Minakshi Prasad

A novel genomic constellation (G10P[3]) of group A rotavirus detected from buffalo calves in northern India

Group A bovine rotaviruses cause gastroenteritis and calf mortality leading to significant economic losses to dairy farmers in India. Due to segmented nature of the RNA genome and wide host range, vast genetic and antigenic diversity exists among different isolates of rotavirus. Molecular characterization of locally prevalent group A rotavirus strains in buffalo population in north India was undertaken. Out of a total of 455 faecal samples, 21 samples (4.61%) were positive for bovine rota virus (BRV) as determined by PAGE and ELISA, whereas of these only 15 isolates yielded specific products for VP4 and VP7 genes by RT-PCR. Genotyping by nested PCR typed G6, G10 and P[11] genotypes but VP4 genes of 11 isolates remained untyped. The phylogenetic and evolutionary analysis of nucleotide and predicted amino acid sequences of the cloned products of VP4 and VP7 genes confirmed typing results obtained by nested PCR for G6, G10 and P[11] and classified the untyped isolates as P[3] genotypes. In this study, it was observed that G6P[11] (26.66%) and G10P[3] (73.34%) group A rotaviruses are circulating in buffalo herds of organized farms in north India. Unusual reassortants G10P[3] of group A rotaviruses isolated from buffalo calves show novel genomic constellations indicative of interspecies reassortment.

Advances in diagnosis of respiratory diseases of small ruminants.

Irrespective of aetiology, infectious respiratory diseases of sheep and goats contribute to 5.6 percent of the total diseases of small ruminants. These infectious respiratory disorders are divided into two groups: the diseases of upper respiratory tract, namely, nasal myiasis and enzootic nasal tumors, and diseases of lower respiratory tract, namely, peste des petits ruminants (PPR), parainfluenza, Pasteurellosis, Ovine progressive pneumonia, mycoplasmosis, caprine arthritis encephalitis virus, caseous lymphadenitis, verminous pneumonia, and many others. Depending upon aetiology, many of them are acute and fatal in nature. Early, rapid, and specific diagnosis of such diseases holds great importance to reduce the losses. The advanced enzyme-linked immunosorbent assays (ELISAs) for the detection of antigen as well as antibodies directly from the samples and molecular diagnostic assays along with microsatellites comprehensively assist in diagnosis as well as treatment and epidemiological studies. The present review discusses the advancements made in the diagnosis of common infectious respiratory diseases of sheep and goats. It would update the knowledge and help in adapting and implementing appropriate, timely, and confirmatory diagnostic procedures. Moreover, it would assist in designing appropriate prevention protocols and devising suitable control strategies to overcome respiratory diseases and alleviate the economic losses.

Nanotherapeutics: An insight into healthcare and multi-dimensional applications in medical sector of the modern world

In recent years nanotechnology has revolutionized the healthcare strategies and envisioned to have a tremendous impact to offer better health facilities. In this context, medical nanotechnology involves design, fabrication, regulation, and application of therapeutic drugs and devices having a size in nano-range (1-100 nm). Owing to the revolutionary implications in drug delivery and gene therapy, nanotherapeutics has gained increasing research interest in the current medical sector of the modern world. The areas which anticipate benefits from nano-based drug delivery systems are cancer, diabetes, infectious diseases, neurodegenerative diseases, blood disorders and orthopedic problems. The development of nanotherapeutics with multi-functionalities has considerable potential to fill the lacunae existing in the present therapeutic domain. Nanomedicines in the field of cancer management have enhanced permeability and retention of drugs thereby effectively targeting the affected tissues. Polymeric conjugates of asparaginase, polymeric micelles of paclitaxel have been recmended for various types of cancer treatment .The advancement of nano therapeutics and diagnostics can provide the improved effectiveness of the drug with less or no toxicity concerns. Similarly, diagnostic imaging is having potential future applications with newer imaging elements at nano level. The newly emerging field of nanorobotics can provide new directions in the field of healthcare. In this article, an attempt has been made to highlight the novel nanotherapeutic potentialities of polymeric nanoparticles, nanoemulsion, solid lipid nanoparticle, nanostructured lipid carriers, dendrimers, nanocapsules and nanosponges based approaches. The useful applications of these nano-medicines in the field of cancer, nutrition, and health have been discussed in details. Regulatory and safety concerns along with the commercial status of nanosystems have also been presented. In summary, a successful translation of emerging nanotherapeutics into commercial products may lead to an expansion of biomedical science. Towards the end of the review, future perspectives of this important field have been introduced briefly.

