During cryopreservation, oxidative damage to sperm biomolecules occurs, ultimately compromising its fertilizing ability. Simple proteome analysis may not accurately reflect the cellular functions that are affected, as most carbonylated proteins lose their functions. The carbonylated proteins act as a good marker of oxidative stress. Choosing buffalo spermatozoa as a model, carbonylated proteins from fresh-extended (FESCL) and frozen-thawed (FTSCL) spermatozoa were enriched by affinity chromatography using biotin hydrazide and a monomeric avidin agarose matrix followed by mass spectrometry. Data were analyzed with Proteome Discoverer (v2.2), post-translational modification analysis was performed using SEQUEST, and affected functional activities were predicted using the FunRich tool (v3.0). The mass spectrometric analysis of sperm proteins led to the identification of 415 carbonylated proteins, 151 in FESCL and 405 in FTSCL. Twelve highly abundant carbonylated proteins identified only in FTSCL can be the major contributors to the cryoinjury. The cryopreservation induces the carbonylation of selective sperm proteins. A total of 98 peptides have been precisely annotated that have oxidative modifications. In comparison to FESCL, more proteins in various cellular components, pathways, and processes in FTSCL were carbonylated. The findings indicate the molecular injury to spermatozoa through carbonylated proteins affecting energy metabolism, free-radical scavenging, cytoskeleton, plasma membrane, capacitation, zona-pellucida binding, and sperm-oocyte interaction and are well correlated with the functional parameters of the spermatozoa. The findings provide strong evidence that protein carbonylation is one of the major factors and molecular basis of cryodamage, which compromises the functions of FTSCL. With cryopreservation becoming an inevitable tool, the study becomes more relevant, and this experimental design would give better-quality frozen-thawed spermatozoa with good molecular profiles.

Bovine tropical theileriosis, caused by Theileria annulata, is an economically important disease that reduces livestock productivity through decreased milk yield, high morbidity, and mortality, particularly in exotic and cross-bred cattle. Timely diagnosis and treatment are crucial for effectively managing susceptible animals. This study aimed to develop a robust serological diagnostic tool targeting the sporozoite and macroschizont 2 antigen (Spm2), a protein expressed across multiple parasite stages including sporozoites, macroschizonts, and piroplasms. The recombinant Spm2 (rSpm2) protein was cloned, expressed in a prokaryotic system, and purified via Ni-NTA chromatography, with its immunoreactivity confirmed through Western blot (WB) analysis using known T. annulata-positive serum samples. An indirect rSpm2-ELISA (Enzyme-linked immunosorbent assay) was developed and standardized using 23 known positive and 30 known negative sera samples. Its diagnostic potential was evaluated by analyzing 360 field serum samples and comparing the results with a previously validated rTaSP-ELISA. The rSpm2-ELISA detected T. annulata infection in 78.9 % of cattle sera, compared to 83.9 % with rTaSP-ELISA. Notably, rSpm2 exhibited no cross-reactivity with reference-positive sera from cattle infected with Babesia bigemina and Anaplasma marginale in WB and ELISA assays. The sensitivity and specificity of the rSpm2-ELISA were determined to be 93.7 % and 98.3 %, respectively, highlighting its strong diagnostic potential. This study is the first comparative serological evaluation of tropical theileriosis using rSpm2-ELISA and rTaSP-ELISA. The high accuracy, specificity, and sensitivity of rSpm2-ELISA make it a promising tool for serological surveillance of tropical theileriosis, particularly in regions where T. parva is absent.

Background: Leishmania infantum, one important intracellular parasite causes most potentially lethal diseases such as leishmaniasis i.e. Visceral leishmaniasis in dogs. Although certain chemical drugs such as pentostam, amphotericin B, miltefosine have been trailed against this disease, but all these drugs induced antibiotic resistance and toxicity in the host. Further, the trypanothione synthetase, a key enzyme of this parasite which catalyzes a reaction, maintaining thiol redox within the cell. Methods: The binding study was carried out with selected natural/synthetic phytocompounds/drugs against the modeled trypanothione synthetase though molecular docking. Result: The generated protein model with lowest discrete potential energy (DOPE) -19960.97 was found good in quality with z score of -5.19 and quality factor of 61.83% and it was found that the natural inhibitors glycyrrhetic acid (GRA) and Theaflavin (TFN) showed highest binding energies of -7.34 and -6.95 Kcal/mol. This study may be concluded that the natural inhibitors glycyrrhetic acid (GRA) and Theaflavin (TFN) would be potential regimen in treatment of canine leishmaniasis.

