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Fulltext Development of Methyl Ester Antibody-Based Competitive Indirect ELISA for Quantitative Detection of Mitragynine in Human Urine

Mustafa RR, Sukor R, Mohd Nor SM, Saari N, Mohsin AZ

ACS Omega, 2023 Dec 19;8(50):47412-47426.
PMID: 38144118 DOI: 10.1021/acsomega.3c02734

Mitragynine is the main psychoactive compound of Mitragyna speciosa Korth. (kratom). This alkaloid could render psychotropic effects and is often misused as a substitute for commercial drugs. Nowadays, the increasing popularity of kratom has led to the development of a rapid and effective detection method. The detection of mitragynine in a biological sample such as urine requires a highly sensitive and specific method due to the complex nature of mitragynine in urine. Enzyme-linked immunosorbent assay (ELISA) is well known as a rapid screening method for biological samples. In this study, a competitive indirect ELISA was successfully developed using MG-22-OCH3 IgG as a detection antibody for mitragynine in human urine. The mitragynine immunoassay showed a limit of detection and a limit of quantification of 0.412 and 1.25 μg/mL, respectively. The measurement range was between 0.01 and 100.0 μg/mL, with a minimal inhibition (IC50) value of 0.152 μg/mL. The developed ELISA was validated using a gold method such as high-performance liquid chromatography-mass spectrometry (HPLC-MS). The percentage of recovery and the coefficient of variation (CV) for the ELISA and LCMS/MS analyses were 84.0-95.70%, 99.20-112.0%, 7.69-9.78%, and 2.86-6.62%, respectively. This indicates that the developed ELISA is a reliable method that can be used as a rapid approach for quantifying mitragynine content in biological samples.
Fulltext Effects of Storage Time and Temperature on Lipid Oxidation and Protein Co-Oxidation of Low-Moisture Shredded Meat Products

Wazir H, Chay SY, Zarei M, Hussin FS, Mustapha NA, Wan Ibadullah WZ, et al.

Antioxidants (Basel), 2019 Oct 16;8(10).
PMID: 31623062 DOI: 10.3390/antiox8100486

Studies on the oxidative changes in meat-based, low-moisture, ready to eat foods are complicated due to complex food system and slow lipid-protein oxidative deterioration. The current study evaluates the oxidative changes over six months of storage on shredded beef and chicken products (locally known as serunding) for physicochemical analysis, lipid oxidation (conjugated dienes and malondialdehydes) and protein co-oxidation (soluble protein content, amino acid composition, protein carbonyl, tryptophan loss and Schiff base fluorescence) at 25 °C, 40 °C and 60 °C. The lipid stability of chicken serunding was significantly lower than beef serunding, illustrated by higher conjugated dienes content and higher rate of malondialdehyde formation during storage. In terms of protein co-oxidation, chicken serunding with higher polyunsaturated fatty acids (PUFA) experienced more severe oxidation, as seen from lower protein solubility, higher protein carbonyl and Schiff base formation compared to beef serunding. To conclude, chicken serunding demonstrates lower lipid and protein stability and exhibits higher rate of lipid oxidation and protein co-oxidation than beef serunding. These findings provide insights on the progression of lipid oxidation and protein co-oxidation in cooked, shredded meat products and could be extrapolated to minimize possible adverse effects arising from lipid oxidation and protein co-oxidation, on the quality of low-moisture, high-lipid, high-protein foods.
Fulltext Anti-obesity effect of ethanolic extract from Cosmos caudatus Kunth leaf in lean rats fed a high fat diet

Rahman HA, Sahib NG, Saari N, Abas F, Ismail A, Mumtaz MW, et al.

