Percent Survival Research Articles

Effect of gamma radiation on percent survival, lethality percentage of axillary buds of apple cv Ambri and determination of Lethal Dose (LD50)

Effect of gamma radiation on percent survival, lethality percentage of axillary buds of apple cv Ambri and determination of Lethal Dose (LD50)

Enhancing aquaponic cultivation in Indonesia: A comparative study of bioball biofilters with different additives

Abstract The development of aquaponic cultivation in Indonesia remains limited despite its potential to significantly boost farmers’ and communities’ income. In aquaponics, fish are regularly fed, while plants solely rely on nutrients derived from leftover feed and fish waste, without external supplementation. Biofilters play a crucial role in aquaponic systems, providing a growth medium for microorganisms that convert fish feces’ ammonia into nitrates, essential for plant nutrition. In general, biofilters use bioball material, so as to optimize the biofilter, this study compares the effectiveness of different materials, namely activated charcoal, shells, and coconut fiber, when incorporated into bioball biofilters within aquaponic systems. The research was conducted on an aquaponic installation featuring an NFT system utilizing PVC pipes. The experiment involved 45 tilapia fish, each measuring 7-11 cm and weighing 10-15 grams, alongside red lettuce plants planted in 10 holes per pipe, with 6 replications. After 45 days from planting, the plants were harvested, and observations were made on both plant and fish growth. The growth of red lettuce plants in the biofilter installation with shells addition (BS) showed the best results based on parameters such as plant height, leaf width, plant dry weight, and shoot:root ratio. The installation with coconut fiber addition (BS) showed the best results in terms of number of leaves. BA installation with activated carcoal addition (BA) showed the best results in terms of number of Leaf chlorophyll. The highest percentage of survival for Nile tilapia was observed in the BS and BA installation with a rate of 88.89%, followed by 78.57% in the BC installation, and 60.46% in the BB installation.

Evaluation of virulence of Aeromonas veronii strain GZ21-2 and development of a highly effective vaccine for grass carp with the potential for industrial application

Bacterial septicemia represents a significant disease affecting cultured grass carp culture, with the primary etiological agent identified as the Gram-negative bacterium Aeromonas veronii. In response to an outbreak of septicemia in Guangzhou, we developed a formaldehyde-inactivated vaccine against an A. veronii strain designated AV-GZ21-2. This strain exhibited high pathogenicity in experimental infections across at all developmental stages of grass carp. Mortality rates for grass carp weighing 15 ± 5 g ranged from 16 % to 92 % at exposure temperatures of 19 °C–34 °C, respectively. The median lethal dose (LD50) for grass carp groups weighing 15 ± 5 g, 60 ± 10 g, 150 ± 30 g and 500 ± 50 g were determined to be 1.43, 2.52, 4.65 and 7.12 × 107(CFU/mL), respectively. We investigated the inactivated vaccine in conbination with aluminum hydroxide gel (AV-AHG), Montanide ISA201VG (AV-201VG), and white oil (AV-WO) adjuvants. This study aimed to optimize inactivation conditions and identify the adjuvant that elicits the most robust immune response. The AV-GZ21-2 inactivated bacterial solution (AV),when combined with various adjuvants, was capable of inducing a strong specific immune response in grass carp. The relative percent survival (RPS) following a lethal challenge with AV-GZ21-2 were 94 % for AV-AHG, 88 % for AV-201VG, 84 % for AV-WO and 78 % for AV alone. The minimum immunization dose of the AV-AHG vaccine was determined to be 6.0 × 107 CFU per fish, providing immunity for a duration of six months with an immune protection level exceeding 75 %. Furthermore, the AV-AHG vaccine demonstrated significant protective efficacy against various epidemic isolates of A. veronii. Consequently, we developed an inactivated vaccine targeting a highly pathogenic strain of A. veronii, incorporating an aluminum hydroxide gel adjuvant, which resulted in high immune protection and a duration of immunity exceeding six months. These findings suggest that the AV-AHG vaccine holds substantial potential for industrial application.