To combat drought stress in rice, a major threat to global food security, three major quantitative trait loci for 'yield under drought stress' (qDTYs) were successfully exploited in the last decade. However, their molecular basis still remains unknown. To understand the role of secondary regulation by miRNA in drought stress response and their relation, if any, with the three qDTYs, the miRNA dynamics under drought stress was studied at booting stage in two drought tolerant (Sahbaghi Dhan and Vandana) and one drought sensitive (IR 20) cultivars. In total, 53 known and 40 novel differentially expressed (DE) miRNAs were identified. The primary drought responsive miRNAs were Osa-MIR2919, Osa-MIR3979, Osa-MIR159f, Osa-MIR156k, Osa-MIR528, Osa-MIR530, Osa-MIR2091, Osa-MIR531a, Osa-MIR531b as well as three novel ones. Sixty-one target genes that corresponded to 11 known and 4 novel DE miRNAs were found to be co-localized with the three qDTYs, out of the 1746 target genes identified. We could validate miRNA-mRNA expression under drought for nine known and three novel miRNAs in eight different rice genotypes showing varying degree of tolerance. From our study, Osa-MIR2919, Osa-MIR3979, Osa-MIR528, Osa-MIR2091-5p and Chr01_11911S14Astr and their target genes LOC_Os01g72000, LOC_Os01g66890, LOC_Os01g57990, LOC_Os01g56780, LOC_Os01g72834, LOC_Os01g61880 and LOC_Os01g72780 were identified as the most promising candidates for drought tolerance at booting stage. Of these, Osa-MIR2919 with 19 target genes in the qDTYs is being reported for the first time. It acts as a negative regulator of drought stress tolerance by modulating the cytokinin and brassinosteroid signalling pathway.

A comprehensive study on the pathogenicity and host immune response was conducted in White Pekin ducklings after experimental infection with an Indian isolate of duck enteritis virus (DEV). The virus was found to be highly pathogenic and pantropic, which rapidly multiplied in various organs, mainly in the spleen and liver showing higher viral load with severe pathological lesions and caused 100% mortality. Expression profiles of immune gene transcripts in tissues (liver, spleen, brain) revealed upregulation of proinflammatory cytokines IFN-α, IFN- β, IL-1β, IL-6 and also iNOS with stimulation of TLRs (TLR-2, 3, 21). IFN-α was robustly upregulated (p < 0.05) especially in liver, might be playing role in antiviral innate immunity. Further, massive upregulation of MHC class-I (p < 0.01), expression of Th1 cytokines (IFN-γ & IL-2) and certain Th2 cytokines (IL-4 & IL-10) suggests stimulation of cell mediated as well as humoral immunity. To our knowledge, we are reporting first time about the robust upregulation of MHC class-I in spleen, liver and brain along with expression of certain cytokines in the peripheral blood mononuclear cells (PBMCs) during experimental DEV infection.

