128.119.84.54

June 2019

Hello, I'm Ifnord. I noticed that you recently removed content from Vibrio anguillarum without adequately explaining why. In the future, it would be helpful to others if you described your changes to Wikipedia with an accurate edit summary. If this was a mistake, don't worry; the removed content has been restored. If you would like to experiment, please use the sandbox. If you think I made a mistake, or if you have any questions, you can leave me a message on my talk page. You have removed referenced material. Please use the article's talk page to discuss such changes first. Ifnord (talk) 16:53, 20 June 2019 (UTC)

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The following post is meant to establish a digital record: I have tried to remove content from Wikipedia 3 times because it directly violated my personal copyright. The content was reinstated by Wikimeida each time, without my consent. The user El_C placed a temporary (7 days) block on my IP Address as a result of my posting the word, "I will sue" to a 'talk' page.

I will lose viewership of my hard work through this plagiarized content. I will lose citations to my published content. I will lose credit for my work. I put time into my work, I put money into its publication. I feel that I have been robbed of professional credit, credit for my time, credit for my effort, credit for my investment in the publication.— Preceding unsigned comment added by 128.119.84.54 (talk • contribs)

Do you have proof for that? See below. You were blocked for violating our no legal threats policy. You are entitled to seek legal remedy, but may not remain a Wikipedia editor while that is outstanding. El_C 17:29, 20 June 2019 (UTC)

I am happy to send my published article to you for your review. The irony is that another Wikipedia user claimed he/she/they did not have access to the content. In other words, they did not want to pay for my content. — Preceding unsigned comment added by 128.119.84.54 (talk • contribs)

That's not necessary. Please just give me a few samples. El_C 17:33, 20 June 2019 (UTC)

Should you prefer to keep your email private, the content can be found here: https://onlinelibrary.wiley.com/doi/full/10.1111/raq.12188 — Preceding unsigned comment added by 128.119.84.54 (talk • contribs)

Again, not necessary. A few samples will do. El_C 17:34, 20 June 2019 (UTC)

Full original text:

