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Contact micropredation may play a more important role than exotoxicity does in the lethal effects of Karlodinium australe blooms: Evidence from laboratory bioassays

Song X, Hu Z, Shang L, Leaw CP, Lim PT, Tang YZ

Harmful Algae, 2020 11;99:101926.
PMID: 33218448 DOI: 10.1016/j.hal.2020.101926

Multiple dinoflagellate species from the genus Karlodinium have been well known to form massive and toxic blooms that consequently cause fish kills in many coastal waters around the world. Karlodinium australe is a mixotrophic and potentially ichthyotoxic species associated with fish kills. Here, we investigated phagotrophy of K. australe (isolate KaJb05) established from a bloom event in the West Johor Strait, Malaysia, using several prey species (phytoplankton, zooplankton, and larval fish). The results showed that K. australe ingested relatively small prey cells of co-occurring microalgae by direct engulfment, while it fed on larger prey cells of microalgae by tube feeding. The results of animal exposure bioassays using rotifer (Brachionus plicatilis), brine shrimp (Artemia salina), and larval fish (Oryzias melastigma) demonstrated that phagotrophy (in terms of the trophic mode of the dinoflagellate), or micropredation (in terms of the mechanism of lethal effects on prey), played a more important role than the toxicity did in causing the lethal effects of K. australe on these aquatic animals under low cell densities of K. australe, while the mortalities of animals observed in the exposure to cell lysates of K. australe were solely caused by the toxicity. A comparison of the lethal effects between K. australe and K. veneficum revealed that the lethal effect of K. australe on rotifers was much stronger than that of K. veneficum at all cell densities applied in the experiments and the more "aggressive" micropredation of K. australe is suggested to explain the difference in lethal effect between K. austale and K. veneficum. Our results may explain why K. australe exhibited fish killings during moderate blooms at cell densities < 2.34 × 106 cells L-1, whereas K. veneficum was observed to cause massive fish kills only if the cell density was above 107 cells L-1. We believe these findings provide new insights into the ecological consequences of phagotrophy exhibited in some mixotrophic and harmful algae such as species of Karlodinium and of HAB events in general.

Matched MeSH terms: Exotoxins*
Fulltext Prevalence and association of Panton-Valentine Leukocidin gene with the risk of sepsis in patients infected with Methicillin Resistant Staphylococcus aureus

Ahmad NI, Yean Yean C, Foo PC, Mohamad Safiee AW, Hassan SA

J Infect Public Health, 2020 Oct;13(10):1508-1512.
PMID: 32653480 DOI: 10.1016/j.jiph.2020.06.018

BACKGROUND: Panton-Valentine Leukocidin (PVL), is one of the virulence gene expressed by Methicillin Resistant Staphylococcus aureus (MRSA) and is known to be associated with severe form of community acquired MRSA infection. The aim of this study is to investigate its prevalence in our setting and patient's clinical outcome.
METHODS: A cross sectional study involve retrospective record review were done involving 90 MRSA positive isolates between November 2016 and October 2017. Multiplex PCR was performed to detect femA, mecA and PVL genes. Clinical presentation and outcomes of patients were reviewed and presented as descriptive analysis.
RESULTS: All of the 90 MRSA isolates included in this study were positive for femA and mecA genes following PCR. PVL gene was detected in 20% (n = 18) of the isolates of which 61.1% (n = 11) were community acquired infections and 38.8% (n = 7) were hospital acquired. Case distribution from community acquired infections include patients with skin and soft tissue infections (33.3%, n = 6), infected diabetic foot ulcers (16.7%, n = 3), and one patient each (5.5%, n = 1) for community acquired pneumonia and meningitis. Half of the PVL positive MRSA cases (50%, n = 9) were having sepsis and four of them succumbed to death due to severe infection.
CONCLUSION: This study shows a high prevalence of PVL positive MRSA infection in our population. Skin and soft tissue infections accounting for the major sources. In addition, the presence of the PVL gene is associated with increased risk for developing sepsis.