Various toll-like receptor (TLR) agonists have shown potential as adjuvants with different vaccines in both human and livestock species, including chickens. Our previous studies on combination of lipopolysaccharide (LPS; TLR4 agonist) and resiquimod (R-848; TLR7 agonist) showed the synergistic up-regulation of pro-inflammatory Th1 and Th2 cytokines in chicken peripheral blood mononuclear cells (PMBCs). Hence, the present study aimed to explore the combined adjuvant effect of LPS and R-848 with inactivated Newcastle disease virus (NDV) vaccine in chickens. Two weeks-old SPF chickens were immunized with inactivated NDV vaccine along with a combination of LPS and R-848 as an adjuvant with suitable control groups. A booster dose was given two weeks later. Antibody responses were assessed by enzyme linked immunosorbent assay (ELISA) and hemagglutination inhibition (HI) test, while cell-mediated immune responses were analyzed by a lymphocyte transformation test (LTT) and flow cytometry following vaccination. Two weeks post-booster, the birds were challenged with a velogenic strain of NDV, and protection against clinical signs, mortality and virus shedding was analyzed. The results indicated that inactivated NDV vaccine with R-848 induced significantly higher humoral and cellular immune responses with 100% protection against mortality and viral shedding following a virulent NDV challenge. However, the combination of LPS and R-848 along with inactivated NDV vaccine produced poor humoral and cellular immune responses and could not afford protection against challenge infection and virus shedding when compared to the vaccine-alone group, indicating the deleterious effects of the combination on antigen-specific immune responses. In conclusion, the combination of LPS and R-848 showed the inhibitory effects on antigen-specific humoral, cellular and protective immune responses when used as an adjuvant with inactivated NDV vaccines in chickens. This inhibitory effect might have occurred due to systemic cytokine storm. A nanoparticle-based delivery of the combination of LPS and R-848 for slow and sustained release could be tried as an alternative method to explore the synergistic effect of the combination as an adjuvant in chickens.

A comprehensive study on the pathogenicity and host immune response was conducted in White Pekin ducklings after experimental infection with an Indian isolate of duck enteritis virus (DEV). The virus was found to be highly pathogenic and pantropic, which rapidly multiplied in various organs, mainly in the spleen and liver showing higher viral load with severe pathological lesions and caused 100% mortality. Expression profiles of immune gene transcripts in tissues (liver, spleen, brain) revealed upregulation of proinflammatory cytokines IFN-alpha, IFN-beta, IL-1 beta, IL-6 and also iNOS with stimulation of TLRs (TLR-2, 3, 21). IFN-alpha was robustly upregulated (p < 0.05) especially in liver, might be playing role in antiviral innate immunity. Further, massive upregulation of MHC class-I (p < 0.01), expression of Th1 cytokines (IFN-gamma & IL-2) and certain Th2 cytokines (IL-4 & IL-10) suggests stimulation of cell mediated as well as humoral immunity. To our knowledge, we are reporting first time about the robust upregulation of MHC class-I in spleen, liver and brain along with expression of certain cytokines in the peripheral blood mononuclear cells (PBMCs) during experimental DEV infection.

Non-typhoidal salmonellosis (NTS) is a food-borne zoonotic disease of human and veterinary importance. Besides enteric infections, most often it leads to invasive systemic bacteremia in immunocompromised patients leading to mortality. Emergence of antimicrobial-resistant species warrant the development of newer preventive approaches against NTS. Here, we adopted an in-silico approach to design a multiepitope-based peptide vaccine which will likely enhance the body’s immune response against most commonly isolated serovars (Salmonella Typhimurium, Salmonella Enteritidis and Salmonella Dublin) causing NTS. Using computational tools, we screened the B- cell, MHC-1 and MHC-2 epitopes of the highly conserved outer membrane proteins (OMPs)- OmpC, OmpD and OmpF. By further immunoinformatic processing, for enhanced vaccine efficacy, nineteen most promising epitopes were selected based on their antigenicity. Epitopes were assembled with help of molecular linkers to create a multivalent recombinant protein and then named as NTSVax. Safety assessment of the construct showed zero allergenicity and toxicity. To increase its immunogenicity, synthetic adjuvants RS09 and TpD were added to the C and N terminal ends respectively. Tertiary structure of the vaccine construct, refined at multiple stages, was shown to have 99.7% of residues in favoured regions of Ramachandran plot. The vaccine’s efficacy was further screened via molecular dynamics and immune simulations by docking the complex against human toll like receptors (TLR-1 & 4). Then the peptide sequence was reverse translated and its nucleotide sequence encoding NTSVax was cloned in a plasmid for bulk expression in E. coli.