BMC Complement Altern Med, 2017 Feb 22;17(1):122.
PMID: 28228098 DOI: 10.1186/s12906-017-1640-4

BACKGROUND: Obesity is a major health concern both in developed and developing countries. The use of herbal medicines became the subject of interest for the management of obesity due to its natural origin, cost effectiveness and minimal side effects. The present study aimed at investigating anti-obesity potential of ethanolic extract from Cosmos caudatus Kunth leaf (EECCL).
METHODS: In this study, the rats were randomly divided into six groups i.e., (1) Normal Diet (ND); (2) Normal Diet and 175 mg/kgBW of EECCL (ND + 175 mg/kgBW); (3) Normal Diet and 350 mg/kgBW of EECCL (ND + 350 mg/kgBW); (4) High Fat Diet (HFD); (5) High Fat Diet and 175 mg/kgBW of EECCL (HFD + 175 mg/kgBW); (6) High Fat Diet and 350 mg/kgBW of EECCL (HFD + 350 mg/kgBW). The anti-obesity potential was evaluated through analyses of changes in body weight, visceral fat weight, and blood biochemicals including total cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c), leptin, insulin, adiponectin, ghrelin and fecal fat content. In addition, metabolite profiling of EECCL was carried out using NMR spectroscopy.
RESULTS: Rats receiving EECCL together with HFD showed significant (p 0.05) different with those of ND rats. Other related obesity biomarkers including plasma lipid profiles, insulin, leptin, ghrelin and adiponectin levels also showed significant improvement (p
Fulltext A comparative study of extraction techniques for maximum recovery of glutamate decarboxylase (GAD) from Aspergillus oryzae NSK

Lee Ying Yeng A, Kadir MS, Ghazali HM, Raja Abd Rahman RN, Saari N

BMC Res Notes, 2013 Dec 10;6:526.
PMID: 24321181 DOI: 10.1186/1756-0500-6-526

BACKGROUND: γ-Amino butyric acid (GABA) is a major inhibitory neurotransmitter of the mammalian central nervous system that plays a vital role in regulating vital neurological functions. The enzyme responsible for producing GABA is glutamate decarboxylase (GAD), an intracellular enzyme that both food and pharmaceutical industries are currently using as the major catalyst in trial biotransformation process of GABA. We have successfully isolated a novel strain of Aspergillus oryzae NSK that possesses a relatively high GABA biosynthesizing capability compared to other reported GABA-producing fungal strains, indicating the presence of an active GAD. This finding has prompted us to explore an effective method to recover maximum amount of GAD for further studies on the GAD's biochemical and kinetic properties. The extraction techniques examined were enzymatic lysis, chemical permeabilization, and mechanical disruption. Under the GAD activity assay used, one unit of GAD activity is expressed as 1 μmol of GABA produced per min per ml enzyme extract (U/ml) while the specific activity was expressed as U/mg protein.
RESULTS: Mechanical disruption by sonication, which yielded 1.99 U/mg of GAD, was by far the most effective cell disintegration method compared with the other extraction procedures examined. In contrast, the second most effective method, freeze grinding followed by 10% v/v toluene permeabilization at 25°C for 120 min, yielded only 1.17 U/mg of GAD, which is 170% lower than the sonication method. Optimized enzymatic lysis with 3 mg/ml Yatalase® at 60°C for 30 min was the least effective. It yielded only 0.70 U/mg of GAD. Extraction using sonication was further optimized using a one-variable-at-a-time approach (OVAT). Results obtained show that the yield of GAD increased 176% from 1.99 U/mg to 3.50 U/mg.
CONCLUSION: Of the techniques used to extract GAD from A. oryzae NSK, sonication was found to be the best. Under optimized conditions, about 176% of GAD was recovered compared to recovery under non optimized conditions. The high production level of GAD in this strain offers an opportunity to conduct further studies on GABA production at a larger scale.
Fulltext HOspitalised Pneumonia Extended (HOPE) Study to reduce the long-term effects of childhood pneumonia: protocol for a multicentre, double-blind, parallel, superiority randomised controlled trial

Chang AB, Fong SM, Yeo TW, Ware RS, McCallum GB, Nathan AM, et al.

