Bacterial diseases, causative organisms, and their crustacean hosts.
\r\n\tTSC involves mutations in chromosomes 9 and 16 encoding for the proteins hamartin and tuberin, respectively. Mutations in these genes cause upregulation of the mTOR pathway and inhibitors of this pathway, such as rapamycin and everolimus, have been shown to be effective in controlling the growth of unresectable tumors. Due to involvement of multiple organ systems, a multidisciplinary treatment plan is necessary and genetic counseling is often part of the management of TSC. Treatment options are quite variable and depended upon symptoms and organ involvement.
\r\n\r\n\tThe aim of this book is to provide the reader with an overview of the tuberous sclerosis complex including its genetic causes, clinical manifestations, and management of its most serious signs and symptoms.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"763892736c7dfc107dc82453265142ad",bookSignature:"Dr. Scott Turner",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10438.jpg",keywords:"Hypomelanotic Macules, Shagreen Patches, Cardiac Rhabdomyoma, Pulmonary Lymphangioleiomyomatosis, Renal Angiomyolipoma, Genetic Testing, Hamartin, Tuberin, Tubers, Subependymal Nodule, Subependymal Giant Cell Astrocytoma, Rapamycin",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"July 2nd 2020",dateEndSecondStepPublish:"July 23rd 2020",dateEndThirdStepPublish:"September 21st 2020",dateEndFourthStepPublish:"December 10th 2020",dateEndFifthStepPublish:"February 8th 2021",remainingDaysToSecondStep:"7 months",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:"Dr. Turner received his medical degree from the Medical College of Wisconsin. He completed a neurology residency at the State University of New York in Stony Brook and a neuro-oncology fellowship at Duke University. He specializes in the treatment of primary and metastatic tumors of the brain and spine. Dr. Turner's undergraduate and master's degree in molecular biology and biochemistry is critical in understanding the complex mechanisms involved with tumor biology.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"181611",title:"Dr.",name:"Scott",middleName:null,surname:"Turner",slug:"scott-turner",fullName:"Scott Turner",profilePictureURL:"https://mts.intechopen.com/storage/users/181611/images/system/181611.png",biography:"Dr. Scott Turner is a clinical Neuro-oncologist and Associate Professor of Neurology at the University of California, Irvine. He attended graduate school at Cornell University and received his medical degree from the Medical College of Wisconsin in 2003. He completed a Neurology residency at SUNY Stony Brook followed by a Neuro-oncology fellowship at Duke University in 2010. He has served as an Assistant Professor of Neurology at both Temple University and the University of Missouri - Kansas City School of Medicine. 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Freshwater finfish represents half of the global aquaculture production (54%), molluscs being the second more produced aquaculture item in the world (24%) [2]. Crustaceans come next in production relevance, represented mostly by penaeid shrimps and grapsid crabs [2, 3]. Aquaculture is the world’s fastest growing segment with a global increase of 5.7% per annum in shrimp production resulting in an increase of 18% by 2020, and the estimated world production of farmed shrimp is 3.5 million metric tons though the diseases, international market prices, and production costs are the main challenges and constrains to the growth and productivity of the shrimp industry on a global level [4]. However, disease outbreaks have caused serious economic losses in several countries, and the estimated global losses due to shrimp diseases are around US$ 6 billion per annum [5, 6]. Such concerns confirm that the bacterial diseases are the most important contracting factors for development of the global aquaculture industry [7].
Bacteria in the aquatic environment and the bacterial diseases, viz. vibriosis, shell diseases (chitinolytic bacteria), and gaffkemia of lobsters, are ubiquitous and are significant for the survival of crustaceans in confined habitats [8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22]. Diseases, causative organisms, and their crustacean hosts are listed in Table 1. Gaffkemia of lobsters is caused by Aerococcus viridans var. homari and is the root cause of mass mortalities of lobsters, while crabs, viz. Cancer borealis and C. irroratus, serve as reservoir hosts of Aerococcus viridans [8, 9, 10, 11, 12, 13]. Vibriosis causes mass mortalities in several crustaceans such as penaeid shrimp Penaeus monodon and P. japonicus, fresh water prawn Macrobrachium, lobster Homarus americanus, blue crabs Callinectes sapidus, rock crabs Cancer irroratus, and shore crab Carcinus maenas. Shell diseases are caused by chitinolytic bacteria which were encountered in English prawn Palaemon serratus; American lobsters Homarus americanus; penaeid shrimp; king crabs, Paralithodes camtschaticus and Paralithodes platypus; and tanner crabs Chionoecetes tanneri, and these crustaceans are affected by rust diseases which are caused by chitin-destroying bacteria [8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22]. Significant mortalities of larval, post-larval, and adult crustaceans, viz. shrimp Penaeus monodon, Litopenaeus vannamei, and Macrobrachium, lobster Homarus americanus, and crab Portunus trituberculatus, are caused by common pathogens such as Vibrio harveyi, V. alginolyticus, V. parahaemolyticus, V. anguillarum, V. furnissii, V. mimicus, V. damsela, Pseudomonas, and Aeromonas [9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22]. Infectious diseases caused by Vibrio species represent the greatest challenges that cause vibriosis with considerable economic losses, and that is the most overwhelming problem in aquaculture, shrimp, and crustaceans [7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101].
Diseases | Hosts | Causative bacterial species |
---|---|---|
Vibriosis | Penaeus monodon, P. merguiensis, and P. indicus (eggs, larvae, postlarvae, juveniles, and adults); Litopenaeus vannamei, Macrobrachium, lobster Homarus americanus, crab Portunus trituberculatus | Vibrio harveyi, V. splendidus; Vibrio harveyi, V. alginolyticus, V. parahaemolyticus, V. anguillarum, V. furnissii, V. mimicus, V. damsela [7, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22] |
Bacterial fouling of surfaces with filamentous bacterial disease | Penaeus monodon, P. merguiensis, P. indicus | Leucothrix sp., Thiothrix sp., Flexibacter sp., Cytophaga sp., Flavobacterium sp. [7] |
Shell disease, brown/black spot, black gill, black rot/erosion, blisters, necrosis of appendages | Crabs and shrimp, white and brown shrimp Penaeus monodon, P. merguiensis, P. indicus | Chitonoclastic bacteria Vibrio spp. infections Vibrio, Aeromonas, Pseudomonas, Flavobacterium [7, 8, 11] |
Chitinolytic bacterial disease, shell disease, box burnt disease, bacterial shell disease | Cancer spp., Callinectes sapidus, other crabs; lobsters, shrimps, and crayfish | Chitinolytic or chitinoclastic bacteria (Gram-negative), viz. Vibrio spp., Pseudomonas spp., and Aeromonas spp. [7, 9, 10, 11, 13, 17, 20, 21, 22] |
Milky hemolymph disease (milky hemolymph syndrome [MHS]) | Spiny lobster Panulirus spp., Panulirus ornatus, P. homarus, and P. stimpsoni; Litopenaeus vannamei (Penaeus monodon, Carcinus maenas) | Rickettsia-like bacterium, a-Proteobacteria, Streptococcus sp., [7, 10, 11, 12] |
Gaffkemia, septicemia | Lobsters Homarus americanus | Gaffkya homari [7, 9, 10, 11] |
Bacteremias | Bacterial diseases of crabs | Vibrio, Aeromonas, Rhodobacteriales-like organism, Vibrio cholerae, Vibrio vulnificus, chitinoclastic bacteria, Rickettsia intracellular organisms, chlamydia-like organism, Spiroplasma, chitinoclastic bacteria, Rickettsia intracellular organisms, chlamydia-like organism, and Spiroplasma [7] |
Fungal diseases | Penaeus monodon, [larval (nauplii, zoea, and mysis) Indian tiger prawn] Macrobrachium rosenbergii | Lagenidium callinectes [16, 17] |
Bacterial diseases, causative organisms, and their crustacean hosts.
