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A solute-binding protein for iron transport in Streptococcus iniae
Lili Zou, Jun Wang, Baofeng Huang, Mingquan Xie, Anxing Li
BMC Microbiology , 2010, DOI: 10.1186/1471-2180-10-309
Abstract: An ABC transporter system, named as mtsABC (metal transport system) was cloned from S. iniae HD-1, and was found to be involved in heme utilization. mtsABC is cotranscribed by three downstream genes, i.e., mtsA, mtsB, and mtsC. In this study, we cloned the first gene of the mtsABC transporter system (mtsA), and purified the corresponding recombinant protein MtsA. The analysis indicated that MtsA is a putative lipoprotein which binds to heme that can serve as an iron source for the microorganism, and is expressed in vivo during Kunming mice infection by S. iniae HD-1.This is believed to be the first report on the cloning the ABC transporter lipoprotein from S. iniae genomic DNA. Together, our data suggested that MtsA is associated with heme, and is expressed in vivo during Kunming mice infection by S. iniae HD-1 which indicated that it can be a potential candidate for S. iniae subunit vaccine.Streptococcus iniae (S. iniae) is a hemolytic Gram-positive coccus that is a major pathogen of culture fish. It has been associated with disease outbreak in several species of freshwater and marine fish cultured worldwide, including tilapia [1,2], barramundi [3], channel catfish [4], hybrid striped bass [1,5], Japanese flounder [6,7], olive flounder [8], rabbitfish [9], and rainbow trout [9,10]. Streptococcal infection can lead to serious symptoms such as meningoencephalitis and generalized septicaemia with high mortality rates of up to 50% [9,11]. S. iniae is also known to be an opportunistic pathogen that can cause fulminant soft tissue infection in humans, such as bacteremic cellulitis, septicarthritis, and endocarditis [12]. Identifying potential virulence determinants of streptococcal infection will eventually help to the control and eradication of the disease.Iron plays a significant role in many biological processes and is vital for several metabolic processes. Moreover, many proteins such as cytochromes and tricarboxylic acid metalloenzymes use iron as a cofactor [13]. I
Identification and molecular characterisation of a fibrinogen binding protein from Streptococcus iniae.
Justice CF Baiano, Reiny A Tumbol, Aarti Umapathy, Andrew C Barnes
BMC Microbiology , 2008, DOI: 10.1186/1471-2180-8-67
Abstract: Fibrinogen binding by S. iniae significantly reduced respiratory burst activity of barramundi peritoneal macrophages in primary cultures compared to BSA-treated or untreated controls, indicating a potentially important role for fibrinogen binding cell-surface proteins in avoiding phagocytic attack in fish. We describe a novel emm-like gene, simA, encoding a 57 kDa fibrinogen binding M-like protein in S. iniae. These SiM proteins and their corresponding tetrameric structures from some sequevar types (~230 kDa) bound fibrinogen in Western blots. simA was most closely related (32% identity) to the demA gene of S. dysgalactiae. Genome walking and sequencing determined the genetic organization of the simA region had similarities to the mgrC regulon in GCS and to S. uberis. Moreover, a putative multigene regulator, mgx was orientated in the opposite direction to the simA gene in common with S. uberis, but contrary to findings in GAS and GCS. In GAS, diversity among emm-genes and consequent diversity of their M-related proteins results in substantial antigenic variation. However, an extensive survey of S. iniae isolates from diverse geographic regions and hosts revealed only three variants of the gene, with one sequevar accounting for all but two of the 50 isolates analysed.These proteins play a role in avoiding oxidative attack by phagocytic cells during infection of fish by S. iniae, but genetic diversity amongst these key surface proteins has not yet arisen. This lack of diversity coupled with a functional role in macrophage resistance suggests that these proteins may constitute important targets for future vaccines against S. iniae in fish.One of the most devastating diseases to warm water finfish aquaculture is caused by Streptococcus iniae. First isolated from a captive Amazon River dolphin, Inia geoffrensis, in 1976, S. iniae has caused outbreaks of disease in Israel, Australia, Japan, and the United States [1]. The economic losses attributed to S. iniae are estimat
Role of an Iron-Dependent Transcriptional Regulator in the Pathogenesis and Host Response to Infection with Streptococcus pneumoniae  [PDF]
Radha Gupta, Minny Bhatty, Edwin Swiatlo, Bindu Nanduri
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0055157
Abstract: Iron is a critical cofactor for many enzymes and is known to regulate gene expression in many bacterial pathogens. Streptococcus pneumoniae normally inhabits the upper respiratory mucosa but can also invade and replicate in lungs and blood. These anatomic sites vary considerably in both the quantity and form of available iron. The genome of serotype 4 pneumococcal strain TIGR4 encodes a putative iron-dependent transcriptional regulator (IDTR). A mutant deleted at idtr (Δidtr) exhibited growth kinetics similar to parent strain TIGR4 in vitro and in mouse blood for up to 48 hours following infection. However, Δidtr was significantly attenuated in a murine model of sepsis. IDTR down-regulates the expression of ten characterized and putative virulence genes in nasopharyngeal colonization and pneumonia. The host cytokine response was significantly suppressed in sepsis with Δidtr. Since an exaggerated inflammatory response is associated with a poor prognosis in sepsis, the decreased inflammatory response could explain the increased survival with Δidtr. Our results suggest that IDTR, which is dispensable for pneumococcal growth in vitro, is associated with regulation of pneumococcal virulence in specific host environments. Additionally, IDTR ultimately modulates the host cytokine response and systemic inflammation that contributes to morbidity and mortality of invasive pneumococcal disease.