Maternal Immunity and Immune Improvement of Koi Cyprinus rubrofuscus Larvae after DNA Vaccine against Hoi Herpesvirus

Vaccination is an efficient step in preventing disease caused by koi herpesvirus in koi fish (Cyprinus rubrofuscus). This study aimed to investigates the impact of maternal broodstock vaccination with ORF81 DNA vaccine on the transmission of maternal immunity to offspring against KHV. The ORF81 DNA vaccine has been shown to increase RPS, but its effect on maternal immunity in koi offspring after vaccination has not been evaluated. The study examined koi broodstocks vaccinated with the ORF81 DNA vaccine 45 and 60 days before spawning (45V and 60V) at a dose of 12.5 μg/100 g. Immunological parameters were assessed in broodstock, eggs, and larvae. Immune gene expression was also analyzed using the qPCR method. The results showed that lysozyme activity and antibody levels in eggs from vaccinated mothers were higher, though not significantly different. Meanwhile, lysozyme activity and antibody levels in mothers were significantly higher (p<0.05) compared to controls. Immunoglobulin-M gene (IgM) and recombination-activating gene 1 (RAG1), were significantly induced. The relative survival percentage increased significantly in larvae produced from broodstocks with 60V. These findings indicate that broodstocks treated with the ORF81 DNA vaccine can enhance the immune response of their offspring.

The role of aroA and ppk1 in Aeromonas veronii pathogenicity and the efficacy evaluation of mutant strain AV-ΔaroA/ppk1 as a live attenuated vaccine

Aeromonas veronii is an opportunistic pathogen that poses great threat to aquaculture and human health, so there is an urgent need for green and efficient methods to deal with its infection. In this study, single and double gene deletion strains (AV-ΔaroA, AV-Δppk1 and AV-ΔaroA/ppk1) that can be stably inherited were constructed. Pathogenicity test showed that the toxicity of AV-ΔaroA and AV-ΔaroA/ppk1 was significantly lower compared to wild-type A. veronii. Biological characterization analysis revealed that the decrease in pathogenicity might be due to the declined growth, motility, biofilm formation abilities and the expression of virulence-related genes in mutants. Subsequently, we evaluated the efficacy of AV-ΔaroA/ppk1 as a live attenuated vaccine (LAV). Safety assessment experiments showed that AV-ΔaroA/ppk1 injected at a concentration of 3 × 107 CFU/mL was safe for C. carassius. The relative percentage survival of AV-ΔaroA/ppk1 was 67.85 %, significantly higher than that of the inactivated A. veronii, which had an RPS of 54.84 %. This improved protective effect was mainly attributed to the increased levels of A. veronii specific IgM antibody, enhanced alkaline phosphatase, lysozyme and superoxide dismutase activities, as well as higher expression levels of several immune related genes. Together, these findings deepen our understanding of the functional roles of aroA and ppk1 in A. veronii pathogenicity, provide a good candidate of LAV for A. veronii.

A naturaly attenuated largemouth bass ranavirus strain provided protection for Micropterus salmoides by immersion immunization

Largemouth bass ranavirus (LMBV) causes disease outbreaks and high mortality at all stages of largemouth bass farming. Therefore, live vaccine development is critical for largemouth bass prevention against LMBV by immersion immunization. Herein, an attenuated LMBV strain with good immunogenicity, designated as LMBV-2007136, was screened from the natural LMBV strains bank through challenge assay and immersion immunization experiment. After determing the safe concentration range of LMBV-2007136, the minimum immunizing dose of immersion immunization was verified. When largemouth bass were vaccinated by immersion at the lowest concentration of 102.0 TCID50/mL, all of fish were survival post virulent LMBV challenge, and the relative percent survival (RPS) was 100 %. And the immune gene expression levels of IL-10, IL-12, IFN-γ, and IgM in the spleen and kidney post-vaccination were significantly up-regulated compared to the control group, but TNF-α expression showed no significant changes. The safety and efficacy of LMBV-2007136 at passages P8, P13, and P18 were futher assessed, and no death of largemouth bass was observed within 21 days post-immunization and RPS of three vaccination groups was 100 %, suggesting that the safety and efficacy of the attenuated strain at different passages was stable. Furthermore, in the virulence reversion test, the attenuated strain was propagated through 5 times in largemouth bass by intraperitoneal injection and no abnormality and mortality were observed, further proving the attenuated vaccine candidate LMBV-2007136 was safe. These results proved that LMBV-2007136 could be a promising candidate for a live vaccine to protect largemouth bass from LMBV disease.

