Enhancing immune and antioxidant responses in Nile tilapia through dietary supplementation with Ulva fasciata extract: A study on gene expression and resistance to Aeromonas hydrophila.

BACKGROUND: The study focused on the impact of Ulva fasciata extract (UFE) supplementation in the diets of Nile tilapia (Oreochromis niloticus) on blood and biochemical markers, immune and oxidative responses, and the expression of related genes, with a specific interest in their condition following exposure to Aeromonas hydrophila. METHODS: Four different levels of UFE were tested in the diets: 0% (0 mg kg- 1) for the control group (U0), and incremental additions of 0.05% (50 mg kg-1), 0.1% (100 mg kg-1), and 0.15% (150 mg kg-1) for the experimental groups U50, U100, and U150 respectively. Groups of 45 fish weighing 3.126 ± 0.120 g were fed these diets over 90 days. RESULTS: The study found that groups treated with UFE showed statistically significant enhancements (p < 0.05) compared to the control group. These improvements included increased red and white blood cell counts, higher haemoglobin concentrations, greater packed cell volume, and elevated enzyme activities-specifically, superoxide dismutase, catalase, alanine aminotransferase, and aspartate aminotransferase. Additionally, lysozyme and phagocytic activities were notably higher, especially in the U100 group after exposure. Before exposure to Aeromonas hydrophila, all levels of UFE supplementation led to increased expression of TNF-α and COXII genes and decreased NFκ-B expression. After the challenge, UFE intake resulted in varied expression levels of immune and antioxidant genes (TNF-α, NFκ-B, SOD, and COXII) in the liver, with the most effective responses observed in the U50, U100, and U150 groups. CONCLUSIONS: The findings underscore the potential of dietary UFE as a natural antioxidant and immune booster for Nile tilapia.

Understanding the nature of blast resistance in combined bacterial leaf blight and blast gene pyramided lines of rice variety tellahamsa.

BACKGROUND: Rice blast and bacterial leaf blight (BLB) are the most limiting factors for rice production in the world which cause yield losses typically ranging from 20 to 30% and can be as high as 50% in some areas of Asia especially India under severe infection conditions. METHODS AND RESULTS: An improved line of Tellahamsa, TH-625-491 having two BLB resistance genes (xa13 and Xa21) and two blast resistance genes (Pi54 and Pi1) with 95% Tellahamsa genome was used in the present study. TH-625-491 was validated for all four target genes and was used for backcrossing with Tellahamsa. Seventeen IBC1F1 plants heterozygous for all four target genes, 19 IBC1F2 plants homozygous for four, three and two gene combinations and 19 IBC1F2:3 plants also homozygous for four, three and two gene combinations were observed. Among seventeen IBC1F1 plants, IBC1F1-62 plant recorded highest recurrent parent genome (97.5%) covering 75 polymorphic markers. Out of the total of 920 IBC1F2 plants screened, 19 homozygous plants were homozygous for four, three and two target genes along with bacterial blight resistance. Background analysis was done in all 19 homozygous IBC1F2 plants possessing BLB resistance (possessing xa13, Xa21, Pi54 and Pi1 in different combinations) with five parental polymorphic SSR markers. IBC1F2-62-515 recovered 98.5% recurrent parent genome. The four, three and two gene pyramided lines of Tellahamsa exhibited varying resistance to blast. CONCLUSIONS: Results show that there might be presence of antagonistic effect between bacterial blight and blast resistance genes since the lines with Pi54 and Pi1 combination are showing better resistance than the combinations with both bacterial blight and blast resistance genes.

Lignocellulose and probiotic supplementation in broiler chicken diet: effect on growth performance, digestive health, litter quality, and genes expression.

