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  1. Fulltext Characterization of Edwardsiella tarda isolated from Asian Seabass, Lates calcarifer
    Nadirah, M., Najiah M., Teng, S. Y.
    International Food Research Journal, 2012;19(3):1247-1252.
    MyJurnal
    This study described the antibiotic and heavy metal resistance pattern of 17 isolates of Edwardsiella tarda obtained from Asian seabass (Lates calcarifer). E.tarda isolates were resistant to oleandomycin, lincomycin, novobiocin and spiramycin. In contrast, most of the isolates showed high level of susceptibility to tetracycline, doxycycline, florfenicol, chloramplenicol, nitrofurantoin, fosfomycin, kanamycin, oxolinic acid and flumequine. MAR value was 0.35 which indicated that the cultured Asian seabass have received high exposure to those tested antibiotics. Besides, very high level of heavy metal resistance among these isolates was observed. Genotypic profile of DNA fingerprintings generated by RAPD-PCR using M13 universal primer and M13 wild type phage primer showed high degree of genetic diversity with percentages similarity and genetic distance among the isolates were ranging from 10.5% to 100% and 0 to 0.895, respectively. This result indicates that strains that belong to the same origin were not always closely related genetically.
  2. Fulltext Differential responses of Vibrio sp. to young and mature leaves extracts of Terminalia catappa L
    Nadirah, M., Wee, T. L., Najiah, M.
    International Food Research Journal, 2013;20(2):961-966.
    MyJurnal
    Young and mature leaves of Terminalia catappa of alcoholic and aqueous extracts were evaluated for in vitro antibacterial activity against Vibrio sp. isolated from aquatic animals. Young leaves of T. catappa showed higher antibacterial activity when compared to mature leaves against Vibrio parahemolyticus, with methanolic and aqueous extracts exhibited the largest inhibition zones, 23 and 24 mm, respectively as determined by disc diffusion technique. Ethanolic extract of young leaves showed the lowest MIC and MBC at 3.13 mg/ml and 6.25 mg/ml, respectively. Both alcoholic and aqueous extracts of young and matures leaves exhibit variations in protein, RNA as well as pyrine and pyrimidines leakage of Vibrio sp. Cell membrane disruption is proposed as the mechanism of action of T. catappa leaves extract against Vibrio sp.
  3. Fulltext Antibiogram and heavy metal tolerance of bullfrog bacteria in Malaysia
    Tee LW, Najiah M
    Open Vet J, 2011;1(1):39-45.
    PMID: 26623279
    Bacterial isolates from 30 farmed bullfrogs (Lithobates catesbeianus) weighing 500-600 g at Johore, Malaysia with external clinical signs of ulcer, red leg and torticollis were tested for their antibiograms and heavy metal tolerance patterns. A total of 17 bacterial species with 77 strains were successfully isolated and assigned to 21 antibiotics and 4 types of heavy metal (Hg(2+), Cr(6+), Cd(2+), Cu(2+)). Results revealed that bacteria were resistant against lincomycin (92%), oleandomycin (72.7%) and furazolidone (71.4%) while being susceptible to chloramphenicol and florfenicol at 97.4%. The multiple antibiotic resistance (MAR) index for C. freundii, E. coli and M. morganii was high with the value up to 0.71. Bacterial strains were found to exhibit 100 % resistance to chromium and mercury. High correlation of resistance against both antibiotics and heavy metals was found (71.4 to 100%) between bullfrog bacteria isolates, except bacteria that were resistant to kanamycin showed only 25% resistance against Cu(2+). Based on the results in this study, bacterial pathogens of bullfrog culture in Johore, Malaysia, were highly resistant to both antibiotics and heavy metals.
  4. Fulltext Differentiation of chromoplasts and other plastids in plants
    Sadali NM, Sowden RG, Ling Q, Jarvis RP
    Plant Cell Rep, 2019 Jul;38(7):803-818.