Histological study of the development of the digestive system during larval feeding in the pike silverside Chirostoma estor.

Chirostoma estor (Jordan, 1879) is an endemic freshwater species with a high potential for aquaculture; however, as in many other fish, larviculture of this species is the most critical stage, in which the higher mortality rates. Therefore, it is necessary to fully describe the development of the digestive system to establish better feeding protocols in the larval culture of C. estor, both for aquaculture and restoration purposes. In the present study, larviculture was carried out from hatching to 20days after hatching (DAH). The organisms were fed with the rotifer Brachionus plicatilis from 2 to 14 DAH, and nauplii of Artemia sp. from 15 to 20 DAH. A total of 12 organisms (0, 3, 5, 10, 15, and 20 DAH) were taken for size and weight growth and histological and histochemical analysis. The histological analysis indicated that after 3 DAH, the opening of the mouth and anus was observed, coinciding with the beginning of exogenous feeding. In addition, the digestive system developed, with differentiation of the oropharyngeal sections, esophagus, and intestine, folding of the intestinal mucosa, as well as associated organs (liver and pancreas) that reach their maximum development at 20 DAH. Thus, C. estor at this stage of development can digest and absorb nutrients despite being an agastric fish. The results obtained in this study will facilitate a better understanding of the ontogenetic morphophysiological development processes, associated with the transition of larvae to exogenous feeding, which ensures a higher percentage of survival during larval development and of course, adds to the diversity ontogenetics of teleostean.

Immune response and protection efficacy of formalin-killed vaccines against Streptococcus iniae in four-finger threadfin Eleutheronema tetradactylum.

Four-finger threadfin, Eleutheronema tetradactylum farming in southern Taiwan has been facing disease problems caused by Streptococcus iniae since 2018. The development of a vaccine against infectious S. iniae in the cultured threadfin industry is necessary. Thus, this study aimed to examine the efficacy of threadfin immunized formalin-killed cells (FKC) from S. iniae GSI-111 for 42 days post-vaccination (dpv) using two doses of FKC alone (a booster at 14 dpv) as group A, and FKC mixed with ISA763A adjuvant using a single dose as group B or double doses as group C. Immunoglobulin (Ig)-M was purified from threadfin, and rabbit anti-threadfin IgM polyclonal antibodies were used to detect antibody level in immunized fish; the vaccinated group A displayed higher levels at 3 dpv and all vaccinated treatments demonstrated high antibody levels between 14 and 42 dpv. All vaccine groups showed significantly higher values of lysozyme activity at 42 dpv compared with the control group; the vaccinated A group peaked at 14 dpv. The expression profiles of pro-inflammatory and immune-related genes, TNF-α, IL-12A, and C2 were upregulated at 3 dpv, while CD8A and chemokine receptor CXCR4 were upregulated at 42 dpv. Finally, the threadfins were challenged with S. iniae at 42 dpv. The average relative percent survival was 96% for vaccination A and B treatments, and 100% for vaccination C treatment. In summary, this study demonstrated that FKC vaccines whether formulated with an adjuvant could stimulate immune response and effective protect threadfins against S. iniae infection.

Shewanella putrefaciens as an emerging pathogen of hepatopancreas necrosis disease in Chinese mitten crab Eriocheir sinensis.