Abstract Vibrio (Listonella) anguillarum is a marine bacteria that is pathogenic to a number of aquatic organisms including several species which are important to the aquaculture industry. Organisms that are infected by V. anguillarum are diagnosed with vibriosis which can be lethal in a matter of days. Vibriosis can be particularly devastating to aquaculture businesses and measures of prevention or treatment are generally quite expensive. Efforts to understand and control V. anguillarum virulence have been of high‐priority among international aquatic research studies. The knowledge that has accumulated as a result of this collective research effort is reviewed in this article. Introduction Vibrio anguillarum is a bacteria that is pathogenic to a variety of fish, crustaceans and bivalves which collectively amount to more than 90 susceptible aquatic organisms in at least 28 different countries. Septicemic host infection with V. anguillarum is known as vibriosis and causes symptoms such as internal and external ulceration, abdominal distension, petechia, flesh rot, lethargy, appetite loss, necrosis, erythema, sheathing of arteries and circulatory haemorrhage, boil formation upon muscle tissue, visual lesions and eventually death. Infection by V. anguillarum progresses rapidly among a host as the pathogen induces infection as quickly as 2 days following initial exposure. Vibrio anguillarum aggressively penetrates a host using a series of genetic virulence factors which can be lethal as soon as 3 days postinfection (Gram et al. 1999) meaning a host could be killed as quickly as 5 days postexposure to the pathogen (Mikkelsen et al. 2007) or as quickly as 2 days in certain larvae cultures (Chair et al. 1994). Imminently lethal levels of V. anguillarum are quantities of at least Log 4 CFU mL−1 in seawater (Thorburn et al. 1987; Dierckens et al. 2009) – or Log 3 CFU mL−1 when exposed for prolonged duration (Lei et al. 2006) – but larval or juvenile organisms can contract vibriosis in levels as low as about Log 3.5 CFU mL−1 (Mikkelsen et al. 2007) and the vibriosis infection renders the host organism more susceptible to cooperative infection by other pathogens as well (Jang et al. 2014). Vibrio anguillarum virulence in nature is rather seldom due to a constant state of stressed survival and conditional variations that result from the environment or seasonal weather. Farming of aquatic organisms under controlled conditions has been expanding now for several decades and V. anguillarum virulence has gained increased prevalence under such circumstances as a result. Aquaculture losses due to V. anguillarum‐induced mortalities have been particularly devastating reaching magnitudes as high as 100% losses in some reports (Foscarini 1988; Larsen et al. 1994; Austin et al. 2005) and such losses in aquaculture volumes have translated to financial deficits as high as $18–30 million in Japan, annually (Austin & Austin 1993). Aquaculture practices are quite sensitive to V. anguillarum, and even the mode of transporting hatchlings to aquaculture farms is known to increase mortality yields when moved later in the growing season rather than earlier (Thorburn 1987). Foundational research and taxonomy Vibrio anguillarum virulence in marine settings has been reported as early as 1718 along the coastline of Continental Europe (Hofer 1904) and was referenced on record as the red disease in eel (Anguilla anguilla) mortality reports as early as 1893 (Canestrini 1893; Feddersen 1896), serving as the root of the name anguillarum. Although misunderstood at the time, a red disease epidemic was described in 1790 in which nearly 40 tons of eel were lost to an unknown inhabiting pathogen, denoting the severity of the septicemic infection (Bergman 1909). In 1932, Bruun and Heiberg conducted in‐depth survey collection and analyses of red disease outbreaks in Scandinavia in which fisherman described significant mortality yields that ultimately hindered the business of several large‐scale fisheries in the late 1800s. As described in a review by Sindermann (1966), there was evident confusion among researchers until the early 1900s as to which bacterium was responsible for red disease, citing similar symptoms related to Aeromonas, Atherynops, Fundulus, Gillichthys and Pseudomonas infection. However, V. anguillarum was the only species that was consistently isolated from the various outbreaks described and so the disease was classified as vibriosis. By 1970, V. anguillarum was known to have devastating effects on several marine organisms such as cod, eel, finnock, flounder, herring, pike, plaice, oyster, salmon and trout (Bagge & Bagge 1956; Smith 1959; Sindermann 1966; Evelyn 1971; Levin et al. 1972; Brown 1973; Egidius & Andersen 1987) and its reputation as the causative agent of vibriosis grew considerable recognition. Epizootiological studies of V. anguillarum were of interest dating back to the earliest known outbreak of red disease in eel but valid microbiological data on the subject was lacking in publication until the 1950s. Hoshina (1956) described a disease in Japanese rainbow trout as an epidemic caused by a bacterium which he identified as Vibrio piscium var. japonicus. Not long after, Smith (1959) reported similar mortalities in wild finnock due to an unknown Vibrio spp., following widespread local distress among fishermen in Scotland. Smith (1959) cited that of Hoshina (1956) as the closest relative study in the identification of the red disease‐causing microbial fish pathogen and the two served as major reference points for further studies throughout the 1960s. Smith (1961) noted that V. anguillarum was excluded from Bergey's Manual of Determinative Bacteriology, 7th ed. and prior, evidently suggesting the absence of the strain's notoriety among the microbiological research community during that time‐period; the 8th edition (Buchanan & Gibbons 1974) and onward were updated to contain information on V. anguillarum and research studies on the subject have accumulated immensely ever since. Until that time, V. anguillarum was empirically identified by four different taxon prior to their reclassification as one and the same species: Beneckea anguillara (Baumann et al. 1978), V. anguillarum Bergman 1909; V. piscium var. japonicus David 1927 and Pseudomonas ichthyodermis Shewan, Hobbs and Hodgkiss 1960 (Hendrie et al. 1971). A fifth classification, Bacillus anguillarum, was used early on but the bacterium was determined to be Gram‐positive, while V. anguillarum is Gram‐negative (Bergman 1909). Vibriosis was identified as the cause of substantial mortalities in the oyster industry of Spain, but the infective agent was identified inconclusively as both V. anguillarum and Vibrio tubiashii (Tubiash et al. 1970; Bolinches et al. 1986). As such, it cannot be known for certain but some early studies believed to be V. anguillarum‐specific might very well have been V. tubiashii‐specific instead (Hada et al. 1984). The bacterium identified as Beneckea anguillara was initially segregated into two biotypes (I & II), but biotype II was reclassified as Vibrio ordalii in acknowledgement of Erling J. Ordal by Schiewe, Trust and Crosa in 1981; both strains are related by approximately ~70% genetic homology (Anderson & Ordal 1972). Studies by Nybelin (1935) and Smith (1961) collectively categorized V. anguillarum under three biotypes: (i) type A being able to produce acid from sucrose or mannitol without gaseous by‐product, as well as producing indole, (ii) type B being unable to react with sucrose or mannitol to produce gaseous by‐product or indole and (iii) type C being able to produce acid from sucrose and mannitol but not gaseous by‐product or indole. In 1984, MacDonell and Colwell screened Vibrionaceae bacteria for genetic variations among the 5S rRNA region that ultimately culminated in the establishment of two novel genera for the phylogenetic classification of true Vibrio spp., Listonella and Shewanella. As a result, V. anguillarum was reclassified as Listonella anguillarum and was officially excluded from the Vibrionaceae family taxon. Only three bacterial species were classified within the genus Listonella – L. anguillarum, Listonella damsel and Listonella pelagius – until they were later reclassified under the Vibrio genus (Dikow 2011; Thompson et al. 2011). At the present time, the accepted classification of V. anguillarum appears to be Vibrio (Listonella) anguillarum as controversy among preference and related studies remain prevalent in publication. Research into the discrimination of V. anguillarum by serotype began expanding in the 1980s when it was observed that V. anguillarum isolates varied among one‐another by cellular sugar compositions when screened using gas liquid chromatography (Yanagihara et al. 1984) and reacted differently when exposed to thermostable O‐antigens on the basis of cross‐agglutination and cross‐absorption trends (Kitao et al. 1983). As such, data collected in these studies expanded the serotyping scheme of V. anguillarum beyond only types A–C to accommodate for previously ‘unknown’ strains which resulted in a novel serotyping scheme consisting of six serotypes A–F (Kitao et al. 1983). Sørensen and Larsen (1986) further refined the V. anguillarum serotyping system not long after based on the detection of O‐antigens using immune‐serum slide‐agglutination procedures developed previously by Pacha and Kiehn (1969). Vibrio anguillarum was then divided into ten serotypes O1–O10 based on O‐antigen content within heat killed broth culture, and it was determined that serotypes O1 and O2 were statistically dominant among vibriosis outbreaks in fish (Sørensen & Larsen 1986). Vibrio anguillarum serotype O2 is divided into two sub‐serogroups O2α and O2β (Knappskog et al. 1993). Serotype O3 has been referenced as the causative virulence strain among vibriosis outbreaks in eel, but the serotype is generally not isolated from other afflicted aquatic organisms (Silva‐Rubio et al. 2008). Grisez and Ollevier (1995) then converged the O1–O10 serotyping scheme with the methods and strains described in research by Kitao et al. (1983, 1984) which culminated in the establishment of six novel serotypes, expanding the V. anguillarum serotyping scheme to O1–O16. Pedersen et al. (1999) are responsible for modernizing the V. anguillarum serotyping scheme by expanding the discrimination system once more to comprise of twenty‐three different serotypes based on serological reactions which result in distinct lipopolysaccharide patterns observable using SDS‐PAGE and western blotting. Vibrio anguillarum is now understood to consist of 23 O‐serogroups, but there is reason to believe that additional serotypes of V. anguillarum exist in nature which are not typable by the methods described in these studies (Pedersen et al. 1999). The general consensus among V. anguillarum virulence research in aquaculture resonates around serotypes O1–O3 being the only virulent strains of the pathogenic species. However, there have been cases in which serotypes O4–O5 have been identified as the causative agents of vibriosis in aquatic organisms and serotype O4 was responsible for the loss of 100% of reared cod in western Denmark (Pazos et al. 1992; Larsen et al. 1994). As such, serotypes O1–O5 should all be emphasized as virulent V. anguillarum strains when developing vaccination or preventative measures against the pathogen in aquaculture systems. Ecology and viability Growth conditions Vibrio anguillarum is a Gram‐negative, arcuate‐rod bacterium with one polar flagella which exists in marine environments as an opportunistic pathogen to a variety of fish and shellfish (Fig. 1). The pathogen grows best between 30 and 34°C with a maximum growth temperature of 38.5°C (Guérin‐Faublée et al. 1995), and the growth rate of V. anguillarum is known to increase with temperature (Groberg et al. 1983). Binary fission of the V. anguillarum cell is pH sensitive being most efficient at neutral pH 7, inhibited entirely over pH 9 and significantly disrupted at pH 6 or below (Gilmour et al. 1976; Larsen 1984). The bacterium is halophilic and thrives within NaCl concentrations between 1% and 2%, but it is understood that temperature serves as a more detrimental factor towards V. anguillarum growth than does salinity as V. anguillarum is known to remain viable when relocated to freshwater environments (Rucker 1959; Hacking & Budd 1971; Rødsæther et al. 1977; Larsen 1984; Laurencin & Germon 1987); however, V. anguillarum motility is enhanced with increased salinity (Kao et al. 2009). Temperature and salinity levels are lethal to V. anguillarum when exceeding 41°C or 7%, respectively (Golten & Scheffers 1975), while V. anguillarum growth below 5°C is quite minimal (Larsen 1984) and exposure of the cell to temperatures exceeding 44°C will sterilize the culture in less than 3 min (Jacobsen & Liltved 1988). The pathogen is able to remain viable in seawater for over 50 months, but variability in the halophilic nature of V. anguillarum is evident when the strain reaches stationary growth as it can no longer tolerate salinities which exceed 5% (Hoff 1989). Vibrio anguillarum is more prevalent in nature among seed‐production environments inhabiting larval fish or rotifer populations (Sugita et al. 2005) and wild‐type V. anguillarum isolates are highly diverse as a result of environmental interactions when screened for their biochemical interactions, such as their reactions to malonate, methyl red, citrate, cellobiose, arabinose, indole and trehalose, which are known to vary regardless of genetic homology (Egidius & Andersen 1977). Vibrio anguillarum thrives in environments which contain divalent cations such as Ca2+ and Mg2+ or when the V. anguillarum population is high as opposed to low and both factors are known to contribute to the survival of the bacteria under conditions of osmotic stress (Miyamoto & Eguchi 1997). Furthermore, copper is an initiating factor of vibriosis in eel (Rødsæther et al. 1977) and aquacultures stressed under elevated copper levels are 50% more susceptible to V. anguillarum infection (Baker et al. 1983). Vibrio anguillarum is stressed for survival during the majority of the cell's life cycle facing seasonal difference in temperature, salinity changes following water flow and depleted nutritional resources making it necessary for the pathogen to adapt to its environment for long‐term survival and ultimate colonization among a viable host. image