Phage therapy is a prospective ideal therapy for vibriosis in aquaculture of crustaceans. Bacteriophages are defined as bacterial viruses that can infect cells, multiply in, cytolyse, and destroy susceptible bacteria. Bacteriophages are viruses of bacteria which have a natural ability to target, infect, and destroy their host cells of a particular bacterial species or groups or even unrelated bacteria, and thus they play a major role in controlling their target bacterial population density in nature [23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60]. They are both omnipresent and copious in the aquaculture environment, especially in seawater, in which the total numbers of viruses normally surpass the bacterial cell concentration by a factor of 10 [25]. “Most phages reproduce upon entering a cell and in that process kill their host. They encode two families of proteins, holins, and lysins, which allow the phage progeny to burst through the bacterial cell wall and go off in search of new hosts. But the cell is already dead by the time that happens in terms of lethality; the lysis is just a matter of burning down the house for good measure.” An estimated 108 strains of phage with approximately 1031–1032 phages are known to occur in the biosphere at any given time [23]. Bacteriophages are used for the isolation and identification of specific bacteria to help in the diagnosis of the bacterial diseases, to kill antibiotic-resistant, virulent bacteria through a natural phenomenon called lysogeny, whereby one of the phage-infected bacteria in a colony kills another uninfected bacterium through phage missiles or antibacterial peptides [24, 25, 36, 37, 38, 39, 41].
Due to their specific antibacterial activities and significance of the phage therapy as an alternative to antibiotics, bacteriophage therapy is re-emerging, and consequently this has become a potentially novel and useful concept to kill even intracellular pathogenic bacteria and warrant future development. Bacteriophage therapy has been extended from medical applications into the fields of agriculture, aquaculture, and the food industry [28, 29, 30, 39]. Bacteriophages specific for Vibrio spp. have been described [36, 37, 38, 39, 40, 41]. Bacteriophages are known to infect >140 bacterial genera, and they are the most valuable and ubiquitous (1031) phage organisms in the world [26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 41, 42, 43, 44, 45]. Earliest description of phages and their antibacterial activity has been independently demonstrated [62]. D’Herelle [33] published a comprehensive account of phages, and hence the International Bacteriophage Institute was established in Tbilisi, Georgia, in 1923, now called as “the George Eliava Institute of Bacteriophages, Microbiology and Virology” [32, 34], which is still involved in researching phage therapy applications and supplies phage for the treatment of various bacterial infections. D’Herelle’s first phage therapy experiments against bacterial dysentery were exceptionally successful and were very promising in the removal of the infectious organisms [32, 34]. However some of the results of the early phage therapy experiments of infectious host organisms were known to be contradictory with reports of both success and failure. Whenever failures occurred in phage therapy, they were attributed to a range of factors. They include unsatisfactory understanding of phage biology, inadequate experimental technical knowledge, poor quality of phage preparations, and a lack of understanding of the causes of illness being treated. The discovery of such specific bacteriophages were at first considered to become powerful beneficial therapeutic agents against pathogenic microorganisms but could not be put into practice because of the dawn of antibiotic era experiencing overuse/misuse of antibacterial medication that resulted in the development of untreatable antibiotic resistance [32, 36]. Commercialization of antibiotics in the Western countries in the 1940s had led to a simultaneous decline in the use of phages as human therapeutics, while in some of the Eastern European countries, exploitation of phage therapy was continued either alone or in combination with antibiotics [19, 34, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73]. The experimental phage therapy could be an alternative to antibiotics and replace them when they fail for the treatment of chronic infections, and such a successful eradication of drug-resistant bacteria was documented [19, 42, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73]. Moreover, the significantly decline costs of phage therapy constitute an important additional battle for its wider application in the current era of a worldwide circumstance in antibiotic resistance. The efficacy of phage therapy is well recognized and demonstrated in a few cases [32, 64, 65, 66]. Even a single dose of phage was reported to be much more effective than multiple doses of antibiotics such as ampicillin, tetracycline, and chloramphenicol [66]. The use of phage therapy to control fish pathogens has also been reported [23, 24, 42, 43]. Phage lysins have been used as potential therapeutics for treatment of bacterial infections [44]. Furthermore, the US Food and Drug Administration has approved commercial phage preparations to prevent bacterial contaminations. Such developments have prompted to explore the possibilities of using bacteriophages to control bacterial infections in crustacean aquaculture [36, 37, 38, 39, 40, 41]. Further cautious phage collection and perfect experimental conditions for phage propagation and purification, route and timing of phage administration, and environmental monitoring of phage for use are needed. Such a focus may help in further development of probiotics, phage therapy applicable to a wide variety of systems, which is considered to signify an emerging alternative to antibiotic therapy and vaccination.
Probiotics are defined as applications of whole or components of microorganisms or “a live microbial adjunct which has a beneficial effect on the host by modifying the host-associated or ambient microbial community, by ensuring improved use of the feed or enhancing its nutritional value, by enhancing the host response towards disease, or by improving the quality of its ambient environment” [74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93]. A probiotic is a live microbial feed supplement which benefits the host animal by improving its intestinal microbial balance [82]. Lactic acid bacterium Bacillus S11 was tested as probiotics and has been proven as antagonists of shrimp pathogens [75]. Bacillus spores were used as biocontrol agents to reduce Vibrio species in shrimp culture facilities [83]. The inhibitory activity of Bacillus subtilis BT23, isolated from shrimp culture ponds, is against pathogenic Vibrio harveyi under in vitro and in vivo conditions [39, 73]. The concept of probiotics is exploited to “augment” naturally occurring pathogenic bacterial population to increase the growth rate and control diseases of organisms in aquaculture such as fish/shellfish [84, 85]. Several microorganisms have been evaluated as probiotics which have been shown to be successful in the larval stages of the aquatic organisms in preventing the diseases, improving digestion and growth [82, 84, 87]. Some of the proposed mechanisms that provide protection against pathogens involve the production of inhibitory compounds, competition for essential nutrients and adhesion sites, enhancement of disease resistance, and modulation of host immune responses [82, 87]. A probiotic containing a combination of several different bacteria has been shown to be more efficient at controlling bacterial pathogens [82, 90, 91]. Probiotics are known to act and inhibit the growth and proliferation of bacterial pathogens both in vivo and in vitro by generating antibacterial compounds such as bacteriocin, siderophores, lysozymes, proteases, hydrogen peroxide, antibiotics, and organic acids [91]. In aquaculture, some microorganisms are more beneficial to host organisms in reducing the incidence of diseases though the factors and mechanisms which mediate the benefits to the host are poorly understood. Future studies should be focused on evaluating the mechanisms by which the probiotics interact with the host and pathogens and their biology.