Shewanella putrefaciens has been recognized as an emerging important pathogen in aquaculture. However, scarce information is available on the characterization and microbial control of S. putrefaciens as a causal agent of hepatopancreas necrosis disease in Chinese mitten crab Eriocheir sinensis. In this study, a multi-resistant S. putrefaciens isolate (DZ-A) was identified as a causal pathogen of hepatopancreas necrosis disease in Chinese mitten crabs. It showed a lethal dose (LD50) of 2.20 × 105 CFU ml-1 in Chinese mitten crabs, and multiple resistance to aminoglycoside, chloramphenicol, macrolide, penicillin, peptide, and tetracycline antimicrobials. In addition, Bdellovibrio powder exhibited a significant antibacterial effect against the pathogenic S. putrefaciens, and conferred significant protection to challenged Chinese mitten crabs with relative percentage survivals of 80.00% to 93.33% via significant improvement in their immune response and antioxidant capability. The findings of this study provide valuable insights into the phenotypic characterization and biological control of pathogenic S. putrefaciens in Chinese mitten crabs.

Growth and Economics of Production of Rooted Black Pepper (Piper nigrum L.) Cuttings as Affected by Drenching Cattle Urine

An experiment on survival and growth of rooted black pepper (Piper nigrum L.) cuttings as affected by drenching cattle urine was conducted at Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Horticulture, Dapoli, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli during the year 2023-24. Eleven treatments and 3 replications in Randomized Block Design viz. T1 Cattle urine 5 %, T2 Cattle urine 7.5 %, T3 Cattle urine 10 %, T4 Cattle urine 12.5 %, T5 Cattle urine 15 %, T6 Cattle urine 17.5 %, T7 Cattle urine 20 %, T8 Cattle urine 22.5 %, T9 Cattle urine 25 %, T10 Keradix powder and T11 Control with the aim to investigate effect of drenching cattle urine on growth and economics of production of rooted black pepper cuttings. The highest length of sprout (65.17 cm), number of leaves per cutting (14.32) and leaf area (78.16 cm2) were recorded in treatment T7 i.e. drenching cattle urine 20 %. The maximum survival percentage (90.33%), net profit (₹ 2515.00) and B: C ratio (1.87) found in treatment T7.

Molecular Diversity Analysis of in vitro and Irradiated Tomato (Lycopersicon esculentum Mill) Grew Under Salt Stress Expressed by SCoT and ISSR Markers

Tomato buds of cv. Idkawy were cultured in vitro on solid MS medium with 0.2 mg-l BAP. The plantlets that were produced were exposed to different doses of gamma radiation, ranging from 100 to 200 Gy. Afterward, single pieces of nodes were cut and moved to a fresh MS medium with 0.2 mg-l of BAP. The gamma radiation caused a mortality rate of 18.75% to 52.5% among the explants. The surviving plantlets were then cut into single node pieces and transferred to an MS medium containing 0.2 mg-l of BAP, with added NaCl concentrations of either 50 or 100 mM. There was increased mortality of the vegetative buds on the explants with increased salt concentrations. It was shown that the all gamma radiation doses caused reduced the percentage of survival at saline levels. The genetic diversity was assessment by using ten primers for each SCoT and ISSR markers to six irradiated treatments grew under salt stress (100 Gy x 50 mM, 150 Gy x 50 mM, 200 Gy x 50 mM, 100 Gy x 100 mM, 150 Gy x 100 mM, 200 Gy x 100 mM). It was showed that the polymorphism percentage mean of SCoT marker (29.56%) is higher than the ISSR marker (26.78%). The average of PIC values for both markers SCoT and ISSR were 0.197 and 0.288 (PIC ˂0.5), as well as, MI values were 0.077 and 0.081, respectively. In contrast, when considering the number of alleles (Ne), Nei's genetic diversity (H), and Shannon's information index (I) parameters, it was observed that the greatest genetic variation was caused by the combined treatment of 200 Gy x 50 mM NaCl using the SCoT marker. On the other hand, with the ISSR marker, the highest induced genetic variation was seen with the combined treatment of 150 Gy x 50 mM NaCl. The obtained results demonstrate that SCoT marker was more accurate and efficient than ISSR marker for distinguishing and genetic variation analysis of irradiated tomato plantlets grew under salt stress. The relationships within treatments were estimated through cluster analysis (UPGMA) based on SCoT and ISSR analysis.