Matrix Metalloproteinases (MMPs)-induced altered proteolysis of extracellular matrix proteins and basement membrane holds the key for tumor progression and metastasis. Matrix metalloproteinases-7 (Matrilysin), the smallest member of the MMP family also performs quite alike; thus serves as a potential candidate for anti-tumor immunotherapy. Conversely, being an endogenous tumor-associated antigen (TAA), targeting MMP-7 for immunization is challenging. But MMP-7-based xenovaccine can surmount the obstacle of poor immunogenicity and immunological tolerance, often encountered in TAA-based conventional vaccine for anti-tumor immunotherapy. This paves the way for investigating the potential of MMP-7-derived major histocompatibility complex (MHC)-binding peptides to elicit precise epitope-specific T-cell responses towards their possible inclusion in anti-tumor vaccine formulations. Perhaps it also ushers the path of achieving multiple epitope-based broad and universal cellular immunity. In current experiment, an immunoinformatics approach has been employed to identify the putative canine matrix matelloproteinases-7 (cMMP-7)-derived peptides with MHC class-I-binding motifs which can elicit potent antigen-specific immune responses in BALB/c mice. Immunization with the cMMP-7 DNA vaccine induced a strong CD8 + cytotoxic T lymphocytes (CTLs) and Th1- type response, with high level of gamma interferon (IFN-γ) production in BALB/c mice. The two identified putative MHC-I-binding nonameric peptides (Peptide 32-40 and Peptide 175-183 ) from cMMP-7 induced significant lymphocyte proliferation along with the production of IFN-γ from CD8 + T-cells in mice immunized with cMMP-7 DNA vaccine. The current observation has depicted the immunogenic potential of the two cMMP-7-derived nonapeptides for their possible exploitation in xenovaccine-mediated anti-tumor immunotherapy in mouse model.

The study consisted of application of anti-ubiquitin antibodies (Abs)-coated iron oxide-nanoparticles (IONPs) for minimisation of oxidative stress to contemporary live spermatozoa from the raw semen. Round-shaped IONPs (12.09 ± 0.91 nm) after two-stage functionalisation (silanisation and pegylation) were conjugated with Abs. Four aliquots from each of the 24 ejaculates (4 buffalo bulls) formed Control (Group I) and treatment (II, III and IV) groups; each containing 150 ± 25 million dead/damaged spermatozoa. IONPs-Abs complex were added at ratio of 1:1 (0.5 µg/ml), 1:2 (1.0 µg/ml) and 1:4 (2.0 µg/ml), respectively, in Groups II, III and IV. The semen quality parameters showed improvement at lag-stage (post-nano-purification before processing for cryopreservation). The mean post-thaw motility (%) in Group IV was found to be greater (p < .05) than Group I. Moreover, the overall DNA integrity (%) at post-thaw stage was improved in the nano-purified semen samples. The value of malondialdehyde was greater (p < .001) in Group I than Groups II, III and IV. The mean total antioxidant capacity and superoxide dismutase (U/mg protein) activity values in Group IV was greater (p < .05) than Group I. The study results show that IONPs conjugated with anti-ubiquitin Abs at 2.0 µg/ml can be an effective dose for depletion of dead/damaged spermatozoa from buffalo ejaculates to minimise oxidative stress.

Bovine herpesvirus 1 (BoHV-1) affects mainly large ruminants (cattle and buffalo), resulting in a substantial negative impact on the industry.The virus causes respiratory symptoms, abortion and a decrease in milk production.Among envelope proteins of the virus, the glycoprotein D (gD) is considered a dominant immunogenic.We expressed the immunogenic portion of gD in the prokaryotic system (E.cloni 10G) and did evaluate its potential as a diagnostic antigen for indirect ELISA.Using the Ni-HRP probe, the expressed recombinant gD protein (rgD, 45kDa) was characterized by SDS-PAGE and western blotting.The immunogenic reactivity of red was optimized for indirect ELISA using known true positive and negative sera (antibodies) of the virus.Sera (n=158) were screened and, with a cut-off value of 0.238, relative diagnostic sensitivity (69%) and specificity (52.3%) of in-house developed indirect ELISA was determined.With these preliminary results, the study suggests the diagnostic potential of recombinant gD protein for serodiagnosis of BoHV-1 infection at a large scale.