BMJ Open, 2019 Apr 24;9(4):e026411.
PMID: 31023759 DOI: 10.1136/bmjopen-2018-026411

INTRODUCTION: Early childhood pneumonia is a common problem globally with long-term complications that include bronchiectasis and chronic obstructive pulmonary disease. It is biologically plausible that these long-term effects may be minimised in young children at increased risk of such sequelae if any residual lower airway infection and inflammation in their developing lungs can be treated successfully by longer antibiotic courses. In contrast, shortened antibiotic treatments are being promoted because of concerns over inducing antimicrobial resistance. Nevertheless, the optimal treatment duration remains unknown. Outcomes from randomised controlled trials (RCTs) on paediatric pneumonia have focused on short-term (usually <2 weeks) results. Indeed, no long-term RCT-generated outcome data are available currently. We hypothesise that a longer antibiotic course, compared with the standard treatment course, reduces the risk of chronic respiratory symptoms/signs or bronchiectasis 24 months after the original pneumonia episode.
METHODS AND ANALYSIS: This multicentre, parallel, double-blind, placebo-controlled randomised trial involving seven hospitals in six cities from three different countries commenced in May 2016. Three-hundred-and-fourteen eligible Australian Indigenous, New Zealand Māori/Pacific and Malaysian children (aged 0.25 to 5 years) hospitalised for community-acquired, chest X-ray (CXR)-proven pneumonia are being recruited. Following intravenous antibiotics and 3 days of amoxicillin-clavulanate, they are randomised (stratified by site and age group, allocation-concealed) to receive either: (i) amoxicillin-clavulanate (80 mg/kg/day (maximum 980 mg of amoxicillin) in two-divided doses or (ii) placebo (equal volume and dosing frequency) for 8 days. Clinical data, nasopharyngeal swab, bloods and CXR are collected. The primary outcome is the proportion of children without chronic respiratory symptom/signs of bronchiectasis at 24 months. The main secondary outcomes are 'clinical cure' at 4 weeks, time-to-next respiratory-related hospitalisation and antibiotic resistance of nasopharyngeal respiratory bacteria.
ETHICS AND DISSEMINATION: The Human Research Ethics Committees of all the recruiting institutions (Darwin: Northern Territory Department of Health and Menzies School of Health Research; Auckland: Starship Children's and KidsFirst Hospitals; East Malaysia: Likas Hospital and Sarawak General Hospital; Kuala Lumpur: University of Malaya Research Ethics Committee; and Klang: Malaysian Department of Health) have approved the research protocol version 7 (13 August 2018). The RCT and other results will be submitted for publication.
TRIAL REGISTRATION: ACTRN12616000046404.
In vitro binding of mutagenic heterocyclic aromatic amines by Bifidobacterium pseudocatenulatum G4

Faridnia F, Hussin AS, Saari N, Mustafa S, Yee LY, Manap MY

Benef Microbes, 2010 Jun;1(2):149-54.
PMID: 21831754 DOI: 10.3920/BM2009.0035

Consumption of probiotics has been associated with decreased risk of colon cancer and reported to have antimutagenic/ anti-carcinogenic properties. One possible mechanism for this effect involves physical binding of the mutagenic compounds, such as heterocyclic amines (HCAs), to the bacteria. Therefore, the objective of this study was to examine the binding capacity of bifidobacterial strains of human origin on mutagenic heterocyclic amines which are suspected to play a role in human cancers. In vitro binding of the mutagens Trp-p-2, IQ, MeIQx, 7,8DiMeIQx and PhIP by three bacterial strains in two media of different pH was analysed using high performance liquid chromatography. Bifidobacterium pseudocatenulatum G4 showed the highest decrease in the total HCAs content, followed by Bifidobacterium longum, and Escherichia coli. pH affects binding capacity; the highest binding was obtained at pH 6.8. Gram-positive tested strains were found to be consistently more effective than the gram-negative strain. There were significant decreases in the amount of HCAs in the presence of different cell concentrations of B. pseudocatenulatum G4; the highest decrease was detected at the concentration of 10(10) cfu/ml. The results showed that HCAs were able to bind with all bacterial strains tested in vitro, thus it may be possible to decrease their absorption by human intestine and increase their elimination via faeces.
Fulltext The improvement of the endogenous antioxidant property of stone fish (Actinopyga lecanora) tissue using enzymatic proteolysis