Evaluation of protective immune response of immersion inactivated vaccine against Singapore grouper iridovirus

Singapore grouper iridovirus (SGIV) always causes high transmission efficiency and mortality in the larval and juvenile stages of grouper in aquaculture industry. Although inactivated virus and recombinant DNA vaccines administered via intraperitoneal injection have shown efficacy in protection against SGIV, their potential applications in field testing were limited due to the vaccine delivery methods. Here, we developed an immersion vaccine containing inactivated virus and Montanide IMS 1312 adjuvant (IMS 1312) and evaluated its protective efficacy against SGIV infection. Compared to the PBS group, fish vaccinated with immersion inactivated vaccine with or without IMS 1312 were significantly protected against SGIV, with a relative percent survival (RPS) of 57.69 % and 38.47 %, respectively. Furthermore, the transcripts of viral core genes were reduced, and the histopathological severity caused by SGIV were relatively mild in multiple tissues of the IMS + V group. The immersion vaccine activated the AKP and ACP activities and increased the mRNA levels of IFN and inflammation-associated genes. The transcriptome analysis showed that a total of 731 and 492 genes were significantly regulated in the spleen and kidney from the IMS + V group compared to the PBS group, respectively. Among them, 129 DEGs were co-regulated, and enriched in the KEGG pathways related to immune and cell proliferation, including MAPK signaling, JAK-STAT signaling and PI3K-Akt signaling pathways. Similarly, the DEGs specially regulated in the kidney and spleen upon vaccine immunization were significantly enriched in the KEGG pathways related to interferon and inflammation response. Together, our results elucidated that the immersion vaccine of inactivated SGIV with IMS 1312 induced a protective immune response of grouper against SGIV.

Production and evaluation of three kinds of vaccines against largemouth bass virus, and DNA vaccines show great application prospects

Largemouth bass virus (LMBV) infections has resulted in high mortality and economic losses to the global largemouth bass industry and has seriously restricted the healthy development of the bass aquaculture industry. There are currently no antiviral therapies available for the control of this disease. In this study, we developed three types of vaccine against LMBV; whole virus inactivated vaccine (I), a subunit vaccine composed of the major viral capsid protein MCP (S) as well as an MCP DNA vaccine(D), These were employed using differing immunization and booster strategies spaced 2 weeks apart as follows: II, SS, DD and DS. We found that all vaccine groups induced humoral and cellular immune responses and protected largemouth bass from a lethal LMBV challenge to varying degrees and DD produced the best overall effect. Specifically, the levels of specific IgM in serum in all immunized groups were elevated and significantly higher than those in the control group. Moreover, the expression of humoral immunity (CD4 and IgM) and cellular immunity (MHCI-α) as well as cytokines (IL-1β) was increased, and the activity of immunity-related enzymes ACP, AKP, LZM, and T-SOD in the serum was significantly enhanced. In addition, the relative percent survival of fish following an LMBV lethal challenge 4 weeks after the initial immunizations were high for each group: DD(89.5 %),DS(63.2 %),SS(50 %) and II (44.7 %). These results indicated that the MCP DNA vaccine is the most suitable and promising vaccine candidate for the effective control of LMBV disease.

Synthesis and Antimicrobial Activity of Newly Synthesized Nicotinamides.