Figure 1 Open in figure viewerPowerPoint Immunogold electron microscopy. Primary antiserum was raised against formalin‐killed whole cells of Vibrio anguillarum 775.17B and has been shown to be highly antigenic for the lipopolysaccharide in the sheath. Bars, 0.5 μm (Milton et al. 1996). Bacteriology and stress survival Vibrio anguillarum contains two chromosomes which have been sequenced in full for serotypes O1, O2 and O3 (Naka et al. 2011; Li et al. 2013a; Busschaert et al. 2014) and may or may not contain plasmids (Skov et al. 1995). Plasmid presence in V. anguillarum is not required for virulence and is independent of biochemical properties of the bacterial culture (Larsen & Olsen 1991), but certain plasmids are understood to contribute to V. anguillarum virulence and resistance to certain antimicrobial substances. The pathogen contains similar genetic, pathogenic islands to Vibrio harveyi clade, Vibrio cholerae and Vibrio alginolyticus specifically among genes nanH, flrA, luxA and vspR which encode for neuraminidase‐released sialic acid utilization, flagellar response to environmental change, bioluminescence and quorum‐sensing, as well as biofilm production, respectively (Gennari et al. 2012). Carbohydrate transport and binary fission in V. anguillarum are deterred in highly osmotic conditions under which V. anguillarum expresses genes ompW and ompU that encode for NaCl efflux porins or contrarily the gene ompV which is expressed under lowly osmotic conditions to draw‐in NaCl concentrations (Kao et al. 2009). Vibrio anguillarum utilizes a proton motive force to inwardly direct nutrients during the infection of a host but when viable in seawater the cell must utilize a sodium motive force to survive under conditions of stressed starvation which is why the pathogen requires Na+ from salt for long‐term survival (Fujiwara‐Nagata & Eguchi 2004). Na+ and K+ enhance nicotinamide adenine dinucleotide hydride (NADH) oxidase activity in V. anguillarum, but the pathogen is not dependent upon these ions for NADH oxidase functionality due to adaptations made in the marine environment which explain why the cell can remain virulent in freshwater settings (Fujiwara‐Nagata et al. 2003, 2004). The gene nqrA is involved in respiratory chain functions specific to NADH quinone oxidoreductase which functions as a result of the promoters nqrP1 and nqrP2. The nqrP1 promoter functions during exponential and stationary growth while the nqrP2 promoter functions only during stationary growth meaning that the pathogen utilizes the nqrP2 promoter of this mechanism to survive under conditions of starvation stress (Fujiwara‐Nagata et al. 2007). The alternative sigma factor rpoS gene in V. anguillarum plays a central, genetic‐regulation role in the cellular response to environmental stress specifically during stationary growth due to high temperature, osmotic stress, UV irradiation and oxidative stress, but the exact mechanisms of rpoS in such survival pathways remain to be elucidated; deletion of the rpoS gene decreases virulence in V. anguillarum by as much as 50% (Ma et al. 2009). The amiB gene in V. anguillarum is highly conserved and encodes for an N‐acetylmuramoyl‐L‐alanine amidase which is believed to contribute to the separation of daughter cells following binary fission reproduction (Ahn et al. 2006). Temperature serves as an important antagonist against protein expression in V. anguillarum as elevated or declining temperature disrupts the desired protein folding patterns which are essential to the survival of the bacterium in nature. Higher temperatures induce a 60‐kDa protein identified as GroEL which is believed to contribute to the assembly of new protein complexes which are essential for environmental adaptation (Mutharia et al. 1998). Vibrio anguillarum produces a peptidyl‐prolyl cis/trans isomerase (PPIase) when exposed to lower temperatures or higher pH which compensates for effectual misfolding in proteins when expressed together with protein disulphide isomerases to ultimately maintain the desired conformation of peptide bonds and overall functions related to protein folding (Kim et al. 2014). The avirulent, self‐regulating toxR gene in V. anguillarum encodes for a transmembrane regulatory protein which is expressed in response to environmental stimuli – most prominently under depleted salinity conditions (Crisafi et al. 2014) – which is believed to exhibit a porin‐like outer‐membrane protein function which responds to pressure stress (Okuda et al. 2001) and the toxR gene is known to some extent to contribute to V. anguillarum motility (Wang et al. 2002). Vibrio anguillarum synthesizes a 40‐kDa protein which functions as an outer‐membrane general diffusion protein and is recognized as ‘OM1’ (Simón et al. 1996, 1998). The gene ompK encodes for another outer‐membrane protein which is dependent upon bile salts and iron‐chelating agents for expression suggesting that the protein plays a role in survival mechanisms within a host (Hamod et al. 2014). The pathogen also contains a 1755‐bp chitinase gene vac which is believed to contribute to virulence in crustaceans, but the notion remains unverified (Hirono et al. 1998). Genes in the ivi region are believed to play a role in virulence mechanisms, phospholipase signalling, genetic regulation, metabolism and outer‐membrane assembly, but such functions are left only to speculation at the present time and more research in this area is likely to emerge in future studies (Zou et al. 2010). Furthermore, the gene recA is expressed to repair damaged DNA and the gene is required for recombination but not survival (Singer 1989; Singer et al. 1996). Quorum‐sensing systems Vibrio anguillarum facilitates communication between cells using four systems of intracellular quorum‐sensing signal transductions (Fig. 2). The first system of communication in V. anguillarum is vanIR which is homologous to that of the luxIR quorum‐sensing system in Vibrio fischeri (Milton et al. 1997). An N‐acylhomoserine lactone (AHL) autoinducer molecule (Eberl 1999) identified as N‐(3‐oxodecanoyl)‐l‐homoserine lactone (3‐oxo‐C100HSL or ODHL) is expressed by the vanI gene in V. anguillarum which is initiated by the transcriptional activator gene vanR and is also regulated by the AHL synthase gene vanM (Milton et al. 1997; Buch et al. 2003; García‐Aljaro et al. 2011). The vanR transcriptional activator of vanI is believed to play a regulatory role of other genes as well (Milton 2006), but our understanding of such other mechanisms remains absent, at the present time. The second system of communication, luxMN, is homologous to that of V. harveyi in which the signal molecules N‐hexanoyl‐l‐homoserine lactone (C6‐HSL) and N‐(3‐hydroxyhexanoyl)‐l‐homoserine lactone (3‐hydroxy‐C6‐HSL) are produced by vanM for sensation by a vanN‐encoded protein receptor; the gene vanM is regulated by uspA ‘universal stress protein A’ (Milton et al. 2001; Lindell 2012). The third system of communication, luxSPQ is also homologous to that of V. harveyi in which a furanosyl borate diester autoinducer‐2 (AI‐2) is synthesized by the gene vanS to act as a signal molecule to be sensed by vanP and hybrid sensor kinase vanQ‐encoded receptors (Croxatto et al. 2004). The fourth quorum‐sensing system, cqsAS is predicted to exist in V. anguillarum in which a cqsA synthase produces an (S)‐3‐hydroxytridecan‐4‐one signal molecule known as the cholerae autoinducer‐1 (CAI‐1) (Miller et al. 2002; Wei et al. 2011) for sensation by a cqsS‐encoded receptor protein (Henke & Bassler 2004). Each of these networks exists in V. anguillarum as parallel phosphorelay quorum‐sensing systems except for luxIR (Lindell 2012) and all three parallel phosphorelay pathways mutually merge as individual regulators of the transcriptional regulator vanT (Milton 2006). image Figure 2 Open in figure viewerPowerPoint Model of the Vibrio anguillarum quorum‐sensing systems. For a detailed description of the model, refer to the text. It is not known whether the low cell population density model functions similarly to that of Vibrio harveyi. However, for simplicity of discussion, it will be predicted to function similarly. Thus, only the model at high cell population density is shown for indicating differences between species. ‘Question marks indicate unknown mechanisms’ (Milton 2006). The gene vanT is a positive regulator of metalloproteases (empA, prtV), pigment (hpdA), serine (serA) and biofilm production (sat, vps73) as well as a negative regulator of the type VI secretion system (hcp) (Croxatto et al. 2002; Weber et al. 2009, 2011; Lindell 2012) which is susceptible to malfunction as a result of interaction with certain RNAs. The genes vanO and sigma factor rpoN σ54 act as response regulators which initiate the production of four quorum regulatory RNAs (qrr1‐4) (Weber et al. 2008, 2011). Each Qrr exists as an RNA which inactivates mRNA produced by vanT when in the presence of an hfq‐encoded chaperone protein (Weber et al. 2011). The alternative sigma factor rpoS σ38 and uspA can repress the hfq‐expressed RNA chaperone protein which stabilizes vanT mRNA functionality (Weber et al. 2008). Quorum‐sensing systems in V. anguillarum are highly similar to those in other vibrios except for the presence of a second response regulator (rr‐2) which belongs to the ntrC protein family and is predicted to regulate qrr1‐4 (Weber et al. 2011; Lindell 2012). This pathway is understood to merge systems which are independent of quorum‐sensing with the phosphorelay quorum‐sensing networks in V. anguillarum (Weber et al. 2011; Lindell 2012). The rr‐2 can be phosphorylated by either the histidine kinase gene hk2 or the vanU phosphotransferase which acts as a branching‐point for the cross‐regulation of each phosphorelay pathway in merging them as one towards vanT (Weber et al. 2011; Bobay et al. 2014). Genes vanN, vanQ and cqsS autophosphorylate when cell densities are low which culminate in the phosphorylation of vanU and vanO which trigger qrr1‐4 destabilization of vanT mRNA functionality; vanT expression is initiated when cell densities are highest causing vanO dephosphorylation and qrr1‐4 in expression (Weber et al. 2011). This complexly layered communication system is not believed to be directly involved in virulence mechanisms but ultimately allows V. anguillarum to monitor its own population density (Milton et al. 1997), regulate stress responses to temperature shock and DNA damage (Lindell 2012), as well as adapt to ever‐changing circumstances for survival in the aquatic environment. Pathology and virulence Host‐adherence and invasion Vibrio anguillarum is able to adhere to fish surface cells, absorb fish mucus (Krovacek et al. 1987a,b), as well as aggressively penetrate host epithelial and vascular tissue (Krovacek et al. 1987a,b). The adhesive nature of V. anguillarum is dependent upon the functionality of the hfq gene which encodes for exopolysaccharide functionality and is involved in a series of quorum‐sensing mechanisms (Weber et al. 2010). The exopolysaccharide transport system enables V. anguillarum to maintain attachment to fish integument during the natural shedding of mucus by the host; operons involved in this mechanism are orf1‐wbfD‐wbfC‐wbfB and wza‐wzb‐wzc which encode for unidentified proteins and secretin, tyrosine kinase, tyrosine phosphatase, respectively (Croxatto et al. 2007). Adhesion of V. anguillarum is independent of the pathogen's flagella, but flagellar function must be maintained in order for the cell to invade host tissue (Ormonde et al. 2000). The highly conserved sigma factor rpoN σ54 gene contributes to environmental stress responses by regulating V. anguillarum protease secretion, exopolysaccharide production, as well as biofilm formation (Hao et al. 2013; Bobay et al. 2014), is essential for survival under iron‐limited conditions (O'Toole et al. 1996) and is essential in the production of flagella serving as a requirement for V. anguillarum virulence (O'Toole et al. 1997). The bacterium's only flagellum is comprised of five flagellin proteins which are encoded by genes flaABCDE and the proteins FlaA, FlaD and FlaE serve as direct contributors to V. anguillarum virulence (McGee et al. 1996; Milton et al. 1996). The flagella contains lipopolysaccharide surface antigens in the outermost flagellar sheath which are encoded by genes virA and virB, resistant to proteinase K, sensitive to periodate anion and serve as essential requirements for virulence in pathogenic V. anguillarum strains (Norqvist & Wolf‐Watz 1993). Vibrio anguillarum maintains ordinal migration preference towards intestinal, fish skin and gill mucus, respectively, by means of cheR encoded chemotactic motility due to the presence of amino acids, bile acids and carbohydrates therein which bait the pathogen towards intestinal colonization following host adhesion (O'Toole et al. 1999) and the process is essential for virulence among a host organism, although avirulent strains also exhibit chemotaxis in motility (O'Toole et al. 1996; Larsen et al. 2001). The same concept of chemotaxis in V. anguillarum holds true even in reference to human intestinal mucus denoting such mechanisms as factors which are host‐independent. Adhesion occurs more frequently within the mucus of fish intestines as opposed to surface skin cells (Chabrillón et al. 2004) as the intestinal lining of fish is rich in glycosphingolipids – critical components of endocytotic functions in fish digestion (Jennemann et al. 2012) – upon which the pathogen is able to utilize as a key attachment site for colonization (Chisada et al. 2013). Research suggests that V. anguillarum adhesion to host cells is facilitated using afimbrial means (Wang & Leung 2000), but the pathogen inherently holds the capability to assemble type IV pili (Rodkhum et al. 2006) which have been investigated for their role in host adhesion and virulence (Frans et al. 2013a), data which remain inconclusive at the present time, in terms of virulence contribution. The initial site of V. anguillarum entry into a host remains open to debate as the mouth (Cisar & Fryer 1969), gills (Evelyn 1984), anus (Chart & Munn 1980; Ringø et al. 2006), nasal cavity (Traxler & Li 1972), epidermis (Spanggaard et al. 2000) and eyes (Yiagnisis et al. 2007) have all been observed as regions of external vibriosis symptoms in fish. Fish gills garner an abundance of V. anguillarum levels during the earliest stages of vibriosis infection (Laurencin & Germon 