Antibacterial compounds from Rutaceae with activities against Flavobacterium columnare and Streptococcus iniae  [PDF]
Kumudini M. Meepagala, Kevin K. Schrader, Charles L. Burandt
Journal of Agricultural Chemistry and Environment (JACEN) , 2013, DOI: 10.4236/jacen.2013.24014
Abstract: From the ethyl acetate extract of Murraya koenegii (Rutaceae) leaves, isomahanine (1) and mahanine (2) were isolated that showed antibacterial activity towards Flavobacterium columnare and Streptococcus iniae which caused columnaris disease and streptococcosis respectively. Isomahanine was found to have the strongest activity against F. columnare (isolate ALM-00-173) and S. iniae (isolate LA94-426) based on 24-h 50% inhibition concentration (IC50) and minimum inhibition concentration (MIC). Although compound (7), a nicotinamide isolated from Amyris texana had the lowest MIC (2.8 ± 0 mg/L) of any of the test compounds against F. columnare, the 24-h IC50 of 14.8 ± 0.6 mg/L was higher than that of isomahanine and subsequently the 24-h IC50 RDC values for (7) were almost a magnitude of order higher than those obtained for isomahanine. Isomahanine also had the strongest activity against S. iniae, with a 24-h IC50 of 1.3 ± 0.1 mg/L and MIC of 3.5 ± 0 mg/L, respectively.
Sil: A Streptococcus iniae Bacteriocin with Dual Role as an Antimicrobial and an Immunomodulator That Inhibits Innate Immune Response and Promotes S. iniae Infection  [PDF]
Mo-fei Li, Bao-cun Zhang, Jun Li, Li Sun
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0096222
Abstract: Streptococcus iniae is a Gram-positive bacterium and a severe pathogen to a wide range of economically important fish species. In addition, S. iniae is also a zoonotic pathogen and can cause serious infections in humans. In this study, we identified from a pathogenic S. iniae strain a putative bacteriocin, Sil, and examined its biological activity. Sil is composed of 101 amino acid residues and shares 35.6% overall sequence identity with the lactococcin 972 of Lactococcus lactis. Immunoblot analysis showed that Sil was secreted by S. iniae into the extracellular milieu. Purified recombinant Sil (rSil) exhibited a dose-dependent inhibitory effect on the growth of Bacillus subtilis but had no impact on the growths of other 16 Gram-positive bacteria and 10 Gram-negative bacteria representing 23 different bacterial species. Treatment of rSil by heating at 50°C abolished the activity of rSil. rSil bound to the surface of B. subtilis but induced no killing of the target cells. Cellular study revealed that rSil interacted with turbot (Scophthalmus maximus) head kidney monocytes and inhibited the innate immune response of the cells, which led to enhanced cellular infection of S. iniae. Antibody blocking of the extracellular Sil produced by S. iniae significantly attenuated the infectivity of S. iniae. Consistent with these in vitro observations, in vivo study showed that administration of turbot with rSil prior to S. iniae infection significantly increased bacterial dissemination and colonization in fish tissues. Taken together, these results indicate that Sil is a novel virulence-associated bacteriostatic and an immunoregulator that promotes S. iniae infection by impairing the immune defense of host fish.