Proprobiotic is defined as a synbiotic comprising pre- and probiotics involving a combination of strains of nonpathogenic bacteria and yeast so as to provide supplementary nutrients and to protect against invading pathogens. Commercially available proprobiotics are known to competitively exclude the potential pathogen through the use of proprobiotics in aquaculture. Effects of Ergosan and Vibromax are used to prevent vibriosis in Litopenaeus vannamei [42, 43]. Probiotics (e.g., Arda-Tek, Australia, viz. DMS1000, S-1100, DMS-2001, DMS-2002, DMS-2004, etc., Wunapuo-15 of Team Aqua Corporation) are used to reduce the number of Gram-negative pathogenic bacteria including Vibrio in the water and to maintain stable water quality, to stabilize plankton, to reduce rate of sludge buildup on the pond bottom, and to digest microbial slime [73, 82, 84, 85, 86, 87, 88, 89, 90, 91]. To assess the actual impact of these probiotic products in the field, it is fundamental to determine the efficacy of the probiotics and the continuance of their application. The potency of the probiotic products against selected pathogenic bacteria (and viruses) also needs to be critically evaluated. The application of some beneficial bacteria has been an option in aquaculture to achieve a number of benefits, viz. to reduce mortality in shrimp, to enhance production and increase harvest yield and eliminate antibiotic use, and to enhance immunity and disease resistance in black tiger shrimp Penaeus monodon by a probiont bacterium (Bacillus S11) [84, 85, 86, 87, 88, 89, 90, 91]. Immunity enhancement occurs in black tiger shrimp Penaeus monodon by a probiont bacterium (Bacillus S11), and the immune stimulator capacity of probiotics may be affected by factors such as source, type, dose, and duration of supplementation [80, 85]. The use of probiotic mixtures consisting of Bacillus tequilensis and B. endophyticus along with commercial probiotics was shown to be beneficial in altering the bacterial community of larval shrimp Litopenaeus vannamei when the hosts were challenged with Vibrio parahaemolyticus. Similarly application of Bacillus sp. promoted cellular and humoral immune resistances and provided disease protection in tiger shrimp P. monodon. The effects of probiotics such as Bacillus cereus, Paenibacillus polymyxa, and Pseudomonas sp. PS-102 as biocontrol agents against pathogens of various Vibrio species in shrimp were evaluated. Growth promoter probiotics have been used in aquaculture to enhance the growth of cultivated species, whereas their side effects if any on the host need to be investigated. The probiotics was shown to increase the survival and growth of white shrimp (Litopenaeus vannamei), and the production increased by 35%, whereas with the use of antimicrobials, it decreased by 94% and thus demonstrated the significance of endemic Bacillus probiotics on the improvement of the health of larval shrimp [71, 73]. The efficacy of the non-hemolytic probiotic Bacillus strains against pathogenic Vibrio species, viz. Vibrio campbellii, V. vulnificus, V. parahaemolyticus, and V. alginolyticus on the growth of the larval shrimp was tested by using a daily concentration of 1 × 105 cfu ml−1 with initial bioassay density of 225 nauplii L−1, and the treatments promoted a considerable increase in survival and growth of the larval shrimp compared to the control, and thus the study has established a significant antagonistic activity of the probiotic Bacillus strains against the Vibrio species [48]. Various strains of Bacillus, or a commercial product made from Bacillus sp., Saccharomyces cerevisiae, Nitrosomonas sp., and Nitrobacter sp., used as probiotics show prevention of infection and promotion of growth rate of all the stages of the white shrimp Litopenaeus vannamei Boone and Fenneropenaeus indicus when the diet was supplemented with 50 g of probiotic kg−1 of food. The use of probiotic Shewanella algae in shrimp farms is found to be safe for the consumer of shrimp [92]. A scientific rational approach for the evaluation of aquaculture probiotics and guidelines is needed for their use and safety.
Pathogenic Vibrio spp. were controlled by cell-free extracts of Bacillus under in vitro and in vivo conditions indicating that probiotic treatment offers a promising alternative to the use of antibiotics in shrimp aquaculture [41, 63, 70, 85]. Though probiotic bacteria are currently used to improve the health of the shrimp ponds and increase productivity and reduce mortality of prawns, little attention has been paid to bacteriophages preying on the probiotic bacteria of crustacean culture such as Vibrio/Bacillus spp. from aquaculture systems. A special focus needs to be given to bacteriophages infecting probiotic bacteria to explore the antibacterial potential of bacteriophages of Bacillus spp., Vibrio spp., and other bacteria. The existence of bacteriophages in the probiotic Bacillus sp. and Vibrio spp. controlling the infectivity of the Vibrio spp. with a significant reduction in the mortality of infected shrimp was demonstrated [37, 41]. Anti-Vibrio activities of Bacillus spp. mainly due to the production of bacteriocin or bacteriocin-like substances have been reported from B. subtilis, B. megaterium, B. stearothermophilus, B. licheniformis, B. thuringiensis, B. thermovorans, and B. cereus [16, 22, 45, 47, 68, 69, 83]. P. monodon larvae fed with Bacillus S11 showed 100% survival after challenge with pathogenic V. harveyi, whereas only 26% control animals survived.
Vibrio species are Gram-negative curved rod-shaped bacteria that belong to the Vibrionaceae family, and they naturally inhabit the estuarine, coastal, and marine environment worldwide. Vibrio species occur as the dominant flora in all developmental stages of Penaeus monodon, and they have been described as the causal pathogens. Vibriosis is a severe bacterial disease in penaeid shrimp and is responsible for large-scale losses in the aquaculture industry, leading to prophylactic as well as therapeutic use of antimicrobials [18, 19, 20, 21, 22, 70, 71, 72, 73]. Antibiotics are used in shrimp farming to control or treat bacterial disease outbreaks with an expected 100,000–200,000 tons of antibiotics being consumed in the world every year [57]. Oxytetracycline, tetracycline, quinolones, sulfonamides, and trimethoprim are among the antimicrobials utilized to control bacterial infections in the aquaculture industry. Indiscriminate prophylactic uses of diverse antibiotics in the shrimp hatcheries/farms have resulted in antibiotic resistance of the bacteria causing mass mortalities of the hosts [73, 74, 75]. The potential negative consequences are the development of drug-resistant bacteria and reduced efficacy of antibiotic treatment. Shrimp farmers are exceedingly dependent on various antibiotics as a preventive measure against shrimp bacterial infections with 14% of farmers using antibiotics on a daily basis in their farms [22].
All of the 121 isolates of Vibrio spp. were found to be 100% resistant to ampicillin, cloxacillin, oxacillin, erythromycin, vancomycin, penicillin G, and furazolidone and partially resistant to cefaclor, streptomycin, rifampicin, oxytetracycline, nalidixic acid, cefotaxime, and chlorotetracycline [14, 16]. Molitoris et al. [14] reported a high degree of resistance to ampicillin and furazolidone, and incidence of resistance to chloramphenicol, neomycin, and gentamycin has also been detected [16, 18, 19, 20, 22, 46, 69]. Multidrug-resistant Vibrio harveyi isolated from black gill-diseased Fenneropenaeus indicus and antibacterial activity against pathogenic Vibrio harveyi and its protective efficacy on juvenile F. indicus were reported. In vitro susceptibility of antibiotics against Vibrio spp. and Aeromonas spp. isolated from Penaeus monodon hatcheries and ponds and the effect of probiotics, antibiotics, and pathogenicity of Listonella anguillarum-like bacteria isolated from Penaeus monodon culture systems and antibiotic-resistant Vibrio spp. were commonly isolated from hatchery-reared postlarvae compared to farm-reared P. monodon [61, 62, 72, 73, 79, 85]. The unrelenting use of antibiotics against diseases in human beings and other life forms may pollute the aquatic system, and their impact on developing antibiotic-resistant Vibrio sp. may be a serious threat in addition to the use of antibiotics in aquaculture farms [63, 70]. Control of the bacterial diseases depends on improvement of management practices to minimize the risk of introduction of infectious agents into aquaculture systems and to reduce predisposing factors such as overcrowding and overfeeding.
A study assessed a variety of antibiotic-resistant bacteria and detected the presence of their resistance genes from mariculture environments. A variety of antibiotics that were used in aquaculture have led to the occurrence of antibiotic-resistant genes in bacteria, and in these bacteria many different ARGs can be found, and they are β-lactam- and penicillin-resistant genes penA and blaTEM-1; chloramphenicol-resistant genes catI, catII, catIII, catIV, and floR; tetracycline-resistant genes tatA, tatB, tatC, tatD, tatE, tatG, tatH, tatJ, tatY, and tatZ; and many more [65, 68]. ARGs can be transferred among bacteria via conjugation, transduction, or transformation. The widespread emergence of antimicrobial resistant bacteria worldwide has become a major therapeutic challenge. Therefore there is a need for development of novel non-antibiotic approach such as phage therapy biocontrol agents to fight against resistant bacterial infections due to the shortage of new antibiotics in developmental pipeline [19, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73]. Thiel [25] stated that phage therapy is “a nearly forgotten therapy that may yet re-emerge as a savior to this accelerating crisis of antibiotic resistance. For every bacterium known on this planet, there are legions of bacteriophages—tiny viruses that seek out bacteria and use them as a breeding ground, almost invariably destroying their prokaryotic host in the process. That makes them harmless to even to nontarget bacteria—distinguishing them from broadspectrum antibiotics, which, when they work, can wipe out beneficial flora along with a troublesome infection.” The phage specificity has its drawbacks as there is a requirement of the right match between phage and bacteria which needs to be determined for the phage therapy to work. There has been renewed concern in the application of bacteriophage as a non-antibiotic approach to control bacterial infections in various fields including human infections, food safety, agriculture, and veterinary applications. However the guideline data on the use of phage therapy applied to invertebrates, like shrimp and other crustaceans, are nonexistent [64, 68, 92, 93].