The synthesis of iron oxide nanoparticles (IONPs)-antiubiquitin antibodies (Abs) complex for depletion of dead/damaged spermatozoa from buffalo semen was done. The IONPs synthesized were round in shape with size of 12.09 ± 0.91 nm. At the end of the two-step functionalization, that is, silanization and pegylation of bare IONPs and bioconjugation of functionalized IOPNs, particles with the sizes of 19.15 ± 1.46, 20.72 ± 0.95, and 73.01 ± 7.56 nm, respectively, were obtained. Twenty-four semen samples from four bulls with mean individual progressive motility (%) and sperm concentration (million/mL) of 77.1 ± 0.9 and 1,321.2 ± 84.7, respectively, were divided into Group I (control), and treatment groups viz. Groups II, III, and IV; with each group containing 150 ± 25 million dead/damaged spermatozoa. The IONPs-Abs complex was added at the ratio of 1:1 (0.5 μg/mL), 1:2 (1.0 μg/mL), and 1:4 (2.0 μg/mL), respectively, in the Groups II, III, and IV. The mean efficiency (%) of nanopurification was estimated to be greater in nanopurified semen with the increasing doses of the IONPs-Abs complex. A reduction of 29.3 ± 6.4%, 48.4 ± 5.3%, and 55.4 ± 4.4% in dead/damaged spermatozoa following nanopurification in Groups II, III, and IV, respectively, was observed. The study shows that in-house synthesized IONPs-Abs complex can be successfully used to deplete dead/damaged spermatozoa from buffalo semen with improvement in quality.

The current study intended to optimize the concentration of Oxyrase in the semen dilutor and to evaluate its effect on freezability of spermatozoa of Sahiwal bulls. Supplementation of Oxyrase at 0.125 IU/mL concentration significantly reduced dissolved oxygen (DO) in the dilutor to 4 ppm in 16-18 min at 35 degrees C. For supplementation studies, a total of 24 ejaculates were categorized into poor and good ejaculates categories (n = 12 each) based on their initial progressive motility. Each ejaculate was further divided into two aliquotes. The first aliquote was diluted with tris-egg yolk extender without Oxyrase (control group) whereas, in the treatment group, Oxyrase was supplemented at the concentration of 0.125 IU/mL of extender. The parameters evaluated include cholesterol and plasma membrane phospholipids (PMP) at fresh, while IPM, acrosomal and plasma membrane integrity, cholesterol, PMP and oxidative stress parameters like lipid peroxidation (LPO), total antioxidant capacity (TAC) and reactive oxygen species (ROS) were evaluated at pre-freeze and post-thaw stages. The IPM and acrosomal intactness were higher (p < 0.05) in treatment group at post-thaw stage in good ejaculates. Oxyrase supplementation resulted in lower (p < 0.05) cholesterol leakage in both categories and lower (p < 0.05) LPO in good ejaculates at post-thaw stage. No statistical difference in ROS was observed between control and treatment groups at all stages whereas, level of TAC was higher (p < 0.05) in the treatment group compared to control group at post-thaw stage of both categories. Therefore, Oxyrase as an oxygen scavenging agent could preserve the post-thaw quality of Sahiwal bull spermatozoa.

The genetic basis of differential host immune response vis-a-vis transcriptome profile was explored in PBMCs of indigenous (Ghurrah) and crossbred pigs after classical swine fever vaccination and in monocyte derived macrophages (MDMs) challenged with virulent classical swine fever (CSF) virus. The humoral immune response (E2 antibody) was higher (74.87%) in crossbred than indigenous pigs (58.20%) at 21st days post vaccination (21dpv). The rate of reduction of ratio of CD4(+)/CD8(+) was higher in crossbred pigs than indigenous pigs at 7th days post vaccination (7dpv). The immune genes IFIT1, IFIT5, RELA, NFKB2, TNF and LA72 were up regulated at 7dpv in RNA seq data set and was in concordance during qRT-PCR validation. The Laminin Subunit Beta 1 (LAMB]) was significantly (p <= 0.05) down-regulated in MDMs of indigenous pigs and consequently a significantly (p <= 0.01) higher copy number of virulent CSF virus was evidenced in macrophages of crossbred pigs than indigenous pigs. Activation of LXR:RXR pathway at 60 h post infection (60hpi) in MDMs of indigenous versus crossbred pigs inhibited nuclear translocation of NF-kappa B, resulted into transrepression of proinflammatory genes. But it helped in maintenance of HDL level by lowering down cholesterol/LDL level in MDMs of indigenous pigs. The key immune genes (TLR2, TLR4, IL10, IL8, CD86, CD54, CASP1) of TREM1 signaling pathway were upregulated at 7dpv in PBMCs but those genes were downregulated at 60hpi in MDMs indigenous pigs. Using qRT-PCR, the validation of differentially expressed, immunologically important genes (LAMB1, OAS1, TLR 4, TLR8 and CD86) in MDMs revealed that expression of these genes were in concordance with RNA-seq data.