Three hundred one-day-old Avian 48 broiler chicks were used to investigate the effect of lignocellulose (LC) and probiotic supplementation in broiler chicken diet on growth performance, digestive health, litter quality, and some gene expression. Experimental treatments consisted of 3 × 2 factorial arrangements with 3 levels of LC without or with probiotics to formulate 6 experimental groups. Groups 1, 2, and 3 were fed on the basal diet with dietary LC inclusion at 0, 0.5, and 1.0%, respectively, while groups 4, 5, and 6 were fed on the previously mentioned design with Bacillus subtilis at 100 gm/ton. The results revealed that Dietary LC inclusion nonsignificantly (P ≥ 0.05) reduced body weight (BW), body weight gain (BWG), and feed intake. Meanwhile, B. subtilis supplementation improved BW and BWG and enhanced the effect of LC on the broilers' weight. The group fed a 0.5% LC and B. subtilis-supplemented diet recorded the best (P ≥ 0.05) BW, BWG, FCR, PER, EEU, and PI. LC and or B. subtilis supplementation improved carcass traits of broiler (higher dressing% with lower abdominal fat% compared with a control group), intestinal health, and absorptive capacity. LC potentiates the effect of B. subtilis supplementation in broilers' diet in modulating intestinal microflora (lowered (P ≥ 0.05) cecal Coliform and increased Lactobacillus counts), the highest Coliform counts were recorded in group fed 0.5 or 1.0% LC plus B. subtilis. LC at 0.5 or 1.0% and or B. subtilis supplementation reduced (P ≥ 0.05) litter moisture% at the 2nd, 4th, and 6th wk compared to the control group. Dietary inclusion of LC and or B. subtilis supplementation significantly (P < 0.001) up-regulated hepatic growth-related genes (growth hormone receptor (GHR) and insulin growth factor1 (IGF-1)) and antioxidant-related genes (superoxide dismutase 1 (SOD1), glutathione peroxidase (GPX1) and uncoupling protein (UCP) and down-regulated (P < 0.001) splenic toll-like receptor 4 (TLRP) gene expression while had no significant effect on splenic interleukin 8 (IL8) and tumor necrosis factor (TNF) with the best-obtained results with 1.0% followed by 0.5% LC with B. subtilis supplementation. We concluded that dietary LC and/or B. subtilis supplementation positively affected the growth performance, feed efficiency, carcass quality, intestinal absorptive capacity and health, litter quality and growth, and antioxidant and immune-related gene expression.

Evaluating estrus synchronization and early pregnancy detection in Ossimi sheep: The influence of fluorogestone acetate treatment duration and dosage.

Estrus synchronization is important for improving sheep reproduction. To enhance sheep reproduction efficiency, this study investigated the impact of different durations (7 vs. 14 days) and fluorogestone acetate (FGA) doses in intravaginal sponges on estrus synchronization and early pregnancy detection in Ossimi sheep. Two hundred ewes were evenly divided into two groups, each receiving a full 40 mg or a halved 20 mg FGA sponge for their respective durations. The study aimed to optimize breeding efficiency by examining the effectiveness of these treatments in synchronizing estrous cycles and by evaluating the use of serum levels of pregnancy-associated glycoprotein 1 (PAG1) and progesterone (P4) as markers for early pregnancy identification. Prostaglandin F2α and equine chorionic gonadotropin were administered to enhance the synchronization process. Results highlighted that the 7-day treatment protocol significantly improved estrus, pregnancy, and lambing rates compared to the 14-day protocol. Furthermore, pregnant ewes demonstrated elevated levels of PAG1 and P4, with PAG1 levels particularly higher in ewes with multiple pregnancies. The findings underscore that the shorter duration of FGA treatment is more effective for reproductive management in Ossimi sheep without significantly affecting PAG1 levels based on the dose or duration of FGA. PAG1 also proved to be a reliable marker for early pregnancy detection, offering a promising approach to identifying fetal numbers early in pregnancy. This research suggests optimizing FGA sponge use could be cost-efficient for improving reproductive efficiency and early pregnancy management in sheep.

Exploration of bioactive compounds from Olea dioica in Western Ghats of Karnataka using GC-MS.

Bioactive compounds in plants are essential for the formation of novel chemotherapeutic drugs, which have been used in Ayurveda to treat a variety of illnesses. Indian medicinal herbs have been used for thousands of years to treat a variety of illnesses, such as fever, cancer, snake bites, rheumatism, skin problems, and neurodegenerative diseases. GC-MS was used to locate and categorize bioactive components in Olea dioica leaves. The results showed that presence of octanoic acid, methyl ester, decanoic acid, methyl ester, desulphosinigrin, l-gala-l-ido-octose, methyl tetradecanoate, Tetradecanoic acid, 6-benzoxazolesulfonamide, N-(2-hydroxyethyl)-2-methyl-, 10-chloro-5-methoxy-5H-dibenzo[a,d][7]annulene, pentadecanoic acid, oleic acid, n-hexadecanoic acid, hexanedioic acid, dioctyl ester, and squalene. The methanol extract of Olea dioica was effective against a wide spectrum of pathogenic bacteria at four different concentrations, with the highest activity against Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Xanthomonas campestris, and Salmonella typhimurium. It also showed moderate activity against Agrobacterium tumefaciens, Pseudomonas aeruginosa, Streptomyces pneumonia, and Pseudomonas syringae. The pharmacological properties of O. dioica, as well as their variety and comprehensive phytochemistry, could be exploited as a potent antimicrobial agent for future therapeutics.

Immunoinformatic exploration of a multi-epitope-based peptide vaccine candidate targeting emerging variants of SARS-CoV-2.