A bacteriophage isolated from a shrimp hatchery was shown to infect V. harveyi, signifying its potential as a biocontrol agent of luminous vibriosis. In vitro treatments of bacteriophages were shown to exhibit a significant reduction (2–3 log units) in the number of V. harveyi host cells [41, 46, 48, 49]. These studies showed that bacteriophages could be used for biocontrol of V. harveyi and that bacteriophage therapy could be effective as an alternative to antibiotics in the control of luminous vibriosis in shrimp hatchery systems [23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69]. In vitro experiments confirmed that bacteriophages could be effectively used in vivo as biological agents to control Vibrio sp. in aquaculture systems [36, 37, 38, 39, 41, 46, 48, 49]. Investigation on the occurrences of luminescent V. harveyi and their bacteriophages in shrimp showed that the presence of low concentrations of bacteriophages in the larval rearing tank waters could not prevent the development of luminous vibriosis [46, 47, 48, 49], and this study showed that the presence of optimal concentration of phages is required for effective reduction of the pathogenic bacteria. A lytic spectrum of bacteriophages (Viha1, Viha2, Viha3, Viha4, Viha6, Viha7) occurring in nature exhibited a wide spectrum of activity against V. harveyi, suggesting their potential as agents for biocontrol of vibriosis in aquaculture environments [51]. A lytic phage (PW2) was isolated and characterized under controlled conditions in the laboratory from the host bacterium V. harveyi CS101 [49], and the useful bacteriophages for the biocontrol of vibriosis have been explored. A probiotic strain V. alginolyticus reduced the diseases and mortality of infected P. monodon with A. salmonicida, V. anguillarum, and V. ordalli [49]. A soil bacterial strain, PM-4, was shown to exhibit in vitro inhibitory effect against V. anguillarum and to promote the growth of P. monodon nauplius [53]. Inoculation of Bacillus S11, a saprophytic strain, resulted in greater survival of the post-larval P. monodon that were challenged with pathogenic luminescent bacterial culture [51]. These works strongly suggest an effective control of microflora in crustaceans can be achieved in aquaculture environments by bacteriophage-producing bacteria. The crustacean host, causative agent, diseases, and source of bacteriophages/bacteria are listed in Table 2.
Crustacean host | Bacterial agent | Infection/disease | Bacteriophage | Source of the bacteriophage | Efficacy of bacteriophage | Outcome of the phage therapy | References |
---|---|---|---|---|---|---|---|
Shrimp larvae Penaeus monodon | Vibrio harveyi | Luminous vibriosis | Myoviridae (VHLM) | Extracted from a toxin-producing strain of V. harveyi isolated from moribund prawn | Vibriolysis | VHML showed a narrow host range with a preference for V. harveyi rather than 63 other Vibrio isolates and 10 other genera | [100] |
Shrimp larvae Penaeus monodon | Vibrio harveyi | Luminous vibriosis | Siphoviridae | Shrimp farm waters from the West coast of India | 18-day-old PL shrimp were challenged with the bacteria (105 cells ml−1, laboratory trial: (1) bacteriophage suspension (109 pfu ml−1) was added initially; after 24 h (another 0.1 ml), (2) only once initially with 0.1 ml of the phage suspension; (3) no addition. Hatchery trial (1) treatment with bacteriophage (109 pfu ml−1) at the rate of 200 ppm daily so that phage concentration in the water was 29–105 pfu ml−1; (2) treatment with antibiotics (oxytetracycline 5 ppm, kanamycin 10 ppm daily); (3) no treatment | Enhanced survival (80%) of P. monodon larvae on treatment with two doses of bacteriophage when compared with the control (25%). Hatchery trial: survival in the control tank was only 17%, while in antibiotic-treated tanks, survival was 40%; in the bacteriophage-treated tank, survival was 86%. Bacteriophage therapy has an excellent potential in management of luminous vibriosis in aquaculture systems | [48] |
Larval shrimp Penaeus monodon | Vibrio harveyi | Luminous vibriosis | Siphoviridae | Three from oyster tissue and one from shrimp hatchery water | Hatchery tanks, with post-larval five-stage larvae, presenting luminescence and mortality, were used. Bacteriophage treatment (two tanks): one suspension (29 106 pfu ml−1) was added by day following the order: Viha10, Viha8, Viha10, and Viha8 chemotherapy (two tanks): oxytetracycline (5 mg L−1), kanamycin (10 mg L−1) | Bacteriophage treatment resulted in over 85% survival of Penaeus monodon larvae. The normal hatchery practice of antibiotic treatment resulted in a survival range from 65 to 68%. This study shows that bacteriophages could be used for biocontrol of V. harveyi | [46] |
Penaeid shrimp P. monodon | Vibrio harveyi | Luminous vibriosis | Seven bacteriophages specific to Vibrio harveyi (Viha1 to Viha7), six from Siphoviridae and one Myoviridae (Viha4) | Coastal aquaculture systems like shrimp farms, hatcheries, and tidal creeks along the East and West coast of India | All the phages were found to be highly lytic for V. harveyi. The phages exhibited a different lytic spectrum for a large number of bacterial isolates tested. Three of the phages (Viha1, Viha3, and Viha7) caused 65% of the strains to lyse, while Viha2, Viha4, and Viha6 caused 40% of the host strains to lyse. Only Viha5 had a narrow spectrum (14%). Six of the seven phages isolated had a broad lytic spectrum and could be potential candidates for biocontrol of V. harveyi in aquaculture | [51] | |
Shrimp P. monodon | Vibrio harveyi | Luminous vibriosis | Siphoviridae (VH1 to VH8) | Shrimp farm | In vitro experiment | All the isolates of bacteriophage (VH1–VH8) caused lysis of the host bacterial cells within 2 h. The propagation curve for each phage showed a burst time from 1 to 10 h. Bacteriophages of Vibrio sp. shall be effectively used in vivo as biological agents to control these pathogenic bacteria in aquaculture systems | [39] |
Shrimp P. monodon | Vibrio harveyi CS101 | Luminous vibriosis | Siphoviridae (phage PW2) | Shrimp pond water | Phage adsorption rate increased rapidly in the first 15 min of infection to 80% and continued to increase to 90% within 30 min of infection. The stability of phage PW2 was dependent on temperature and pH. It was inactivated by heating at 90°C for 30 min and by treating at pH 2, 3, 11, and 12. One-step growth curve and latent and burst periods were 30 and 120 min, respectively, with a burst size of 78 pfu per infected center. Six structural proteins were detected | [52] | |
Larval shrimp P. monodon | V. harveyi | Luminous vibriosis | ϕH17-5c, ϕH17-7b, ϕH17-8b, and ϕH17-9b | Secluded from shrimp farm water from the West coast of India and demonstrated to exhibit a broad lytic activity against V. harveyi isolates | In a set of laboratory experiments, post-larval Penaeus monodon was exposed to 106 cfu ml−1 cells of Vibrio harveyi and was treated with 100 ppm phage which has led to a drastic reduction of Vibrio harveyi counts with 86% survival of the infected larvae, while the survival of the phage-untreated larvae was 25% | In the antibiotic (oxytetracycline 5 ppm, kanamycin 100 ppm/day)-treated hatchery tanks, an initial reduction of luminous bacterial counts were shown, and after 48 h the bacterial count increased to 106 ml−1 showing the luminous vibriosis and mortality of the nauplii of Penaeus monodon with 40% larval survival. In contrast, in the bacteriophage-treated tank, the larval survival was 86%, and the survival rate in the control tank was only 17% | [48, 49] |
Shrimp, P. monodon | V. vulnificus | Vibriosis | Phages VV1, VV2, VV3, and VV4 from V. vulnificus | VV1, VV2, VV3, and VV4 phages were detected from shrimp aquaculture system | In vitro experiments show successful potential phage therapy; lytic V. vulnificus phages infect a wide variety of other Vibrio spp./isolates | V. vulnificus phages exhibited a broad lytic spectrum and potential biocontrol of luminous vibriosis in the shrimp aquaculture system | [36, 37, 38, 39] |