Bordbar S, Ebrahimpour A, Abdul Hamid A, Abdul Manap MY, Anwar F, Saari N

Biomed Res Int, 2013;2013:849529.
PMID: 23586061 DOI: 10.1155/2013/849529

The stone fish (Actinopyga lecanora) ethanolic and methanolic tissue extracts were investigated for total phenolic contents (TPCs) as well as antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH(•)) radical scavenging activity and ferric reducing antioxidant power (FRAP) assays. Both extracts showed low amount of phenolics (20.33 to 17.03 mg of gallic acid equivalents/100 g dried sample) and moderate antioxidant activity (39% to 34% DPPH(•) radical scavenging activity and 23.95 to 22.30 mmol/100 mL FeSO4 FRAP value). Enzymatic proteolysis was carried out in order to improve the antioxidant activity using six commercially available proteases under their optimum conditions. The results revealed that the highest increase in antioxidant activity up to 85% was obtained for papain-generated proteolysate, followed by alcalase (77%), trypsin (75%), pepsin (68%), bromelain (68%), and flavourzyme (50%) as measured by DPPH(•) radical scavenging activity, whilst for the FRAP value, the highest increase in the antioxidant activity up to 39.2 mmol/100 mL FeSO4 was obtained for alcalase-generated proteolysate, followed by papain (29.5 mmol/100 mL FeSO4), trypsin (23.2 mmol/100 mL FeSO4), flavourzyme (24.7 mmol/100 mL FeSO4), bromelain (22.9 mmol/100 mL FeSO4), and pepsin (20.8 mmol/100 mL FeSO4). It is obvious that proteolysis of stone fish tissue by proteolytic enzymes can considerably enhance its antioxidant activity.
Fulltext Angiotensin Converting Enzyme (ACE)-Peptide Interactions: Inhibition Kinetics, In Silico Molecular Docking and Stability Study of Three Novel Peptides Generated from Palm Kernel Cake Proteins

Zarei M, Abidin NBZ, Auwal SM, Chay SY, Haiyee ZA, Sikin AM, et al.

Biomolecules, 2019 10 04;9(10).
PMID: 31590308 DOI: 10.3390/biom9100569

Three novel peptide sequences identified from palm kernel cake (PKC) generated protein hydrolysate including YLLLK, WAFS and GVQEGAGHYALL were used for stability study against angiotensin-converting enzyme (ACE), ACE-inhibition kinetics and molecular docking studies. Results showed that the peptides were degraded at different cleavage degrees of 94%, 67% and 97% for YLLLK, WAFS and GVQEGAGHYALL, respectively, after 3 h of incubation with ACE. YLLLK was found to be the least stable (decreased ACE-inhibitory activity) compared to WAFS and GVQEGAGHYALL (increased ACE-inhibitory activity). YLLLK showed the lowest Ki (1.51 mM) in inhibition kinetics study when compared to WAFS and GVQEGAGHYALL with Ki of 2 mM and 3.18 mM, respectively. In addition, ACE revealed the lowest Kmapp and Vmaxapp and higher catalytic efficiency (CE) in the presence of YLLLK at different concentrations, implying that the enzyme catalysis decreased and hence the inhibition mode increased. Furthermore, YLLLK showed the lowest docking score of -8.224 and seven interactions with tACE, while peptide GVQEGAGHYALL showed the higher docking score of -7.006 and five interactions with tACE.
The structural reconformation of peptides in enhancing functional and therapeutic properties: Insights into their solid state crystallizations