Many countries around the world are facing severe challenges due to the recently emerging variants of SARS-CoV-2. Over the last few months, scientists have been developing treatments, drugs, and vaccines to subdue the virus and prevent its transmission. In this context, a peptide-based vaccine construct containing pathogenic proteins of the virus known to elicit an immune response was constructed. An analysis of the spike protein-based epitopes allowed us to design an "epitope-based subunit vaccine" against coronavirus using the approaches of "reverse vaccinology" and "immunoinformatics." Computational experimentation and a systematic, comprehensive protocol were followed with an aim to develop and design a multi-epitope-based peptide (MEBP) vaccine candidate. Our study attempted to predict an MEBP vaccine by introducing mutations of SARS-CoV-2 (Delta, Lambda, Iota, Omicron, and Kappa) in Spike glycoprotein and predicting dual-purpose epitopes (B-cell and T-cell). This was followed by screening the selected epitopes based on antigenicity, allergenicity, and population coverage and constructing them into a vaccine by using linkers and adjuvants. The vaccine construct was analyzed for its physicochemical properties and secondary structure prediction, and a 3D structure was built, refined, and validated. Furthermore, the peptide-protein interaction of the vaccine construct with Toll-like receptor (TLR) molecules was performed. Immune profiling was performed to check the immune response. Codon optimization of the vaccine construct was performed to obtain the GC content before cloning it into the E. coli genome, facilitating its progression it into a vector. Finally, an in-silico simulation of the vaccine-protein complex was performed to comprehend its stability and conformational behavior.

Exploration of Tilmicosin Cardiotoxicity in Rats and the Protecting Role of the Rhodiola rosea Extract: Potential Roles of Cytokines, Antioxidant, Apoptotic, and Anti-Fibrotic Pathways.

Tilmicosin (TIL) is a common macrolide antibiotic in veterinary medicine. High doses of TIL can have adverse cardiovascular effects. This study examined the effects of Rhodiola rosea (RHO) that have anti-inflammatory, antioxidant, and anti-fibrotic effects on tilmicosin (TIL)-induced cardiac injury targeting anti-inflammatory, antioxidant, apoptotic, and anti-apoptotic signaling pathways with anti-fibrotic outcomes. Thirty-six male Wistar albino rats were randomly divided into groups of six rats each. Rats received saline as a negative control, CARV 1 mL orally (10 mg/kg BW), and RHO 1 mL orally at 400 mg/kg BW daily for 12 consecutive days. The TIL group once received a single subcutaneous injection (SC) dose of TIL (75 mg/kg BW) on the sixth day of the experiment to induce cardiac damage. The standard group (CARV + TIL) received CARV daily for 12 consecutive days with a single TIL SC injection 1 h after CARV administration only on the sixth day of study and continued for another six successive days on CARV. The protective group (RHO + TIL) received RHO daily for the same period as in CARV + TIL-treated rats and with the dosage mentioned before. Serum was extracted at the time of the rat's scarification at 13 days of study and examined for biochemical assessments in serum lactate dehydrogenase (LDH), cardiac troponin I (cTI), and creatine phosphokinase (CK-MB). Protein carbonyl (PC) contents, malondialdehyde (MDA), and total antioxidant capacity (TAC) in cardiac homogenate were used to measure these oxidative stress markers. Quantitative RT-PCR was used to express interferon-gamma (INF-γ), cyclooxygenase-2 (COX-2), OGG1, BAX, caspase-3, B-cell lymphoma-2 (Bcl-2), and superoxide dismutase (SOD) genes in cardiac tissues, which are correlated with inflammation, antioxidants, and apoptosis. Alpha-smooth muscle actin (α-SMA), calmodulin (CaMKII), and other genes associated with Ca2+ hemostasis and fibrosis were examined using IHC analysis in cardiac cells (myocardium). TIL administration significantly increased the examined cardiac markers, LDH, cTI, and CK-MB. TIL administration also increased ROS, PC, and MDA while decreasing antioxidant activities (TAC and SOD mRNA) in cardiac tissues. Serum inflammatory cytokines and genes of inflammatory markers, DNA damage (INF-γ, COX-2), and apoptotic genes (caspase-3 and BAX) were upregulated with downregulation of the anti-apoptotic gene Bcl-2 as well as the DNA repair OGG1 in cardiac tissues. Furthermore, CaMKII and α-SMA genes were upregulated at cellular levels using cardiac tissue IHC analysis. On the contrary, pretreatment with RHO and CARV alone significantly decreased the cardiac injury markers induced by TIL, inflammatory and anti-inflammatory cytokines, and tissue oxidative-antioxidant parameters. INF-γ, COX-2, OGG1, BAX, and caspase-3 mRNA were downregulated, as observed by real-time PCR, while SOD and Bcl-2 mRNA were upregulated. Furthermore, the CaMKII and α-SMA genes' immune reactivities were significantly decreased in the RHO-pretreated rats.

