Displaying publications 21 - 40 of 128 in total

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  1. Fulltext Relationship Between Adherence to Opioid Analgesics and Pain Beliefs Among Patients with Cancer Pain at Tertiary Care Hospitals in Malaysia
    Kan E, Mustafa S, Chong WW, Premakumar CM, Mohamed Shah N
    Patient Prefer Adherence, 2020;14:1411-1419.
    PMID: 32848370 DOI: 10.2147/PPA.S255289
    Context: Pain is a common and distressing symptom among cancer patients. Opioid analgesics are the mainstay of cancer pain management, and adequate adherence plays an important role in achieving good pain control.

    Purpose: To determine the level of adherence to opioid analgesics in patients with cancer pain and to identify factors that may influence the adherence.

    Patient and Methods: This was a cross-sectional study conducted from March to June 2018 at two tertiary care hospitals in Malaysia. Study instruments consisted of a set of validated questionnaires; the Medication Compliance Questionnaire, Brief Pain Inventory and Pain Opioid Analgesic Beliefs─Cancer scale.

    Results: A total of 134 patients participated in this study. The patients' adherence scores ranged from 52-100%. Factors with a moderate, statistically significant negative correlation with adherence were negative effect beliefs (rs= -0.53, p<0.001), pain endurance beliefs (rs = -0.49, p<0.001) and the use of aqueous morphine (rs = -0.26, p=0.002). A multiple linear regression model on these predictors resulted in a final model which accounted for 47.0% of the total variance in adherence (R2 = 0.47, F (7, 126) = 15.75, p<0.001). After controlling for other variables, negative effect beliefs were the strongest contributor to the model (β = -0.39, p<0.001) and uniquely explained 12.3% of the total variance.

    Conclusion: The overall adherence to opioid analgesics among Malaysian patients with cancer pain was good. Negative effects beliefs regarding cancer pain and opioids strongly predicted adherence.

  2. Immunomodulatory effects of zerumbone isolated from roots of Zingiber zerumbet
    Keong YS, Alitheen NB, Mustafa S, Abdul Aziz S, Abdul Rahman M, Ali AM
    Pak J Pharm Sci, 2010 Jan;23(1):75-82.
    PMID: 20067871
    In this study, the immunomodulatory effects of zerumbone isolated from Zingiber zerumbet were investigated by evaluating the effects of this compound towards the lymphocytes proliferation (mice thymocytes, mice splenocytes and human human peripheral blood mononuclear cells, PBMC), cell cycle progression and cytokine (interleukin 2 and 12) induction. Lymphocyte proliferation assay showed that zerumbone was able to activate mice thymocytes, splenocytes and PBMC at dosage dependent pattern where the best concentration was 7.5 microg/ml. Flow cytometry analysis showed the highest population of PBMC entered into G2/M phase after treatment for 72 h with 7.5 microg/ml zerumbone. The production of human interleukin-2 and human interleukin-12 cytokines in culture supernatant from zerumbone activated lymphocytes was prominently upregulated at 24 hour and decreased from 48 h to 72 h. The above results indicate that zerumbone can be used as immunomodulatory agent which can react toward the immune cell cytokine production in dosage dependent pattern.
  3. Fulltext The Relationship of Orofacial Pain and Dental Health Status and Oral Health Behaviours in Facial Burn Patients
    Chaudhary FA, Ahmad B, Javed MQ, Yakub SS, Arjumand B, Khan AM, et al.
    Pain Res Manag, 2021;2021:5512755.
    PMID: 34055118 DOI: 10.1155/2021/5512755
    This study aims to examine the association of orofacial pain and oral health status and oral health behaviours in facial burn patients. The participants in this cross-sectional study were randomly recruited from the Burn Care Center, Institute of Medical Sciences, Islamabad, Pakistan. An intraoral evaluation was carried out to record the DMFT and OHI-S. A self-administered questionnaire was used to collect information on sociodemographic status, brushing frequency, and dental visits. Orofacial pain during mandibular movement was assessed using the Visual Analogue Scale (VAS). Psychological status was assessed using the Generalized Anxiety Disorder Scale and Impact of Events Scale. ANOVA and simple and multiple linear regression tests were used to analyse the data. From the 90 facial burn patients included, the majority were below 34 years of age, female, single or divorced, and unemployed. The mean DMFT was 10.7, and 71% had poor oral hygiene. 56% of the participants had moderate-to-severe anxiety, and 68% had posttraumatic stress disorder. 53% of the participants had moderate-to-severe pain during mouth opening or moving the mandible with a mean score of 41.5. Analyses showed that orofacial pain was associated with less frequent brushing, irregular dental visits, greater DMFT score, and more plaque accumulation (OHI-S). It was also associated with employment status, the severity of a burn, anxiety, and stress. The treatment and management of dental and oral conditions in burn patients need judicious balance in controlling and accurate assessment of the pain and improving psychological problems in burn patients.