    PMID: 31079194 DOI: 10.1007/s00299-019-02420-2
    Plant cells are characterized by a unique group of interconvertible organelles called plastids, which are descended from prokaryotic endosymbionts. The most studied plastid type is the chloroplast, which carries out the ancestral plastid function of photosynthesis. During the course of evolution, plastid activities were increasingly integrated with cellular metabolism and functions, and plant developmental processes, and this led to the creation of new types of non-photosynthetic plastids. These include the chromoplast, a carotenoid-rich organelle typically found in flowers and fruits. Here, we provide an introduction to non-photosynthetic plastids, and then review the structures and functions of chromoplasts in detail. The role of chromoplast differentiation in fruit ripening in particular is explored, and the factors that govern plastid development are examined, including hormonal regulation, gene expression, and plastid protein import. In the latter process, nucleus-encoded preproteins must pass through two successive protein translocons in the outer and inner envelope membranes of the plastid; these are known as TOC and TIC (translocon at the outer/inner chloroplast envelope), respectively. The discovery of SP1 (suppressor of ppi1 locus1), which encodes a RING-type ubiquitin E3 ligase localized in the plastid outer envelope membrane, revealed that plastid protein import is regulated through the selective targeting of TOC complexes for degradation by the ubiquitin-proteasome system. This suggests the possibility of engineering plastid protein import in novel crop improvement strategies.
  5. Bacteria associated with wild mud crab (Scylla serrata) from Setiu Wetland, Malaysia with emphasis on antibiotic resistances
    Najiah M, Nadirah M, Sakri I, Shaharom-Harrison F
    Pak J Biol Sci, 2010 Mar 15;13(6):293-7.
    PMID: 20506717
    A study was carried out to investigate the presence of bacteria flora in wild mud crab (Scylla serrata) from Setiu Wetland as well as their antibiotic resistances. A total of 91 bacterial isolates consisting of 12 bacterial species were successfully isolated from mud crab. Oxolinic acid was found to be effective against all the bacterial isolates whilst the highest percentage of antibiotic resistance was shown by lincomycin (94.5%) followed by ampicillin (90.1%), amoxicillin (86.8%) and oleandomycin (78.0%). The study is very useful to evaluate the safety of mud crab for human consumption based on wild mud crab-associated bacteria as well as their antibiotic resistances.
  6. Phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. in Malaysia
    Najiah M, Lee KL, Noorasikin H, Nadirah M, Lee SW
    Res Vet Sci, 2011 Dec;91(3):342-5.
    PMID: 20971487 DOI: 10.1016/j.rvsc.2010.09.010
    Mycobacteriosis due to mycobacteria is one of the most common bacterial diseases in ornamental fish. We describe here the phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. using ATCC Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium chelonae as references. A total of four isolates (M1, M2, M3, M4) were obtained from four out of 106 fish samples using selective agar, and identified to Mycobacterium genus using acid-fast staining and 16s rRNA gene-based genus specific polymerase chain reaction. DNA sequencing and NCBI-BLAST analysis further identified isolate M1 as M. marinum and isolates M2, M3, M4 as M. fortuitum. Morphological, physiological and biochemical tests were carried out for phenotypic characterizations. Universal M13 and wild-type phage M13 RAPD dendogram was generated to illustrate the genetic relationship of the isolates and reference strains.
  7. Occurrence of Plesiomonas shigelloides in cultured red hybrid tilapia (Oreochromis niloticus) from tropical rivers, east coast Malaysia
    Nadirah M, Ruhil HH, Jalal KC, Najiah M
    Pak J Biol Sci, 2012 Jun 15;15(12):600-3.
    PMID: 24191623
    A total of 182 isolates of Plesiomonas shigelloides were identified from 40 healthy red hybrid tilapia, Oreochromis niloticus cultured at two important rivers in Terengganu, Malaysia namely Como River and Terengganu River from east coast Malaysia. P. shigelloides count in Digestive Tract Content (DTC) and Muscle (MUS) of red hybrid tilapia cultured at Terengganu River was 1000-fold higher than Como River. Antibiotic susceptibility test was also performed on Plesiomonas shigelloides isolates. The incidence of antibiotic resistance was higher in Plesiomonas shigelloides isolated from red hybrid tilapia cultured at Terengganu River compared to Como river. Thus, the findings of the study indicate that P. shigelloides from tilapia muscle and an intestine could be an alarming for serious public health risk to consumers.