AbstractLive intermediate plus infectious bursal disease virus (IBDV) vaccines (hot vaccines) are used for protection against the virulent IBDV strains in young chickens. We evaluated the potential of Toll-like receptor (TLR) agonists to alleviate hot vaccine-induced immunosuppression. The combination of Pam3CSK4 and poly I:C synergistically upregulated IFN-β, IFN-γ, IL-12, IL-4, and IL-13 transcripts and cross-inhibited IL-1β, IL-10, and iNOS transcripts in the chicken peripheral blood mononuclear cells (PBMCs) as analyzed by quantitative real-time PCR. Further, four-week old specific pathogen free White Leghorn chickens (n = 60) were randomly divided into six groups and either immunized with hot IBDV vaccine with or without Pam3CSK4 and/or poly I:C or not vaccinated to serve as controls. The results indicated that poly I:C alone and in combination with Pam3CSK4 alleviated vaccine-induced immunosuppression, as evidenced by greater weight gain, increased overall antibody responses to both sheep erythrocytes and live infectious bronchitis virus vaccine, upregulated IFN-γ transcripts and nitric oxide production by PBMCs (P < 0.05), and lower bursal lesion score in the experimental birds. In conclusion, poly I:C alone and its combination with Pam3CSK4 reduced the destruction of B cells as well as bursal damage with restoration of function of T cells and macrophages when used with a hot IBDV vaccine.

Toll-like receptors (TLRs) are pattern recognition receptors that are critical for the functions of the host innate immunity. This study reports complete coding sequences of TLR-7 and TLR-8 of cattle and goat of Himalayan Kumaon (HK) region. Further, predicted 26 leucine rich repeats (LRRs), two terminal LRR modules and putative common and differential ligand binding amino acids in TLR-7 and -8 of ruminants. The differential ligand binding amino acids between TLR-7 and TLR-8 of HK-cattle were Val396, Arg422, Val513, Asp538 and Val566 in TLR-8; Thr406, Lys432, Thr533, Leu558 and Ile586 in TLR-7. Codon selection analysis revealed co-localization of positively selected codons 377 and 408 of cattle, and 402 and 859 of goat with already reported SNPs in ruminants. The comparative analysis of deduced electrostatic surface potential of the extracellular domain of TLR-7 and -8 resulted in grouping of different species within ruminants.

The present study was conducted to identify the differentially expressed miRNAs (DE miRNAs) in the peripheral blood mononuclear cells of crossbred pigs in response to CSF vaccination on 7 and 21 days of post vaccination as compared to unvaccinated control (0 dpv). Simultaneously, set of miRNA was predicted using mRNA seq data at same time point. The proportion of CD4−CD8+ and CD4+CD8+ increased after vaccination, and the mean percentage inhibition was 86.89% at 21 dpv. It was observed that 22 miRNAs were commonly expressed on both the time points. Out of predicted DE miRNAs, it was found that 40 and 35 DE miRNAs were common, obtained from miRNA seq analysis and predicted using mRNA seq data on 7 dpv versus 0 dpv and 21 dpv versus 0 dpv respectively. Two DE miRNAs, ssc-miR-22-5p and ssc-miR-27b-5p, were selected based on their log2 fold change and functions of their target genes in immune process/pathway of viral infections. The validations of DE miRNAs using qRT-PCR were in concordance with miRNA seq analysis. Two set of target genes, CD40 and SWAP70 (target gene of ssc-miR-22-5p) and TLR4 and Lyn (target gene of ssc-miR-27b-5p), were validated and were in concordance with results of RNA seq analysis at a particular time point (except TLR4). The first report of genome-wide identification of differentially expressed miRNA in response to live attenuated vaccine virus of classical swine fever revealed miR-22-5p and miR-27b-5p were differentially expressed at 7 dpv and 21 dpv.