  4. Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
    Ali ME, Hashim U, Mustafa S, Man YB, Yusop MH, Bari MF, et al.
    Nanotechnology, 2011 May 13;22(19):195503.
    PMID: 21430321 DOI: 10.1088/0957-4484/22/19/195503
    We used 40 ± 5 nm gold nanoparticles (GNPs) as colorimetric sensor to visually detect swine-specific conserved sequence and nucleotide mismatch in PCR-amplified and non-amplified mitochondrial DNA mixtures to authenticate species. Colloidal GNPs changed color from pinkish-red to gray-purple in 2 mM PBS. Visually observed results were clearly reflected by the dramatic reduction of surface plasmon resonance peak at 530 nm and the appearance of new features in the 620-800 nm regions in their absorption spectra. The particles were stabilized against salt-induced aggregation upon the adsorption of single-stranded DNA. The PCR products, without any additional processing, were hybridized with a 17-base probe prior to exposure to GNPs. At a critical annealing temperature (55 °C) that differentiated matched and mismatched base pairing, the probe was hybridized to pig PCR product and dehybridized from the deer product. The dehybridized probe stuck to GNPs to prevent them from salt-induced aggregation and retained their characteristic red color. Hybridization of a 27-nucleotide probe to swine mitochondrial DNA identified them in pork-venison, pork-shad and venison-shad binary admixtures, eliminating the need of PCR amplification. Thus the assay was applied to authenticate species both in PCR-amplified and non-amplified heterogeneous biological samples. The results were determined visually and validated by absorption spectroscopy. The entire assay (hybridization plus visual detection) was performed in less than 10 min. The LOD (for genomic DNA) of the assay was 6 µg ml(-1) swine DNA in mixed meat samples. We believe the assay can be applied for species assignment in food analysis, mismatch detection in genetic screening and homology studies between closely related species.
  5. Fulltext [Zinc induces normoxic accumulation of transcriptionally active hypoxia-inducible factor 1-alpha in mammary epithelial cells]
    Lee SY, Mustafa S, Ching YW, Shafee N
    Mol Biol (Mosk), 2017 3 3;51(1):104-110.
    PMID: 28251972 DOI: 10.7868/S0026898417010116
    Both zinc and the α-subunit of hypoxia-inducible factor (HIF-1α) play important roles in the remodelling of mammary gland tissues. In the present study, we examined the level and the transcriptional activity of HIF-1α in mammary cells upon zinc treatment. In MCF-7 mammary adenocarcinoma and MCF-10A mammary epithelial cell lines, the toxicity levels of zinc differ. Interestingly, both cell lines overexpress HIF-1α following zinc treatment. As it was evident from an up-regulation of its specific target gene CA9 that encodes carbonic anhydrase IX, the stabilized HIF-1α translocated to the nucleus and was transcriptionally active. Hence, we conclude that zinc causes normoxic accumulation of transcriptionally active HIF-1α by interfering with its post-translational regulation.
  6. Fulltext Prebiotic activity of polysaccharides extracted from Gigantochloa levis (Buluh beting) shoots
    Azmi AF, Mustafa S, Hashim DM, Manap YA
    Molecules, 2012 Feb 07;17(2):1635-51.
    PMID: 22314383 DOI: 10.3390/molecules17021635
    Bamboo shoot crude polysaccharides (BSCP) extracted from the shoots of Gigantochloa levis gave about 3.27 ± 0.18% on dry basis and a very minute percentage of protein (0.02 ± 0.01%). The molecular weight of BSCP estimated by gel chromatography was found to be around 7.49 × 103 Da, while the molecular weights of purified fractions (F1 to F5) were around 1550.96, 1471.63, 1685.78, 1691.61 and 1551.67 Da, respectively. The FTIR spectrum of BSCP revealed the possibility that the extract contains β-glucan, which can be considered a valuable compound for the medical and food industries. These relate to the resistance of BSCP towards artificial human gastric juice which is more than 99%. Prebiotic activity tested using BSCP as a carbon source showed significant increase in the growth of B. animalis ATCC 1053, B. longum BB 536 and L. acidophilus ATCC 4356 as compared to the use of FOS. Survivality of S. choleraesuis JCM 6977 was found to be slower in both BSCP and FOS. Study conducted reflects a good sign for the BSCP to be exploited as a promising prebiotic.