Phyllosoma larvae of the tropical rock lobster Panulirus ornatus | V. harveyi | Luminous vibriosis | Six bacteriophages from Siphoviridae (VhCCS-01, VhCCS-02, VhCCS-04, VhCCS-06, VhCCS-17, and VhCCS-20) and two from Myoviridae (VhCCS-19 and VhCCS-21) | Isolated from an epizootic in aquaculture-reared larval phyllosomas of the ornate spiny lobster Panulirus ornatus water samples from discharge channels and grow-out ponds of a prawn farm | Exhibited a clear lytic activity against V. harveyi (1) Addition of phage VhCCS-06 (1 ml) 2 h after inoculation; (2) addition of phage VhCCS-06 (1 ml) 6 h after Bacteria-free supernatants were obtained by centrifugation at 10,000 g for 15 min and by filtration of the aliquots of the enriched water samples; supernatants (10 ll) were inoculated onto NAMS plates, grown at 28°C for 24 h, and the bacteria-free supernatants were stored at 4°C | Exhibited a clear lytic activity against V. harveyi with no apparent transducing properties. Phages of VhCCS-19 and VhCCS-21 are Myoviridae bacteriophages and lysogenic and induce bacteriocin production in the host bacteria (V. harveyi strain 12); Siphoviridae phage (VhCCS-06) delayed the entry of a broth culture of V. harveyi strain 12 into exponential growth, though it could not prevent the overall growth of the bacterial strain. This effect was due to the multiplication of phage-resistant cells of V. harveyi. Phage resistance is an obstacle to the use of phage as therapeutic agents. The isolated phages exhibited lytic activity against strains of V. harveyi, a primary pathogen of phyllosoma of the tropical rock lobster, P. ornatus. These phages can be used as a biocontrol agent to combat vibriosis in the rearing system of phyllosoma of the tropical rock lobster, P. ornatus | [55] |
Live prey Artemia salina | V. alginolyticus strain V1 | Vibriosis | Two novel bacteriophages φSt2 and φGrn1 | Vibrio alginolyticus strain V1, isolated during a vibriosis outbreak in cultured seabream | In vitro cell lysis experiments against the bacterial host V. alginolyticus strain V1 and also against 12 Vibrio strains originating from live prey Artemia salina cultures, viz. V. anguillarum, V. harveyi, V. alginolyticus, V. ordalii, V. parahaemolyticus, V. splendidus, and V. owensii). It indicated a strong lytic efficacy of the 2 phages | In vivo administration of the phage cocktail consisting of φSt2 and φGrn1, directly on live prey A. salina cultures, has led to a 93% decrease of Vibrio population, viz. V. anguillarum, V. harveyi, V. alginolyticus, V. ordalii, V. parahaemolyticus, V. splendidus, and V. owensii, after 4 h of treatment | [98, 99] |
Brine shrimp nauplii Artemia franciscana | V. parahaemolyticus | Vibriosis of Artemia franciscana | VPMS1 phage | VPMS1 is a lytic phage of Vibrio parahaemolyticus, isolated from a marine clam | V. parahaemolyticus-infected brine shrimp nauplii were treated with a single dosage of VPMS1 phage, which was effective enough to eliminate the adverse effects of V. parahaemolyticus in brine shrimp. Efficacy was not affected by the reduction in the dosage | The phage therapy was successful in preventing vibriosis; a single dosage of VPMS1 phage was effective enough to get rid of the adverse effects of V. parahaemolyticus in brine shrimp; the beneficial effects of the therapy were compromised if the application of phages was delayed | [97] |
Penaeus monodon rearing waters in shrimp ponds in Palk Strait, South East coast of India | Vibrio parahaemolyticus | Vibrio parahaemolyticus and its potential lytic phage from Penaeus monodon | Lytic phage (VVP1) belongs to the Myoviridae family | Lytic phage (VVP1) able to infect strains of N1A and N7A, V. parahaemolyticus, and strains of N3B and N13B Vibrio alginolyticus | One-step growth experiments, multiplication and host range, and pH and temperature stability of the lytic phage (VVP1) were shown; the phage showed protective biocontrol effects in reducing the pathogenic V. parahaemolyticus in infected shrimp larvae | P. monodon larvae infected with V. parahaemolyticus showed enhanced survival of the larvae in the presence of phage treatment at lytic phage (VVP1), 2.3 × 1010 PFU ml−1, when compared with the control and showed that the application of phage therapy is a useful strategy to prevent and eliminate or reduce shrimp pathogenic V. parahaemolyticus in the aquaculture system | [101] |
V. alginolyticus phages were isolated from seawater samples after enrichment | V. alginolyticus, Vibrio alginolyticus, a zoonotic pathogen that causes mass mortality in aquatic animals and infects humans | Vibriosis, a zoonotic pathogen causing mass mortality in aquatic animals and infecting humans | Phage pVa-21, Myoviridae | Phage pVa-21 infects bacteria belonging to the family Vibrionaceae, viz. V. alginolyticus and V. harveyi, and could not infect V. parahaemolyticus, V. anguillarum, V. campbellii, and V. vulnificus | Bacteriophage pVa-21 belongs to Myoviridae, characterized as a candidate biocontrol agent against V. alginolyticus. It exhibits planktonic or biofilm lytic activity and showed stability under various conditions. It has latent period and burst size approximately 70 min and 58 plaque-forming units/cell, respectively. Phage pVa-21 can inhibit bacterial growth in both the planktonic and biofilm states. The phage is related to the giant phiKZ-like phages, classified as a new member of the phiKZ-like bacteriophages that infect bacteria belonging to the family Vibrionaceae | Infect bacteria, viz. V. alginolyticus and V. harveyi in both the planktonic and biofilm states | [101] |
Crustacean host, bacteria, disease, and source of bacteriophages/bacteria for bacteriotherapy.
In vitro and in vivo antagonistic effects of Bacillus against the pathogenic Vibrio species were evaluated [40]. Cell-free extracts of Bacillus subtilis BT23 were shown to exhibit inhibitory effects against the growth and proliferation of Vibrio harveyi isolated from black gill-diseased Penaeus monodon. The probiotic effect of Bacillus was tested by exposing shrimp to B. subtilis BT23 at a density of 106/108 cfu ml−1 for 6 days before a challenge with V. harveyi at 103/104 cfu ml−1 for 1 h infection (Figure 1a-f). Probiotic treatment of B. subtilis BT23 showed a 90% reduction in accumulated mortality of P. monodon. Pathogenic Vibrio species were controlled by Bacillus under in vitro and in vivo conditions, and the results indicated that probiotic treatment offers a promising alternative to the use of antibiotics in shrimp aquaculture.
(a) Light micrograph of uninfected Bacillus sp.; (b) TEM micrograph showing phages on the surface of the infected Bacillus. (c) TEM micrograph of phage-lysed bacterial cell. (d) TEM micrograph of uninfected Vibrio cells. (e) TEM micrograph of phage-infected Vibrio cell. (f) TEM micrograph of phage-lysed Vibrio cell.
The occurrence of phages in infected cells of Bacillus sp. is illustrated (Figure 1a-c). Each bacteriophage particle of the bacterium Bacillus sp. presented as electron-dense objects comprising a distinctive head with a flexible noncontractile tail. The preparations consisted of 30 min–2 h exposures of Bacillus cells to in vitro phage infection, and the TEM of such experimentally phage-infected cells showed actively adsorbing and infecting phages onto the surface of bacterial cell membrane (Figure 1b), while phage particles occurred within the completely lysed cells in 8–24 h (Figure 1c).
TEM analysis of the phages of Vibrio host cells showed the occurrence of tailless phages with a double-layer membrane covering the icosahedral head. Phage-uninfected, phage-infected, and completely lysed Vibrio cells are shown (Figure 1d–f).