Multispecies reassortant bovine rotavirus strain carries a novel simian G3-like VP7 genotype

Rotavirus-A (RVAs), are the major cause of severe gastroenteritis in the young of mammals and birds. RVA strains possessing G6, G8, and G10 genotypes in combination with P[1] or P[11] have been commonly detected in cattle. During a routine surveillance for enteric viruses in a bovine population on North-Western temperate Himalayan region of India, an uncommon bovine RVA strain, designated as RVA/Cow-wt/IND/M1/09/2009 was detected in a diarrhoeic crossbred calf. The examination of nearly complete genome sequence of this RVA strain revealed an unusual G-P combination (G3P[11]) on a typical bovine RVA genotype backbone (I2-R2-C2-M2-A11-N2-T6-E2-H3). The VP7 gene of M1/09 isolate displayed a maximum nucleotide sequence identity of 73.8% with simian strain (RVA/Simian-tc/USA/RRV/1975/G3P[3]). The VP4 and NSP5 genes clustered with an Indian pig strain, RVA/Pig-wt/IND/AM-P66/2012/G10P[11] (99.6%), and a caprine strain, RVA/Goat-tc/BGD/GO34/1999/G6P[1] (98.9%) from Bangladesh, respectively, whilst the, VP6, NSP1, NSP3 and NSP4 genes were identical or nearly identical to Indian bovine strains (RVA/Cow-wt/IND/B-72/2008/G10P[X], RVA/Cow-wt/IND/B85/2010/GXP[X], and RVA/Cow-wt/IND/C91/2011/G6P[X]). The remaining four genes (VP1, VP2, VP3 and NSP2) were more closely related to RVA/Human-wt/ITA/PAI11/1996/G2P[4] (93.5%), RVA/Sheep-wt/CHN/LLR/1985/G10P[15] (88.8%), RVA/Human-tc/SWE/1076/1983/G2P2A[6] (93.2%) and RVA/Human-wt/AUS/CK20003/2000/G2P[4] (91.2%), respectively. Altogether, these findings are suggestive of multiple independent interspecies transmission and reassortment events between co-circulating bovine, porcine, ovine and human rotaviruses. The complete genome sequence information is necessary to establish the evolutionary relationship, interspecies transmission and ecological features of animal RVAs from different geographical regions.

BATS: CARRIERS OF ZOONOTIC VIRAL AND EMERGING INFECTIOUS DISEASES

Bats are reported as reservoir host for several viruses, which cause significant illness in human and animals. Some of the bat transmitted zoonotic viral diseases such as Ebola, Hendra, Nipah and rabies may cause severe human casualties. They also harbor several other viruses such as MERS and SARS corona viruses, which may cause disease in human through direct spillover to human or through an intermediate host or vectors. Being reservoir hosts bats do not get affected by these viruses. This probably may happen due to the specificity of bat immune system, which reacts differently with viral pathogens in comparison to their other mammalian counterparts. Although bats are important reservoir hosts for several zoonotic viruses, very little information is available regarding host/virus relationships as only few experimental studies have been done on bat colonies, lack of expertise for study of bat immunology and antiviral responses and difficulty in conducting field work. However, with the advancement in epidemiology and molecular biology, these problems can be addressed, which will provide the insight into interactions of bats and zoonotic viruses. It may also clarify regarding virus persistence in nature and various associated risk factors which might facilitate viral transmission to animals and humans. Koushlesh Ranjan 1,* , Minakshi Prasad 2 and Gaya Prasad 3

Development of a rapid test for detection of foot-and-mouth disease virus specific antibodies using gold nanoparticles

Foot and mouth disease is an economically important transboundary disease of wildlife and cloven hoofed animals including ruminants. In the absence of vaccination, detection of antibodies against structural proteins (SPs) of foot-and-mouth disease virus is an indicator of infection. In the present study, a rapid dot blot assay using gold nanoparticlees was developed for the detection of antibodies against SPs of FMDV. Commercially available FMD vaccine was used as a source of FMD antigen. After the synthesis of gold nanoparticles (GNPs), the GNP-dot blot assay was tested and was found very sensitive, as the detection of antibody was up to 10−4 of serum dilution. The GNP-dot assay was found specific as it didn’t give dot with normal horse sera, fetal bovine sera and neonatal bovine calf serum samples when tested at 10−3 working dilution. When 30 serum samples from post-vaccinated buffaloes were tested at dilution of 10−3, all the samples were found positive with the variable intensity of dot. The synthesized GNPs and conjugated GNPS with antibody were characterized for their absorption limit, for their stability and for their approximate size. These characterized conjugated and non-conjugated GNPs were also analyzed by Transmission electron microscopy and Scanning electron microscopy. The GNP dot blot assay developed in this work gave promising results using vaccine antigen and can form an important tool for rapid diagnosis of FMD in FMD free countries, zones free of FMD and during last stage of FMD eradication when FMD vaccination will be withdrawn.