  8. A new bacterial white spot syndrome (BWSS) in cultured tiger shrimp Penaeus monodon and its comparison with white spot syndrome (WSS) caused by virus
    Wang YG, Lee KL, Najiah M, Shariff M, Hassan MD
    Dis Aquat Organ, 2000 May 25;41(1):9-18.
    PMID: 10907134
    This paper describes a new bacterial white spot syndrome (BWSS) in cultured tiger shrimp Penaeus monodon. The affected shrimp showed white spots similar to those caused by white spot syndrome virus (WSSV), but the shrimp remained active and grew normally without significant mortalities. The study revealed no evidence of WSSV infection using electron microscopy, histopathology and nested polymerase chain reaction. Electron microscopy indicated bacteria associated with white spot formation, and with degeneration and discoloration of the cuticle as a result of erosion of the epicuticle and underlying cuticular layers. Grossly the white spots in BWSS and WSS look similar but showed different profiles under wet mount microscopy. The bacterial white spots were lichen-like, having perforated centers unlike the melanized dots in WSSV-induced white spots. Bacteriological examination showed that the dominant isolate in the lesions was Bacillus subtilis. The occurrence of BWSS may be associated with the regular use of probiotics containing B. subtilis in shrimp ponds. The externally induced white spot lesions were localized at the integumental tissues, i.e., cuticle and epidermis, and connective tissues. Damage to the deeper tissues was limited. The BWS lesions are non-fatal in the absence of other complications and are usually shed through molting.
  9. Betanodavirus: Dissection of the viral life cycle
    Low CF, Syarul Nataqain B, Chee HY, Rozaini MZH, Najiah M
    J Fish Dis, 2017 Nov;40(11):1489-1496.
    PMID: 28449248 DOI: 10.1111/jfd.12638
    Progressive research has been recently made in dissecting the molecular biology of Betanodavirus life cycle, the causative pathogen of viral encephalopathy and retinopathy in economic important marine fish species. Establishment of betanodavirus infectious clone allows the manipulation of virus genome for functional genomic study, which elucidates the biological event of the viral life cycle at molecular level. The betanodavirus strategizes its replication by expressing anti-apoptosis/antinecrotic proteins to maintain the cell viability during early infection. Subsequently utilizes and controls the biological machinery of the infected cells for viral genome replication. Towards the late phase of infection, mass production of capsid protein for virion assembly induces the activation of host apoptosis pathway. It eventually leads to the cell lysis and death, which the lysis of cell contributes to the accomplishment of viral shedding that completes a viral life cycle. The recent efforts to dissect the entire betanodavirus life cycle are currently reviewed.
  10. Fulltext Editorial: Molecular mechanisms of bacterial disease in cultured fishes
    Najiah M, Huang L, Pang H, Ebani VV
    Front Vet Sci, 2022;9:1073631.
    PMID: 36467662 DOI: 10.3389/fvets.2022.1073631
  11. Efficacy of live attenuated vaccine derived from the Streptococcus agalactiae on the immune responses of Oreochromis niloticus
    Laith AA, Abdullah MA, Nurhafizah WWI, Hussein HA, Aya J, Effendy AWM, et al.
    Fish Shellfish Immunol, 2019 Jul;90:235-243.