An antagonism assay consisting of cell-free extract of Bacillus BT21, Bacillus BT22, and B. subtilis BT23 showed inhibitory activity against several pathogenic species of Vibrio [40]. They reported that B. subtilis BT23 exhibits a relatively higher inhibitory activity than the other two Bacillus BT21 and Bacillus BT22. Moreover B. subtilis BT23 was shown to inhibit the growth and proliferation of 112 isolates of Vibrio spp. consisting of V. harveyi (39 isolates), V. anguillarum (24 isolates), V. vulnificus (30 isolates), and V. damsela (19 isolates) which were obtained from P. monodon culture hatcheries and ponds. The growth and proliferation of pathogenic V. harveyi was inhibited by B. subtilis BT23 culture inoculated at a preliminary level of 105–109 cfu ml−1, whereas lower concentrations of B. subtilis BT23 (105 and 107 cfu ml−1) allowed early growth and proliferation followed by a decrease in the total viable counts of V. harveyi. Co-culture experiments demonstrated that the growth and proliferation of V. harveyi was controlled under in vitro conditions when the concentrations of B. subtilis BT23 were increased. Experiments of cell-free extracts of Bacillus subtilis BT23 established the inhibitory effects on the growth and proliferation of V. harveyi in liquid culture under aerobic conditions. The inhibitory efficacy was higher in B. subtilis BT23 cell-free extracts of 108 cfu ml−1 and low in 104 cfu ml−1. Cell-free extracts of Bacillus subtilis BT23 initially could not limit the growth and proliferation of V. harveyi for 2 days, and afterward the growth and proliferation of V. harveyi was extremely inhibited when compared with the growth of V. harveyi without B. subtilis BT23. The studies on the probiotic treatment V. harveyi-infected P. monodon revealed a substantial reduction in the mortality of shrimp which were treated with B. subtilis BT23 strains under in vivo conditions. The cumulative mortality of V. harveyi post-infection and probiotic B. subtilis BT23-untreated P. monodon was 50% on the ninth day and 100% on the seventeenth day of post-infection. In contrast, in the probiotic treatment groups, the cumulative mortality of shrimp was 10% in the combined treatment after 5 days post-infection. Long-term and short-term treatments of V. harveyi-infected P. monodon with B. subtilis BT23 showed a decrease in cumulative mortality of 32 and 60%, respectively. In control groups of V. harveyi-uninfected P. monodon, there was no mortality. The growth of pathogenic V. harveyi was inhibited by nonpathogenic B. subtilis BT23 under in vivo and in vitro conditions. Co-culture experiments showed that the inhibitory activity of B. subtilis BT23 increased with increasing density of the antagonist. A high concentration of B. subtilis BT23 (antagonist) was essential to obstruct the growth and multiplication of V. harveyi in the co-culture experiments. The antagonist required being present at significantly higher levels than the pathogen, and the degree of inhibition increased with the level of antagonist. During the co-culture, 107/108 cfu ml−1 was required to inhibit the growth of the pathogen V. harveyi. Therefore, a potential probiotic co-culture must either be supplied on a regular basis or be able to colonize and multiply on or in the host. A similar control of pathogenic Vibrio in fish and shellfish, by the use of nonpathogenic bacterial strains and disease prevention, has received much attention during the last decade [42, 43, 56, 81]. Purification and characterization of the antibacterial substances from the host bacteria and the phages could help to understand the mechanism of antibacterial activity of Bacillus and other strains. Probiotic treatment offers a very promising alternative to the use of antibiotics in fish and shrimp aquaculture. Further study is needed to elucidate the exact mode of action of the observed beneficial effects of the probiotics and to understand the possibilities and limitations of microbial control in aquaculture.
The usefulness of a phage lysate preparation and treatment method adopted for long-term storage of phages was elucidated in a study that demonstrated the infectivity of the phages of Vibrio spp. that remained unaffected with chloroform and DMSO treatments and storage at −40°C for 30 days [37, 38, 39, 41]. Similarly, phages were shown to be highly stable under normal storage conditions and also stable in NaCl and MgSO4 due to its stabilizing effect. Substantial amounts of viable phages were reported to occur even after storage even in distilled water. Phage isolates were found to be stable upon storage at 4°C, and a rapid loss of phage infectivity was encountered with repeated freezing and thawing at −70°C. Bacillus phage was stable to a 1-hour exposure to chloroform, indicating that it probably does not contain chloroform-soluble lipids and lipoproteins. V. vulnificus phage infectivity could not be inhibited with trypsin, protease, and ribonuclease treatments, while the infectivity of the Vibrio phages was inhibited with lysozyme and SDS treatments, perhaps demonstrating that the enzyme has interfered with the adsorbing of the phages [38, 39, 41]. The enzymatic treatments and inhibition of phage infectivity of several phages were reported. Proteinase K treatment could not alter the adsorption ability of phage particles. These studies show that binding of the phages to its host cell membrane is the first step of lytic cycle and this can be an irreversible mechanism, and a similar mechanism may exist in the infection of vibriophages as the infectivity of the phages were inhibited by lysozyme and SDS treatments. Treatment with 1% SDS did not affect the adsorption ability of phage particles. Similarly Mycoplasma arthritidis virulent 1 (MAV1) phage infectivity was reported to be unaffected by treatment with Triton X-100 and was resistant to nonionic detergents. Vibrio phages were found to be fairly resistant to chloroform [46, 101]. Further studies on the proteins and lipids of V. vulnificus phages may help us to understand their role in the life cycle and infectivity of phages. V. vulnificus phages survived 100% at pH 7 and exhibited infectivity, while none of the phages survived at extreme pH conditions (pH 3 and pH 12) [41]. V. harveyi phages were inactivated at pH 3 or less and at pH 12 or greater. In contrast, VPP97 phage was totally inactivated at pH below 5 or over 10. All V. vulnificus phages from the shrimp Penaeus monodon exhibited optimal survival at 37°C, but infectivity occurred, and plaques were observed up to a maximum temperature of 50°C [41]. JSF9 phage was shown to be stable at a temperature below 37°C, and the phages were rapidly inactivated above 50°C temperature. V. parahaemolyticus phage (VPP97) has been shown to be stable up to a temperature of 65°C and was totally inactivated at 70°C [41, 54, 58, 77, 101]. Phages were reported to survive extremes of temperature up to 95°C [54]. Phages, viz. T-ϕD0, T-ϕD2S, T-ϕHSIC, and T-ϕD1B, exhibited a latent period ranging from 90 to 180 min [77]. The results have clearly shown the physicochemical parameters are very important for the survival and infectivity of phages [37, 38, 46, 49]. Additional studies related to infectivity, stage specific expression of proteins, and specific effects of the purified phage proteins are needed to better understand their functions and applications in the control and process of infectivity of V. vulnificus phages.