Protective impacts of Withania somnifera leaf extract from Taif area against diclofenac induced hepato-renal toxicity: role of antioxidants, inflammation, apoptosis, and anti-oxidative stress biomarkers.

Current study examined the boosting impacts of Withania somnifera leaf extract from Taif area (high-altitude area) against hepatic and renal toxicity induced by diclofenac in experimental rats. Withania is highly grown on Taif area as environmental herb with multiple functions. Diclofenac is non-steroidal medication used for treatment of pain but over dose has severe side effects. Thirty-two adult Wistar rats of male type were subdivided into 4 groups. The control rats (group 1) received saline. Second group received diclofenac (50 mg/kg BW intraperitoneally) at days 4 and 5. Third group received W. somnifera leaf extract (250 mg /kg body weight) for 6 days. The fourth protective group, received W. somnifera leaf extract plus diclofenac for 6 days as shown in groups 2 and 3. Diclofenac significantly increased serum AST, ALT, and decreased albumin and total proteins levels. It also increased serum concentrations of uric acid and creatinine. In addition, it increased lipid peroxidation, and decreased reduced glutathione and superoxide dismutase levels. Diclofenac increased inflammatory cytokines secretion and up-regulated hepatic oxidative stress genes (HO-1; hemoxygenase-1 and Nrf2nuclear factor erythroid 2-related factor 2 (Nrf2) and renal inflammatory transcriptional markers (TGF-ß1; transforming growth factor-beta1 and COX-2; cycloxygenas-2). In parallel, hepatic caspase-3 expression was up-regulated as an apoptotic marker, while Bcl2; (B-cell lymphoma 2) mRNA expression was down regulated as anti-apoptotic marker. W. somnifera pre-administration in the protective group ameliorated the altered parameters induced by diclofenac. In conclusion, W. somnifera leaf extract has the potential to antagonize side effects of diclofenac by regulating the pathways of oxidative stress, inflammation, and apoptosis/antiapoptosis.

Insecticidal Activity of Tannins from Selected Brown Macroalgae against the Cotton Leafhopper Amrasca devastans.

Seaweeds, also known as marine macroalgae, are renewable biological resources that are found worldwide and possess a wide variety of secondary metabolites, including tannins. Drifted brown seaweed (DBSW) is particularly rich in tannins and is regarded as biological trash. The cotton leaf hopper Amrasca devastans (Distant) has caused both quantitative and qualitative losses in cotton production. Drifted brown seaweeds (DBSWs) were used in this study to extract, qualitatively profile, and quantify the levels of total tannins, condensed tannins, hydrolyzable tannins, and phlorotannins in the seaweeds; test their insecticidal activity; and determine the mechanism of action. The largest amount of tannin extract was found in Sargassum wightii Greville (20.62%) using the Soxhlet method (SM). Significantly higher amounts of hydrolyzable tannins (p = 0.005), soluble phlorotannins (p = 0.005), total tannins in the SM (p = 0.003), and total tannins in the cold percolation method (p = 0.005) were recorded in S. wightii. However, high levels of condensed tannins (CTAs) were observed in Turbinaria ornata (Turner) J. Agardh (p = 0.004). A. devastans nymphs and adults were examined for oral toxicity (OT) and contact toxicity (CT) against DBSW tannin crude extract and column chromatographic fractions 1 (Rf = 0.86) and 2 (Rf = 0.88). Stoechospermum polypodioides (J.V. Lamouroux) J. Agardh crude tannin was highly effective against A. devastans using the OT method (LC50, 0.044%) when compared with the standard gallic acid (LC50, 0.044%) and tannic acid (LC50, 0.122%). Similarly, S. wightii fraction 2 (LC50, 0.007%) showed a greater insecticidal effect against A. devastans adults in OT than gallic acid (LC50, 0.034%) and tannic acid (LC50, 0.022%). The mechanism of action results show that A. devastans adults treated with crude tannin of T. ornata had significantly decreased amylase, protease (p = 0.005), and invertase (p = 0.003) levels when compared with the detoxification enzymes. The levels of glycosidase, lactate dehydrogenase, esterase, lipase, invertase, and acid phosphate activities (p = 0.005) of S. wightii were reduced when compared with those of the Vijayneem and chemical pesticide Monocrotophos. In adult insects treated with LC50 concentrations of S. wightii tannin fraction 1, the total body protein (9.00 µg/µL) was significantly reduced (OT, LC50-0.019%). The SDS-PAGE analysis results also show that S. wightii tannin fraction 1 (OT and CT), fraction 2 (OT), and S. polypodioides fraction 2 (CT) had a significant effect on the total body portion level, appearance, and disappearance of some proteins and polypeptides. This study shows that the selected brown macroalgae can be utilized for the safer management of cotton leaf hoppers.