The public concern over the harmful effects of chemical pesticides on the environment and human her as enhanced the searchfor safer, environment friendly controlealternatives . Control of the plant pests the application of biological agents holds great promise as an alternative to the use of chemicals. It is genrally recognized that biological control agents are safer and more environmentally sound than is reliance on the use of high volume of pesticides. . It is widely distributed in nature as an integument of insect and crustaceans and as a cell wall component of fungi and algae. Chitinases poly ( 1,4- ( N; acetyl ; B; lucosaminide) glycahodrolase are a group of enzymes that able to degrade chitin directly into low molecular weight products. In this study 18 soil samples were randomly collected from the rhizosphere of wheat from different villages of district, Faridabad. These soil samples were used for the isolation of antifungal chitinase producing bacterial strains. A total of 40 chitinase producing bacterial isolates were obtained. Antagonistic activity of all the isolates was tested against fugal pathogen Fusariumoxysporium using standard dual culture technique. Out of the forty isolates eight isolatesproved to be positive antagonists of this fungal pathogen i.e Fusariumoxysporium. The inhibition of tested fungal pathogen by bacterial isolates varied from 32.00%- 65.00% and average percentage inhibition was calculated as 56.00% R6 has been found as the best isolate showed thein terms of percentage inhibition of fussariumoxysporiumi.e 65.00a%. Due to the importance of cytinolytic enzymes in insects, Nematode, and fungal groth and development, they are receiving attention in regard to their development as biopesticides or chemical defence proteins in transgenic plants and micronbial biocontrol agents. In this sence, biological control of some soil – brone fungal disease has been correlated with chitinase production. This study will lead to isolatic antagonistic Rizobacterial strains having chitinolytic activity against the prevalent fungal athogenes effecting crops, such isolates may be developed into successful soil inoculums strains.

The present study was undertaken to evaluate the effect of kaempferol in isoprenaline (ISP)-induced myocardial injury in rats. ISP was administered subcutaneously for two subsequent days to induce myocardial injury. Assessment of myocardial injury was done by estimation of hemodynamic functions, myocardial infarcted area, cardiac injury markers, lipid profile, oxidative stress, pro-inflammatory cytokines and histopathology of heart and liver. Rats pretreated with kaempferol showed reduction in the myocardial infarcted area and heart rate. However, no improvement was observed in change in body weight, mean arterial, systolic and diastolic blood pressure. Kaempferol showed significant decrease in serum LDH, CK-MB, troponin-I and lipid profile. However, highest dose of kaempferol did not reduce the serum triglyceride level. Further, antioxidant enzymes, SOD and catalase, were also higher. However, reduced glutathione, serum SGOT and creatinine did not show any improvement. Kaempferol showed reduction in MDA level. Kaempferol at highest dose showed reduction in pro-MMP-2 expression and MMP-9 level. mRNA expression level of TNF-α was not different in kaempferol-pretreated myocardial injured rats with ISP-alone group. Pretreatment with kaempferol at highest dose showed mild mononuclear infiltration and degenerative changes in heart tissue section of myocardial injured rats. Rats pretreated with kaempferol at higher concentration showed normal cordlike arrangement of hepatocytes with moderate swelling of hepatocytes (vacuolar degeneration) around the central vein. Study suggests that kaempferol attenuated lipid profile, infarcted area and oxidative stress in ISP-induced myocardial injury in rats.

Only three immediate early genes (IE) BICP0, BICP4 and BICP22 of Bovine herpesvirus 1 (BoHV-1) are known. These genes are expressed coordinately and their promoters are well characterized. We provide evidence for expression of three additional IE genes of BoHV-1 i.e. UL21, UL33 and UL34. These genes are expressed in the presence of cycloheximide (CH) at the same time as known IE genes. Surprisingly, the promoters of newly identified IE genes (UL21, UL33, UL34) lack the OCT-1 binding site, a considered site of transactivation of the BoHV-1 IE genes. The other difference in the promoters of the newly identified IE genes is the presence of TATA box at near optimal site. However, all the IE genes have similar spatial placements of C/EBPα, DPE and INR elements.