Antioxidants are promising compounds with antimicrobial activity against drug-resistant pathogens, especially when combined with conventional antimicrobials. Our study aimed to characterize the structure of nicotinamides synthesized from nicotinic acid and thiocarbohydrazones and to evaluate their antibacterial and antifungal activity. Seven nicotinic acid hydrazides (NC 1-7) were synthesized using mono-thiocarbohydrazones with hydroxyl group substituents, along with quinolone, phenolic, and pyridine rings known for their antimicrobial activity. The in vitro antimicrobial activity of NC 1-7, at concentrations ranging from 0.001 to 1 mM, was tested against Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 29212), Pseudomonas aeruginosa (ATCC 27853), Klebsiella pneumoniae (NCIMB 9111), and Candida albicans (ATCC 24433) using the broth microdilution method per EUCAST 2024 guidelines. Microorganism survival percentages were calculated based on optical density, and target fishing using the PharmMapper database identified potential molecular targets. The results showed that P. aeruginosa was most susceptible to the compounds, while C. albicans was the least susceptible. NC 3 significantly inhibited P. aeruginosa and K. pneumoniae growth at 0.016 mM, while higher concentrations were required for S. aureus, E. faecalis, and C. albicans. NC 5 was most effective against gram-positive bacteria at 0.03 mM. Only NC 4 completely inhibited C. albicans below 1 mM. NC 3, with the lowest concentration for 50% growth inhibition (0.016-0.064 mM), showed promising antibacterial potential against specific AMR-related proteins (bleomycin resistance protein, HTH-type transcriptional regulator QacR, and streptogramin A acetyltransferase), suggesting that this class of compounds could enhance or restore the activity of established antibiotics.

Autochthonous probiotic bacteria improve intestinal pathology and histomorphology, expression of immune and growth-related genes and resistance against Vibrio alginolyticus in Asian seabass (Lates calcarifer).

The study isolated two strains of intestinal autochthonous bacteria Lactiplantibacillus plantarum1 (MH155966.1) (L1) and Lactiplantibacillus plantarum2 (MH105076.1) (L2) from the Choobdeh Abadan region. The aim of this study was to investigate the effects of different strains of probiotic bacteria on the growth performance, digestive enzyme activity, histopathologic and histomorphometric characterization of the intestine, expression of immune and growth related genes, and evaluate Lates calcarifer resistance against Vibrio alginolyticus. To achieve this, for each treatment 60 L. calcarifer juveniles (75 ± 12g) were randomly distributed in three fiberglass tanks (300L) and fed for 45 days. The treatments were established as Diet 1 (control diet); L1 (diet with Lb. plantarum isolated 1); L2 (diet with Lb. plantarum isolated 2) with a bacterial concentration of 1 × 109 CFU/g. Nine fish from each treatment were sampled and examined, after euthanasia. The fish were placed 2cm from the beginning of the intestine for microscopic sampling of villi height, villi width and thickness of the epithelium, with 3 treatments: The result showed differences in the mean values of total weight were found at the end of the experiment. After 45 days of culture, the fish fed with L1 had higher (P < 0.05) growth performance than the other treatment groups. But at the end of the trial, in L2, the digestive enzyme activities were higher (P < 0.05) than the other treatment groups. The fishes fed diets supplemented with the L2 group, like the digestive enzyme activities test, presented an increase in the thickness of the epithelium of the intestine, and villus height, and villus width were greatest in L2. Fish feeding with L1 and L2 probiotics induced higher transcription levels of interleukin-10 (IL-10), granulocyte-macrophage colony-forming cells (GMCFC), epidermal growth factor (EGF), and Transforming Growth Factor Beta (TGF-β) genes in the gut, which may correlate with better immune and hematological parameters in these groups. The results of the challenge test revealed that the percentage of survival was significantly higher in L1 (76.2%) and L2 (80.95%) treatments than in the control (P < 0.05). These results indicate that host-derived probiotics (Lb. plantarum) have significant potential as important probiotics to enhance nutrient utilization, Digestive enzymes, and metabolism by increasing the gut surface area of Lates calcarifer juveniles at 45 days of culture.

Optimalisasi Pengelolaan Kualitas Air Terhadap Kelangsungan Hidup Udang Vaname (Litopenaeus Vannamei) di Farm Vannamei Reborn Probolinggo Jawa Timur

Water quality management means efforts made to maintain water parameters so that they are in a safe condition, the standard of cultivation mediab required. Poor water quality will be the start of disease for shrimp and cn cause stress. Poor water quality can affect growth, metabolic processes and survival. Shrimp become low. The aim of this research is to find out whether the optimal quality management for the survival of vannamei shrimp (litopenaeus vannamei) at the reborn vannamei farm is optimal. The data alalysis method in this research uses quantitative descriptive analiysis techniques.The survival percentage of vaname shirmp from plot A 93,78%, plot B 92,68%, plot C 92,40% and plot D 90,38%. All plot showed a high survival rate from the standard value set by PT. Proteina Central Prima or CP Prima.