The Effect of Cerebrolysin in an Animal Model of Forebrain Ischemic-Reperfusion Injury: New Insights into the Activation of the Keap1/Nrf2/Antioxidant Signaling Pathway.

Forebrain ischemia-reperfusion (IR) injury causes neurological impairments due to decreased cerebral autoregulation, hypoperfusion, and edema in the hours to days following the restoration of spontaneous circulation. This study aimed to examine the protective and/or therapeutic effects of cerebrolysin (CBL) in managing forebrain IR injury and any probable underlying mechanisms. To study the contribution of reperfusion to forebrain injury, we developed a transient dual carotid artery ligation (tDCAL/IR) mouse model. Five equal groups of six BLC57 mice were created: Group 1: control group (no surgery was performed); Group 2: sham surgery (surgery was performed without IR); Group 3: tDCAL/IR (surgery with IR via permanently ligating the left CA and temporarily closing the right CA for 30 min, followed by reperfusion for 72 h); Group 4: CBL + tDCAL/IR (CBL was given intravenously at a 60 mg/kg BW dose 30 min before IR); and Group 5: tDCAL/IR + CBL (CBL was administered i.v. at 60 mg/kg BW three hours after IR). At 72 h following IR, the mice were euthanized. CBL administration 3 h after IR improved neurological functional recovery, enhanced anti-inflammatory and antioxidant activities, alleviated apoptotic neuronal death, and inhibited reactive microglial and astrocyte activation, resulting in neuroprotection after IR injury in the tDCAL/IR + CBL mice group as compared to the other groups. Furthermore, CBL reduced the TLRs/NF-kB/cytokines while activating the Keap1/Nrf2/antioxidant signaling pathway. These results indicate that CBL may improve neurologic function in mice following IR.

Promising bioactive metabolites of mangrove inhabitant Streptomyces tauricus and prostate cancer PC3 cell inhibition by antimicrobial peptides.

Streptomyces is a group of microbes known for antibiotic production and has contributed to more than 70% of present commercially available antibiotics. These antibiotics are important in the management, protection, and treatment of chronic illnesses. In the present study, the isolated S. tauricus strain from mangrove soil in Mangalore, India (GenBank accession number: MW785875) was subjected for differential cultural characterization, phenotype involving brown pigmentation, filamentous mycelia, and ash-colored spore production was observed using field emission scanning electron microscopy (FESEM) analysis revealing filamentous mycelia possessing a straight spore chain. Spores were visualized as elongated, rod-shaped, smooth surfaces with curved edges. After optimized growth conditions for S. tauricus on starch-casein agar medium, the GC/MS analysis of S. tauricus intracellular extract detected bioactive compounds reported for pharmacological applications. Analyzed using the NIST library, most of the bioactive compounds identified in intracellular extract had molecular weights of less than 1 kDa. On the PC3 cell line, the Sephadex G-10 partially purified eluted peak protein fraction demonstrated significant anticancer activity. The LCMS analysis revealed the presence of Tryprostatin B, Fumonisin B1, Microcystin LR, and Surfactin C with molecular weights below 1 kDa. This study found that small molecular weight microbial compounds are more effective in a variety of biological applications.

Multigene Sequence-Based and Phenotypic Characterization Reveals the Occurrence of a Novel Entomopathogenic Nematode Species, Steinernema anantnagense n. sp.

Three entomopathogenic nematode populations were isolated from agricultural fields in the Anantnag district of Jammu and Kashmir (India). Sequences of multiple gene regions and phenotypic features show that they are conspecific and represent a novel species. Molecular and morphological features provided evidence for placing the new species into the "Kushidai" clade. Within this clade, analysis of sequence data of the internal transcribed spacer (ITS) gene, the D2D3 region of the 28S rRNA gene, the mitochondrial cytochrome oxidase I (mtCOI) gene, and the mitochondrial 12S (mt12S) gene depicted the novel species as a distinctive entity closely related to Steinernema akhursti, S. kushidai, and S. populi. Phylogenetic analyses also show that the new species is a sister species to S. akhursti, and these two species are closely related to S. kushidai and S. populi. Additionally, the new species does not mate or produce fertile progeny with any of the closely related species, reinforcing its uniqueness from a biological species concept standpoint. The new species is further characterized by the third-stage infective juveniles with almost straight bodies (0.7-0.8 mm length), poorly developed stoma and pharynx, and conoid-elongate tail (49-66 µm) with hyaline posterior part. Adult females are characterized by short and conoid tails bearing a short mucron in the first generation and long conoid tails with thin mucron in the second generation. Adult males have ventrally curved spicules in both generations. Moreover, the first-generation male has rounded manubrium, fusiform gubernaculum, conoid and slightly ventrally curved tails with minute mucron, and the second generation has rhomboid manubrium anteriorly ventrad bent, and tails with long and robust mucron. The morphological, morphometrical, molecular, and phylogenetic analyses support the new species status of this nematode, which is hereby described as Steinernema anantnagense n. sp. The bacterial symbiont associated with S. anantnagense n. sp. represents a novel species, closely related to Xenorhabdus japonica. These findings shed light on the diversity of entomopathogenic nematodes and their symbiotic bacteria, providing valuable information for future studies in this field.