Matched MeSH terms: Exotoxins/genetics
Bacterial expression of the scFv fragment of a recombinant antibody specific for Burkholderia pseudomallei exotoxin

Su YC, Lim KP, Nathan S

J. Biochem. Mol. Biol., 2003 Sep 30;36(5):493-8.
PMID: 14536033

The scFv antibody towards the Burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. We report here the optimization of the scFv expression in an E. coli expression system. Four different E. coli strains (ER2537, TG1, HB2151, and XL1-Blue) were examined for optimal expression of the scFv protein. Two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scFv expression. Cells that carried the scFv construct were grown at 30 degrees C and induced with 0.05 mM IPTG. The expression was then monitored by SDS-PAGE, Western blotting, and indirect ELISA. The Western blot profile showed different levels of the scFv expression among the host strains; XL1-Blue exhibited the highest level of the scFv protein expression. Glycerol at a concentration of 0.2% (v/v) significantly increased the scFv protein expression level when compared to 0.2% (w/v) glucose. Further optimization demonstrated that the scFv protein expression in XL1-Blue was the most optimal with a glycerol concentration as low as 0.05%. However, by indirect ELISA, only the scFv protein that was expressed in 0.2% (v/v) glycerol exhibited high specificity towards the Burkholderia pseudomallei exotoxin.

Matched MeSH terms: Exotoxins/biosynthesis; Exotoxins/genetics*; Exotoxins/immunology
Fulltext Predominance of SCCmec type IV in community-acquired meticillin-resistant Staphylococcus aureus (MRSA) in multi-centre Malaysian hospitals

Nor Amdan NA, Zamri HF, Mohd Ali MR, Dahalan NA, Anak Maling DR, Wan Hamdan WAF, et al.

J Hosp Infect, 2024 Jan;143:113-114.
PMID: 37979625 DOI: 10.1016/j.jhin.2023.10.023
Matched MeSH terms: Exotoxins
Fulltext Assessment of blood changes post-challenge with Corynebacterium pseudotuberculosis and its exotoxin (phospholipase D): A comprehensive study in goat

Mahmood ZK, Jesse FF, Saharee AA, Jasni S, Yusoff R, Wahid H

Vet World, 2015 Sep;8(9):1105-17.
PMID: 27047206 DOI: 10.14202/vetworld.2015.1105-1117

There is very little information regarding blood changes during the challenge of phospholipase D (PLD) in goats. Therefore, this experiment was conducted to study the changes in blood after the challenge with Corynebacterium pseudotuberculosis and its exotoxin, PLD to fill in the gap of caseous lymphadenitis (CLA) research.

Matched MeSH terms: Exotoxins
Fulltext Dissemination of streptococcal pyrogenic exotoxin G (spegg) with an IS-like element in fish isolates of Streptococcus dysgalactiae

Abdelsalam M, Chen SC, Yoshida T

FEMS Microbiol Lett, 2010 Aug 1;309(1):105-13.
PMID: 20528946 DOI: 10.1111/j.1574-6968.2010.02024.x

The Lancefield group C alpha-hemolytic Streptococcus dysgalactiae ssp. dysgalactiae (GCSD) causes systemic granulomatous inflammatory disease and high mortality rates in infected fish. Superantigen and streptolysin S genes are the most important virulence factors contributing to an invasive streptococcal infection. PCR amplification revealed that all strains isolated from moribund fish harbored the streptolysin S structural gene (sagA). GCSD fish isolates were PCR negative for emm, speA, speB, speC, speM, smeZ, and ssa. However, the size of the streptococcal pyrogenic exotoxin G (spegg) locus, a superantigen, in positive S. dysgalactiae fish and pig strains was variable. The ORF of the spegg locus of 26 GCSD fish strains and one GCSD pig strain was inserted with IS981SC. Interestingly, the ORF of the spegg locus of two fish strains of GCSD collected in Malaysia was inserted with an IS981SC-IS1161 hybrid IS element. The hybrid IS element was found in all of the GCSD fish isolates and one GCSD pig through PCR screening. Although no insertion sequence (IS) was detected in the spegg locus of S. dysgalactiae ssp. equisimilis (GCSE) strains, a five-nucleotide deletion mutation was detected in the ORF of the spegg locus of one GCSE strain at the supposed site of IS981SC insertion, resulting in a frameshift mutation.

