Antiprotease Activity Research Articles

Probing of the reactive center loop region of alpha-1-antitrypsin by mutagenesis predicts new type-2 dysfunctional variants.

Lung disease in alpha-1-antitrypsin deficiency (AATD) mainly results from insufficient control of the serine proteases neutrophil elastase (NE) and proteinase-3 due to reduced plasma levels of alpha-1-antitrypsin (AAT) variants. Mutations in the specificity-determining reactive center loop (RCL) of AAT would be predicted to minimally affect protein folding and secretion by hepatocytes but can impair anti-protease activity or alter the target protease. These properly secreted but dysfunctional 'type-2' variants would not be identified by common diagnostic protocols that are predicated on a reduction in circulating AAT. This has potential clinical relevance: in addition to the dysfunctional Pittsburgh and Iners variants reported previously, several uncharacterized RCL variants are present in genome variation databases. To prospectively evaluate the impact of RCL variations on secretion and anti-protease activity, here we performed a systematic screening of amino acid substitutions occurring at the AAT-NE interface. Twenty-three AAT variants that can result from single nucleotide polymorphisms in this region, including 11 present in sequence variation databases, were expressed in a mammalian cell model. All demonstrated unaltered protein folding and secretion. However, when their ability to form stable complexes with NE was evaluated by western blot, enzymatic assays, and a novel ELISA developed to quantify AAT-NE complexes, substrate-like and NE-binding deficient dysfunctional variants were identified. This emphasizes the ability of the RCL to accommodate inactivating substitutions without impacting the integrity of the native molecule and demonstrates that this class of molecule violates a generally accepted paradigm that equates circulating levels with functional protection of lung tissue.

Type 1 diabetes contributes to combined pulmonary fibrosis and emphysema in male alpha 1 antitrypsin deficient mice.

Type 1 diabetes (T1D) is a metabolic disease characterized by hyperglycemia and can affect multiple organs, leading to life-threatening complications. Increased prevalence of pulmonary disease is observed in T1D patients, and diabetes is a leading cause of comorbidity in several lung pathologies. A deficiency of alpha-1 antitrypsin (AAT) can lead to the development of emphysema. Decreased AAT plasma concentrations and anti-protease activity are documented in T1D patients. The objective of this study was to determine whether T1D exacerbates the progression of lung damage in AAT deficiency. First, pulmonary function testing (PFT) and histopathological changes in the lungs of C57BL/6J streptozotocin (STZ)-induced T1D mice were investigated 3 and 6 months after the onset of hyperglycemia. PFT demonstrated a restrictive pulmonary pattern in the lungs of STZ-injected mice, along with upregulation of mRNA expression of pro-fibrotic markers Acta2, Ccn2, and Fn1. Increased collagen deposition was observed 6 months after the onset of hyperglycemia. To study the effect of T1D on the progression of lung damage in AAT deficiency background, C57BL/6J AAT knockout (KO) mice were used. Control and STZ-challenged AAT KO mice did not show significant changes in lung function 3 months after the onset of hyperglycemia. However, histological examination of the lung demonstrated increased collagen accumulation and alveolar space enlargement in STZ-induced AAT KO mice. AAT pretreatment on TGF-β-stimulated primary lung fibroblasts reduced mRNA expression of pro-fibrotic markers ACTA2, CCN2, and FN1. Induction of T1D in AAT deficiency leads to a combined pulmonary fibrosis and emphysema (CPFE) phenotype in male mice.

A novel approach in valorization of spent mushroom substrate of Cordyceps militaris as in-feed antibiotics in Labeo rohita against Aeromonas hydrophila infection.