Identification and Characterization of Glutathione S-transferase Genes in Spodoptera frugiperda (Lepidoptera: Noctuidae) under Insecticides Stress.

Insect glutathione S-transferases (GSTs) serve critical roles in insecticides and other forms of xenobiotic chemical detoxification. The fall armyworm, Spodoptera frugiperda (J. E. Smith), is a major agricultural pest in several countries, especially Egypt. This is the first study to identify and characterize GST genes in S. frugiperda under insecticidal stress. The present work evaluated the toxicity of emamectin benzoate (EBZ) and chlorantraniliprole (CHP) against the third-instar larvae of S. frugiperda using the leaf disk method. The LC50 values of EBZ and CHP were 0.029 and 1.250 mg/L after 24 h of exposure. Moreover, we identified 31 GST genes, including 28 cytosolic and 3 microsomal SfGSTs from a transcriptome analysis and the genome data of S. frugiperda. Depending on the phylogenetic analysis, sfGSTs were divided into six classes (delta, epsilon, omega, sigma, theta, and microsomal). Furthermore, we investigated the mRNA levels of 28 GST genes using qRT-PCR under EBZ and CHP stress in the third-instar larvae of S. frugiperda. Interestingly, SfGSTe10 and SfGSTe13 stood out with the highest expression after the EBZ and CHP treatments. Finally, a molecular docking model was constructed between EBZ and CHP using the most upregulated genes (SfGSTe10 and SfGSTe13) and the least upregulated genes (SfGSTs1 and SfGSTe2) of S. frugiperda larvae. The molecular docking study showed EBZ and CHP have a high binding affinity with SfGSTe10, with docking energy values of -24.41 and -26.72 kcal/mol, respectively, and sfGSTe13, with docking energy values of -26.85 and -26.78 kcal/mol, respectively. Our findings are important for understanding the role of GSTs in S. frugiperda regarding detoxification processes for EBZ and CHP.

In vitro evaluation of the neuroprotective potential of Olea dioica against Aß peptide-induced toxicity in human neuroblastoma SH-SY5Y cells.

Alzheimer's disease (AD) is a type of neurodegenerative disease, associated with the hastening of ROS, acetylcholinesterase (AChE) activity, and amyloid ß peptides plaques in the brain. The limitations and side effects of existing synthetic drugs incline toward natural sources. In the present communication active principles of methanolic extract of Olea dioica Roxb, leaves are explored as an antioxidant, AChE inhibitor, and anti-amyloidogenic. Furthermore, neuroprotection against the amyloid beta-peptide has been studied. The bioactive principles were identified by GC-MS and LC-MS and further subjected to antioxidant (DPPH and FRAP) and neuroprotection (AChE inhibition, ThT binding, and MTT assay, DCFH-DA and lipid peroxidation (LPO) assay using neuroblastoma (SHSY-5Y) cell lines) assays. Methanolic extract of O. dioica Roxb, leaves was found to contain polyphenols and flavonoids. In vitro assays exhibited potential antioxidant and anti-AChE (˃50%) activities. ThT binding assay indicated protection against amyloid-beta aggregation. MTT assay, Aß1-40 (10 µM) with extract increase the cell viability (˃50%) and showed significant cytotoxicity to SHSY-5Y cells. ROS level (˃25%) significantly decreased in the Aß1-40 (10 µM) + extract (15 and 20 µM/mL) and LPO assay (˃50%) suggesting prevention of cell damage. Results advocate that O. dioica leaves are a good source of antioxidants, anti-AChE, and anti-amyloidogenic compounds which may be further evaluated as a natural medicine for the treatment of AD.

Insecticidal Mechanism of Botanical Crude Extracts and Their Silver Nanoliquids on Phenacoccus solenopsis.