    PMID: 31009810 DOI: 10.1016/j.fsi.2019.04.052
    Streptococcus agalactiae species have been recognized as the main pathogen causing high mortality in fish leading to significant worldwide economical losses to the aquaculture industries. Vaccine development has become a priority in combating multidrug resistance in bacteria; however, there is a lack of commercial live attenuated vaccine (LAV) against S. agalactiae in Malaysia. The aim of this study is to compare two methods using attenuated bacteria as live vaccine and to evaluate the efficacy of selected LAV on the immune responses and resistance of Oreochromis niloticus (tilapia) against S. agalactiae. The LAV derived from S. agalactiae had been weakened using the chemical agent Acriflavine dye (LAV1), whereas the second vaccine was weakened using serial passages of bacteria on broth media (LAV2). Initial immunization was carried out only on day one, given twice-in the morning and evening, for the 42 day period. Serum samples were collected to determine the systemic antibody (IgM) responses and lysozymal (LSZ) activity using ELISA. On day 43 after immunization, the fish were injected intraperitoneally (i.p) with 0.1 mL of S. agalactiae at LD50 = 1.5 × 105 (CFU)/fish. Fish were monitored daily for 10 days. Clinical signs, mortality and the relative percent of survival (RPS) were recorded. Trial 1 results showed a significant increased (P 
  12. Histopathological and immunological changes in green mussel, Perna viridis, challenged with Vibrio alginolyticus
    Laith AA, Ros-Amira MK, Sheikh HI, Effendy AWM, Najiah M
    Fish Shellfish Immunol, 2021 Nov;118:169-179.
    PMID: 34487829 DOI: 10.1016/j.fsi.2021.08.032
    Understanding of pathogenicity and immunity is crucial in producing disease-resistant cultured mollusk varieties. This study aimed to isolate pathogenic Vibrio alginolyticus from naturally infected Perna viridis, and to determine histopathological and immunological changes after challenge test with the same bacteria. Biochemical tests and 16S rDNA identified the pathogen as V. alginolyticus (99%). Antibiotic susceptibility test showed ampicillin resistance of the pathogen. Pathogenicity assay was conducted by immersing P. viridis in 1.5 × 106 CFU mL-1V. alginolyticus for 60 min and observed for 5 days. Clinical signs, histopathological and immunological alterations were observed and monitored. Infected groups showed 60% mortality and decreased immunity factors, including total hemocyte count and lysozymes activity. Histopathological examination revealed pathological lesions in the hepatopancreas at 24 h post-challenge and hemocyte proliferation as part of a severe inflammatory reaction. Karyomegaly in the hepatopancreas tissue, concomitant with necrosis demolition of tubules cells, was also observed. V. alginolyticus was determined to be pathogenic to P. viridis, causing mortality as a result of multiple organ lesions and dysfunction in digestive gland and immune organs. This study demonstrated the role of histopathological and immunological parameters as potential biomarkers in assessing vibriosis caused by Vibrio species in green mussel, P. viridis.
  13. Fulltext Effect of Excoecaria agallocha on non-specific immune responses and disease resistance of Oreochromis niloticus against Streptococcus agalactiae
    Laith AA, Mazlan AG, Effendy AW, Ambak MA, Nurhafizah WWI, Alia AS, et al.
    Res Vet Sci, 2017 Jun;112:192-200.
    PMID: 28499213 DOI: 10.1016/j.rvsc.2017.04.020
    The current study was designed to evaluate the effects of Excoecaria agallocha leaf extracts on immune mechanisms and resistance of tilapia, Oreochromis niloticus, after challenge with Streptococcus agalactiae. Fish were divided into 6 groups; groups 1-5 fed with E. agallocha leaf extracts at 10, 20, 30, 40 and 50mgkg(-1) level, respectively. Group 6 were fed without extract addition and acted as control. E. agallocha extracts were administered as feed supplement in fish diet for 28days and the hematological, immunological, and growth performance studies were conducted. Fish were infected with S. agalactiae at a dose of 15×105CFUmL(-1) and the total white blood cell (WBC), phagocytosis and respiratory burst activities of leukocytes, serum bactericidal activity, lysozyme, total protein, albumin, and globulin levels were monitored and mortalities recorded for 15days post infection. Results revealed that feeding O. niloticus with 50mgkg(-1) of E. agallocha enhanced WBC, phagocytic, respiratory burst, serum bactericidal and lysozyme activities on day 28 pre-challenge and on 3rd, 6th, 9th, 12th and 15th day post-challenge as compared to control. Total protein and albumin were not enhanced by E. agallocha diet. E. agallocha increased the survival of fish after challenge with S. agalactiae. The highest mortality rate (97%) was observed in control fish and the lowest mortality (27%) was observed with group fed with 50mgkg(-1) extract. The results indicate that dietary intake of E. agallocha methanolic leaf extract in O. niloticus enhances the non-specific immunity and disease resistance against S. agalactiae pathogen.