1987) and are a major region of pathogenic uptake by fish hosts (Kato et al. 2013) but V. anguillarum exposure to the gills alone is not enough to trigger systemic infection (Kanno et al. 1989) or V. anguillarum invasion of internal organs (Olsson et al. 1996). Vibrio anguillarum is consumed orally by fish in the marine setting and is able to survive in the acidic environment of the stomach for several hours prior to intestinal relocation and faecal expulsion (Olsson et al. 1998) but oral consumption of the pathogen alone rarely results in vibriosis infection (Laurencin & Germon 1987). Our current understanding of the V. anguillarum portal of entry in a host resonates around chemotactic attraction of the pathogen to mucosal surfaces (Bordas et al. 1996, 1998) with surface skin and fins being the immediate site of attachment (Spanggaard et al. 2000) and it is generally understood that V. anguillarum is able to penetrate a host through epithelial tissue or the gastrointestinal tract via oral or anal entry (O'Toole et al. 2004). Transparent zebra fish appear at present to be the most suitable fish sample for monitoring V. anguillarum invasion when the pathogen is labelled with a fluorescent marker (O'Toole et al. 2004; Oyarbide et al. 2013). Invasion of a host by V. anguillarum ultimately results in the adherence to and colonization of the gut; an anatomical location in which the bacterial cell exhibits strong replication yields (Horne & Baxendale 1983) and serves as a sort of enrichment site for V. anguillarum (Olsson et al. 1998). It is believed that V. anguillarum might possess outer‐membrane vesicles which transport virulence factors onto enterocytes for interaction and pathogenesis when colonized in the host intestine (Rekecki et al. 2013). Vibrio anguillarum does not cause damage to saline‐rinsed intestinal tissue ex vivo (Martinsen et al. 2011) but virulence interactions with enterocytes are evident when the pathogen is in contact with mucin, bile salts and cholesterol as these intestinal factors trigger an increase in protease activity, flagellar motility, biofilm formation and exopolysaccharide production; respective expressions of genes empA (metalloprotease), fleQ (flagellar transcriptional regulator), flaA (flagellin protein), cheR (chemotaxis methyltransferase), wbfD (exopolysaccharide synthesis) and wza (exopolysaccharide export) are specifically enhanced under these conditions (Li et al. 2015). The virulence factors that are initiated following contact with such host‐endogenous cues result in the growth and release of hypervirulent V. anguillarum cells which are easily transmitted by contact among fish populations that are densely contained in high aquaculture quantities therein (Li et al. 2015). Blue oysters have been shown to accumulate and concentrate such virulent V. anguillarum cells which are released via faecal matter as a vector of enhanced pathogenicity in the marine environment (Pietrak et al. 2012). Host gills, anterior and posterior intestinal tissue, as well as oral and anal tissue commonly exude vibriosis symptoms within several hours of the onset of infection but V. anguillarum invasion of internal organs such as the kidney or spleen can take as long as 24 h (Ransom et al. 1984; Laurencin & Germon 1987). Upon exposure to the host kidney, spleen, or lymphocytes, V. anguillarum induces mitosis in such host cells due to antigen content in the bacterial cell and the presence of certain lipopolysaccharides among the bacterial cell wall which are inflammatory to such host tissues (Shimizu et al. 1981; Yui & Kaattari 1987; Boltaña et al. 2014). Fish skin mucus often contains concentrations of antimicrobial substances such as lysozyme or specific peptide components which block bacterial access to the epithelial cellular membrane (Ellis 2001). Vibrio anguillarum secretes extracellular compounds that render the pathogen resistant to such antimicrobial mechanisms which aid the bacteria in degrading fish skin (Weber et al. 2010) and the pathogen does not require phagocytotic uptake by fish macrophages for survival in a host (Boesen et al. 1999a,b; Larsen & Boesen 2001). Vibrio anguillarum is resistant to bile and produces biofilm which enhances the ability of the cell to remain viable under stressful conditions, in both seawater and within a host (Wang et al. 2003); lipopolysaccharide side chains in V. anguillarum are essential for resistance to antibacterial serum produced by host fish (Welch & Crosa 2005). Scallops that are infected by V. anguillarum are unable to carry out normal metabolic functions when stressed by heat and thus cannot synthesize essential proteins to sustain life (Sun et al. 2014). Shellfish are susceptible to oxidative stress and related physiological damages following V. anguillarum infection (Liu et al. 2013) and the pathogen also holds potential to interfere with osmolarity regulation in a host which has been proven lethal in mussels (Wu et al. 2013). Vibrio anguillarum secretes extracellular substances, which are lethal to a variety of fish at low dosage (Inamura et al. 1984, 1985), are thermo‐stable (DiSalvo et al. 1978), acid‐stable, proteolytic and functional in the presence of trypsin (Kodama et al. 1985), a pancreatic enzyme that is actively involved in digestion within the small intestine. These extracellular secretions comprise of a range of virulence factors including metalloproteolytic caseinase, aminopeptidase and collagenase (Stensvåg et al. 1993), haemolytic substances (Munn 1978), as well as water‐soluble iron‐chelating siderophores (Crosa 1989). Certain extracellular secretions by V. anguillarum have been shown to reduce white blood cell quantities in a host (leukopenia), reduce lymphocytes (lymphopenia), increase monocytes which are able to phagocytotically absorb whole‐cell bacteria (Lamas et al. 1994), as well as inhibit leucocyte respiratory function and apoptosis which allow the pathogen to survive and thrive in a fish host (Sepulcre et al. 2007). Each extracellular component contributes to a different task in the survival of V. anguillarum within a host and offers researchers the necessary evidence to plot the course of the pathogen from initial host contamination to ultimate lethal infection. Vibrio anguillarum secretes a zinc metalloprotease which is encoded by the highly conserved (Yang et al. 2007) empA gene and contributes to virulence by degrading tissue within a host resulting in visible lesions among infected fish (Norqvist et al. 1990; Milton et al. 1992). The zinc metalloprotease is expressed as an inactive (Staroscik et al. 2005) 46‐kDa proenzyme (Varina et al. 2008) when V. anguillarum is in contact with intestinal mucus (Garcia et al. 1997; Denkin & Nelson 1999) at which point the gene epp is expressed producing a ‘pro‐EmpA processing’ protease which activates zinc‐metalloproteolytic function by cleaving a ~10‐kDa propeptide from the empA‐encoded proenzyme thus initiating EmpA function (Varina et al. 2008). EmpA expression is dependent upon rpoS encoded σ38 expression during stationary growth and vanT‐based quorum‐sensing mechanisms in addition to mucosal contact among the intestinal tissue of a host (Croxatto et al. 2002; Denkin & Nelson 2004). The expression of empA zinc metalloprotease by V. anguillarum in a host results in higher colonization yields, stronger growth rate, as well as increased invasion and survival by the pathogen which ultimately encourages an increase in host mortality rates (Denkin & Nelson 1999); recent data suggest that empA functionality also contributes to stronger motility and haemolytic activity but the subject requires further verification (Han et al. 2011). Recently, the gene mltD was determined to encode for a putative membrane‐bound lytic murein transglycosylase D protein which is involved in haemolysis, phospholipase, gelatinase and diastase activities and is believed to be linked to V. anguillarum virulence (Xu et al. 2011). Extracellular products of V. anguillarum collectively cause severe damage to vascular tissue, muscular tissue and are able to degrade vital internal organs of a host such as the liver and spleen (de la Cruz & Muroga 1989). Iron appropriation and haemolysis Vibrio anguillarum harbours a 50‐megadalton (Mdal) plasmid class that is known to enhance the bacterium's pathogenicity and its discovery provided researchers with the first genetic marker of V. anguillarum virulence elucidation in the pathogen's history (Crosa et al. 1977). Fish naturally possess immunological defence mechanisms in which iron ions are bound within transferrin and lactoferrin proteins making iron inaccessible to invading pathogenic microbes (Crosa 1980). Vibrio anguillarum facilitates a ‘plasmid‐mediated iron‐sequestering system,’ identified as the 67‐kilobasepair (kbp) ‘pJM1 plasmid series,’ that is highly efficient in dislocating protein‐bound iron in a host for uptake within its cell (Crosa et al. 1980). The pJM1 plasmid has been sequenced in full by Di Lorenzo et al. (2003). Inadequate iron concentration within the V. anguillarum cell promotes pJM1 plasmid syntheses of the diffusible, iron‐chelating siderophore ‘anguibactin’ (Actis et al. 1985, 1986) and its respective iron–siderophore complex outer‐membrane protein receptor ‘OM2’ (Crosa & Hodges 1981) for iron acquisition into the cell (Walter et al. 1983; Actis et al. 1985). Vibrio anguillarum is able to maintain the assembly of its outer‐membrane proteins under viable conditions regardless of growth medium or environmental circumstances (Buckley et al. 1981). It is essential that V. anguillarum express genes fatABCD and ORF1‐5 in order for the pJM1 iron‐sequestering system to function in acquiring iron that is bound within anguibactin complexes using the outer‐membrane receptor protein OM2 (Actis et al. 1988; Köster 2001; Köster et al. 1991). Ferric‐siderophore anguibactin complexes are bound and transported into the V. anguillarum periplasm by the fatA gene‐encoded OM2 protein where a lipoprotein encoded by the gene fatB internalizes the iron with the assistance of integral proteins encoded by genes fatCD (Köster et al. 1991); ORF1‐5 serve as regulating genes of OM2 expression (Actis et al. 1988). An angB chromosome‐encoded 2,3‐dihydroxybenzoic acid is required as a precursor to anguibactin meaning the pJM1 iron‐sequestering system is not entirely plasmid‐based (Chen et al. 1994; Welch et al. 2000) and the additional functionality of the genes angA and angM are essential for the biosynthesis of anguibactin in V. anguillarum (Alice et al. 2005). The gene hdc encodes for a 44.26‐kDa histidine decarboxylase enzyme that is useful in the synthesis of anguibactin from histidine which also serves as a valid anguibactin precursor (Tolmasky et al. 1995). The gene angR serves as a trans‐acting factor (Salinas & Crosa 1995) that is necessary for pJM1 functionality and is regulated by Fe2+ at the transcriptional level encoding for a DNA‐binding protein which also positively modulates other Fe2+ regulated transcription (Farrell et al. 1990; Chen et al. 1996); the protein is homologous to that of the pathogen Yersinia enterocolitica suggesting a possible genetic relationship between the two species (Guilvout et al. 1993). The gene fur is expressed when iron levels among V. anguillarum are sufficient to conserve resources and halt iron‐uptake mechanisms (Tolmasky et al. 1994; Zheleznova et al. 2000) and the gene angT can intervene in the facilitation of angR encoded anguibactin synthesis at any time (Wertheimer et al. 1999). A regulating antisense RNAα is activated under iron‐rich conditions which initiates the gene fatA and inhibits the expression of the gene fatB; the RNA sequence is composed partially of fatA and fatB sequences as it extends from one to the other (Salinas et al. 1993; Waldbeser et al. 1995). A 427‐nucleotide antisense RNAβ then acts as a transcriptional terminator which stops the iron‐transport operon in V. anguillarum (Salinas et al. 1993; Stork et al. 2007). A variety of genes involved in the pJM1 plasmid series have been identified and their chronological progression among the expression of the virulence plasmid is shown in Figure 3. Related derivatives of the pJM1 plasmid series exist in V. anguillarum isolates as siderophore inducers but are present in both virulent and avirulent strains deeming them ineligible for pathogenic profiling (Pederson et al. 1997; Wu et al. 2004). image Figure 3 Open in figure viewerPowerPoint Schematic representation of the pJM1 plasmid (Stork et al. 2002). A second iron‐uptake system is present in V. anguillarum which functions independently of the pJM1 iron‐sequestering system and its respective OM2 receptor protein, based on chromosomally encoded mediation (Lemos et al. 1988). The pJM1 plasmid series is present in most V. anguillarum O1 strains but is absent in serotype O2 while the chromosome‐mediated iron‐uptake system exists in both the O1 and O2 serotype (Conchas et al. 1991). Vibrio anguillarum biotypes which exhibit chromosome‐mediated iron uptake do so using the siderophore vanchrobactin as opposed to anguibactin which is used in plasmid‐mediated iron uptake (Mackie & Birkbeck 1992; Soengas et al. 2006). Vanchrobactin synthesis is encoded by the genes vabABC which are regulated by the activating gene vabE and the non‐ribosomal peptide synthetase gene vanF (Balado et al. 2006). The receptor protein which is responsible for binding and acquiring iron into the cell using the vanchrobactin siderophore is encoded by the gene fvtA (Balado et al. 2009) and the genes responsible for vanchrobactin synthesis are present in all V. anguillarum strains suggesting that the vanchrobactin iron‐uptake system is ancestral to the anguibactin‐based system which was likely acquired later via horizontal gene transfer (Lemos et al. 2010). Evidence suggests that V. harveyi is the likely ancestor of the anguibactin‐based iron‐uptake system which is present in V. anguillarum, although the transition could have also been the reverse (Naka et al. 2013a,b). The anguibactin siderophore is far more efficient in V. anguillarum iron acquisition than is vanchrobactin (Naka et al. 2008) and V. anguillarum‐produced siderophores compete strongly against neighbouring pathogens thus inhibiting their growth under iron‐deficient conditions as a result (Pybus et al. 1994). Genes fatBCDE and fvtBCDE act as ATP‐binding cassette transporters in ferric‐siderophore uptake mechanisms across the V. anguillarum cytoplasmic