The study was carried out to investigate Cordyceps militaris spent mushroom substrate (CMSMS), a renewable biomass, for its effect on fish as an alternative source of in-feed antibiotics. For meticulous examination, four different iso-nitrogenous and iso-caloric diets were prepared containing CMSMS at 0%, 1%, 2%, and 3%. A total of 360 Labeo rohita fingerlings with an average initial weight and length of 12.6 ± 0.04g and 10 ± 0.1 cm, respectively, were blocked into 4 treatments with 3 replicates, i.e., T1 (control), T2 (10 g/kg), T3 (20 g/kg), and T4 (30 g/kg). The feeding trial was conducted for 60 days with samplings on the 7th, 15th, 30th, and 60th days, and blood samples were collected to determine different immunological (NBT, TIg, lysozyme, and antiprotease activity) and biochemical (glucose, total protein, SGPT) parameters. A challenge study was conducted against Aeromonas hydrophila at the end of the feeding trial. The results showed that CMSMS at 30g/kg showed a better immune response over time with increasing doses. All the biochemical readings fall within normal ranges. Fish fed with CMSMS at 30g/kg (3%) have better survival after being challenged with Aeromonas hydrophila. Therefore, Cordyceps militaris spent mushroom substrate (CMSMS) can be biosafely used as in-feed antibiotics in Labeo rohita and can reduce the concern of by-product disposal from mushroom cultivation.

Chitosan Microparticles Enhance the Intestinal Release and Immune Response of an Immune Stimulant Peptide in Oncorhynchus mykiss.

The aquaculture industry is constantly increasing its fish production to provide enough products to maintain fish consumption worldwide. However, the increased production generates susceptibility to infectious diseases that cause losses of millions of dollars to the industry. Conventional treatments are based on antibiotics and antivirals to reduce the incidence of pathogens, but they have disadvantages, such as antibiotic resistance generation, antibiotic residues in fish, and environmental damage. Instead, functional foods with active compounds, especially antimicrobial peptides that allow the generation of prophylaxis against infections, provide an interesting alternative, but protection against gastric degradation is challenging. In this study, we evaluated a new immunomodulatory recombinant peptide, CATH-FLA, which is encapsulated in chitosan microparticles to avoid gastric degradation. The microparticles were prepared using a spray drying method. The peptide release from the microparticles was evaluated at gastric and intestinal pH, both in vitro and in vivo. Finally, the biological activity of the formulation was evaluated by measuring the expression of il-1β, il-8, ifn-γ, Ifn-α, and mx1 in the head kidney and intestinal tissues of rainbow trout (Oncorhynchus mykiss). The results showed that the chitosan microparticles protect the CATH-FLA recombinant peptide from gastric degradation, allowing its release in the intestinal portion of rainbow trout. The microparticle-protected CATH-FLA recombinant peptide increased the expression of il-1β, il-8, ifn-γ, ifn-α, and mx1 in the head kidney and intestine and improved the antiprotease activity in rainbow trout. These results suggest that the chitosan microparticle/CATH-FLA recombinant peptide could be a potential prophylactic alternative to conventional antibiotics for the treatment of infectious diseases in aquaculture.

Nutritional supplement of FOS enhances growth and immune system in tilapia larvae (Oreochromis niloticus)

Aquaculture is an essential economic activity worldwide, supplying the requirements of various nations. The larval stage is one of the most critical periods during the production cycle in aquaculture. To contribute to amplifying the knowledge related to viable methods to enhance this activity, the present study investigated the effects of prebiotic fructooligosaccharide (FOS) and the synthetic peptide GHRP-6 in the growth of fish, as well as its repercussions on a series of digestive enzyme activities and immune response parameters. Tilapias (Oreochromis niloticus) larvae were divided into three groups: group 1, fed with pre-formulated feed; group 2, fed with feed + FOS; group 3, fed with feed + GHRP-6. We measured the gain in size and weight at 72 and 87 days after starting the diets. At the end of treatment, we continue feeding the animals with a standard diet using the pre-formulated feed. We measured immune system parameters and digestive enzyme activities on larvae homogenate, focusing on lysozyme and antiprotease activity and α-amylase, trypsin and esterase activity, respectively. Results show a significant increment in size and weight in those fish fed with feed + FOS. Meanwhile, the primary enzyme activity was detected in those fish fed with feed + GHRP-6. On rough terms, both food combinations stimulated the immune system. Our study suggests that FOS and GHRP-6 could be desirable additives to incorporate into the tilapia larvae feeding and increase production efficiency. Keywords: aquaculture, digestive enzymes activity, FOS, GHRP-6, immune system, larvae

Haematological and Innate Immunological Reference Intervals for Farmed Yellow Catfish <i>Horabagrus brachysoma</i> and Their Seasonal Variations