Saadi S, Ghazali HM, Saari N, Abdulkarim SM

Biophys Chem, 2021 06;273:106565.
PMID: 33780688 DOI: 10.1016/j.bpc.2021.106565

Therapeutic peptides derived proteins with alpha-reconformation states like antibody shape have shown potential effects in combating terrible diseases linked with earlier signs of angiogensis, mutagenesis and transgenesis. Alpha reconformation in material design refers to the folding of the peptide chains and their transitions under reversible chemical bonds of disulfide chemical bridges and further non-covalence lesions. Thus, the rational design of signal peptides into alpha-helix is intended in increasing the defending effects of peptides into cores like adjuvant antibiotic and/or vaccines. Thereby, the signal peptides are able in displaying multiple eradicating regions by changing crystal-depositions and deviation angles. These types of molecular structures could have multiple advantages in tracing disease syndromes and impurities by increasing the host defense against the fates of pathogens and viruses, eventually leading to the loss in signaling by increasing peptide susceptibility levels to folding and unfolding and therefore, formation of transgenic peptide models. Alpha reconformation peptides is aimed in triggering as well as other regulatory functions such as remodulating metabolic chain disorders of lipolysis and glucolysis by increasing the insulin and leptin resistance for best lipid storages and lipoprotein density distributions.
Degradation of veterinary antibiotics and hormone during broiler manure composting

Ho YB, Zakaria MP, Latif PA, Saari N

Bioresour Technol, 2013 Mar;131:476-84.
PMID: 23384781 DOI: 10.1016/j.biortech.2012.12.194

The fate of nine veterinary antibiotics and one hormone in broiler manure during 40 days of composting was investigated. Results showed that composting can significantly reduce the concentration of veterinary antibiotics and hormone in broiler manure, making application of the post-compost manure safer for soil application. More than 99% of the nine antibiotics and one hormone involved in this study were removed from the manure during 40 days of composting. The target antibiotics and hormone showed short half-life in broiler manure composting, ranging from 1.3 to 3.8 days. The relationship between the physico-chemical properties of soil, manure and manure compost and its veterinary antibiotic and hormone concentration was statistically evaluated by Pearson correlation matrix. The concentration of veterinary antibiotics and hormone in manure compost was suggested to be affected by physico-chemical properties such as pH, temperature, total organic carbon (TOC), total nitrogen (TN), total phosphorus (TP) and metal contents.
Recent advances in food biopeptides: production, biological functionalities and therapeutic applications

Saadi S, Saari N, Anwar F, Abdul Hamid A, Ghazali HM

Biotechnol Adv, 2014 12 12;33(1):80-116.
PMID: 25499177 DOI: 10.1016/j.biotechadv.2014.12.003

The growing momentum of several common life-style diseases such as myocardial infarction, cardiovascular disorders, stroke, hypertension, diabetes, and atherosclerosis has become a serious global concern. Recent developments in the field of proteomics offering promising solutions to solving such health problems stimulates the uses of biopeptides as one of the therapeutic agents to alleviate disease-related risk factors. Functional peptides are typically produced from protein via enzymatic hydrolysis under in vitro or in vivo conditions using different kinds of proteolytic enzymes. An array of biological activities, including antioxidative, antihypertensive, antidiabetic and immunomodulating has been ascribed to different types of biopeptides derived from various food sources. In fact, biopeptides are nutritionally and functionally important for regulating some physiological functions in the body; however, these are yet to be extensively addressed with regard to their production through advance strategies, mechanisms of action and multiple biological functionalities. This review mainly focuses on recent biotechnological advances that are being made in the field of production in addition to covering the mode of action and biological activities, medicinal health functions and therapeutic applications of biopeptides. State-of-the-art strategies that can ameliorate the efficacy, bioavailability, and functionality of biopeptides along with their future prospects are likewise discussed.
Fulltext The morphology of Ganoderma lucidum mycelium in a repeated-batch fermentation for exopolysaccharide production

Wan-Mohtar WA, Ab Kadir S, Saari N

Biotechnol Rep (Amst), 2016 Sep;11:2-11.
PMID: 28352534 DOI: 10.1016/j.btre.2016.05.005