In recent years, intensive studies have been carried out on the management of agricultural insect pests using botanical insecticides in order to decrease the associated environmental hazards. Many studies have tested and characterized the toxic action of plant extracts. Four plant extracts (Justicia adhatoda, Ipomea carnea, Pongamia glabra, and Annona squamosa) containing silver nanoparticles (AgNPs) were studied for their effects on Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) using the leaf dip method. The effects were estimated based on assays of hydrolytic enzyme (amylase, protease, lipase, acid phosphatase, glycosidase, trehalase, phospholipase A2, and invertase) and detoxification enzyme (esterase and lactate dehydrogenase) levels; macromolecular content (total body protein, carbohydrate, and lipid); and protein profile. The results show that the total body of P. solenopsis contains trypsin, pepsin, invertase, lipase, and amylase, whereas J. adathoda and I. carnea aqueous extracts considerably decreased the protease and phospholipase A2 levels, and A. squamosa aqueous extract dramatically increased the trehalase level in a dose-dependent manner. The enzyme levels were dramatically decreased by P. glabura-AgNPs (invertase, protease, trehalase, lipase, and phospholipase A2); I. carnea-AgNPs (invertase, lipase, and phospholipase A2); A. squamosa-AgNPs (protease, phospholipase A2); and J. adathoda-AgNPs (protease, lipase, and acid phosphatase). Plant extracts and their AgNPs significantly reduced P. solenopsis esterase and lactate dehydrogenase levels in a dose-dependent manner. At higher concentrations (10%), all of the investigated plants and their AgNPs consistently decreased the total body carbohydrate, protein, and fat levels. It is clear that the plant extracts, either crude or together with AgNPs, may result in the insects having inadequate nutritional capacity, which will impact on all critical actions of the affected hydrolytic and detoxication enzymes.

Risk Assessment of Fluxametamide Resistance and Fitness Costs in Fall Armyworm (Spodoptera frugiperda).

The fall armyworm (FAW), Spodoptera frugiperda, is one of the most devastating invasive polyphagous pests, which has attracted recent global attention by developing resistance to various insecticidal active ingredients with independent mode of action. Fluxametamide, a newly commercialized isoxazoline insecticide, is exceptionally selective towards several lepidopteran pests. The present study aimed to evaluate resistance risk in FAW to fluxametamide and the fitness costs associated with fluxametamide resistance. A field-collected and genetically mixed population of FAW was artificially selected through continuous exposure to fluxametamide. After successive selection of 10 generations, there was no obvious increase in the LC50 (RF: 2.63-fold). The realized heritability (h2) of fluxametamide resistance was estimated as h2 = 0.084 using a quantitative genetic approach. Compared with the susceptible F0 strain, the Flux-SEL (F10) strain of FAW displayed no significant cross-resistance to broflanilide, chlorantraniliprole, fipronil, indoxacarb, lambda cyhalothrin, spinetoram, and tetraniliprole, except emamectin benzoate (RF: 2.08-fold). Increased activity of glutathione S-transferase (ratio 1.94) was observed in the Flux-SEL (F10) strain of FAW, while the cytochrome P450 and carboxylesterase activities were not altered. The fluxametamide-selection significantly affected the development and reproductive traits of FAW with a lower R0, T and relative fitness (Rf = 0.353). The results alluded that the risk of fluxametamide resistance evolution in FAW is relatively lower; however, proactive implementation of resistance management approaches should be done to maintain the field efficacy of fluxametamide against FAW.

Endophytic Colonization by Beauveria bassiana and Metarhizium anisopliae in Maize Plants Affects the Fitness of Spodoptera frugiperda (Lepidoptera: Noctuidae).

The fall armyworm, Spodoptera frugiperda (Noctuidae; Lepidoptera), is a serious threat to food security as it has the potential to feed on over 353 plant species. To control this insect pest, endophytic colonization of entomopathogenic fungi (EPF) in plants is being considered as a safer and more effective alternative. This study evaluated the efficacy of two EPFs, Beauveria bassiana and Metarhizium anisopliae, for endophytic colonization using foliar spray and seed treatment methods on maize plants, and their impact on the survival, development, and fecundity of S. frugiperda. Both EPF effectively colonized the maize plants with foliar spray and seed treatment methods, resulting in 72-80% and 50-60% colonization rates, respectively, 14 days after inoculation. The EPF negatively impacted the development and fecundity of S. frugiperda. Larvae feeding on EPF-inoculated leaves had slower development (21.21 d for M. anisopliae and 20.64 d for B. bassiana) than the control treatment (20.27 d). The fecundity rate was also significantly reduced to 260.0-290.1 eggs/female with both EPF applications compared with the control treatment (435.6 eggs/female). Age-stage-specific parameters showed lower fecundity, life expectancy, and survival of S. frugiperda when they fed on both EPF-inoculated leaves compared with untreated leaves. Furthermore, both EPFs had a significant effect on population parameters such as intrinsic (r = 0.127 d-1 for B. bassiana, and r = 0.125 d-1 for M. anisopliae) and finite rate (λ = 1.135 d-1 for B. bassiana, and λ = 1.1333 d-1 for M. anisopliae) of S. frugiperda compared with the control (r = 0.133 d-1 and λ = 1.146 d-1). These findings suggest that EPF can be effectively used for the endophytic colonization of maize plants to control S. frugiperda. Therefore, these EPFs should be integrated into pest management programs for this pest.