  7. Fulltext Corn silk (Stigma maydis) in healthcare: a phytochemical and pharmacological review
    Hasanudin K, Hashim P, Mustafa S
    Molecules, 2012 Aug 13;17(8):9697-715.
    PMID: 22890173 DOI: 10.3390/molecules17089697
    Corn silk (Stigma maydis) is an important herb used traditionally by the Chinese, and Native Americans to treat many diseases. It is also used as traditional medicine in many parts of the world such as Turkey, United States and France. Its potential antioxidant and healthcare applications as diuretic agent, in hyperglycemia reduction, as anti-depressant and anti-fatigue use have been claimed in several reports. Other uses of corn silk include teas and supplements to treat urinary related problems. The potential use is very much related to its properties and mechanism of action of its plant's bioactive constituents such as flavonoids and terpenoids. As such, this review will cover the research findings on the potential applications of corn silk in healthcare which include its phytochemical and pharmacological activities. In addition, the botanical description and its toxicological studies are also included.
  8. Fulltext Direct purification of pectinase from mango (Mangifera Indica Cv. Chokanan) peel using a PEG/salt-based Aqueous Two Phase System
    Mehrnoush A, Sarker MZ, Mustafa S, Yazid AM
    Molecules, 2011 Oct 10;16(10):8419-27.
    PMID: 21986520 DOI: 10.3390/molecules16108419
    An Aqueous Two-Phase System (ATPS) was employed for the first time for the separation and purification of pectinase from mango (Mangifera Indica Cv. Chokanan) peel. The effects of different parameters such as molecular weight of the polymer (polyethylene glycol, 2,000-10,000), potassium phosphate composition (12-20%, w/w), system pH (6-9), and addition of different concentrations of neutral salts (0-8%, w/w) on partition behavior of pectinase were investigated. The partition coefficient of the enzyme was decreased by increasing the PEG molecular weight. Additionally, the phase composition showed a significant effect on purification factor and yield of the enzyme. Optimum conditions for purification of pectinase from mango peel were achieved in a 14% PEG 4000-14% potassium phosphate system using 3% (w/w) NaCl addition at pH 7.0. Based on this system, the purification factor of pectinase was increased to 13.2 with a high yield of (97.6%). Thus, this study proves that ATPS can be an inexpensive and effective method for partitioning of pectinase from mango peel.
  9. Fulltext 'Heat-treatment aqueous two phase system' for purification of serine protease from Kesinai (Streblus asper) leaves
    Mehrnoush A, Mustafa S, Yazid AM
    Molecules, 2011 Dec 08;16(12):10202-13.
    PMID: 22158589 DOI: 10.3390/molecules161210202
    A 'Heat treatment aqueous two phase system' was employed for the first time to purify serine protease from kesinai (Streblus asper) leaves. In this study, introduction of heat treatment procedure in serine protease purification was investigated. In addition, the effects of different molecular weights of polyethylene glycol (PEG 4000, 6000 and 8000) at concentrations of 8, 16 and 21% (w/w) as well as salts (Na-citrate, MgSO₄ and K₂HPO₄) at concentrations of 12, 15, 18% (w/w) on serine protease partition behavior were studied. Optimum conditions for serine protease purification were achieved in the PEG-rich phase with composition of 16% PEG6000-15% MgSO₄. Also, thermal treatment of kesinai leaves at 55 °C for 15 min resulted in higher purity and recovery yield compared to the non-heat treatment sample. Furthermore, this study investigated the effects of various concentrations of NaCl addition (2, 4, 6 and 8% w/w) and different pH (4, 7 and 9) on the optimization of the system to obtain high yields of the enzyme. The recovery of serine protease was significantly enhanced in the presence of 4% (w/w) of NaCl at pH 7.0. Based on this system, the purification factor was increased 14.4 fold and achieved a high yield of 96.7%.
  10. Fulltext Optimization of the conditions for extraction of serine protease from kesinai plant (Streblus asper) leaves using response surface methodology
    Mehrnoush A, Mustafa S, Sarker MZ, Yazid AM
    Molecules, 2011 Nov 03;16(11):9245-60.