The morphology of Ganoderma lucidum BCCM 31549 mycelium in a repeated-batch fermentation (RBF) was studied for exopolysaccharide (EPS) production. RBF was optimised for time to replace and volume to replace. G. lucidum mycelium showed the ability to self-immobilise and exhibited high stability for repeated use in RBF with engulfed pellets. Furthermore, the ovoid and starburst-like pellet morphology was disposed to EPS production in the shake flask and bioreactor, respectively. Seven RBF could be carried out in 500 mL flasks, and five repeated batches were performed in a 2 L bioreactor. Under RBF conditions, autolysis of pellet core in the shake flask and shaving off of the outer hairy region in the bioreactor were observed at the later stages of RBF (R4 for the shake flask and R6 for the bioreactor). The proposed strategy showed that the morphology of G. lucidum mycelium can withstand extended fermentation cycles.
Identification of Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus fermentum from honey stomach of honeybee

Tajabadi N, Mardan M, Saari N, Mustafa S, Bahreini R, Manap MY

Braz J Microbiol, 2013;44(3):717-22.
PMID: 24516438

This study aimed to isolate and identify Lactobacillus in the honey stomach of honeybee Apis dorsata. Samples of honeybee were collected from A. dorsata colonies in different bee trees and Lactobacillus bacteria isolated from honey stomachs. Ninety two isolates were Gram-stained and tested for catalase reaction. By using bacterial universal primers, the 16S rDNA gene from DNA of bacterial colonies amplified with polymerase chain reaction (PCR). Forty-nine bacterial 16S rDNA gene were sequenced and entrusted in GenBank. Phylogenetic analysis showed they were different phylotypes of Lactobacillus. Two of them were most closely relevant to the previously described species Lactobacillus plantarum. Other two phylotypes were identified to be closely related to Lactobacillus pentosus. However, only one phylotype was found to be distantly linked to the Lactobacillus fermentum. The outcomes of the present study indicated that L. plantarum, L. pentosus, and L. fermentum were the dominant lactobacilli in the honey stomach of honeybee A. dorsata collected during the dry season from Malaysia forest area - specifically "Melaleuca in Terengganu".
Discovery and Development of Novel Anti-fungal Peptides Against Foodspoiling Fungi

Muhialdin BJ, Algboory HL, Mohammed NK, Kadum H, Hussin ASM, Saari N, et al.

Curr Drug Discov Technol, 2020;17(4):553-561.
PMID: 31309892 DOI: 10.2174/1570163816666190715120038

BACKGROUND: Despite the extensive research carried out to develop natural antifungal preservatives for food applications, there are very limited antifungal agents available to inhibit the growth of spoilage fungi in processed foods. Scope and Approach: Therefore, this review summarizes the discovery and development of antifungal peptides using lactic acid bacteria fermentation to prevent food spoilage by fungi. The focus of this review will be on the identification of antifungal peptides, potential sources, the possible modes of action and properties of peptides considered to inhibit the growth of spoilage fungi. Key Findings and Conclusions: Antifungal peptides generated by certain lactic acid bacteria strains have a high potential for applications in a broad range of foods. The mechanism of peptides antifungal activity is related to their properties such as low molecular weight, concentration and secondary structure. The antifungal peptides were proposed to be used as bio-preservatives to reduce and/or replace chemical preservatives.
Fulltext Mitigation of antinutritional factors and protease inhibitors of defatted winged bean-seed proteins using thermal and hydrothermal treatments: Denaturation/unfolding coupled hydrolysis mechanism

Saadi S, Saari N, Ghazali HM, Abdulkarim MS

Curr Res Food Sci, 2022;5:207-221.
PMID: 35106485 DOI: 10.1016/j.crfs.2022.01.011