Antibacterial activity of Iranian medicinal plants against Streptococcus iniae isolated from rainbow trout (Oncorhynchus mykiss)
Pirbalouti Ghasemi Abdollah,Broujeni Nikobin Vahid,Momeni Manouchehr,Poor Malek Fatemeh
Archives of Biological Sciences , 2011, DOI: 10.2298/abs1101059p
Abstract: Streptococcus iniae is among the major pathogens of a large number of fish species cultured in fresh and marine recirculating and net pen production systems. Ten Iranian medicinal plants were assessed for their antimicrobial activity against Streptococcus iniae isolates obtained from diseased Oncorhynchus mykiss (Salmonidae; Walbaum, 1972) collected from fish farms in Iran. The antibacterial activity of ethanol extracts of Punica granatum, Quercus branti, Glycyrrhiza glabra and essential oils of Heracleum lasiopetalum, Satureja bachtiarica, Thymus daenensis, Myrtus communis, Echinophora platyloba, Kelussia odoratissima and Stachys lavandulifolia against Steptococcus iniae was evaluated by disc diffusion and serial dilution assays. Most of the extracts and essential oils showed a relatively high antibacterial activity against Streptococcus iniae. Of the plants studied, the most active extracts were those obtained from the essential oils of Satureja bachtiarica, Echinophora platyloba, Thymus daenensis and the ethanol extract of Quercus branti. Some of the extracts were active against Streptococcus iniae. Two essential oils showed lower MIC values; Heracleum lasiopetalum (78 μg/ml) and Satureja bachtiarica (39 μg/ml). The essential oil of Satureja bachtiarica could be an important source of antibacterial compounds against the Streptococcus iniae isolated from rainbow trout.
Streptococcus iniae M-Like Protein Contributes to Virulence in Fish and Is a Target for Live Attenuated Vaccine Development  [PDF]
Jeffrey B. Locke, Ramy K. Aziz, Mike R. Vicknair, Victor Nizet, John T. Buchanan
PLOS ONE , 2008, DOI: 10.1371/journal.pone.0002824
Abstract: Background Streptococcus iniae is a significant pathogen in finfish aquaculture, though knowledge of virulence determinants is lacking. Through pyrosequencing of the S. iniae genome we have identified two gene homologues to classical surface-anchored streptococcal virulence factors: M-like protein (simA) and C5a peptidase (scpI). Methodology/Principal Findings S. iniae possesses a Mga-like locus containing simA and a divergently transcribed putative mga-like regulatory gene, mgx. In contrast to the Mga locus of group A Streptococcus (GAS, S. pyogenes), scpI is located distally in the chromosome. Comparative sequence analysis of the Mgx locus revealed only one significant variant, a strain with an insertion frameshift mutation in simA and a deletion mutation in a region downstream of mgx, generating an ORF which may encode a second putative mga-like gene, mgx2. Allelic exchange mutagenesis of simA and scpI was employed to investigate the potential role of these genes in S. iniae virulence. Our hybrid striped bass (HSB) and zebrafish models of infection revealed that M-like protein contributes significantly to S. iniae pathogenesis whereas C5a peptidase-like protein does not. Further, in vitro cell-based analyses indicate that SiMA, like other M family proteins, contributes to cellular adherence and invasion and provides resistance to phagocytic killing. Attenuation in our virulence models was also observed in the S. iniae isolate possessing a natural simA mutation. Vaccination of HSB with the ΔsimA mutant provided 100% protection against subsequent challenge with a lethal dose of wild-type (WT) S. iniae after 1,400 degree days, and shows promise as a target for live attenuated vaccine development. Conclusions/Significance Analysis of M-like protein and C5a peptidase through allelic replacement revealed that M-like protein plays a significant role in S. iniae virulence, and the Mga-like locus, which may regulate expression of this gene, has an unusual arrangement. The M-like protein mutant created in this research holds promise as live-attenuated vaccine.