Phage therapy is a re-emerging field, and the bacteriophages represent potential biocontrol agents for the control of virulent and drug-resistant bacteria [19, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73]. However the use of phage therapy in shrimp is still in its early years, and these are highlighted especially the need for using more than one kind of bacteriophage in aquaculture to evade development of bacterial resistance [47, 48, 49, 58, 59, 60]. Phage therapy has been effectively used to protect against Vibrio diseases in a shrimp and prawn hatchery. Inhibitory effects of bacteriophages against shrimp pathogenic Vibrio spp., efficacy of potential phage cocktails against Vibrio harveyi and closely related Vibrio species isolated from shrimp, morphological characterization and biocontrol effects of Vibrio vulnificus phages against vibriosis in the shrimp, and a phage therapy in aquaculture system have been described [37, 38, 39, 41]. Four new V. vulnificus phages were detected from shrimp aquaculture system, named VV1, VV2, VV3, and VV4 [39]. All lytic V. vulnificus phages belonged to Tectiviridae family with typical double-layered elongated icosahedral head and tailless morphology [39, 101]. Lytic V. vulnificus phages which infect other Vibrio isolates were further characterized for long-term storage by enzyme treatment, organic solvent treatment, detergent treatment, pH stability, temperature stability, and agar bioassay method and one-step growth experiment. The infectivity, growth, and multiplication of VV1, VV2, VV3, and VV4 phages were unaffected by the treatment effects of chloroform, acetone, ethyl alcohol, methyl alcohol, ribonuclease (RNase), trypsin, protease, and Triton-X100. The phages (VV1–VV4) were inactivated completely with temperature (over 60°C), pH (below 3 and above 12), and lysozyme and sodium dodecyl sulfate (SDS) treatment. One-step growth experiments indicated a latent period of 3 h and a burst size at 37°C. Agar bioassay method indicated that the percentage inhibition of the bacteria Vibrio was 75 (VV1) and 70 (VV2, VV3, and VV4), respectively. V. vulnificus phages had a broad lytic spectrum and potential biocontrol of luminous vibriosis in the shrimp aquaculture system [39, 41, 101]. The lytic Vibrio vulnificus phages may provide a better understanding of phage-host interactions and development of phage therapy in the aquaculture system. Besides, 12 V. harveyi phages showing broad host ranges were recovered from seawater samples. Further, some of the phages of ϕH17-5c, ϕH17-7b, ϕH17-8b, and ϕH17-9b were reported to show inhibitory activity against V. harveyi based on various tests, viz. heat stability, chloroform stability, adsorption rate, and one-step growth experiment [58]. A bacteriophage of Vibrio harveyi was secluded from shrimp farm water from the West coast of India and demonstrated to exhibit a broad lytic activity against V. harveyi isolates [48, 49]. V. harveyi-infected larval shrimp exhibited a higher rate of survival in the presence of the bacteriophage than the uninfected control larval shrimp. Bacteriophage treatment of the vibriosis of shrimp in hatchery tanks enhanced the survival of the V. harveyi-infected shrimp larvae and reduced the bacterial counts [50, 51, 52, 53, 54, 55, 57, 58]. Bacteriophages secluded from a shrimp hatchery and farmed P. monodon samples exhibited lytic activity against V. harveyi and also controlled the population of V. harveyi and improved the survival of Penaeus monodon larvae [41, 48, 49, 50, 51, 52, 58]. Purified bacteriophages exhibited lytic activity, indicating they are appropriate for phage therapy application [29, 53]. A few bacteriophages that infected V. harveyi in a shrimp hatchery were unable to control the outbreak of luminescent vibriosis disease in a shrimp culture system [75]. The potential of the bacteriophages of Vibrio harveyi was reported to control population of pathogenic Vibrio harveyi in a hatchery setting [48]. In a set of laboratory experiments, post-larval Penaeus monodon was exposed to 106 cfu ml−1 cells of Vibrio harveyi and was treated with 100 ppm phage which has led to a drastic reduction of Vibrio harveyi counts with 86% survival of the infected larvae, while the survival of the phage-untreated larvae was 25%. In the antibiotic (oxytetracycline 5 ppm, kanamycin 100 ppm/day)-treated hatchery tanks, an initial reduction of luminous bacterial counts was shown, and after 48 h the bacterial count increased to 106 ml−1, showing the luminous vibriosis and mortality of the nauplii of Penaeus monodon with 40% larval survival. In contrast, in the bacteriophage-treated tank, the larval survival was 86%, and the survival rate in the control tank was only 17% [48]. The presence of V. mimicus (15 isolates), exhibiting a typica profile of Vibrio [49] (two isolates), was obtained from diseased tissues of penaeid shrimp. A prominent occurrence of V. harveyi, V. furnissii, V. mimicus, V. damsela, and V. anguillarum in the hatchery tank water besides MBV- and WSSV-uninfected P. monodon, sea sediment, seawater, and shrimp culture pond sediment and shrimp culture pond water has been recorded [30, 53]. Vibriosis (V. alginolyticus)-associated mortality has been recorded in several invertebrates such as Penaeus monodon and Macrobrachium rosenbergii. Phage therapy has been shown to inhibit the growth and multiplication of V. harveyi, V. parahaemolyticus, and V. anguillarum. Bacteriophages belonging to Siphoviridae family are positive to control Vibrio species as the Siphoviridae phages are considered to have a specific host range consisting of species of Vibrio harveyi, Vibrio parahaemolyticus, and Vibrio campbellii [41, 44, 46, 48, 49, 50, 51, 52, 53, 58, 60, 69]. A lytic phage PW2 was obtained from shrimp pond water in Songkhla Province, Thailand, and the morphological characteristics of the phage consisted of an icosahedral head and a long noncontractile tail categorized under the order Caudovirales and family of Siphoviridae. The phage PW2 showed lytic properties against Vibrio harveyi [52]. Most of the Vibrio harveyi phages were found to be siphophages [25, 26, 29, 41, 51]. However, Vibrio harveyi phages which were from other families such as Myoviridae and Podoviridae were also reported [41, 51]. Selection of a suitable bacteriophage or a cocktail of phages is the key issue in the success of phage therapy of Vibrio species. Moreover phage cocktails have been demonstrated to be more effective than individual phages in treatment of Vibrio infection. By making a phage cocktail, it would become easier to treat a wide range of drug-resistant bacterial infections [58, 68]. Moreover phage-resistant mutants are exceptional, and hence the use of a multiphage therapy might decrease the resistance mechanism [61, 62, 63, 64, 65, 81, 82]. The limitations of the use of bacteriophages have been recognized, and the phages cannot be used if they (1) have toxin gene insertion; (2) have endotoxin release due to their lytic effect on the host bacterial cells; (3) have a risk of genetic material exchange, i.e., transduction or phage conversion; (4) have propagation and continuous maintenance; (5) have development of anti-bacteriophage bodies against them; (6) have the cost temperate phages contributing to bacterial virulence; (7) have emergence of phageresistant bacteria; (8) have disease outbreak of unknown bacteria when phage specificity is a problem; and (9) are continuously removed by the immune system [80, 81, 82]. Vibriophages themselves may have some protective effects though they can be candidates as therapeutic agents in bacterial infections in the aquaculture system, and therefore there is a need to be further investigating them in their natural environment. Future work may involve obtaining consistent credible results which can substantiate the application of bacteriophages to control vibriosis in shrimp and extending such research to other organisms. The usefulness of phage therapy to control microbial infections that occur in dissimilar organisms at various stages of from eggs to brood stock, as well as in laboratory, tanks, or field applications, was shown in experiments made with shrimp larvae, showing a promising potential for phage therapy [46]. The life-saving antibiotics and the new-generation antibiotics cannot be used in aquaculture, and therefore the bacteriophages have the natural advantages and potential to be used in management of the vibriosis in aquaculture.
Significant (71%) mortalities of larval, post-larval, and adult lobsters were caused by shell diseases where Vibrio spp., besides Pseudomonas, and Aeromonas have also been isolated. A strain of Vibrio owensii (DY05) was isolated from an epizootic in aquaculture-reared larval phyllosomas of the ornate spiny lobster Panulirus ornatus [10, 11, 12, 55]. Bacteriophage therapy was one of the techniques that controlled and removed the pathogenic Vibrio spp. from the larval cultures of the tropical rock lobster, Panulirus ornatus. V. harveyi has been found to be associated with diseases in spiny lobster [55].
Crothers-Stomps and colleagues [55] demonstrated that from eight bacteriophages (six phages belonged to the family Siphoviridae, and two belonged to the family Myoviridae), only one bacteriophage from the family Siphoviridae was shown to exhibit a clear lytic activity against V. harveyi with no apparent transducing properties. They have identified the occurrence of phage resistance as a major constraint to the use of phage therapy in aquacultures as the pathogenic bacteria were not completely eliminated [29, 41], and a similar approach on other phages is essential for understanding of the mechanism of development of phage resistance.
Bacterial and fungal diseases are accountable for the high mortality and profound economic loss encountered in the giant freshwater prawn M. rosenbergii aquaculture industry [19, 20, 21]. Vibriosis (V. alginolyticus)-associated mortality was recorded in M. rosenbergii [19, 20, 21]. They have shown the occurrence of a very high load of total heterotrophic bacterial count and a total presumptive Vibrio count in the 11 different larval stages of M. rosenbergii, whereas the total heterotrophic bacterial count was higher in the larval tank water throughout the cycle of the M. rosenbergii larval culture. The control treatment methods for the vibriosis consisted of chlorination, application of antibiotics, UV radiation alone, and a combination of sequential treatments starting from chlorination (2 ppm) followed by sand filtration, dechlorination, UV radiation (10 s) and microfiltration (5 μm size), and the sequential treatments in M. rosenbergii, and these had resulted in the gradual reduction of V. alginolyticus counts, and these were shown to be the best methods to control the pathogenic bacterial population in hatcheries of freshwater prawn M. rosenbergii. Antimicrobial resistance profile of Vibrio species isolated from the hatchery system of M. rosenbergii has been reported [19, 20, 21]. Vibriosis of M. rosenbergii can be controlled through phages φSt2 and φGrn1 of V. alginolyticus from live feed A. salina [97, 98, 99].