The inactivation of antinutritional factors, protease inhibitors within winged bean protein was induced by two respective method treatments. The physical method based on steam vapor that was conducted using an autoclave and chemical method consisting on pH-gradients of buffer solutions prepared at respective acidic pH, neutral pH and alkaline pH ranges. The activity of remaining protease inhibitors of bowman birk inhibitor (BBI), and kunitz-trypsin inhibitor (KTI) after and before treatments was enzymatically confirmed using relevant antagonistic trypsin and combined trypsin-α-chymotrypsin digests. The resulting molecular assembly indicating an interval molecular relaxation range of °0.16 < °DA < °0.2 corresponding to reconformation in protein units with volume-mass changes of -2.17 < ∂v' < +2.17 and with denaturation/unfolding efficiency based on heat capacity ΔCp of 36.36 < DE/UF% < 54.67. These structural changes had a great benefit in determining and producing functional protein hydrolysates.
Use of prebiotics in oral delivery of bioactive compounds: a nanotechnology perspective

Heidarpour F, Mohammadabadi MR, Zaidul IS, Maherani B, Saari N, Hamid AA, et al.

Pharmazie, 2011 May;66(5):319-24.
PMID: 21699064

The oral route is considered the most patient-convenient means of drug administration. In recent years there has been a tendency to employ smart carrier systems that enable controlled or timed release of a bioactive material, thereby providing a better dosing pattern and minimizing side effects. Nano-encapsulation systems (nanocarriers) offer important advantages over conventional drug delivery techniques. Nanocarriers can protect the drug from chemical/enzymatic degradation and enhance bioavailability. Prebiotics are ideal ingredients for the nano-encapsulation and oral drug delivery due to their natural ability to protect the encapsulated compound in the upper gasterointestinal (GI) tract. Here the potential of prebiotics for oral delivery of drugs and other bioactives is reviewed.
Effects of skin abrasion in immersion challenge with Vibrio harveyi in Asian seabass Lates calcarifer fingerlings

Chin YK, Ina-Salwany MY, Zamri-Saad M, Amal MNA, Mohamad A, Lee JY, et al.

Dis Aquat Organ, 2020 Jan 16;137(3):167-173.
PMID: 31942862 DOI: 10.3354/dao03435

Skin abrasions often occur in farmed fish following handling by labourers, injury by farm facilities, cannibalism and ectoparasites. Vibrio spp. are opportunistic pathogens that can invade host fish through damaged tissues and cause outbreaks of vibriosis. This study describes the effect of skin abrasions on the infectivity of V. harveyi using Asian seabass Lates calcarifer (Bloch, 1790) fingerlings as a case example and compares bacterial load and fish survival following immersion challenge with different doses. In total, 315 fish (6.67 ± 1.8 g) were divided into 3 treatments: skin abrasion followed by immersion infection, immersion infection only and an uninfected, uninjured control. Fish in the infection treatments were divided into 3 subgroups and exposed in triplicate to a 7 d immersion challenge with 106, 107 and 108 CFU ml-1 of live V. harveyi. No mortalities were observed in the control and immersion infection groups. However, fish in the skin abrasion treatment group that were infected with 108 CFU ml-1 of live V. harveyi showed signs of progressing disease throughout the experiment, which resulted in mortalities. Significantly higher bacterial loads (p < 0.05) were recorded in the intestine, liver and gills of the fish in this group. Fish in the skin abrasion treatment that were exposed to 107 and 108 CFU ml-1 of V. harveyi showed 100% mortality by Days 5 and 4, respectively. These findings confirm that skin injuries increase the susceptibility of seabass fingerlings to V. harveyi infection.
Acute oral toxicity study on Wistar rats fed microalgal protein hydrolysates from Bellerochea malleus

Barkia I, Ketata Bouaziz H, Sellami Boudawara T, Aleya L, Gargouri AF, Saari N

Environ Sci Pollut Res Int, 2020 Jun;27(16):19087-19094.
PMID: 30612348 DOI: 10.1007/s11356-018-4007-6