membrane, although the fat genes encode for anguibactin‐based systems on the pJM1 plasmid and the fvt genes encode for vanchrobactin systems on the chromosome (Naka et al. 2013a,b). Iron appropriation is also possible when virulent V. anguillarum strains secrete haemolysins that disrupt erythrocyte stability allowing for post‐rupture‐abandoned iron ions to be absorbed readily into the cell and this occurs most frequently during stationary‐phased growth (Fig. 4) (Munn 1978). Vibrio anguillarum can utilize haeme‐compounds as an adequate source of iron only when the genes huvAZBCD are collectively functional (Mouriño et al. 2004) and it is now understood that V. anguillarum serotypes O1‐O10 can each utilize haeme‐compounds as an iron source regardless of geographic origin (Mouriño et al. 2005, 2006). Vibrio anguillarum haeme‐based iron‐uptake outer‐membrane proteins are normally encoded by the huvA gene (Mouriño et al. 2005). However, serotype O3 – a serotype of higher virulence under conditions which are high in iron concentration – harbours a haeme‐based iron‐uptake outer‐membrane protein unlike that of any other V. anguillarum serotype (Muiño et al. 2001) which is encoded by the gene huvS, aiding in its discrimination (Mouriño et al. 2005). Vibrio anguillarum contains several virulence gene clusters that encode for haemolysin secretion including llpA, vah1 (Hirono et al. 1996), vah2‐5 and rtx (Rodkhum et al. 2006; Li et al. 2008). The gene plp negatively regulates haemolytic expression of llpA and vah1 (Rock & Nelson 2006) while the gene hlyU – which is dependent upon its own positive regulation by the gene hns (Mou et al. 2013) – is responsible for the positive regulation of both vah1 and rtx (Li et al. 2011); genetic regulation of the vah2‐5 genes are unclear at the present time. Negative regulation by plp is only partial to its role in V. anguillarum haemolysis as the gene additionally encodes for a specific phospholipase secretion which degrades phosphatidylcholine, a class of phospholipids that comprise approximately 58% of fish erythrocytes (Lee et al. 1989; Li et al. 2013a,b). The plp gene‐encoded phospholipase does not affect V. anguillarum virulence but is now understood to impact fish erythrocytes more significantly than vah1 or rtx and operates at a wide‐range of both pH (5–9) as well as temperature (34–64°C); vah1 and rtx are known virulence factors of V. anguillarum but do not encode for phosphatidylcholine degradation (Li et al. 2013a,b). Vibrio anguillarum is able to lyse human, sheep and fish erythrocytes (Toranzo et al. 1987), but the plp‐encoded phospholipase is only compatible for haemolysis in fish erythrocytes and does so more strongly in fish than other erythrocyte samples while vah1‐ and rtx‐encoded haemolysins disrupt sheep erythrocytes more strongly than plp (Li et al. 2013a,b). Serotypes O1 through O10 all maintain the ability to utilize haemin and haemoglobin as sources of iron (Mouriño et al. 2004) through the expression of either genes huvA or huvS which encode for outer‐membrane reception and uptake of haeme‐compounds into the cell (Mouriño et al. 2005); only serotypes O1 through O3 are considered pathogenic by statistical majority (Larsen et al. 1994; Tiainen et al. 1997). image Figure 4 Open in figure viewerPowerPoint Vibrio anguillarum adopts iron into the cell by means of (a) siderophore acquisition, (b) haemolysin‐based iron release and (c) haeme‐binding mechanisms which are understood to share either chromosome or plasmid genetic bases. Fish erythrocytes are particularly vulnerable to V. anguillarum disruption by plp gene‐based phosphatidylcholine degradation in the red blood cell wall. Haeme‐binding and haemagglutination Vibrio anguillarum contains cellular membrane proteins that bind specifically to haeme or haemoglobin regardless of iron‐concentration vicinal to the cell (Mazoy & Lemos 1996; Mazoy et al. 1997b) and is able to produce haemagglutinins (Oishi et al. 1979) that theoretically cause red blood cells to aggregate into erythrocyte‐adhesive masses. It has been determined that V. anguillarum haemagglutinins are ineffective in the agglutination of striped bass (Toranzo et al. 1983) and rainbow trout (Larsen & Mellergaard 1984) erythrocytes in vivo, however, V. anguillarum haemagglutinins were confirmed to exhibit agglutination of human, poultry, guinea pig, trout, horse and pig erythrocytes, more consistently by serotype O2 than O1 (Larsen et al. 1988). Johnsen (1977) used V. anguillarum‐derived lipopolysaccharides as an in vitro method of vibriosis diagnoses in infected fish as the lipopolysaccharides exhibited haemagglutination in fish tissue extract. The impact of such mechanisms towards the virulence of the pathogen remains undetermined and insufficient when considering the wide‐range of marine organisms that are also susceptible to vibriosis. The notion that V. anguillarum haemagglutinins could competitively inhibit the binding of neighbouring microbes and microbial products to animal cells or combine with latent physiological substances to independently drive their activation has been suggested (Oishi et al. 1979) but research in the area appears to have been neglected entirely since that time. The research possibilities in this area are quite tempting to speculate given our current knowledge of V. anguillarum interaction with marine organisms and the ability of the pathogen to aggressively invade fish tissue, specifically circulatory matter and related haemolytics. Modern research into the haeme‐binding properties and haemagglutinins involved in V. anguillarum virulence is important towards developing a more universal understanding of the Vibrionaceae Proteobacterial family taxon, V. anguillarum as an invasive marine pathogen, and towards the development of a viably natural‐aquaculture industry free of chemical treatments in the form of vaccinations or antibiotics. Detection strategies and diagnostic procedures Vibrio anguillarum can be cultured using brain heart infusion, trypticase soy supplemented with 1–2% NaCl (Crosa et al. 2006) and VAM growth medium (Alsina et al. 1994), but no one media is specialized for the growth of V. anguillarum alone. In certain cases, V. anguillarum can also be isolated using thiosulphate citrate bile salt sucrose agar (TCBS) (Kobayashi et al. 1963), salt‐starch agar (Baross & Liston 1968) and bromothymol blue‐teepol‐salt agar (Bartley & Slanetz 1971) but V. anguillarum is not usually culturable by such means and these methods are more commonly applicable to the isolation of other Vibrio species. Methods of V. anguillarum detection have shifted away from culture‐based methods to accelerate diagnostic and preventative measures in reference to vibriosis. Medium cultivation of V. anguillarum has its benefits when studying the pathogen in a laboratory setting but can take several days to identify rendering the procedure obsolete when attempting to detect the strain or diagnose an infection. Furthermore, the virulence capacity of a V. anguillarum isolate is highly complex and the genetic expression of certain virulence factors varies among pathogenic strains as a result of environmental interactions which are isolate‐specific (Kühn et al. 1996; Frans et al. 2013b; Busschaert et al. 2014). Detection measures have focused less on the bacterial isolate itself but rather the antigen content or genetic material that is contained within the cell. The early transition between culture‐based detection and genetic detection consisted of a trn9 gene‐specific DNA hybridization probe which was combined with a growth colony to establish a dot‐blot DNA hybridization procedure which could detect as little as Log 4 V. anguillarum CFU colony−1 (Aoki et al. 1989). Other methods consisted of immunoassays based on enzyme‐linked immune sorbent assays (ELISA) which were proven to be specific in detecting V. anguillarum but lacked competitive sensitivity being able to detect only as little as Log 5 V. anguillarum CFU assay−1 (Gonzalez et al. 2004). Using the polymerase chain reaction (PCR), V. anguillarum detection can be detected in a matter of hours and several protocols have been established serving as benchmark detection methods which targeted genes such as amiB, rpoS, 16S, toxR, rpoA, rpoN, empA, virA/B and vah1‐5 (Martínez‐Picado et al. 1994; Rodkhum et al. 2005; Demircan & Candan 2006; Hong et al. 2007; Kim et al. 2008; Prol et al. 2008; Xiao et al. 2009; Zhang et al. 2014ab). The virulence or species specificity and detection sensitivity of each molecular assay varies by study but overall allow technologists to diagnose vibriosis or detect V. anguillarum in rapid fashion even when V. anguillarum isolates are dormant or non‐culturable. Isothermal methods such as loop‐mediated isothermal amplification (LAMP) (Gao et al. 2010; Yu et al. 2013) and upconversion fluorescent strip sensor detection (Zhao et al. 2014) have also been configured to detect V. anguillarum and novel procedures to detect and quantify the pathogen are likely to continue to emerge with increased accessibility to technological advancements over time. Some diagnostic methods consist of measuring immune responses among an infected host such as prophenoloxidase which is expressed by shrimp within hours of being infected with V. anguillarum (Gao et al. 2009). Research into primary immune responses which are respective to certain aquaculture species infected with V. anguillarum could serve as vectors of novel diagnostic measures which are financially less expensive than cutting‐edge molecular assays. Furthermore, the identification of V. anguillarum by use of culture‐based methods still holds value today if time is not a factor of concern and it is certainly valid to identify a V. anguillarum isolate using the API 20E system (BioMerieux S.A, Marcy‐rEtoile, France; Grisez et al. 1991). Vaccination and prevention Sterilization and dietetic prevention Preventative procedures are taken very seriously by aquaculture farmers in terms of time commitment, application efficacy, as well as financial costs and such measures are the first steps in averting vibriosis in fish (Lillehaug 1989b). A great deal of research has been conducted over the past few decades which helped to elucidate measures of V. anguillarum growth inhibition and to diminish mortality rates that result from V. anguillarum infection. For example, the soluble mixture halquinol, which results from the chlorination of 8‐hydroxyquinoline, and the compounds sulphamethoxazol and trimethoprim have been shown to inhibit viable growth of V. anguillarum in the presence of fish (Austin et al. 1981). The V. anguillarum cell is inactivated following exposure to 44°C for as little as 3 min or 47.5°C for as little as 2 min and aquaculture water temperature can be elevated prior to the transfer of fish to each containing pool (Jacobsen & Liltved 1988). Ozone treatment as little as 1 mg L−1 is sufficient eradicating aquaculture presence of V. anguillarum entirely (Sugita et al. 1992) and the continuous application of antimicrobial peptides (cecropin‐bee melittin hybrid peptide, CEME and pleurocidin amide) is known to reduce vibriosis mortalities by up to 50% (Jia et al. 2000). The modification of aquaculture feed has shown promise as a consistent vector of disease prevention in fish and the inclusion of Selenium (Le & Fotedar 2014), hen‐derived egg yolk immunoglobulins (Li et al. 2014a) and oregano‐derived carvacrol (Volpatti et al. 2014) are proven feed additives able to reduce V. anguillarum mortalities in fish. The adhesive nature of V. anguillarum is a critical virulence factor which can be targeted by prophylactic compounds or modified aquaculture feed to prevent the onset of vibriosis in a host (Chabrillón et al. 2006). Certain bacterial species can be applied to feed as antagonists which regularly maintain fish digestive tracts that are free of V. anguillarum‐attached enterocytes (Olsson et al. 1998). Recently, probiotic supplementation of Pediococcus pentosaceus was shown to increase resistance of orange‐spotted grouper to V. anguillarum primarily due to lactic acid production as a result of host digestion (Huang et al. 2014). Vibrio anguillarum mortalities have been reduced by probiotic administration of V. alginolyticus by as much as 16% (Austin et al. 1995), Vagococcus fluvialis by over 40% (Sorroza et al. 2012), Pseudomonas fluorescens AH2 by as much as 50% (Gram et al. 1999; Holmstrøm & Gram 2003) and extracellular proteins of probiotics Kocuria spp. and Rhodococcus spp. by as much as 75% (Sharifuzzaman et al. 2011). Vaccine administration and efficacy Vaccines against vibriosis are administered to fish cultures using several different exposure methods and each vaccine composition is variable to accommodate the immunological requirements of a given fish species. Initial vaccination procedures against vibriosis involved the exposure of heat‐ (Fletcher & White 1973) or formaldehyde‐killed V. anguillarum cultures to fish (Rombout et al. 1986) but have since expanded to a variety of delivery methods such as feed in the form of wet‐packed whole‐cell or lyophilized whole‐cell bacterin, injections for instalment into the host peritoneal cavity (Fryer et al. 1978) or muscle and vaccine‐containing immersion solutions in which fish are submerged prior to relocation into aquaculture pools (Miyazaki 1987). Vibriosis vaccines can also be delivered by injection in the form of monoclonal anti‐idiotype antibodies as surrogate antigens which induce protective immunity against V. anguillarum in fish (Yongjuan et al. 2002) and outer‐membrane antigens derived from adhesion proteins of Aeromonas hydrophila are also applicable as immunization factors against vibriosis (Fang et al. 2000). Live‐attenuated vaccines have been developed in which V. anguillarum cultures are mutated but remain viable for effective immunization in fish (Norqvist et al. 1989). Plasmid‐free V. anguillarum strains are strong candidates for attenuated vaccines and can be genetically modified to produce foreign proteins which enhance immunization in fish depending on the expressed secretion (Shao et al. 2005). Attenuated vaccines can also be freeze‐dried as lyophilized pellets of viable cells but require protection using skim milk (Yang et al. 2005). Live‐attenuated vaccines (Ma et al. 2010) induce mucosal immune responses within host intestines (Zhang et al. 2014a) when administered by bath submersion and the anus is believed to be the main