Metabolomics to investigate the effect of preconditioned mesenchymal stem cells with crocin on pulmonary epithelial cells exposed to 2-chloroethyl ethyl sulfide

Metabolomics significantly impacts drug discovery and precise disease management. This study meticulously assesses the metabolite profiles of cells treated with Crocin, Dexamethasone, and mesenchymal stem cells (MSCs) under oxidative stress induced by 2-chloroethyl ethyl sulfide (CEES). Gas chromatography/mass spectrometry (GC/MS) analysis unequivocally identified substantial changes in 37 metabolites across the treated groups. Notably, pronounced alterations were observed in pathways associated with aminoacyl-tRNA biosynthesis and the metabolism of aspartate, serine, proline, and glutamate. These findings demonstrate the potent capacity of the analyzed treatments to effectively reduce inflammation, mitigate reactive oxygen species production, and enhance cell survival rates. Significance•Crocin, Dexamethasone, and the metabolites of the conditioned media of mesenchymal stem cells to decline the injury caused by CEES.•Metabolites can acquaint treatment groups in diminishing inflammation and ROS production and expanding the percentage of cell survival.•Aminoacyl-tRNA biosynthesis, nitrogen metabolism, glyoxylate and dicarboxylate metabolism, and propanoate metabolism were significant pathways involved among groups.

Interleukin-17B in common carp (Cyprinus carpio L.): Molecular cloning and immune effects as immune adjuvant of Aeromonas veronii formalin-killed vaccine

The interleukin-17 (IL-17) family of cytokines is critical for host defense responses and mediates different pro- or anti-inflammatory mediators through different signaling pathways. However, the function of the related family member, IL-17B, in teleosts is poorly understood. In the present study, an IL-17B homolog (CcIL-17B) in common carp (Cyprinus carpio) was identified, and sequence analysis showed that CcIL-17B had eight conserved cysteine residues, four of which could form two pairs of disulfide bonds, which in turn formed a ring structure composed of nine amino acids (aa). The deduced aa sequences of CcIL-17B shared 35.79–92.93 % identify with known homologs. The expression patterns were characterized in healthy and bacteria-infected carp. In healthy carp, IL-17B mRNA was highly expressed in the spleen, whereas Aeromonas veronii effectively induced CcIL-17B expression in the liver, head, kidney, gills, and intestine. The recombinant protein rCcIL-17B could regulate the expression levels of inflammatory cytokines (such as IL-1β, IL-6, TNF-α, and IFN-γ) in primary cultured head kidney leukocytes in vitro. As an adjuvant for the formalin-killed A. veronii (FKA) vaccine, rCcIL-17B induced the production of specific antibodies more rapidly and effectively than Freund's complete adjuvant (FCA). The results of the challenge experiments showed that the relative percent survival (RPS) after vaccination with rCcIL-17B was 78.13 %. This percentage was significantly elevated compared to that observed in the alternative experimental groups (62.5 % and 37.5 %, respectively). Additionally, the bacterial loads in the spleen of the rCcIL-17B + FKA group were significantly lower than those in the control group from 12 h to 48 h after bacterial infection. Furthermore, histological analysis showed that the epithelial cells were largely intact, and the striated border structure was complete in the intestine of rCcIL-17B + FKA group. Collectively, our results demonstrate that CcIL-17B plays a crucial role in eliciting immune responses and evokes a higher RPS against A. veronii challenge compared to the traditional adjuvant FCA, indicating that rCcIL-17B is a promising vaccine adjuvant for controlling A. veronii infection.

Sustainable management of peanut damping-off and root rot diseases caused by Rhizoctonia solani using environmentally friendly bio-formulations prepared from batch fermentation broth of chitinase-producing Streptomyces cellulosae.