    PMID: 22051935 DOI: 10.3390/molecules16119245
    Response surface methodology (RSM) using a central composite design (CCD) was employed to optimize the conditions for extraction of serine protease from kesinai (Streblus asper) leaves. The effect of independent variables, namely temperature (42.5,47.5, X₁), mixing time (2-6 min, X₂), buffer content (0-80 mL, X₃) and buffer pH (4.5-10.5, X₄) on specific activity, storage stability, temperature and oxidizing agent stability of serine protease from kesinai leaves was investigated. The study demonstrated that use of the optimum temperature, mixing time, buffer content and buffer pH conditions protected serine protease during extraction, as demonstrated by low activity loss. It was found that the interaction effect of mixing time and buffer content improved the serine protease stability, and the buffer pH had the most significant effect on the specific activity of the enzyme. The most desirable conditions of 2.5 °C temperature, 4 min mixing time, 40 mL buffer at pH 7.5 was established for serine protease extraction from kesinai leaves.
  11. Fulltext A Novel Aqueous Two Phase System Composed of Surfactant and Xylitol for the Purification of Lipase from Pumpkin (Cucurbita moschata) Seeds and Recycling of Phase Components
    Amid M, Manap MY, Hussin M, Mustafa S
    Molecules, 2015 Jun 17;20(6):11184-201.
    PMID: 26091076 DOI: 10.3390/molecules200611184
    Lipase is one of the more important enzymes used in various industries such as the food, detergent, pharmaceutical, textile, and pulp and paper sectors. A novel aqueous two-phase system composed of surfactant and xylitol was employed for the first time to purify lipase from Cucurbita moschata. The influence of different parameters such as type and concentration of surfactants, and the composition of the surfactant/xylitol mixtures on the partitioning behavior and recovery of lipase was investigated. Moreover, the effect of system pH and crude load on the degree of purification and yield of the purified lipase were studied. The results indicated that the lipase was partitioned into the top surfactant rich phase while the impurities partitioned into the bottom xylitol-rich phase using an aqueous two phase system composed of 24% (w/w) Triton X-100 and 20% (w/w) xylitol, at 56.2% of tie line length (TLL), (TTL is one of the important parameters in this study and it is determined from a bimodal curve in which the tie-line connects two nodes on the bimodal, that represent concentration of phase components in the top and bottom phases) and a crude load of 25% (w/w) at pH 8.0. Recovery and recycling of components was also measured in each successive step process. The enzyme was successfully recovered by the proposed method with a high purification factor of 16.4 and yield of 97.4% while over 97% of the phase components were also recovered and recycled. This study demonstrated that the proposed novel aqueous two phase system method is more efficient and economical than the traditional aqueous two phase system method for the purification and recovery of the valuable enzyme lipase.
  12. Fulltext Comprehensive Review on Application of FTIR Spectroscopy Coupled with Chemometrics for Authentication Analysis of Fats and Oils in the Food Products
    Rohman A, Ghazali MAB, Windarsih A, Irnawati, Riyanto S, Yusof FM, et al.
    Molecules, 2020 Nov 23;25(22).
    PMID: 33238638 DOI: 10.3390/molecules25225485
    Currently, the authentication analysis of edible fats and oils is an emerging issue not only by producers but also by food industries, regulators, and consumers. The adulteration of high quality and expensive edible fats and oils as well as food products containing fats and oils with lower ones are typically motivated by economic reasons. Some analytical methods have been used for authentication analysis of food products, but some of them are complex in sampling preparation and involving sophisticated instruments. Therefore, simple and reliable methods are proposed and developed for these authentication purposes. This review highlighted the comprehensive reports on the application of infrared spectroscopy combined with chemometrics for authentication of fats and oils. New findings of this review included (1) FTIR spectroscopy combined with chemometrics, which has been used to authenticate fats and oils; (2) due to as fingerprint analytical tools, FTIR spectra have emerged as the most reported analytical techniques applied for authentication analysis of fats and oils; (3) the use of chemometrics as analytical data treatment is a must to extract the information from FTIR spectra to be understandable data. Next, the combination of FTIR spectroscopy with chemometrics must be proposed, developed, and standardized for authentication and assuring the quality of fats and oils.