portal of live‐attenuated vaccinal entry in reference to these circumstances (Liu et al. 2014). The role of V. anguillarum flagellins is believed to vary in reference to immunization mechanisms in fish as the protein FlaB induces stronger protection against vibriosis infection while the V. anguillarum FlaE protein has been shown to enhance vaccination against Edwardsiella tarda (Jia et al. 2013). The body mucus of vibriosis‐immunized fish is in some cases antimicrobial to V. anguillarum (Harrell et al. 1976; Boesen et al. 1999a,b) and the kidney cells or plasma from such fish can be used as intraperitoneal vaccine injections for other fish as well (Viele et al. 1980); however, immunization efficacy of such host serum varies by fish species (Bøgwald et al. 1994). Various fish disease immunizations can also be combined as one simultaneous vaccine (Amend & Johnson 1984) and the practice is encouraged as vaccination of fish against V. anguillarum alone can result in an increase in fish infection by other marine pathogens such as A. hydrophila, Vibrio pelagius, Vibrio splendidus and Vibrio vulnificus (Austin 1984; Pazos et al. 1993), contrary to the theory that antigenic interference would prevent multi‐vaccine efficacy (Pross & Eidinger 1974). Furthermore, the extracellular products and cell‐surface components of each V. anguillarum serotype are often different and there is reason to believe that each serotype should be included in V. anguillarum vaccines to fully ensure efficacy in immunization (Espelid et al. 1991; Santos et al. 1995). Oral vaccines have proven to be the least effective when immunizing fish against vibriosis, in comparison to injection or immersion procedures, and this seems to be attributable to gastric fluid in the upper intestine in which digestive enzymes inactivate critical components of the active ingredients among vaccines which render the compounds less‐effective (Johnson & Amend 1983; Hart et al. 1988). Additives such as sodium salicylate, sodium caprate and vitamin E can enhance absorption of oral vaccines and feed pellets can be coated to protect against degradation during digestion (Vervarcke et al. 2004). Anal administration of formalin‐killed (Rombout et al. 1986) V. anguillarum or related vibriosis vaccines have proven to be 30% more effective than vaccines administered orally but vaccines administered by injection (Li et al. 2005) or immersion have proven to be effective in preventing vibriosis by as much as 100% (Bowden et al. 2002; Angelidis et al. 2006). The efficacy of vaccines delivered by immersion is enhanced by ultrasonic pulsation of the exposed vaccination solution (Zhou et al. 2002) and when fish are submerged in a hyperosmotic solution prior to vaccinal exposure (Croy & Amend 1977; Antipa et al. 1980) but the risk of vibriosis between vaccination and transfer to the marine setting increases with delays in duration (Lillehaug 1989b). Delivery of vaccines by immersion is most efficient in warmer temperature using water‐soluble compounds as opposed to whole‐cell bacterin and uptake by fish is enhanced when the solution contains concentrations of potassium aluminium sulphate (Tatner & Horne 1983a). Vaccines are not full‐proof, and in some cases, efficacy is reduced by over 20% as soon as 2–4 weeks postvaccination requiring fish to receive additional booster shots to maintain aquaculture yields and prevent mortalities (Zhang et al. 2014b). Injectable vaccines are the least expensive immunization measure in protecting fish against vibriosis but the cost of labour and consumption of time associated with vaccinating aquaculture masses greatly enhances its financial impact on aquaculture companies (Lillehaug 1989a). Vaccination by oral administration and immunization by immersion are more financially costly in terms of materials but the most decisive factor in purchasing vaccinations for aquaculture is efficacy. Aquaculture farmers are more likely to pay more for vaccines which are at least 99% effective against vibriosis or if the aquaculture farmer has experienced a vibriosis outbreak in the past (Thorburn & Carpenter 1987). Fish vaccinated by immersion must each be submerged in an immunizing solution for at least 5 s to achieve optimal efficacy and, rather than age, the size of the fish is the most important factor when determining an appropriate dose to vaccinate fish (Johnson et al. 1982b). Larger fish take‐in vaccines better than smaller fish when administered by immersion (Tatner & Horne 1983) while smaller fish are more susceptible than larger fish in contracting vibriosis and vaccination doses require alteration to compensate as the duration of immunity following vaccination is understood to vary by fish species (Johnson et al. 1982a). In some cases, fish cannot be vaccinated against vibriosis until 6–8 weeks after hatching and the immunization of juvenile or larval aquaculture has proven to be challenging as a result (Tatner & Horne 1983ab). Larvae cultures that are pre‐exposed to vaccines prior to maturation have no effect on immunization against vibriosis, and in some cases, hatchlings must grow to at least 0.4 g prior to valid vaccination utilization (Tatner & Horne 1984). Rather than attempt to vaccinate juvenile aquacultures, prevention measures against vibriosis infection have proven to be the best alternative in protecting hatchlings during early stages of growth. For example, the application of polyb‐hydroxybutyrate (PHB) is known to protect crab larvae against V. anguillarum‐induced mortalities and promotes strong growth yields among the larvae culture (Sui et al. 2012) while mannan oligosaccharides included in feed can prevent V. anguillarum‐induced mortalities by over 50% in juvenile fish by protecting the cultures against pathogenic intestinal colonization (Torrecillas et al. 2012). Cultures of Roseobacteriae can completely eradicate V. anguillarum in larval aquaculture settings due to the production of tropodithietic acid by the microbe and it has been suggested that Roseobacteriae inclusion among aquaculture circulation filters could improve larvae culture growth yields in protection against V. anguillarum infection (D'Alvise et al. 2010). Formalin‐killed V. anguillarum has proven to be suitable for the protection of post‐larvae shrimp cultures against vibriosis (Patil et al. 2014). Rotifers fed Crataegi fructus or a mixture of C. fructus, Artemisia capillaries and Cnidium officinale can develop resistance to V. anguillarum which can promote growth yields of larvae culture when used as feed (Takaoka et al. 2011). Vibrio anguillarum treated with indole prior to larvae exposure decreases vibriosis mortalities as indole decreases V. anguillarum biofilm production and inhibits the expression of the gene wbfD which is a virulence factor responsible for exopolysaccharide production; the same effects can be induced by deleting the rpoS gene (Li et al. 2014b). Aquaculture farmers generally do not purchase surplus juvenile fish quantities to compensate for potential V. anguillarum‐induced losses but are willing to pay more for younger fish cultures which are resistant to vibriosis infection (Thorburn & Carpenter 1987). Vibriosis treatment and resistance Procedures to treat vibriosis infection in fish are less efficient than vaccination methodologies in producing high aquaculture yields but are often necessary to preserve fish production quantities and financial stature. Terramycin can be administered in fish feed as a treatment of V. anguillarum infection but mortality yields are not entirely suspended (Sawyer & Strout 1977). The nitrofuran‐derivative compound nifurpirinol is able to kill V. anguillarum and eliminate the pathogen from the bloodstream of infected fish in as little as 2 h when administered in the form of immersion bathing, serving as a useful therapeutic to treat vibriosis in fish (Pearse et al. 1974). Treatment can be administered orally using antimicrobial oxalic acid (Samuelsen et al. 2003) and florfenicol but mortality rates increase with delayed treatment (Seljestokken et al. 2006). Antibiotics such as tetracycline, sulfaisozole and sulphamonomethioxine (Sano & Fukuda 1987) do not always cure vibriosis in fish and some argue that V. anguillarum infection progresses too rapidly to treat aquacultures based solely on symptomatic diagnoses (Flücher 1979). Vibrio anguillarum strains often harbour genes such as tet which allows the pathogen to remain resistant to tetracycline (Zhao & Aoki 1992b), cat which encodes for chloramphenicol acetyltransferase among a transferrable r‐plasmid (Zhao & Aoki 1992a), or gyrA and parC which contribute to quinolone resistance (Rodkhum et al. 2008) in V. anguillarum which contribute to the pathogens resistance of various chemical treatments (Palm et al. 2003). Vaccinations by injection of immunized fish‐derived serum extract is also susceptible to V. anguillarum resistance as serotype O2 develops high‐molecular‐weight O‐antigen (Bøgwald et al. 1990) lipopolysaccharide side chains that prevent serum damage to the cell (Boesen et al. 1999a,b). Resistance of V. anguillarum to antibiotics and vaccines varies by strain and the pathogen is known to transfer r‐plasmids among compatible bacterial neighbours in nature. Genetic expression by liver and spleen cells of live‐attenuated immunized fish vary in comparison to killed V. anguillarum vaccines and it has been suggested that such genetic variations can be explored to identify genetic V. anguillarum resistance in fish cultures (Pan et al. 2011). Future vaccination and prevention measures against V. anguillarum in aquaculture appear to focus on genetic modifications which allow for the manipulation of protein expression in whole‐cell vaccines (Yang et al. 2008), modification of virulence in V. anguillarum strains by genetic transformation with different bacterial species (Cutrín et al. 1995) and the directed expression of specific surface antigens from various bacteria in live‐attenuated V. anguillarum cells to universally protect fish against marine microbial pathogens (Wang et al. 2009). Closing remarks Future research Research studies into V. anguillarum have historically focused on three major categories: (i) further understanding the organism's viability in nature, (ii) determining the conditions or related mechanisms at which such factors contribute to virulence or infection, and (iii) identifying points of vulnerability that allow for the prevention or control of such pathogenic presence in aquaculture settings. There is sufficient evidence among available literature that the comprehension of V. anguillarum viability and virulence is well established to the point that private companies are able to profit on such findings via the commercialization of preventative cocktails or vaccinations. Considerable opportunity remains for additional research in vibriosis prevention as vaccination in aquacultures can be specifically optimized for each respective fish species (Joosten et al. 1997). However, because vibriosis vaccination of aquaculture has proven successful (Joosten et al. 1997), research efforts towards V. anguillarum vaccination will effectively diminish over the coming years. Furthermore, a new generation of research is now at its infancy in which natural methods of high‐throughput food production are in high demand by health‐conscious consumers and legislative officials will continue to rule out the ways of the old (growth hormones, antibiotics, vaccinations, etc.) in the coming decades for new, natural preventative measures that are proven effective in application. Although future microbiological studies will certainly revolve around next‐generation sequencing (NGS) technologies and the prevalence of probiotic studies will surely sustain for some time, the future of vibrio research is without question phage therapy in aquaculture. Whether delivered in the form of feed pellets or by direct release into the environment, bacteriophage have shown substantial promise in agricultural sciences and their application in aquaculture has proven successful in long‐term efficacy even at the nanogram scale of a single dose (Xia et al. 2005; Wright et al. 2009; Monk et al. 2005). Research into vibrio‐phage has recently gained attention in the field after each of the three major phage families (Myoviridae, Podoviridae and Siphoviridae) were proven to contain at least one species that is effectively specific to the lysis of the V. anguillarum cell wall (Tan et al. 2014). However, studies related to the environmental application of biocontrol agents such as antibiotics and vaccinations face the ever‐present challenge of bacterial resistance or mutation and phage therapy in aquaculture is no exception. Even slight variances in wild‐strain V. anguillarum genotypes have been proven to evade infection among artificial vibrio‐phage challenge, and so research in the area remains widely open‐ended. Alongside this, many research opportunities exist in the area of Moritella viscosa – a bacterium that is closely similar to V. anguillarum – which is gaining increased attention for its vibriosis‐resembling fish infection winter ulcer disease that is causing devastating losses in fisheries located in geographies of colder climates. Many V. anguillarum researchers will shift their focus to the lesser understood M. viscosa because, in addition to its similarity to V. anguillarum, measures of its control are nearly absent at the present time and their establishment is critical to the industry (Colquhoun & Lillehaug 2014). Ultimately with time, aquaculture will stray from its present practices and evolve to more resemble natural, functional marine habitats. Romero et al. (2014) demonstrated that Chilean mussels naturally produced V. anguillarum‐specific Podoviridae‐phage CHOED in situ and in vitro (Higuera et al. 2013). If facilities cultured such submerged organisms as Chilean mussels in the bottom of each aquaculture tank, healthy fish masses could be produced naturally without the need for commercialized chemicals. In any case, pathogenic detection is necessary at all levels of production to ensure quality aquaculture, food safety, pathogenic surveillance and to avoid unethical malpractices. Acknowledgements This study was supported by the 1890 Capacity Building Grants Program (Grant no. 2013‐38821‐21456 and 2011‐38821‐30923) from the USDA National Institute of Food and Agriculture. — Preceding unsigned comment added by 128.119.84.54 (talk • contribs)