The reference intervals for haematological and innate immune response variables of healthy adult yellow catfish Horabagrus brachysoma raised in captivity were determined. Reference ranges were established for all parameters and significant (P < 0.05) seasonal variations in most of the haematological and innate immune parameters were observed. Total erythrocyte count, packed cell volume, serum lysozyme and myeloperoxidase activities were found to be higher during winter season whereas most of the red blood cell indices, superoxide production by phagocytes, serum ceruloplasmin and anti-protease activities were marked to be significantly higher during rainy season of the year. Except high total leucocyte count and mean corpuscular haemoglobin, other parameters were found to be significantly lower during summer, possibly indicating a higher disease risk in summer months for this species. However, no significant variations in these variables were obtained between male and female catfish during breeding season. The information generated would be indirectly helpful for determining health status of this endangered species.

Dietary tributyrin supplementation enhances the immune and antioxidant responses of rainbow trout (Oncorhynchus mykiss) without changes in fish performance

Dietary supplementation with tributyrin (TBT), a source of butyric acid, has been shown to improve growth performance and health status when high levels of plant ingredients are included in aquafeeds. Here, we investigated the response of rainbow trout (Oncorhynchus mykiss) to a plant-based diet supplemented with TBT aiming to improve fish performance and welfare. Juvenile rainbow trout (19.0 ± 0.19 g, mean ± SEM) were divided into 12 tanks (210 L, 40 fish/tank) connected to a RAS system and fed four experimental diets in triplicate to apparent satiety. Diets tested comprised a basal diet (CTR) including 10% fishmeal and 10% fish oil, plus 83% plant-derived ingredients (44% crude protein, 18% crude fat, gross energy 21.8 MJ kg−1) with increasing inclusion levels of TBT (55% purity), supplying 0.05% (TBT1), 0.1% (TBT2) and 0.2% (TBT4) of this compound. Fish performance was evaluated after 44 days. Following the growth trial, the effect of the diets on selected plasma innate immune and hepatic antioxidant parameters of trout was investigated at 3 h and 24 h after feeding. TBT supplementation had no significant impact on growth performance, feed efficiency, feed intake, or proximal composition in rainbow trout (P > 0.05). However, the plasma lysozyme activity was higher in fish fed the TBT2 diet than in those fed CTR or TBT4 diets after 3 h (P = 0.021). In addition, the plasma antiprotease activity in fish fed the TBT2 and TBT4 diets was higher than those fed the CTR diet after 24 h (P = 0.010 and 0.013, respectively). Catalase activity in the liver of trout fed the TBT4 diet was higher than in fish fed CTR or TBT2 diets after 3 h (P = 0.016 and 0.019). Results showed that effective levels of dietary TBT supplementation at 0.1% and 0.2% could result in improved immune and antioxidant responses in trout, respectively. Therefore, dietary TBT supplementation in this carnivorous species may result in enhanced welfare when high levels of plant-derived ingredients are used in aquafeeds.

Phycocyanobilin is a new binding partner of human alpha-2-macroglobulin that protects the protein against oxidative stress

Under simulated physiological conditions, this study investigates the interaction between nutraceutical phycocyanobilin (PCB) and the universal anti-protease protein human alpha-2-macroglobulin (α2M). Extensive molecular docking analyses on multiple α2M conformations, spectroscopic techniques, and α2M activity assays were utilized to examine the complex formation. The results revealed that for every protein conformation, two high energy binding sites exist: the first, conformationally independent, at the interface region between two monomer chains and the second, conformationally dependent, in the pocket composed of amino acids from four distinct domains (TED, RBD, CUB, and MG2) of the single protein chain. Spectrofluorimetric measurements indicated a moderate affinity between α2M and PCB with a moderately high binding constant of 6.3 × 105 M−1 at 25 °C. The binding of PCB to α2M resulted in minor changes in the secondary structure content of α2M. Furthermore, PCB protected α2M from oxidation and preserved its anti-protease activity in the oxidative environment. These findings suggest that PCB binding could indirectly impact the body’s response to oxidative stress by influencing α2M's role in controlling enzyme activity during the inflammatory process. Communicated by Ramaswamy H. Sarma

In Vitro Evaluation of Antioxidant and Anti-inflammatory Potentials of Herbal Formulation Containing Marigold Flower (Calendula officinalis L.) Tea.