Protein hydrolysates and bioactive peptides from various protein sources have demonstrated their effectiveness for the prevention of illness and the improvement of symptoms from several diseases. In particular, the use of microalgae to generate bioactive peptides has received a growing interest because of their potential to be cultivated on non-arable land and high nutritional value. However, scant research is available on the toxicity of peptide-based preparations. The present study aims to evaluate the toxicity of microalgal protein hydrolysates (MPH) from one marine species of microalgae (Bellerochea malleus) to determine the feasibility of their use for functional food applications. Results showed that the oral administration of MPH at three doses (D1, 100 mg kg-1 BW; D2, 400 mg kg-1 BW; and D3, 2000 mg kg-1 BW) to male Wistar rats did not induce any adverse effects or mortality up to13 days of treatment. Data analysis of relative organ weights and biochemical and hematological parameters did not show any significant differences between control and treated groups at the three doses investigated. Data from histopathological observations did not reveal any signs of major toxicity at the doses D1 and D2. However, mild signs of inflammation and necrosis were observed in the kidney of rats fed MPH at D3. All together, these results reveal the overall safety of MPH and provide new evidence for advocating their use for functional food or nutraceutical applications.
Production of glycerolamines based conjugated γ-aminobutyric acids using microbial COX and LOX as successor enzymes to GAD

Yavarzadeh M, Anwar F, Saadi S, Saari N

Enzyme Microb Technol, 2023 Sep;169:110282.
PMID: 37393814 DOI: 10.1016/j.enzmictec.2023.110282

Gamma-aminobutyric acid (γ-ABA) can be produced by various microorganisms including bacteria, fungi and yeasts using enzymatic bioconversion, microbial fermentation or chemical hydrolysis. Regenerating conjugated glycerol-amines is valid by the intervention of microbial cyclooxygenase [COX] and lipooxygenase [LOX] enzymes produced via lactobacillus bacteria (LAB) as successor enzymes to glutamate decarboxylases (GAD). Therefore, the aim of this review is to provide an overview on γ-ABA production, and microbiological achievements used in producing this signal molecule based on those fermenting enzymes. The formation of aminoglycerides based conjugated γ-ABA is considered the key substances in controlling the host defense against pathogens and is aimed in increasing the neurotransmission effects and in suppressing further cardiovascular diseases.
Fulltext Enzyme Hydrolysates from Stichopus horrens as a New Source for Angiotensin-Converting Enzyme Inhibitory Peptides

Forghani B, Ebrahimpour A, Bakar J, Abdul Hamid A, Hassan Z, Saari N

Evid Based Complement Alternat Med, 2012;2012:236384.
PMID: 22927875 DOI: 10.1155/2012/236384

Stichopus horrens flesh was explored as a potential source for generating peptides with angiotensin-converting enzyme (ACE) inhibitory capacity using 6 proteases, namely alcalase, flavourzyme, trypsin, papain, bromelain, and protamex. Degree of hydrolysis (DH) and peptide profiling (SDS-PAGE) of Stichopus horrens hydrolysates (SHHs) was also assessed. Alcalase hydrolysate showed the highest DH value (39.8%) followed by flavourzyme hydrolysate (32.7%). Overall, alcalase hydrolysate exhibited the highest ACE inhibitory activity (IC(50) value of 0.41 mg/mL) followed by flavourzyme hydrolysate (IC(50) value of 2.24 mg/mL), trypsin hydrolysate (IC(50) value of 2.28 mg/mL), papain hydrolysate (IC(50) value of 2.48 mg/mL), bromelain hydrolysate (IC(50) value of 4.21 mg/mL), and protamex hydrolysate (IC(50) value of 6.38 mg/mL). The SDS-PAGE results showed that alcalase hydrolysate represented a unique pattern compared to others, which yielded potent ACE inhibitory peptides with molecular weight distribution lower than 20 kDa. The evaluation of the relationship between DH and IC(50) values of alcalase and flavourzyme hydrolysates revealed that the trend between those parameters was related to the type of the protease used. We concluded that the tested SHHs would be used as a potential source of functional ACE inhibitory peptides for physiological benefits.

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