What I'm looking for are short passages. Can you demonstrate copyrights violation using that method? El_C 17:42, 20 June 2019 (UTC)

Copyrights claim

Please feel free to substantiate, by quoting passages directly. El_C 17:16, 20 June 2019 (UTC)

I would like to say for the record that the overall story matters just as much as the language.

examples:

Brief Specific Examples: _ Original: The bacterium is halophilic and thrives within NaCl concentrations between 1% and 2%, but it is understood that temperature serves as a more detrimental factor... Wiki: Vibrio anguillarum are halophiles that prefer warmer temperatures and neutral pH conditions. Original: V. anguillarum was known to have devastating effects on several marine organisms such as cod, eel, finnock, flounder, herring, pike, plaice, oyster, salmon and trout Wiki: Vibriosis has been observed in salmon, bream, eel, mullet, catfish, oysters, tilapia, and shrimp amongst others. Original: Vibrio anguillarum virulence in marine settings has been reported as early as 1718 along the coastline of Continental Europe (Hofer 1904) and was referenced on record as the red disease... Wiki: As early as 1718, Vibrio anguillarum was found on the coastline of continental Europe with mortality reports as early as 1893 referenced as the red disease in eels. Original: ..V. anguillarum under three biotypes: (i) type A being able to produce acid from sucrose or mannitol without gaseous by‐product, as well as producing indole, (ii) type B being unable to react with sucrose or mannitol to produce gaseous by‐product or indole and (iii) type C being able to produce acid from sucrose and mannitol but not gaseous by‐product or indole. Wiki:In the 1900s, Vibrio anguillarum was classified under three biotypes, A, B, and C.[1][3] Type A are capable of producing acid without gaseous by-product from sucrose or mannitol and producing indole.[1] Type B are incapable of reacting with sucrose or mannitol to produce gaseous by-product or indole.[1] Type C are capable of producing acid from sucrose or mannitol but incapable of producing gaseous by-product or indole. Original: The pathogen grows best between 30 and 34°C with a maximum growth temperature of 38.5°C (Guérin‐Faublée et al. 1995), and the growth rate of V. anguillarum is known to increase with temperature (Groberg et al. 1983). Binary fission of the V. anguillarum cell is pH sensitive being most efficient at neutral pH 7, inhibited entirely over pH 9 and significantly disrupted at pH 6 or below (Gilmour et al. 1976; Larsen 1984) Wiki:Vibrio anguillarum have optimal growth temperatures between 30 °C and 34 °C.[1][3] Growth rates are found to be increasing with temperature with a maximum growth temperature at 38.5 °C.[1] They are Halophiles but growth is more dependent on temperature than salinity. Orignal:The initial site of V. anguillarum entry into a host remains open to debate as the mouth (Cisar & Fryer 1969), gills (Evelyn 1984), anus (Chart & Munn 1980; Ringø et al. 2006), nasal cavity (Traxler & Li 1972), epidermis (Spanggaard et al. 2000) and eyes (Yiagnisis et al. 2007) have all been observed as regions of external vibriosis symptoms in fish Wiki:The host's mouth, gills, anus, nasal cavity, epidermis, and eyes all have been regions of external vibriosis symptoms which causes the initial site of entry of Vibrio anguillarum to still be up for debate. Original:Furthermore, copper is an initiating factor of vibriosis in eel (Rødsæther et al. 1977) and aquacultures stressed under elevated copper levels are 50% more susceptible to V. anguillarum infection (Baker et al. 1983) Wiki:The presence of copper can be an initiating factor in the infection of eels and aquaculture are 50% more susceptible to infection when stressed by copper. Original:Septicemic host infection with V. anguillarum is known as vibriosis and causes symptoms such as internal and external ulceration, abdominal distension, petechia, flesh rot, lethargy, appetite loss, necrosis, erythema, sheathing of arteries and circulatory haemorrhage, boil formation upon muscle tissue, visual lesions and eventually death. Wiki:As stated by Hickey et al., symptoms may include: flesh rot, internal and external ulceration, lethargy, appetite loss, necrosis, erythema, boil formation upon muscle tissue, visual lesions, abdominal distension, petechia, sheathing of arteries and circulatory haemorrhage, and eventually death Original:Ozone treatment as little as 1 mg L−1 is sufficient eradicating aquaculture presence of V. anguillarum entirely Wiki:The addition of 1 mg/L ozone has been shown to completely destroy the presence of Vibrio anguillarum. I would also like to point out that the original reviewers for the page objected to information from Canada. Describing that the info did not fit the storyline... That is because the Canadian story is not part of our story - it did not fit the story naturally because it is not part of our story.

