diverse chemical substances released in root exudates affect the structural and physical heterogeneity of the soil. Moreover, chemical compounds released in root exudates trigger various signaling pathways in microbial populations that influence rhizosphere biology. Therefore, deciphering the language of interkingdom communication and understanding the mechanisms involved is innovative and promising approach for improving crop production in sustainable agriculture. This chapter describes briefly the shaping of the rhizomicrobiome in response to released root exudates. Moreover, predicting and controlling the microbiome structure and its function in the rhizosphere by understanding of rhizomicrobiome communication through different secreted compounds will allow us to better harness beneficial plant-microbe interactions. The recent progress in understanding interkingdom communication and interactions between plants and microbes is discussed in relation to plant growth, gene expression, nutrient uptake and resistance to pests and diseases along with mitigation of abiotic stresses in plants to improve plant ecosystem productivity for sustainable agriculture.