Matched MeSH terms: Exotoxins/genetics*; Exotoxins/metabolism
Fulltext Prevalence of Panton-Valentine leukocidin genes among carriage and invasive Staphylococcus aureus isolates in Malaysia

Neela V, Ehsanollah GR, Zamberi S, Van Belkum A, Mariana NS

Int J Infect Dis, 2009 May;13(3):e131-2.
PMID: 18955004 DOI: 10.1016/j.ijid.2008.07.009
Matched MeSH terms: Exotoxins/genetics*; Exotoxins/isolation & purification
Community-acquired necrotising pneumonia caused by Panton-Valentine leucocidin-producing methicillin-resistant Staphylococcus aureus

Leong CL, Norazah A, Azureen A, Lingam R

Med J Malaysia, 2017 12;72(6):378-379.
PMID: 29308781

A 61-year-old male presented with community-onset pneumonia not responding to treatment despite given appropriate antibiotics. Computed tomography scan of the thorax showed large multiloculated pleural effusion with multiple cavitating foci within collapsed segments; lesions which were suggestive of necrotising pneumonia. Drainage of the effusion and culture revealed methicillin-resistant Staphylococcus aureus, which had the same antibiotic profile with the blood isolate and PVL gene positive.

Matched MeSH terms: Exotoxins/adverse effects*; Exotoxins/biosynthesis*
Fulltext Draft Genome Sequence of Methicillin-Resistant Staphylococcus aureus KT/Y21, a Sequence Type 772 (ST772) Strain Isolated from a Pediatric Blood Sample in Terengganu, Malaysia

Suhaili Z, Lean SS, Yahya A, Mohd Desa MN, Ali AM, Yeo CC

Genome Announc, 2014;2(2).
PMID: 24723714 DOI: 10.1128/genomeA.00271-14

Here, we report the draft genome sequence of a methicillin-resistant Staphylococcus aureus (MRSA) strain, KT/Y21, isolated from a blood sample of a pediatric patient. This strain belongs to sequence type 772 (ST772), harbors the staphylococcal cassette chromosome mec element (SCCmec) type V, and is positive for the Panton-Valentine leukocidin (PVL) pathogenic determinant.

Matched MeSH terms: Exotoxins
Prevalence of antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens in military personnel in Sabah and Sarawak, Malaysia

Embi N, Suhaimi A, Mohamed R, Ismail G

Microbiol. Immunol., 1992;36(8):899-904.
PMID: 1474938

Sera from 420 military personnel serving in Sabah and Sarawk, Malaysia, were tested for antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens by enzyme-linked immunosorbent assay procedure (ELISA). Data showed that 54.4% of serum samples were positive for antibodies to P. pseudomallei exotoxin and 65.7% were positive for antibodies to the whole cell antigens. Samples gave much lower titers for anti-exotoxin antibodies compared to titers against crude whole cell antigens. The incidence of antibody to exotoxin was highest in the age groups ranging from 26 to 32 years, where the positive rates were higher than 40% and 30% for military personnel serving in Sarawak and Sabah, respectively.