  14. Bacteria flora and heavy metals in cultivated oysters Crassostrea iredalei of Setiu Wetland, East Coast Peninsular Malaysia
    Najiah M, Nadirah M, Lee KL, Lee SW, Wendy W, Ruhil HH, et al.
    Vet Res Commun, 2008 Jun;32(5):377-81.
    PMID: 18369732 DOI: 10.1007/s11259-008-9045-y
    Slipper oyster Crassostrea iredalei is a species of good demand for its sweet flavor and white coloured flesh. The filter feeding nature predisposes oysters to accumulation of pathogenic and heavy metals in waters impacted by sewage pollutions and may thus render the oysters unfit for human consumption. A study was undertaken to investigate the presence of bacteria flora and heavy metal concentrations in cultivated oysters Crassostrea iredalei at Setiu Wetland, Terengganu, the only source of cultivated oysters in East Coast of Malaysia. A total of 200 slipper oyster samples were analyzed. The bacteria were isolated using non selective agar such as TSA agar and selective agars before they were then identified using conventional methods in combination with BBL Crystal identification kit. Heavy metals such as zinc (Zn), copper (Cu), cadmium (Cd), and lead (Pb) concentrations were determined using atomic absorption spectrophotometry. Results showed that the oysters harbor predominantly Shewanella putrifaciens followed by Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio cholerae, Enterobacter cloacae, Escherichia coli and Chromobacterium violaceum. They also contain high concentration of Zn (785.68 +/- 285.88 microg/g) with the lowest heavy metal was Pb (0.17 +/- 0.15 microg/g), whilst the concentrations of other heavy metals were Cu (38.9 +/- 13.2 microg/g) and Cd (1.60 +/- 0.28 microg/g). The study is very useful to evaluate the type of bacteria and heavy metal present in oyster meat for human consumption.
  15. Virulence properties and pathogenicity of multidrug-resistant Vibrio harveyi associated with luminescent vibriosis in Pacific white shrimp, Penaeus vannamei
    Nurhafizah WWI, Lee KL, Laith A AR, Nadirah M, Danish-Daniel M, Zainathan SC, et al.
    J Invertebr Pathol, 2021 11;186:107594.
    PMID: 33878330 DOI: 10.1016/j.jip.2021.107594
    Global high demand for Pacific white shrimp Penaeus vannamei has led to intensified cultivation and a wide range of disease problems, including bacterial diseases due to vibrios. Three presumptive luminescent Vibrio harveyi strains (Vh5, Vh8 and Vh10) were isolated from the hepatopancreas (Vh5) and haemolymph (Vh8 and Vh10) of diseased growout Pacific white shrimp from a farm in Setiu, Terengganu, Malaysia, using Vibrio harveyi agar (VHA) differential medium. All three strains were identified as V. harveyi by biochemical characteristics. 16S rRNA gene-based phylogenetic analyses by neighbour-joining, maximum likelihood and maximum parsimony methods showed all three strains in the V. harveyi cluster. All three strains were β-haemolytic and positive for motility, biofilm formation and extracellular products (caseinase, gelatinase, lipase, DNase, amylase and chitinase). Vh10 was subjected to pathogenicity test in Pacific white shrimp by immersion challenge and determined to have a LC50 of 6.0 × 108 CFU mL-1 after 168 h of exposure. Antibiotic susceptibility tests showed that all strains were resistant to oxytetracycline (OXT30), oleandomycin (OL15), amoxicillin (AML25), ampicillin (AMP10) and colistin sulphate (CT25) but sensitive to doxycycline (DO30), flumequine (UB30), oxolinic acid (OA2), chloramphenicol (C30), florfenicol (FFC30), nitrofurantoin (F5) and fosfomycin (FOS50). Each strain was also resistant to a slightly different combination of eight other antibiotics, with an overall multiple antibiotic resistance (MAR) index of 0.40, suggesting prior history of heavy exposure to the antibiotics. Vh10 infection resulted in pale or discoloured hepatopancreas, empty guts, reddening, necrosis and luminescence of uropods, as well as melanised lesions in tail muscle. Histopathological examination showed necrosis of intertubular connective tissue and tubule, sloughing of epithelial cells in hepatopancreatic tubule, haemocytic infiltration, massive vacuolation and loss of hepatopancreatic tubule structure.