Occurrence of Neglected Picobirna Virus in Buffalo Calves Associated with Diarrhoea

Picobirna viruses (PBVs) thought to be neglected viruses, are emerging as causative agents of viral gastrointestinal infections in human and other animal species with enteritis. In the present study, a total of 408 fecal samples of diarrheic buffalo calves of under age of 2 months were collected from different locations in Haryana. These samples were preliminarily screened for PBV by RNApolyacrylamide gel electrophoresis (RNA-PAGE). The RNA-PAGE positive samples were further confirmed by RT-PCR using RdRp gene specific primer pair PicoB25 (+) and PicoB43 (-) for amplification of genogroup I PBV that generated an amplicon of 201 bp. Out of 408 buffalo calves fecal samples, 52 (13%) samples were found positive for genogroup 1(GGI) PBV using RT-PCR. Genogroup II (GGII) specific primer generating 369 bp product size did not show amplification in any of the samples. The results suggested occurrence of PBV in Haryana State with predominance of GGI PBV. To the best of our knowledge, this is the first report of detection as well as occurrence of GGI picobirna virus among diarrheic buffalo calves in Haryana State.

Advances in Designing and Developing Vaccines, Drugs and Therapeutic Approaches to Counter Human Papilloma Virus

Human papillomavirus (HPV) is a viral infection with skin-to-skin based transmission mode. HPV annually caused over 500,000 cancer cases including cervical, anogenital and oropharyngeal cancer among others. HPV vaccination has become a public-health concern, worldwide, to prevent the cases of HPV infections including precancerous lesions, cervical cancers, and genital warts especially in adolescent female and male population by launching national programs with international alliances. Currently, available prophylactic and therapeutic vaccines are expensive to be used in developing countries for vaccination programs. The recent progress in immunotherapy, biotechnology, recombinant DNA technology and molecular biology along with alternative and complementary medicinal systems have paved novel ways and valuable opportunities to design and develop effective prophylactic and therapeutic vaccines, drugs and treatment approach to counter HPV effectively. Exploration and more researches on such advances could result in the gradual reduction in the incidences of HPV cases across the world. The present review presents a current global scenario and futuristic prospects of the advanced prophylactic and therapeutic approaches against HPV along with recent patents coverage of the progress and advances in drugs, vaccines and therapeutic regimens to effectively combat HPV infections and its cancerous conditions.

Virus-Host Interactions: New Insights and Advances in Drug Development Against Viral Pathogens

BACKGROUND Viruses are the most devastating pathogens of almost all life forms including humans and animals. Viruses can replicate very fast and may affect any metabolic and physiological function of the host cell. Therefore, it has been a challenge to develop a universal and common treatment against viral pathogens, in contrast to bacterial pathogens. Virus-host interaction is a complex phenomenon and often is virus- and host cell-specific. Exciting new insights into the molecular pathogenesis and host-virus interactions have been gained over the past few decades. These advances have enabled researchers to design better antiviral drugs. METHODS The literature related to various aspects of virus-host interactions: new insights and advances in drug development was collected from several scientific research related databases such as Science Direct, Google Scholar, Scopus, PubMed, AGRICOLA, and Medline, etc. Total number of 319 research papers was used to compile the information regarding drug development against viral pathogens. RESULTS Clinical adequacy of antiviral drugs and their bioavailability are important parameters for effective treatment of viral infections. The problems associated with effective delivery of a drug in a safe and desired quantity have led to the search for (and design of) better drug delivery systems. In recent past, several new antiviral drugs have been developed, which have high therapeutic effectiveness against life-threatening viral diseases such as HIV, hepatitis B virus, herpes virus, dengue virus and influenza virus infections. The majority of recent advances in antiviral drug discovery were possible due to the developments in allied fields such as in vitro virus cultivation technology, molecular biology of viral-genome-encoded enzymes, complete-genome-sequence-based studies of viruses and identification of suitable targets for antiviral drugs in viral genomes. Recently, several novel drug delivery approaches including small interfering RNAs (siRNAs) have emerged to aid antiviral therapy. CONCLUSION The present review is aimed at providing an update on research and development efforts being made to create effective antiviral chemotherapeutic agents and approaches to their delivery to appropriate targeted cells or tissues.

Synthesis, characterization and anticancer activity of vincristine loaded folic acid-chitosan conjugated nanoparticles on NCI-H460 non-small cell lung cancer cell line

Abstract Vincristine is used to treat different type of cancers. But unwanted side effects limit their applications in medicine. To overcome these side effects by targeted drug delivery approach, we synthesized vincristine loaded folic acid-chitosan conjugated nanoparticles by ionic gelation method. The nanoparticles were characterized using Dynamic Light Scattering (DLS), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy. DLS confirmed small sized nanoparticles (<200 nm) while FTIR confirmed different functional groups associated with synthesized nanoparticles. SEM showed spherical shaped nanoparticles with smooth surface, whereas, TEM confirmed loading of vincristine in folic acid-chitosan conjugated nanoparticles. Anticancer activity of vincristine loaded folic acid-chitosan conjugated nanoparticles was checked by MTT assay on NCI-H460 cells followed by reactive oxygen species level, mitochondrial transmembrane potential and apoptotic morphological changes, which confirmed the cytotoxicity of nanoparticles. Erythrocyte aggregation assay confirmed non-toxicity of nanoparticles. Hence, these nanoparticles can be used to treat NCI-H460 cells.