Matched MeSH terms: Exotoxins/immunology*
Inhibition of macromolecular synthesis in cultured macrophages by Pseudomonas pseudomallei exotoxin

Mohamed R, Nathan S, Embi N, Razak N, Ismail G

Microbiol. Immunol., 1989;33(10):811-20.
PMID: 2615673

Pseudomonas pseudomallei exotoxin was found to be a potent inhibitor of protein and DNA synthesis in cultured macrophages. Inhibition of DNA synthesis occurred at toxin concentrations as low as 1-2 micrograms/ml and inhibition of 3H-thymidine uptake was almost complete at concentrations of 8 micrograms/ml or more. A close correlation between cell damage and inhibition by DNA synthesis was observed. For protein synthesis, inhibition was obtained at much lower doses (0.06-2.0 micrograms/ml) of the toxin. At similar toxin concentrations, DNA synthesis was marginally affected. Further, it was shown that protein synthesis inhibition occurred almost immediately after incubation, reaching its maximal inhibitory effect of 70% after 6 hr. DNA synthesis, however, was minimally affected by a similar toxin concentration even after 10 hr of incubation. The inhibition of macromolecular synthesis in macrophages by P. pseudomallei exotoxin may be relevant to its modulatory effect on the host defense mechanism.

Matched MeSH terms: Exotoxins/pharmacology*
Antibody to Pseudomonas pseudomallei exotoxin in sheep exposed to natural infection

Ismail G, Mohamed R, Rohana S, Sharifah HS, Embi N

Vet Microbiol, 1991 May;27(3-4):277-82.
PMID: 1882505

Specific antibody to Pseudomonas pseudomallei exotoxin was detected in sheep sera exposed to natural infection. An enzyme-linked immunosorbent assay (ELISA) was used. Serum antitoxin was present in 49.3% of sera obtained from a flock of sheep naturally exposed to P. pseudomallei infection. Among these sera, 17.0% gave titers of 10,000. In contrast, serum antitoxin was present in only 6.0% of sera collected from sheep kept on a melioidosis-free farm. The ELISA reactivity of all positive sera could be completely absorbed with purified P. pseudomallei exotoxin. Similarly, preincubation of the exotoxin-coated wells with specific antiserum inhibited the ELISA reactivity of sheep sera. The results indicate that exotoxin is produced in vivo during infection by P. pseudomallei.

Matched MeSH terms: Exotoxins/immunology*
Fulltext A pentaplex PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus and Panton-Valentine Leucocidin

Al-Talib H, Yean CY, Al-Khateeb A, Hassan H, Singh KK, Al-Jashamy K, et al.

BMC Microbiol, 2009;9:113.
PMID: 19476638 DOI: 10.1186/1471-2180-9-113

Staphylococcus aureus is a major human pathogen, especially methicillin-resistant S. aureus (MRSA), which causes a wide range of hospital and community-acquired infections worldwide. Conventional testing for detection of MRSA takes 2-5 days to yield complete information of the organism and its antibiotic sensitivity pattern.

Matched MeSH terms: Exotoxins/isolation & purification*
Exotoxin profiles of clinical isolates of Aeromonas hydrophila

Vadivelu J, Puthucheary SD, Navaratnam P

J Med Microbiol, 1991 Jun;34(6):363-7.
PMID: 2056519

Eighty-six clinical isolates of Aeromonas hydrophila were studied for their ability to produce four exotoxins: a haemolysin active against rabbit erythrocytes, cytotoxin and enterotoxin detectable with Vero cell cultures, and the cholera toxin-like factor detected by an enzyme-linked immunosorbent assay. At least one exotoxin was produced by 80% of enteric and 96% of non-enteric isolates. The exotoxin profiles of non-enteric isolates were more restricted than those of enteric isolates, with haemolysin and cytotoxin producers preponderant. Although haemolysin and cytotoxin were produced by isolates from all sources, the enterotoxin and cholera toxin-like factor were more common amongst enteric isolates. The production of haemolysin and cytotoxin were closely related but the association between the enterotoxin and the cholera toxin-like factor was not significant.