Soil-borne plant diseases represent a severe problem that negatively impacts the production of food crops. Actinobacteria play a vital role in biocontrolling soil-borne fungi. The target of the present study is to test the antagonistic activity of chitinase-producing Streptomyces cellulosae Actino 48 (accession number, MT573878) against Rhizoctonia solani. Subsequently, maximization of Actino 48 production using different fermentation processes in a stirred tank bioreactor. Finally, preparation of bio-friendly formulations prepared from the culture broth of Actino 48 using talc powder (TP) and bentonite in a natural as well as nano forms as carriers. Meanwhile, investigating their activities in reducing the damping-off and root rot diseases of peanut plants, infected by R. solani under greenhouse conditions. Actino 48 was found to be the most significant antagonistic isolate strain at p ≤ 0.05 and showed the highest inhibition percentage of fungal mycelium growth, which reached 97%. The results of scanning electron microscope (SEM) images analysis showed a large reduction in R. solani mycelia mass. Additionally, many aberrations changes and fungal hypha damages were found. Batch fermentation No. 2, which was performed using agitation speed of 200rpm, achieved high chitinase activity of 0.1163 U mL- 1 min- 1 with a yield coefficient of 0.004 U mL- 1 min- 1 chitinase activity/g chitin. Nano-talc formulation of Actino 48 had more a significant effect compared to the other formulations in reducing percentages of damping-off and root rot diseases that equal to 19.05% and 4.76% with reduction percentages of 60% and 80%, respectively. The healthy survival percentage of peanut plants recorded 76.19%. Furthermore, the nano-talc formulation of Actino 48 was sufficient in increasing the dry weight of the peanut plants shoot, root systems, and the total number of peanut pods with increasing percentages of 47.62%, 55.62%, and 38.07%, respectively. The bio-friendly formulations of actinobacteria resulting from this investigation may play an active role in managing soil-borne diseases.

Cytotoxicity Assessment of α Helix Antarctic Yeast Oriented Antifreeze Peptide (Afp1m) on &lt;i&gt;M. dunni&lt;/i&gt; (Clone III8C) Cells

In order to assess the cytotoxic effects of the cryoprotectant helix Antarctic yeast-orientated antifreeze peptide Afp1m on normal mouse skin fibroblasts, an &lt;i&gt;in vitro&lt;/i&gt; model was developed for cytotoxicity assessment. In order to evaluate the cytotoxic effects of Afp1m, the cells of M. dunni (Clone III8C) were subjected to various amounts of Afp1m. The cell viability was assessed using MTT Assay (Tetrazolium dye MTT 3-(4, 5 dimethylthiazol-2-yl)-2, 5-10 diphenyltetrazolium bromide) against the positive control cells (Clone III8C) that were cultured with 10% FBS (Foetal Bovine Serum) using an Elisa reader and in medium containing various amounts (10, 5, 2, 1 and 0.5 mg/mL) of Afp1m, the control group (10% FBS) displayed varying survival percentages (78.86 ± 10.17%, 88.38 ± 3.19%, 88.75 ± 7.19 %, 90.61 ± 7.11%, 91.19 ± 4.52%, and 100.00 ± 0.0 %) throughout 24 hr. At 72 hr of treatment, the cell viability scores of Afp1m at 5, 2, 1, and 0.5 mg/mL were significantly higher (p&lt;0.05) than those of 10mg/mL, which showed 86.73 ± 6.92 % viability of cells (103.9 ± 6.56 %, 104.3 ± 5.13%, 100.9 ± 1.71%, 102.8 ± 1.24%, and 100.00 ± 0.0%). At 24, 48, and 72 hr, retarded development was noted in 10 mg/mL Afp1m. Development was observed, albeit more slowly than in the positive control and treated with lesser concentrations. The findings of this work indicate that Afp1m exhibits cryoprotective properties without inducing toxicity when used for the cryopreservation of &lt;i&gt;M. dunni&lt;/i&gt; (Clone III8C) cells.