  13. Fulltext Probiotic Properties of Exopolysaccharide-Producing Lactobacillus Strains Isolated from Tempoyak
    Khalil ES, Abd Manap MY, Mustafa S, Alhelli AM, Shokryazdan P
    Molecules, 2018 Feb 13;23(2).
    PMID: 29438288 DOI: 10.3390/molecules23020398
    Tempoyak is a functional Malaysian food (an acid-fermented condiment) which is produced from the pulp of the durian (Durio zibethinus) fruit. The current study aimed to isolate and identify potential exopolysaccharide (EPS)-producing Lactobacillus strains from tempoyak for potential use as probiotics. Seven isolates (DUR2, DUR4, DUR5, DUR8, DUR12, DUR18, and DUR20) out of 44 were able to produce EPS, and exhibited resistance to acid and bile salt compared to the reference strains Lactobacillus rhmnosus (ATCC53103) and L. plantarum (ATCC8014). The seven isolated strains belonged to five different species-L. plantarum, L. fermentum, L. crispatus, L. reuteri, and L. pentosus-which were identified using API 50 CHL and 16S rRNA gene sequences (Polymerase chain reaction, PCR - based). The seven strains displayed different ability to produce EPS (100-850 mg/L). Isolates exhibited a high survivability to acid (pH 3.0), bile salts (0.3%), and gastrointestinal tract model (<70%). Results showed that the auto-aggregation and cell surface hydrophobicity ranged from 39.98% to 60.09% and 50.80% to 80.53%, respectively, whereas, the highest co-aggregation value (66.44%) was observed by L. fermentum (DUR8) with Pseudomonas aeruginosa. The isolates showed good inhibitory activity against tested pathogens, high antioxidant activity (32.29% to 73.36%), and good ability to reduce cholesterol (22.55% to 75.15%). Thus, the seven tested strains have value as probiotics.
  14. Fulltext Tentative identification of volatile flavor compounds in commercial Budu, a Malaysian fish sauce, using GC-MS
    Mohamed HN, Man YC, Mustafa S, Manap YA
    Molecules, 2012 May 03;17(5):5062-80.
    PMID: 22555296 DOI: 10.3390/molecules17055062
    Budu is a famous Malaysian fish sauce, usually used as seasoning and condiment in cooking. Budu is produced by mixing fish and salt at certain ratio followed by fermentation for six months in closed tanks. In this study, four commercial brands of Budu were analyzed for their chemical properties (pH, salt content and volatile compounds). The pH of Budu samples ranged from 4.50-4.92, while the salt (NaCl) content ranged between 11.80% and 22.50% (w/v). For tentative identification of volatile flavor compounds in Budu, two GC columns have been used, DB-WAX and HP-5MS. A total of 44 volatile compounds have been detected and 16 were common for both columns. 3-Methyl-1-butanol, 2-methylbutanal, 3-methylbutanal, dimethyl disulfide, 3-(methylthio)-propanal, 3-methylbutanoic acid and benzaldehye have been identified as the aroma-active compounds in Budu due to their lower threshold values.
  15. Fulltext Evaluation of the Estimation Capability of Response Surface Methodology and Artificial Neural Network for the Optimization of Bacteriocin-Like Inhibitory Substances Production by Lactococcus lactis Gh1
    Jawan R, Abbasiliasi S, Tan JS, Kapri MR, Mustafa S, Halim M, et al.
    Microorganisms, 2021 Mar 12;9(3).
    PMID: 33809201 DOI: 10.3390/microorganisms9030579
    Bacteriocin-like inhibitory substances (BLIS) produced by Lactococcus lactis Gh1 had shown antimicrobial activity against Listeria monocytogenes ATCC 15313. Brain Heart Infusion (BHI) broth is used for the cultivation and enumeration of lactic acid bacteria, but there is a need to improve the current medium composition for enhancement of BLIS production, and one of the approaches is to model the optimization process and identify the most appropriate medium formulation. Response surface methodology (RSM) and artificial neural network (ANN) models were employed in this study. In medium optimization, ANN (R2 = 0.98) methodology provided better estimation point and data fitting as compared to RSM (R2 = 0.79). In ANN, the optimal medium consisted of 35.38 g/L soytone, 16 g/L fructose, 3.25 g/L sodium chloride (NaCl) and 5.40 g/L disodium phosphate (Na2HPO4). BLIS production in optimal medium (717.13 ± 0.76 AU/mL) was about 1.40-fold higher than that obtained in nonoptimised (520.56 ± 3.37 AU/mL) medium. BLIS production was further improved by about 1.18 times higher in 2 L stirred tank bioreactor (787.40 ± 1.30 AU/mL) as compared to that obtained in 250 mL shake flask (665.28 ± 14.22 AU/mL) using the optimised medium.