Aim To assess the anti-inflammatory and antioxidant properties of Calendula officinalis tea formulation. Materials and methods In this study, a formulation of 2 grams of dried marigold flower petals and 100 milliliters (ml) of distilled water was subjected to anti-inflammatory testing using albumin denaturation assay and anti-protease activity and antioxidant testing by DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay. An independent sample t-test was done to compare the anti-inflammatory and antioxidant potentials of marigold tea formulation and control using SPSS version 22.0 software (IBM Corp., Armonk, NY), and any p-value less than 0.05 was considered statistically significant. Results The highest anti-inflammatory and antioxidant activities of marigold extract were exhibited at 10 microliters (µl) and 20 µl (p-value = 0.002 and 0.000), respectively. The anti-inflammatory activity was higher than the control at all concentrations, whereas the antioxidant activity was higher at lower concentrations when compared to higher concentrations. Conclusion Marigold flower tea formulation exhibited better anti-inflammatory and antioxidant activities than the controls and therefore could be evaluated as a potential therapeutic agent.

Using Azadirachta indica protein hydrolysate as a plant protein in Nile tilapia (Oreochromis niloticus) diet: Effects on the growth, economic efficiency, antioxidant-immune responseand resistance to Streptococcus agalactiae.

A feeding trial for 90 days was conducted on Nile tilapia (Oreochromis niloticus) (average weight: 25.50 ± 0.05 g) to evaluate the effect of dietary inclusion of Azadirachta indica seed protein hydrolysate (AIPH). The evaluation included the impact on the growth metrics, economic efficiency, antioxidant potential, hemato-biochemical indices, immune response, and histological architectures. A total of 250 fish were randomly distributed in five treatments (n = 50) and received diets included with five levels of AIPH (%): 0 (control diet, AIPH0), 2 (AIPH2), 4 (AIPH4), 6 (AIPH6)or 8 (AIPH8), where AIPH partially replace fish meal by 0, 8.7%, 17.4%, 26.1%, and 34.8%, respectively. After the feeding trial, a pathogenic bacterium (Streptococcus agalactiae, 1.5 × 108 CFU/mL) was intraperitoneally injected into the fish and the survival rate was recorded. The results elucidated that AIPH-included diets significantly (p < 0.05) enhanced the growth indices (final body weight, total feed intake, total body weight gain,and specific growth rate) and intestinal morpho-metrics (villous width, length, muscular coat thickness, and goblet cells count) in comparison to the control diet, with the AIPH8 diet recording the highest values. Dietary AIPH inclusion significantly improved (p < 0.05) the economic efficacy indicated by reduced feed cost/kg gain and increased performance index. The fish fed on the AIPH diets had noticeably significantly higher (p < 0.05) protein profile variables (total proteinsand globulin)and antioxidant capabilities (superoxide dismutase and total antioxidant capacity) than the AIPH0 group. The dietary inclusion of AIPH significantly (p < 0.05) boosted the haematological parameters (haemoglobin, packed cell volume %, and counts of red blood cells and white blood cells) and immune indices (serum bactericidal activity %, antiprotease activity, and immunoglobulin Mlevel) in a concentration-dependent manner. The blood glucose and malondialdehyde levels were significantly (p < 0.05) lowered by dietary AIPH (2%-8%). The albumin level and hepatorenal functioning parameters (aspartate aminotransferase, alanine aminotransferase, and creatinine) were not significantly (p > 0.05) altered by AIPH diets. Additionally, AIPH diets did not adversely alter the histology of the hepatic, renalor splenic tissues with moderately activated melano-macrophage centres. The mortality rate among S. agalactiae-infected fish declined as dietary AIPH levels rose, where the highest survival rate (86.67%) was found in the AIPH8 group (p < 0.05). Based on the broken line regression model, our study suggests using dietary AIPH at the optimal level of 6%. Overall, dietary AIPH inclusion enhanced the growth rate, economic efficiency, health status, and resistance of Nile tilapia to the S. agalactiae challenge. These beneficial impacts can help the aquaculture sector to be more sustainable.

Serum α1-Proteinase Inhibitor, Calprotectin, and S100A12 Concentrations in the Characterization of Pancreatitis in Dogs.