Matched MeSH terms: Exotoxins/biosynthesis*
An ELISA-disc procedure for antibodies toPseudomonas pseudomallei: application for a serological study of melioidosis in an endemic area

Embi N, Devarajoo D, Mohamed R, Ismail G

World J Microbiol Biotechnol, 1993 Jan;9(1):91-6.
PMID: 24419848 DOI: 10.1007/BF00656525

The optimization and development of an ELISA-disc procedure for the detection of antibodies to whole cell surface antigens and purified exotoxin ofPseudomonas pseudomallei is described. Comparison of the serum agglutination test (SAT), the serum based enzyme-linked immunosorbent assay (ELISA) and the ELISA-disc procedures used on goat and human sera demonstrated a high correlation in their ability to detect antibodies specific forP. pseudomallei antigens. A serological survey using the ELISA-disc method was carried out on a normal human population in Sabah, Malaysia, an area known to be endemic for melioidosis. The prevalances of antibodies towards cell surface antigens and exotoxin ofP. pseudomallei were 28% and 8%, respectively. As a procedure, the ELISA-disc technique reported here is technically simple and provides savings in costs and is thus deemed suitable for seroepidemiological surveillance of melioidosis in remote areas of South-East Asia.

Matched MeSH terms: Exotoxins
Fulltext Development of loop-mediated isothermal amplification (LAMP) for the detection of speB gene in invasive Streptococcus pyogenes clinical isolates

Azi Simon Onyema, Leslie Than Thian Lung, Suresh Kumar, Rukman Awang Hamat

Malaysian Journal of Medicine and Health Sciences, 2019;15(106):1-1.
MyJurnal

Introduction: Group A streptococcus (GAS) is responsible for high morbidity and mortality globally. Hence, the need to develop sensitive, reliable and cost- effective method of detection is crucial. In this study, we developed a visual detection method for the common virulence gene, streptococcal pyrogenic exotoxin B (speB) involved in invasive GAS diseases using loop-mediated isothermal amplification (LAMP) with fluorescent detection dye (calcein). Meth-ods: The LAMP reaction was optimized at 63°C for 35 minutes using five sets of primer designed with LAMP primer V5 software. When the dye was added prior to amplification, samples with speB DNA developed a characteristic green color after the reaction, but no color reactions were observed in samples with DNAs of non-GAS isolates. De-tection of speB by LAMP assay was done among 43 clinical isolates of blood, pus, wound, tissue and throat samples and ATCCs for controls. Our findings were further reconfirmed by subjecting the LAMP products to 0.5% gel electro-phoresis. Results: The detection limit of this LAMP assay for speB was 10-7 ng/μl of genomic DNA per reaction, which was 10,000-fold more sensitive than conventional PCR 10-3 ng/μl. All 100 % samples were positive for speB gene by LAMP, and 93% by conventional PCR method. Conclusion: LAMP assay could offer remarkably high sensitivity, specificity, repeatability, reliability, affordability, and visibility; it is appropriate for rapid detection of speB in Group A streptococci (GAS) as a point of care testing.

Matched MeSH terms: Exotoxins
Fulltext Necrotising Fasciitis of the Lower Limb caused by Community-Acquired Methicillin-Resistant Staphylococcus aureus

Chan, C.K., Merican, A.M., Nawar, A.M., Hanifah, Y.A., Thong, K.L.

Malays Orthop J, 2010;4(3):36-38.
MyJurnal

Necrotising fasciitis caused by Community-Acquired Methicillin-resistant Staphylococcus aureus (CA-MRSA) has emerged as a new entity. Although it is recognised worldwide, there have been no reported cases to date in Malaysia. We report a case of necrotising fasciitis of the left lower limb in an otherwise healthy 20-year-old man. He presented with septic shock and despite the paucity of clinical signs in the limb, the infection was aggressive. Methicillin-Resistant Staphylococcus aureus (MRSA) was isolated from the deep fascia of the leg. Panton-Valentine leucocidin gene (PVL), which is a stable genetic marker for CA-MRSA strain, was positive in this case. This case of community acquired MRSA necrotising fasciitis is of concern and may herald the emergence of this resistant organism in Malaysia. Vigilant surveillance and microbiological monitoring is needed to follow this CA-MRSA trend.