  16. Influence of Culture Conditions and Medium Compositions on the Production of Bacteriocin-Like Inhibitory Substances by Lactococcus lactis Gh1
    Jawan R, Abbasiliasi S, Tan JS, Mustafa S, Halim M, Ariff AB
    Microorganisms, 2020 Sep 23;8(10).
    PMID: 32977375 DOI: 10.3390/microorganisms8101454
    Antibacterial peptides or bacteriocins produced by many strains of lactic acid bacteria have been used as food preservatives for many years without any known adverse effects. Bacteriocin titres can be modified by altering the physiological and nutritional factors of the producing bacterium to improve the production in terms of yield and productivity. The effects of culture conditions (initial pH, inoculum age and inoculum size) and medium compositions (organic and inorganic nitrogen sources; carbon sources) were assessed for the production of bacteriocin-like inhibitory substances (BLIS) by Lactococcus lactis Gh1 in shake flask cultures. An inoculum of the mid-exponential phase culture at 1% (v/v) was the optimal age and size, while initial pH of culture media at alkaline and acidic state did not show a significant impact on BLIS secretion. Organic nitrogen sources were more favourable for BLIS production compared to inorganic sources. Production of BLIS by L. lactis Gh1 in soytone was 1.28-times higher as compared to that of organic nitrogen sources ((NH4)2SO4). The highest cell concentration (XmX = 0.69 ± 0.026 g·L-1) and specific growth rate (μmax = 0.14 h-1) were also observed in cultivation using soytone. By replacing carbon sources with fructose, BLIS production was increased up to 34.94% compared to BHI medium, which gave the biomass cell concentration and specific growth rate of 0.66 ± 0.002 g·L-1 and 0.11 h-1, respectively. It can be concluded that the fermentation factors have pronounced influences on the growth of L. lactis Gh1 and BLIS production. Results from this study could be used for subsequent application in process design and optimisation for improving BLIS production by L. lactis Gh1 at larger scale.
  17. Evaluation of intestinal damage caused by V. cholerae O139, an in vivo study
    Murugaiah C, Noor NZ, Mustafa S, Manickam R, Pattabhiraman L
    Microb Pathog, 2017 Apr;105:25-29.
    PMID: 28179117 DOI: 10.1016/j.micpath.2017.02.002
    Cholera, a severe form of gastroenteritis, is one of the most widespread diseases in developing countries. The mechanism of intestinal infection caused by V. cholerae O139 remains unclear. In order to explore some morphological aspects of its infection in the intestine including Peyer's patches, we investigated the V. cholerae O139 infection at intestinal site of the rabbit gut-loop model. The electron microscopic analysis revealed denuded mucosal surface with loss of microvilli and integrity of the surface epithelium. Infection of the intestine with V. cholerae O139 induces destruction of villi, microvilli and lining epithelium with exposure of crypts of Lieberkuhn.
  18. Immunohistochemical, histological and ultrastructural evaluation of protection provided by cholera vaccine against V. cholerae O139
    Murugaiah C, Nik Mohd Noor NZ, Al-Talib H, Mustafa S, Manickam R, Pattabhiraman L
    Microb Pathog, 2020 Mar;140:103964.
    PMID: 31904450 DOI: 10.1016/j.micpath.2020.103964
    In our previous study, complete protection was observed in rabbit immunized with 1 × 1010 CFU of live attenuated VCUSM21P vaccine against challenge with 1 × 109 CFU Vibrio cholerae O139. In the present study, we investigated whether the vaccines can effectively protect immunized animals from any pathologic changes using histological, immunohistochemical and ultrastructural techniques. Severe pathology is evident in wild type injected ileum in non-immunized, showing extensive villous destruction, edema, necrosis and inflammation with infiltration of large numbers of inflammatory cells, extensive damage to the villi and microvilli with pore formation. Histology of ileum injected with wild type in immunized rabbit shows no significant pathological changes except for a few inflammatory cells in lamina propria with mild edema in mucosa and submucosa. immunohistochemical staining revealed O139 antigens of wild type are seen in the lamina propria of edematous villi, muscularis mucosa and submucosa with weak presence in the muscle coat in non-immunized rabbit after challenged with wild type in non-immunized rabbits, but in immunized rabbit localisation of the O139 LPS antigen is seen at the tips of the intact villi, within lamina propria and muscularis mucosa only. These observations suggest that the vaccine can effectively protect animals from any pathologic changes and eliminate V. cholerae O139 from the immunized animals.