Miniature Schnauzers are predisposed to develop pancreatitis, with familial hypertriglyceridemia (HTG) described as a potential risk factor. Diagnosing pancreatitis in dogs is based on the integration of serum canine-specific pancreatic lipase (cPLI) concentration, clinical presentation, and diagnostic imaging findings. However, markers of systemic inflammation and antiprotease activity have not been extensively investigated in the characterization and prognostication of pancreatitis in dogs. Serum concentrations of alpha1-proteinase inhibitor (α1PI; as a marker of systemic antiprotease response) and calprotectin and S100A12 (as markers of systemic inflammation) were measured in serum samples from 35 Miniature Schnauzers diagnosed with pancreatitis (serum cPLI concentration >400 μg/L, clinical signs, abdominal imaging findings). These markers were evaluated for possible associations with patient characteristics, clinical presentation, risk factors for pancreatitis, and outcome. The study showed that biomarkers of systemic inflammation and antiprotease activity are commonly increased in Miniature Schnauzers with pancreatitis. Whereas serum calprotectin and S100A12 concentrations were found to have limited utility in differentiating pancreatitis presentations, serum α1PI concentrations and potentially also the serum calprotectin-to-S100A12 ratio might be non-invasive surrogate markers of disease severity in dogs with pancreatitis.

Effects of yeast hydrolysate supplementation in low-fish meal diets for pikeperch.

Plant proteins have been increasingly used as sustainable substitutes for fish meal (FM) in aquafeeds; however, their high inclusion level compromises fish performance. The objective of this study was to examine whether yeast hydrolysate (YH) supplementation can improve the utilisation of high soybean meal (SM) diet and ameliorate its potential deteriorating impacts in pikeperch (Sander lucioperca). A basal diet was formulated using 44% FM, and four additional diets were produced by replacing 30 or 60% of FM with SM with or without the addition of 2% YH (FM, SM30, SM60, SM30+YH, and SM60+YH diets). Each diet was fed to three groups of fish (35.3±0.10g, 150 fish per group) to visual satiety four times daily for 70days. Fish growth was not impacted by FM replacement level or YH application. However, SM60 group exhibited markedly higher feed conversion ratio and lower survival rate than those fed the FM- and YH-supplemented diets (P<0.05). The highest and the lowest protein efficiency ratio values were obtained for the SM30+YH and SM60 groups, respectively. Whole-body lipid content decreased in SM60 and SM60+YH groups, and muscle lipid decreased in all the replacement groups. Serum triglyceride and glucose concentrations tended to decrease as FM replacement level increased. The highest alanine aminotransferase, aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities were detected in the SM60 group, and YH addition significantly decreased the AST and LDH activities. Serum lysozyme activity decreased in SM30, SM60 and SM60+YH groups. Serum myeloperoxidase and antiprotease activities decreased in SM60 group, and YH supplementation improved their activities. No effects of diets were observed on serum antioxidant parameters such as catalase activity and malondialdehyde concentration, and gut morphological indices. Number of goblet cells in midgut decreased by increasing the SM inclusion level and a slight improvement was observed by YH application. These findings suggest that YH supplementation has the potential to support the replacement of up to 60% FM with defatted SM in pikeperch feed without deteriorating growth, feed utilisation, and survival rate. Further, YH incorporation mitigated the damaging impacts of high SM diet on liver function and non-specific immune response.

Isolation, identification and pathogenicity study of emerging multi-drug resistant fish pathogen Acinetobacter pittii from diseased rohu (Labeo rohita) in India