Matched MeSH terms: Exotoxins
Fulltext Single-molecule sequencing reveals the molecular basis of multidrug-resistance in ST772 methicillin-resistant Staphylococcus aureus

Steinig EJ, Andersson P, Harris SR, Sarovich DS, Manoharan A, Coupland P, et al.

BMC Genomics, 2015;16:388.
PMID: 25981586 DOI: 10.1186/s12864-015-1599-9

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-associated infection, but there is growing awareness of the emergence of multidrug-resistant lineages in community settings around the world. One such lineage is ST772-MRSA-V, which has disseminated globally and is increasingly prevalent in India. Here, we present the complete genome sequence of DAR4145, a strain of the ST772-MRSA-V lineage from India, and investigate its genomic characteristics in regards to antibiotic resistance and virulence factors.

Matched MeSH terms: Exotoxins/genetics
Fulltext Comparison of methicillin-resistant Staphylococcus aureus strains isolated in 2003 and 2008 with an emergence of multidrug resistant ST22: SCCmec IV clone in a tertiary hospital, Malaysia

Lim KT, Hanifah YA, Mohd Yusof MY, Ito T, Thong KL

J Microbiol Immunol Infect, 2013 Jun;46(3):224-33.
PMID: 23523045 DOI: 10.1016/j.jmii.2013.02.001

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) continue to be a problem for clinicians worldwide. The objective of this study was to determine the changes in antibiograms of MRSA and their genotypic characteristics.

Matched MeSH terms: Exotoxins/genetics
Fulltext A persistent antimicrobial resistance pattern and limited methicillin-resistance-associated genotype in a short-term Staphylococcus aureus carriage isolated from a student population

Mat Azis N, Pung HP, Abdul Rachman AR, Amin Nordin S, Sarchio SNE, Suhaili Z, et al.

J Infect Public Health, 2017 Mar-Apr;10(2):156-164.
PMID: 27033676 DOI: 10.1016/j.jiph.2016.02.013

The aim of the present study was to assess and compare the antimicrobial susceptibility pattern against a panel of antibiotics and molecular and methicillin resistance-associated genotypes of 120 carriage S. aureus isolates previously isolated from a student population at two isolation events within a one-month interval. The antibiotic susceptibility of isolates was determined using the Kirby-Bauer disc-diffusion method (cefoxitin by Etest). The MRSA was screened using polymerase chain reaction for the presence of the mecA gene. The mecA-positive isolates were subjected to staphylococcal cassette chromosome (SCC) mec typing, multilocus sequence typing (MLST) and eBURST analysis. All isolates were characterized for the presence of the Panton-Valentine leukocidin (PVL) gene, an enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) pattern and the spa type. For the two occasions where S. aureus was isolated, the highest frequency of resistance was observed for penicillin (70% and 65%, respectively), with a lower rate against erythromycin and tetracycline (<12%). All isolates were susceptible to ciprofloxacin and gentamycin. As for methicillin resistance, eight isolates had minimum inhibitory concentrations (MIC) of resistant categories, but 10 isolates (8.33%) were positive for the mecA gene. The mecA-positive isolates belonged to SCCmec types I (n=9) and V (n=1). MLST was resolved for only three MRSAs, ST508 (n=1), ST88 (n=1) and ST96 (n=1). The results of the eBURST analysis showed that the MRSA isolates analyzed in the present study were potentially related to MRSA identified in other countries. Approximately half of the persistent S. aureus carriers harbored S. aureus of a similar spa type in the respective individuals during both isolation events. A persistent antimicrobial pattern and limited distinct MRSAs were observed over the short study period. The latter frequently exhibited SCCmec type I, commonly associated with hospital-acquired (HA) characteristics, but further delineation is needed to justify the origins of these bacteria.

Matched MeSH terms: Exotoxins/genetics

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