Antiproliferative activity of antimicrobial peptides and bioactive compounds from the mangrove Glutamicibacter mysorens.

The Glutamicibacter group of microbes is known for antibiotic and enzyme production. Antibiotics and enzymes produced by them are important in the control, protection, and treatment of chronic human diseases. In this study, the Glutamicibacter mysorens (G. mysorens) strain MW647910.1 was isolated from mangrove soil in the Mangalore region of India. After optimization of growth conditions for G. mysorens on starch casein agar media, the micromorphology of G. mysorens was found to be spirally coiled spore chain, each spore visualized as an elongated cylindrical hairy appearance with curved edges visualized through Field Emission Scanning Electron Microscopy (FESEM) analysis. The culture phenotype with filamentous mycelia, brown pigmentation, and ash-colored spore production was observed. The intracellular extract of G. mysorens characterized through GCMS analysis detected bioactive compounds reported for pharmacological applications. The majority of bioactive compounds identified in intracellular extract when compared to the NIST library revealed molecular weight ranging below 1kgmole-1. The Sephadex G-10 could result in 10.66 fold purification and eluted peak protein fraction showed significant anticancer activity on the prostate cancer cell line. Liquid Chromatography-Mass Spectrometry (LC-MS) analysis revealed Kinetin-9-ribose and Embinin with a molecular weight below 1 kDa. This study showed small molecular weight bioactive compounds produced from microbial origin possess dual roles, acting as antimicrobial peptides (AMPs) and anticancer peptides (ACPs). Hence, the bioactive compounds produced from microbial origin are a promising source of future therapeutics.

Physicochemical and adulteration study of fresh milk collected from different locations in Pakistan.

The present research was carried out to assess raw milk's quality as collected from the commercial markets of the Mardan district (Khyber Pakhtunkhwa). The locality from which milk samples were collected included; Bijligar (BG), Manga (M), Chamthara (CM), Main Bazar Mardan (MB), Mahidherai (MD), and Sharif Abad (SB), located in district Mardan, Khyber Pakhtunkhwa (KP). A total of 36 milk samples were collected at the rate of 6 samples per location. The outcome of the data exhibited that the percentage of protein content was highest (3.34%) in MB and SA (3.30%), while lower percentages were recorded in sample M (3.03%) and CM 93.06%). Maximum pH were shown in M and BG to be 7.55 and 7.33, respectively. For fats content, the highest percentage of fats was witnessed in MB as 4.04%, and minimum fats content was noted in Sample M as 3.57%. Water content was highest in Sample SA and BG at 15.85% and 15.64%, respectively. Qualitative analysis of adulterants like detergents, Formalin, starch, and Hydrogen peroxide was also carried out for all the collected samples. Adulteration results were positive for all the milk samples, with the highest being in samples MB (30%), while all the remaining samples had adulteration at 20% each. Both MB and CM samples were adulterated with urea, while the remaining 4 were adulterated with neutralizers. Thus, it may be summarized from the whole analysis that the milk available in commercial markets of district Mardan was adulterated with different adulterants and is not recommended for consumption.

Metabolomics approach reveals high energy diet improves the quality and enhances the flavor of black Tibetan sheep meat by altering the composition of rumen microbiota.

This study aims to determine the impact of dietary energy levels on rumen microbial composition and its relationship to the quality of Black Tibetan sheep meat by applying metabolomics and Pearson's correlation analyses. For this purpose, UHPLC-QTOF-MS was used to identify the metabolome, whereas 16S rDNA sequencing was used to detect the rumen microbiota. Eventually, we observed that the high energy diet group (HS) improved the carcass quality of Black Tibetan sheep and fat deposition in the longissimus lumborum (LL) compared to the medium energy diet group (MS). However, HS considerably increased the texture, water holding capacity (WHC), and volatile flavor of the LL when compared to that of MS and the low energy diet group (LS). Metabolomics and correlation analyses revealed that dietary energy levels mainly affected the metabolism of carbohydrates and lipids of the LL, which consequently influenced the content of volatile flavor compounds (VOCs) and fats. Furthermore, HS increased the abundance of Quinella, Ruminococcus 2, (Eubacterium) coprostanoligenes, and Succinivibrionaceae UCG-001, all of which participate in the carbohydrate metabolism in rumen and thus influence the metabolite levels (stachyose, isomaltose, etc.) in the LL. Overall, a high-energy diet is desirable for the production of Black Tibetan sheep mutton because it improves the mouthfeel and flavor of meat by altering the composition of rumen microbiota, which influences the metabolism in the LL.