Growing numbers of disease outbreaks and a wider variety of pathogens have emerged as a result of aquaculture's expansion, intensification, and commercialization. In the current work, we isolated and identified a new multi-drug resistant pathogenic bacteria, Acinetobacter pittii strain COF_AHE14, that causes mass death in rohu (Labeo rohita) during a post-monsoon disease outbreak in the aquaculture farm of Tripura, India. The isolate was initially identified based on morphological and biochemical information, and the species was validated through molecular characterization of the 16S rRNA gene sequence and phylogenetic analysis. An experimental infectivity study in healthy rohu (Labeo rohita), haemato-biochemical and immune-pathophysiological analysis, histopathological study, and fulfilment of the Koch’s postulate all confirmed the pathogenicity of the isolated bacteria. The LD50 of A. pittii strain COF_AHE14 in an experimental infection was 4.1 × 106 CFU fish-1, and the clinical signs observed, such as ulcerative lesions and haemorrhages on the skin and tail and fin rot, were similar to those of the farm-diseased fish. Total RBC count, Hb, PCV, MCV, glucose, Na+, albumin, respiratory burst and anti-protease activity were significantly (p < 0.05) lowered in the infected group as compared to the control group. Whereas, the total WBC count, total protein, globulin, ALP, SGOT, SGPT, and K+ levels were significantly (p < 0.05) higher in the infected group as compared to the control group. Histopathological examination of the infected fish's kidney and liver revealed significant changes, indicating bacterial septicaemia. The antibiotic resistance pattern showed A. pittii strain COF_AHE14 was a multi-drug resistance, showing resistance to cephalexin, cefoxitin, nitrofurantoin, ampicillin, oxacillin, penicillin-G, bacitracin, and trimethoprim. This is the first account of A. pittii as a cause of mass death in rohu (L. rohita), and the current study revealed that, in addition to causing human clinical infections, A. pittii poses a potential threat to farmed fish.

Dietary ARA, DHA, and Carbohydrate Ratios Affect the Immune Status of Gilthead Sea Bream Juveniles upon Bacterial Challenge.

This study aims to assess the effects of different dietary n-6/n-3 long-chain polyunsaturated fatty acid ratios and CHO content in the immune response of gilthead seabream. For that purpose, gilthead sea bream juveniles (initial body weight = 47.5 g) were fed for 84 days with four isoproteic (47% crude protein) and isolipidic (18% crude lipids) diets with high (20%) or low (5%) level of gelatinized starch (HS or LS diets, respectively) and included approximately 2.4% ARA or DHA. At the end of the trial, the DHA-enriched groups presented increased red blood cell (RBC) count, hemoglobin, plasmatic nitric oxide (NO) content, and antiprotease and alternative complement activities. The ARA groups had increased thrombocyte count, and plasmatic bactericidal activity against Vibrio anguillarum was lower in the fish fed the ARA/LS diet. After the feeding trial, the fish were challenged with an intraperitoneal injection (i.p.) of killed Photobacterium damselae subsp. piscicida (Phdp) and sampled at 4 and 24 h after the challenge. At 4 h after i.p., the ARA groups presented increased plasma total immunoglobulins (Ig) and bactericidal activity against V. anguillarum. In addition, the fish fed the ARA/LS diet presented lower white blood cell (WBC) and alternative complement activity. At 24 h after i.p., the ARA groups presented increased RBC, WBC, and thrombocyte numbers, total IG, plasma peroxidase activity, and casp3 expression in the distal intestine. The HS groups presented increased plasma NO content and bactericidal activity against Phdp and decreased protease, antiprotease activity, and bactericidal activity against V. anguillarum. In conclusion, high dietary DHA levels seemed to improve the immune status of unchallenged gilthead sea bream juveniles, while high dietary ARA levels improved the fish immune response to a bacterial challenge. The energy provided by dietary starch seems to be important to promote a fast response by the fish immune system after a challenge.

Ethnomedicinal Use, Phytochemistry, Pharmacology, and Toxicology of Euphorbia resinifera O. Berg. (B): A Review

Euphorbia resinifera (Zaggûm or Tikiut) is an endemic species of Morocco that grows in the Middle Atlas Mountain range. It is used in folk medicine to treat several diseases, especially various types of cancer. Aim of the review: In this review, we critically highlighted and discussed previous reports on E. resinifera, concerning its botanical description, taxonomy, geographical distribution, and medicinal use. In addition, bioactive compounds, toxicology, and pharmacological effects were reported. Materials and methods: We searched various scientific databases, such as Scopus, PubMed, Web of Science, SpringerLink, SciFinder, Wiley Online, and Google Scholar, to collect data on E. resinifera. Studies involving E. resinifera or its bioactive compounds with regards to antitumor, anti-inflammatory, antileishmanial, antiprotease, immunomodulatory, irritant, and lysosomal activities are discussed here. Results: E. resinifera has been widely used in folk medicine practice in Morocco to treat several diseases such as diabetes, cancer, and inflammatory skin conditions, as well as to heal wounds. In phytochemistry studies, biochemical compounds have been identified from E. resinifera belonging specially to terpenoids. Indeed, in vitro and in vivo pharmacological investigations showed that extracts and compounds from the latex of E. resinifera exhibited a wide spectrum of biological properties, particularly antioxidant, antimicrobial, antitumor, anti-inflammatory, antileishmanial, and immunomodulatory and neuroprotective activities. Conclusion: The use of E. resinifera in conventional medicine is supported by processes founded on biological evidence. However, in-depth research is necessary to prove the safety and efficacy of E. resinifera latex extracts and their compounds and to clarify their pharmacological mechanisms. In addition, pharmacokinetic and pharmacodynamics studies are required for extracts and bioactive compounds.