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You neglected the other half. The corresponding passages from the Wikipedia article(!). El_C 18:01, 20 June 2019 (UTC)

Never mind, I see it now. I've gone to a version from March 2018 — please let me know if it's okay. El_C 18:04, 20 June 2019 (UTC)

More examples:

I am still seeing the same issue with the content on the public page. Short example? What does the article still plagiarizes, please show in the form of two passages: original passage alongside passage from the Wikipedia article. El_C 18:11, 20 June 2019 (UTC) Notice the following content from the wikipedia page - entire sections from our paper: [1]...[1]...[1]...[1]...[1]...[1] Treatment measures such as the addition of soluble mixture halquinol have been shown to inhibit growth of Vibrio anguillarum in the presence of fish.[1] Another treatment includes increasing the temperature to 44 °C for 3 minutes or 47.5 °C for 2 minutes.[1] The addition of 1 mg/L ozone has been shown to completely destroy the presence of Vibrio anguillarum.[1] Mortality from Vibrio anguillarum infection can be reduced with the continual addition of anti-microbial peptides (cecropin-bee melittin hybrid peptide, CEME and pleurocidin amide).[1] Mortality can also be reduced with changes to aquaculture feed such as adding selenium, antibodies from hen egg yolks, or oregano-derived carvacrol as feed additives.[1] The addition of antagonistic bacteria can be used to prevent the attachment of Vibrio anguillarum enterocytes.[1] The use of probiotics such as V. alginolyticus, Vagococcus fluvialis, Pseudomonas fluorescens AH2, and extracellular proteins from Kocuria spp., and Rhodococcus spp. has shown to reduce mortality by Vibrio anguillarum infection.[1] Vaccinations have also shown to work by administering Vibrio anguillarum killed by heat or formaldehyde into fish cultures.[1] Other ways to administer dead Vibrio anguillarum is by adding it into feed as wet-packed whole cell or lyophilized whole-cell bacterin.[1] Another way is to place fish cultures into muscle and vaccine containing immersion solution before sending off to aquaculture pools. Monoclonal anti-idiotype antibodies as surrogate antigen can be used instead of administering dead Vibrio anguillarum.[1] Another type of vaccine is through the use of adhesion proteins from the outer-membrane of Aeromonas hydrophila.[1] Other vaccine types include the addition of live-attenuated Vibrio anguillarum that are mutated or plasmid free Vibrio anguillarum.[1] Oral vaccines have been proven to be ineffective due to the fact that the gastric fluid in the upper intestine will inactivate components in the vaccine.[1] _ [1]...[1]...[1]...[1]...[1]...[1] (We are completely 'ripped-off' and plastered on Wikipedia) Vibrio anguillarum can either attach to the host surface cells by absorbing their mucus or by penetrating through the epithelial and vascular tissue.[1] The adhesiveness of V. anguillarum is dependent on the functionality of their exopolysaccharide which is encoded by the hfq gene.[1] Exopolysaccharide transport system utilizes the fish's (host) natural mucus shedding mechanism by its integument system to remain attached to the host.[1] The adhesive ability is not dependent on the flagella but in order for V.anguillarum, the flagella's function must be maintained.[1] The presence of a flagellum enables species as the Vibrio anguillarum to infect other organisms with greater efficiency.[1] Research has found that flagellum may assist with adhesion and/or mobility which are important in virulence as they may enable infective species to dominate.[1] Within the sheath of a V. anguillarum's flagellum, antigens, such as lipopolysaccharides (LPS), may be stored. LPS, in turn, may assist V. anguillarum with infection following entry into the target species.[1] After invasion, V. anguillarum will adhere and colonize in the host's gut where bacteria has the highest replication yield.[1] Vibrio anguillarum rarely infects organisms due to the environment or seasonal weather creating a constant state of stressed survival and conditional variations.[1] Infection by V. anguillarum will rapidly progress and cause infection as fast as 2 days after initial exposure.[1] It does so by using a series of genetic virulence factors to aggressively penetrate a host which could be lethal 5 days after infection or 2 days in certain larvae cultures.[1] Susceptibility of host organism to be cooperatively infected by other pathogens increases when infected by V. anguillarum. [1] The host's mouth, gills, anus, nasal cavity, epidermis, and eyes all have been regions of external vibriosis symptoms which causes the initial site of entry of Vibrio anguillarum to still be up for debate.[1] During the earliest stages of infection, gills contain the most abundance of Vibrio anguillarum but only being exposed to the gills is not enough to cause a systemic infection.[1] V. anguillarum have been shown to survive the acidic environments in fish's stomach for a couple of hours but oral consumption rarely causes infection.[1] It is generally understood for the pathogen to enter the host through the mucosal surfaces such as surface skin and fins and penetrating through the epithelial or vascular tissue.[1]

Too much text. I'm still not seeing it. Are you able to condense at all? Just a sentence or two will do. El_C 18:17, 20 June 2019 (UTC)

Yet more examples

Each sentence is cited "[1}" - Someone has admittedly taken our story. Per your request, though: Original: Vibrio anguillarum virulence in marine settings has been reported as early as 1718 along the coastline of Continental Europe (Hofer 1904) and was referenced on record as the red disease in eel (Anguilla anguilla) mortality reports as early as 1893... Wiki: As early as 1718, Vibrio anguillarum was found on the coastline of continental Europe with mortality reports as early as 1893 referenced as the red disease in eel. - Original:Smith (1961) noted that V. anguillarum was excluded from Bergey's Manual of Determinative Bacteriology, 7th ed. and prior, evidently suggesting the absence of the strain's notoriety among the microbiological research community during that time‐period; the 8th edition (Buchanan & Gibbons 1974) and onward were updated to contain information on V. anguillarum and research studies on the subject have accumulated immensely ever since. Until that time, V. anguillarum was empirically identified by four different taxon prior to their reclassification as one and the same species: Beneckea anguillara (Baumann et al. 1978), V. anguillarum Bergman 1909; V. piscium var. japonicus David 1927 and Pseudomonas ichthyodermis Shewan, Hobbs and Hodgkiss 1960 (Hendrie et al. 1971). A fifth classification, Bacillus anguillarum, was used early on but the bacterium was determined to be Gram‐positive, while V. anguillarum is Gram‐negative (Bergman 1909). Wiki:It wasn't until the 8th edition of the Bergey's Manual of Determinative Bacteriology that the strain Vibrio anguillarum was included.[1] Before it was added to the manual, Vibrio anguillarum was identified as the same species as Beneckea anguillara, Vibrio piscism, and Pseudomonas ichthyodermis.[ - Original:Although misunderstood at the time, a red disease epidemic was described in 1790 in which nearly 40 tons of eel were lost to an unknown inhabiting pathogen, denoting the severity of the septicemic infection (Bergman 1909) Wiki:In 1790, a red disease epidemic was described as nearly 40 tons of eel lost to an unknown pathogen (before the discovery of Vibrio anguillarum).

What about your own version from 26 March 2018 — is that okay? El_C 18:30, 20 June 2019 (UTC)

Absolutely not - wrote "this is not true" here: It is widely distributed across the world. Vibrio anguillarum have been noted as one of the primary causes of bacterial illnesses linked to seafood ingestion (this is not true)

I do not approve of any copy-written content on the site.

 Done. El_C 18:41, 20 June 2019 (UTC)

Thank you for your time.

Sorry it took a little while. I have also unblocked this ip, now that the issue has been resolved. El_C 18:47, 20 June 2019 (UTC)

Thank you again.

Further Thoughts It is unfortunate that a group of interested students who created a Wikipedia page on a topic they were interested in, have been accused of plagiarism. Many of the examples that have been identified have clearly been sourced from the review article, and as far as I can see the source of that information has been appropriately credited. Rather than redacting the entire article, which contains appropriately cited material from a variety of sources, it would be appropriate to reach out to the students that created the page with suggestions on how they might improve it. Given that the purpose of Wikipedia is to share information in an open and transparent way, it is too bad the decision was made to redact the entire page, which served not only to highlight a very important marine microbe, but also draw attention to the important contribution of user talk:128.119.84.54. I think that scrubbing the entire page is inappropriate, and that user talk:128.119.84.54 will instead restore and improve it, given the evident expertise on the topic El_C. --talk:Curt99 "keeper of the virosphere" 04:19, 30 August 2019 (UTC) — Preceding unsigned comment added by Curt99 (talk • contribs)

Not just accused, but found to be in breach of. We take copyright infringement very seriously, here on Wikipedia, and I stand firmly behind my decision to redact the copyrights violation to the uttermost, an action which you are free to bring to review in any forum you see fit. El_C 04:27, 30 August 2019 (UTC)

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