  16. Fulltext Molecular identification and histopathological study of natural Streptococcus agalactiae infection in hybrid tilapia (Oreochromis niloticus)
    Laith AA, Ambak MA, Hassan M, Sheriff SM, Nadirah M, Draman AS, et al.
    Vet World, 2017 Jan;10(1):101-111.
    PMID: 28246454 DOI: 10.14202/vetworld.2017.101-111
    AIM: The main objective of this study was to emphasize on histopathological examinations and molecular identification of Streptococcus agalactiae isolated from natural infections in hybrid tilapia (Oreochromis niloticus) in Temerloh Pahang, Malaysia, as well as to determine the susceptibility of the pathogen strains to various currently available antimicrobial agents.

    MATERIALS AND METHODS: The diseased fishes were observed for variable clinical signs including fin hemorrhages, alterations in behavior associated with erratic swimming, exophthalmia, and mortality. Tissue samples from the eyes, brain, kidney, liver, and spleen were taken for bacterial isolation. Identification of S. agalactiae was screened by biochemical methods and confirmed by VITEK 2 and 16S rRNA gene sequencing. The antibiogram profiling of the isolate was tested against 18 standard antibiotics included nitrofurantoin, flumequine, florfenicol, amoxylin, doxycycline, oleandomycin, tetracycline, ampicillin, lincomycin, colistin sulfate, oxolinic acid, novobiocin, spiramycin, erythromycin, fosfomycin, neomycin, gentamycin, and polymyxin B. The histopathological analysis of eyes, brain, liver, kidney, and spleen was observed for abnormalities related to S. agalactiae infection.

    RESULTS: The suspected colonies of S. agalactiae identified by biochemical methods was observed as Gram-positive chained cocci, β-hemolytic, and non-motile. The isolate was confirmed as S. agalactiae by VITEK 2 (99% similarity), reconfirmed by 16S rRNA gene sequencing (99% similarity) and deposited in GenBank with accession no. KT869025. The isolate was observed to be resistance to neomycin and gentamicin. The most consistent gross findings were marked hemorrhages, erosions of caudal fin, and exophthalmos. Microscopic examination confirmed the presence of marked congestion and infiltration of inflammatory cell in the eye, brain, kidney, liver, and spleen. Eye samples showed damage of the lens capsule, hyperemic and hemorrhagic choroid tissue, and retina hyperplasia accompanied with edema. Brain samples showed perivascular and pericellular edema and hemorrhages of the meninges. Kidney samples showed hemorrhage and thrombosis in the glomeruli and tubules along with atrophy in hematopoietic tissue. Liver samples showed congestion of the sinusoids and blood vessel, thrombosis of portal blood vessel, and vacuolar (fatty) degeneration of hepatocytes. Spleen samples showed large thrombus in the splenic blood vessel, multifocal hemosiderin deposition, congestion of blood vessels, and multifocal infiltration of macrophages.

    CONCLUSION: Therefore, it can be concluded that pathological changes in tissues and organs of fish occur proportionally to the pathogen invasion, and because of their high resistance, neomycin and gentamicin utilization in the prophylaxis or treatment of S. agalactiae infection should be avoided.

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