Innate Immune Response Assessment in Cyprinus carpio L. upon Experimental Administration with Artemia salina Bio-Encapsulated Aeromonas hydrophila Bacterin.

The present study aimed to analyze the enhancement of innate immune responses in juvenile-stage common carp (Cyprinus carpio L.), upon the administration of heat-killed Aeromonas hydrophila at a dosage of 1 × 107 CFU ml-1 through bio-encapsulation in the aquatic crustacean, Artemia salina. This work emphasizes the modulation of innate immune response when administered with the bio-encapsulated heat-killed antigen that acts as an inactivated vaccine against Motile Aeromonas Septicemia disease. Bio-encapsulated oral administration of antigens promotes innate immunity in juvenile-stage fishes. The optimization of effective bio-encapsulation of bacterin in Artemia salina nauplii was carried out and the best optimal conditions were chosen for immunization. The functional immune parameters such as myeloperoxidase, lysozyme, alkaline phosphatase, antiprotease and respiratory burst activity in serum, blood and intestinal tissue samples were analyzed along with blood differential leukocyte count and tissue histopathology studies. Both humoral and cellular immune responses analyzed were substantially induced or enhanced in the treatment groups in comparison with the control group. The results showed a significant variation in the bio-encapsulation group than the control group and also were comparable to the protection conferred with immersion route immunization under similar conditions. Thus, most of the innate non-specific immune responses are inducible, despite being constitutive of the fish immune system, to exhibit a basal level of protection and a road to better vaccination strategy in Cyprinus carpio L. aquaculture worldwide.

Studies on the growth, immunomodulation and gut morphometry of Labeo rohita fed pectin

<br />The present study was designed to evaluate the effect of pectin supplemented feeds on the growth, gut morphology and immunology of <i>Labeo rohita</i>. Experimental feeds containing various concentrations of pectin including control (0%), T1 (0.5%), T2 (1.0%), T3 (2.0%), and T4 (4.0%) were administered to <i>L. rohita</i> twice per day at a rate of 3% body weight. There was a significant (<i>P</i> < 0.05) decrease in average final weight, weight gain%, net weight gain and specific growth rate observed in treatment groups in order of control > T1 > T2 > T3 > T4 while feed conversion ratio was significantly (<i>P </i>< 0.05) higher in T4 compared to the other groups T3 > T2 > T1 > control. Immunological parameters of peroxidase, protease and antiprotease (trypsin) showed significant (<i>P </i>< 0.05) results between control and treated groups. Peroxidase activity was significantly (<i>P </i>< 0.05) higher in T4 group fish compared to T3 > T2 > T1 > control group. Total protein, protease and antiprotease activity were significantly (<i>P </i>< 0.05) higher in the control compared to treatment groups T1 > T2 > T3 > T4. An increased level of the non-starch polysaccharide pectin in feed caused acute effects on gut morphometry. Villus length, villus width, and an increase in mucosal folds were abnormal but no significant (<i>P</i> > 0.05) abnormality was observed in the control group. In the submucosal part of the intestine, hyperplasia was shown in loose connective tissues and in adipose tissue cells in the treated groups. In conclusion, dietary pectin supplementation from 0.5% to 4% had negative effects on the growth, immunological parameters and gut histomorphology of <i>L. rohita</i>.

Immune profiling of rainbow trout (Oncorhynchus mykiss) exposed to Lactococcus garvieae: Evidence in asymptomatic versus symptomatic or vaccinated fish.