  19. Evaluation of three Bacillus spp. isolated from the gut of giant freshwater prawn as potential probiotics against pathogens causing Vibriosis and Aeromonosis
    Sam-On MFS, Mustafa S, Yusof MT, Mohd Hashim A, Abbasiliasi S, Zulkifly S, et al.
    Microb Pathog, 2022 Jan 24;164:105417.
    PMID: 35085717 DOI: 10.1016/j.micpath.2022.105417
    Pathogens from the Vibrio and Aeromonas genera often cause detrimental effects to the aquaculture sector. Previously, antibiotics was used to resolve the infections, but this caused the spread of antibiotic resistant bacteria and antibiotic resistance genes into the environment. As an approach to address this issue, probiotic bacteria were introduced to improve the hosts' microbiome, disease protection, health condition, growth efficiency, feed consumption, stress response and general vigour. However, reports showed that some commercially available probiotics were restricted to a small number of microbial species and there are inconsistencies concerning its effectiveness. Hence, the aim of this study was to isolate and evaluate new Bacillus spp. from the gut of giant freshwater prawn as potential probiotics. Three Bacillus spp. isolates, Bacillus subtilis FS6 (MZ960135), Bacillus velezensis FS26 (MZ960133) and Bacillus pumilus FS97 (MZ960136) were characterised, and in vitro testing showed good probiotic properties which can help in dealing with diseases in aquaculture. Among the Bacillus spp., Bacillus velezensis FS26 showed higher antimicrobial activity towards Aeromonas hydrophila LMG 13658 and Aeromonas veronii clone DK-A. veronii-27 at 23.7 mm and 25 mm, respectively. Bacillus subtilis FS6 and Bacillus velezensis FS26 resulted in good adherence to both xylene and chloroform hydrocarbons. The Bacillus spp. isolated displayed high survivability towards 0.3% bile salt and exhibited amylase, protease, and lipase activities. Thus, the isolated Bacillus spp. are considered safe based on the sensitivity analysis towards antibiotics and γ-haemolytic activity.
  20. Mining the genome of Bacillus velezensis FS26 for probiotic markers and secondary metabolites with antimicrobial properties against aquaculture pathogens
    Sam-On MFS, Mustafa S, Mohd Hashim A, Yusof MT, Zulkifly S, Malek AZA, et al.
    Microb Pathog, 2023 Aug;181:106161.
    PMID: 37207784 DOI: 10.1016/j.micpath.2023.106161
    Bacillus velezensis FS26 is a bacterium from the genus Bacillus that has been proven as a potential probiotic in aquaculture with a good antagonistic effect on Aeromonas spp. and Vibrio spp. Whole-genome sequencing (WGS) allows a comprehensive and in-depth analysis at the molecular level, and it is becoming an increasingly significant technique in aquaculture research. Although numerous probiotic genomes have been sequenced and investigated recently, there are minimal data on in silico analysis of B. velezensis as a probiotic bacterium isolated from aquaculture sources. Thus, this study aims to analyse the general genome characteristics and probiotic markers from the B. velezensis FS26 genome with secondary metabolites predicted against aquaculture pathogens. The B. velezensis FS26 genome (GenBank Accession: JAOPEO000000000) assembly proved to be of high quality, with eight contigs containing 3,926,371 bp and an average G + C content of 46.5%. According to antiSMASH analysis, five clusters of secondary metabolites from the B. velezensis FS26 genome showed 100% similarity. These clusters include Cluster 2 (bacilysin), Cluster 6 (bacillibactin), Cluster 7 (fengycin), Cluster 8 (bacillaene), and Cluster 9 (macrolactin H), which signify promising antibacterial, antifungal, and anticyanobacterial agents against pathogens in aquaculture. The probiotic markers of B. velezensis FS26 genome for adhesion capability in the hosts' intestine, as well as the acid and bile salt-tolerant genes, were also detected through the Prokaryotic Genome Annotation System (Prokka) annotation pipeline. These results are in agreement with our previous in vitro data, suggesting that the in silico investigation facilitates establishing B. velezensis FS26 as a beneficial probiotic for use in aquaculture.
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