Streptococcus iniae infection in cultured Asian sea bass (Lates calcarifer) and red tilapia (Oreochromis sp.) in southern Thailand
Naraid Suanyuk,Nirut Sukkasame,Nopparat Tanmark,Terutoyo Yoshida
Songklanakarin Journal of Science and Technology , 2010,
Abstract: Streptococcal infections are becoming an increasing problem in aquaculture and have been reported worldwide in avariety of fish species. Here we describe the isolation and characterization of Streptococcus iniae from Asian sea bass (Latescalcarifer) and red tilapia (Oreochromis sp.) cultured in southern Thailand. Conventional and rapid identification systems,as well as the polymerase chain reaction (PCR), were used to determine that the isolate was S. iniae. The virulence of thisS. iniae was higher in Asian sea bass than in red tilapia, as shown by the 10 day-LD50 in Asian sea bass and red tilapia, being1.08x104 and 1.14x107 CFU, respectively. Histopathological changes in Asian sea bass are more severe than those observedin red tilapia. The changes can be found in several organs including liver, pancreas, heart, eye and brain. Histopathologicalfindings included cellular necrosis, infiltration of lymphocytes and granuloma formation in the infected organs.
Streptococcus iniae SF1: Complete Genome Sequence, Proteomic Profile, and Immunoprotective Antigens  [PDF]
Bao-cun Zhang, Jian Zhang, Li Sun
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0091324
Abstract: Streptococcus iniae is a Gram-positive bacterium that is reckoned one of the most severe aquaculture pathogens. It has a broad host range among farmed marine and freshwater fish and can also cause zoonotic infection in humans. Here we report for the first time the complete genome sequence as well as the host factor-induced proteomic profile of a pathogenic S. iniae strain, SF1, a serotype I isolate from diseased fish. SF1 possesses a single chromosome of 2,149,844 base pairs, which contains 2,125 predicted protein coding sequences (CDS), 12 rRNA genes, and 45 tRNA genes. Among the protein-encoding CDS are genes involved in resource acquisition and utilization, signal sensing and transduction, carbohydrate metabolism, and defense against host immune response. Potential virulence genes include those encoding adhesins, autolysins, toxins, exoenzymes, and proteases. In addition, two putative prophages and a CRISPR-Cas system were found in the genome, the latter containing a CRISPR locus and four cas genes. Proteomic analysis detected 21 secreted proteins whose expressions were induced by host serum. Five of the serum-responsive proteins were subjected to immunoprotective analysis, which revealed that two of the proteins were highly protective against lethal S. iniae challenge when used as purified recombinant subunit vaccines. Taken together, these results provide an important molecular basis for future study of S. iniae in various aspects, in particular those related to pathogenesis and disease control.
Differential Transcriptomic Response in the Spleen and Head Kidney Following Vaccination and Infection of Asian Seabass with Streptococcus iniae  [PDF]
Junhui Jiang, Masato Miyata, Candy Chan, Si Yan Ngoh, Woei Chang Liew, Jolly M. Saju, Kah Sing Ng, Fong Sian Wong, Yeng Sheng Lee, Siow Foong Chang, László Orbán
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0099128
Abstract: Vaccination is an important strategy in the protection of aquaculture species from major diseases. However, we still do not have a good understanding of the mechanisms underlying vaccine-induced disease resistance. This is further complicated by the presence of several lymphoid organs that play different roles when mounting an immune response. In this study, we attempt to elucidate some of these mechanisms using a microarray-based approach. Asian seabass (Lates calcarifer) were vaccinated against Streptococcus iniae and the transcriptomic changes within the spleen and head kidney at one and seven days post-vaccination were profiled. We subsequently challenged the seabass at three weeks post-vaccination with live S. iniae and similarly profiled the transcriptomes of the two organs after the challenge. We found that vaccination induced an early, but transient transcriptomic change in the spleens and a delayed response in the head kidneys, which became more similar to one another compared to un-vaccinated ones. When challenged with the pathogen, the spleen, but not the head kidneys, responded transcriptomically at 25–29 hours post-challenge. A unique set of genes, in particular those involved in the activation of NF-κB signaling, was up-regulated in the vaccinated spleens upon pathogen challenge but not in the un-vaccinated spleens. A semi-quantitative PCR detection of S. iniae using metagenomic DNA extracted from the water containing the seabass also revealed that vaccination resulted in reduction of pathogen shedding. This result indicated that vaccination not only led to a successful immune defense against the infection, but also reduced the chances for horizontal transmission of the pathogen. In conclusion, we have provided a transcriptomic analysis of how the teleost spleen and head kidneys responded to vaccination and subsequent infection. The different responses from the two organs are suggestive of their unique roles in establishing a vaccine-induced disease resistance.
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