Lactococcosis, caused by the Gram-positive bacterium Lactococcus garvieae, is a major concern in rainbow trout (Oncorhynchus mykiss) farms, which are regularly affected by outbreaks especially during the summer/fall months. In these farms, unvaccinated healthy and symptomatic fish can coexist with vaccinated fish. In the present study, innate (leukogram, serum lysozyme activity, peroxidase activity, antiprotease activity, bactericidal activity, total IgM and total proteins), and specific immune parameters (serum antibodies to L.garvieae) were assessed in unvaccinated adult rainbow trout naturally exposed to the pathogen, with or without evidence of clinical signs, or subjected to vaccination. Blood was drawn from all three groups, and blood smears were prepared. Bacteria were found in the blood smears of 70% of the symptomatic fish but not in any of the asymptomatic fish. Symptomatic fish showed lower blood lymphocytes and higher thrombocytes than asymptomatic fish (p ≤ .05). Serum lysozyme and bactericidal activity did not vary substantially among groups; however, serum antiprotease and peroxidase activity were significantly lower in the unvaccinated symptomatic group than in the unvaccinated and vaccinated asymptomatic groups (p ≤ .05). Serum total proteins and total immunoglobulin (IgM) levels in vaccinated asymptomatic rainbow trout were significantly higher than in unvaccinated asymptomatic and symptomatic groups (p ≤ .05). Similarly, vaccinated asymptomatic fish produced more specific IgM against L.garvieae than unvaccinated asymptomatic and symptomatic fish (p ≤ .05). This preliminary study provides basic knowledge on the immunological relationship occurring between the rainbow trout and L.garvieae, potentially predicting health outcomes. The approach we proposed could facilitate infield diagnostics, and several non-specific immunological markers could serve as reliable indicators of the trout's innate ability to fight infection.

Effects of dietary arachidonic acid supplementation in high plant protein diets on growth, feed utilization, and immunity of olive flounder, Paralichthys olivaceus

This study was designed to evaluate the effects of graded levels of arachidonic acid (ARA) in low fish meal diets (30% fish meal, 30% plant protein blends) on growth, feed utilization, and immunity of olive flounder (Paralichthys olivaceus). The control diet was formulated with 3% fish oil and 5.1% soybean oil (ARA0.0). Five other diets were formulated incorporating different levels of ARA into the ARA0.0 diet at the expense of soybean oil; that is, 0.3, 0.6, 0.9, 1.2, and 1.5% ARA (ARA0.3, ARA0.6, ARA0.9, ARA1.2, and ARA1.5), respectively. Three replicate groups of fish (88.24 ± 0.06 g) were placed into 18 polyvinyl circular tanks (215 L), with 24 fish each and fed an experimental diet twice a day for nine weeks. Overall, significantly improved growth performance was observed in the ARA0.6 group. Liver C18:2n-6 was significantly reduced, and liver ARA content was significantly increased with the increase in dietary ARA levels. Nitroblue tetrazolium activity was significantly higher in the ARA0.6 group, and significantly higher superoxide dismutase and myeloperoxidase activities were observed in the ARA0.9 group. Further, immunoglobulin level and lysozyme activity were significantly improved in the ARA0.3 group, antiprotease activity was increased in the ARA0.6 and ARA0.9 groups, and the ARA0.3 group showed lower liver lipid content than the ARA0.0 group. Serum prostaglandin E2 level was significantly increased with 1.5% ARA inclusion. Pro-inflammatory gene expressions of interleukin (IL)-6, IL-8, IL-10, perforin, and tumor necrosis factor in the intestine were significantly downregulated, and anti-inflammatory gene expression of transforming growth factor-β2 gene expression was significantly upregulated with higher dietary ARA supplementation (0.9–1.5%). These results indicate that 0.3–0.9% supplemented ARA may improve the growth, immune, and hematological parameters of olive flounder.

349 - HSCT Recipients That Develop Bronchiolitis Obliterans Syndrome Exhibit Significant Proteomic Changes in Complement Activation, Angiotensin Maturation, and Antiprotease Activity Prior to Clinical Diagnosis

349 - HSCT Recipients That Develop Bronchiolitis Obliterans Syndrome Exhibit Significant Proteomic Changes in Complement Activation, Angiotensin Maturation, and Antiprotease Activity Prior to Clinical Diagnosis