Shell diseases are caused by chitinolytic bacteria which were encountered in English prawn Palaemon serratus; American lobsters Homarus americanus; penaeid shrimp; and king crabs, Paralithodes camtschaticus and Paralithodes platypus; and the tanner crabs Chionoecetes tanneri are affected by rust diseases caused by chitin-destroying bacteria [7]. Biocontrol method against the infection of V. anguillarum for rearing the swimming crab larvae Portunus trituberculatus in the aquaculture water has been described [94, 95, 96]. A bacterial strain PM-4 Thalassobacter utilis isolated from a crustacean culturing pond was shown to inhibit the growth of pathogenic Vibrio anguillarum in seawater and to improve the growth of larval crab. The cells of the bacterial strain PM-4 T. utilis were cultured in a large quantity and were added daily for 63 of seawater used for culturing larval crab Portunus trituberculatus. Initial V. anguillarum bacterial density in the crustacean culture water was 106 cells ml−1, while at the crab larval growth stage zoea II, the bacterial density was found to increase to more than 107 cells ml−1 and reduced the pathogenic V. anguillarum bacteria to 106 cells ml−1. When the bacterial strain PM-4 Thalassobacter utilis dominated the bacterial populations, the numbers of pathogenic bacteria Vibrio spp. were reduced or could not be detectable in seawater. The growth and production of the larval crab P. trituberculatus were found to be increased by the addition of the bacterial strain PM-4 to the culture water. Investigations on the bacteriophages and phage therapy specific for aquaculture of crabs are desirable.
The occurrences of V. harveyi, V. furnissii, V. mimicus, V. damsela, and V. anguillarum in the nauplii of Artemia and the presence of V. harveyi and V. mimicus in the Artemia-reared water have been recorded [12, 18, 22, 39]. A prominent occurrence of Vibrio in the Artemia nauplii, Artemia-reared water, egg samples, hatchery tank water besides MBV- and WSSV-uninfected P. monodon, sea sediment, seawater, and shrimp culture pond sediment and shrimp culture pond water was recorded. Vibriosis (V. alginolyticus)-associated mortality has been recorded in several invertebrates such as Penaeus monodon and Macrobrachium rosenbergii [18, 19, 20, 21, 34, 97, 98, 99]. Vibrio species normally reside in the live feed organism such as Artemia salina which serves as means of carrying and establishing the bacteria into the hatchery and aquaculture system where V. alginolyticus has been reported as the prevailing member of the cultivable bacterial community of Artemia. There are a number of studies indicative that Artemia nauplii are vectors of potentially harmful bacteria such as Vibrio spp. Goulden et al. [12] reported that Vibrio owensii (DY05)-infected Artemia (brine shrimp) was responsible for 84–89% mortality of the aquacultured larval phyllosomas of the ornate spiny lobster Panulirus ornatus, and an understanding of the infection processes can help to improve targeted biocontrol strategies. The existing disinfection techniques such as filters, chlorination, ozone, UV, etc. could not prevent the occurrence of bacterial pathogens in the hatcheries and may perhaps help in the growth, proliferation, and multiplication of the opportunistic pathogen application of broad-spectrum antibiotics in the feed and hatchery water which has been the most natural strategy to control bacterial infections. The practice of applying antibiotics in aquaculture has become unattractive as many important bacterial pathogens belonging to the genera Aeromonas and Vibrio evolve antibiotic resistance. The excessive and indiscriminate use of antibiotics has resulted in the development of multidrug-resistant bacterial strains and public health problems. In shrimp hatcheries, the use of bacteriophages that infect bacteria is an alternative method to selectively eliminate their bacterial hosts while leaving normal microbiome unaltered. Phage therapy has been shown to inhibit the growth and multiplication of V. harveyi, V. parahaemolyticus, and V. anguillarum [39, 97, 98, 99].
In vitro cell lysis experiment of phage (a) φSt2 and (b) φGrn1 was carried by infecting the fresh cultures of the host bacteria V. alginolyticus strain V1 which was collected from live feed A. salina culture [97, 98, 99]. The lysis of the host bacteria V. alginolyticus strain V1 grown in TCBS was proportional to the multiplicity of infection (MOI) (which is defined as the ratio between the number of viruses in an infection and the number of the host cells which can be resolved by correcting the relative concentration of virus and host) used. The lowest (MOI = 1) showed no effect, while the highest (MOI = 100) showed a complete inhibition of bacterial growth. The phages were also tested in vitro against 12 different bacterial isolates grown in TCBS which originated from live feed A. salina culture. A phage mixture of φSt2 and φGrn1 at MOI = 100 was shown to affect the growth of all 12 bacterial strains tested, and the study showed that in all cases, there was a delay in the exponential phase, and even when the cultures reached a plateau of growth, the density of phage-treated bacteria was lower than their corresponding controls.
The effect of the phage mixture (φSt2 and φGrn1 at MOI = 100) was examined by administrating the phages in vivo in the live prey cultures of A. salina. After 4 h of administration of the phage mixture, Vibrio count was not altered in the phage-untreated control cultures, while the Vibrio count drastically reduced in the phage-treated cultures of A. salina [97]. The total Vibrio counts in the phage-treated cultures of A. salina was 5.3 × 103 ± 3.1 × 103 cfu ml−1, which was 93% lesser than that of the initial total Vibrio counts. The φSt2 and φGrn1 phages affected the growth of a range of Vibrio spp. and exhibited lytic activity in eight different V. alginolyticus strains [97]. The phages also exhibited infection in 10 out of the 25 Vibrio strains (40%) tested and lysis in bacterial strains such as V. harveyi and V. parahaemolyticus species and were known to affect the growth of several strains of bacteria V. harveyi strains isolated from an Artemia live feed organism. A broad host range of φSt2 and φGrn1 phages indicated the occurrences of a very similar host structures as receptors, viz. LPS phage receptors on the outer membrane of many of Gram-negative bacteria and the genetic similarity contributing to this broad lytic spectrum of the phages φSt2 and φGrn1 phages which exhibited a strong lytic effect against V. alginolyticus-type strain DSM 2171 [98]. The bacteriophages, φSt2 and φGrn1, are having a broad host range, and such biological attributes make them the potential candidates for phage therapy application, and these advocate that phage therapy can be an alternative to antibiotics in aquaculture. The phages, viz. φSt2 and φGrn1 and a giant bacteriophage pVa-21, can effectively be used in the biological control of pathogenic Vibrio species in marine hatcheries [97, 98, 99]. These studies indicate the potential applications of the phages for various purposes outside aquaculture, and further research on the specificity, phage-host interactions, proteomics, and genomics of phages are needed to establish their usefulness and utilization. Efficacy of phage therapy was shown to prevent mortality of the vibriosis-infected brine shrimp Artemia franciscana [97, 98, 99]. Application of single-type/cocktails of phages against V. parahaemolyticus- and V. harveyi-infected cysts and nauplii Artemia franciscana showed a drastic improvement in the survival rate (from 85 to 89%) and hatching success (100% in both cases) in groups treated with phages, whereas the control groups exhibited a survival rate of 40–50% and hatching success (50%). These studies indicate that the phage cocktails offer an alternative to chemotherapeutic agents and can be used in brine shrimp production.
The callous use of antibiotics against bacterial diseases in the culture of crustaceans in the aquatic system has led to the development of antibiotic-resistant bacterial infections which can be a serious threat to all life forms and public health, and the phage therapy may help to overcome such complex problems. Control of bacterial diseases in the future may depend on development of novel drugs, innovative approaches, and management practices to minimize the risk of introduction of infectious agents into aquaculture systems and to reduce predisposing factors. The occurrences of lytic bacteriophages of Bacillus spp./Vibrio spp. as well as their efficacy were established. Phage therapy has been shown to inhibit the growth and multiplication of many different pathogenic bacteria, viz. V. harveyi, V. parahaemolyticus, V. alginolyticus, V. furnissii, V. mimicus, V. damsel, and V. anguillarum, and promote better survival and production of the crustaceans in aquaculture. Infected live feed Artemia is responsible for the establishment of the bacterial infections in the hatchery and aquaculture system causing mass mortality of the larval crustaceans in culture, and such bacterial infections can be controlled through lytic phages, viz. ϕH17-5c, ϕH17-7b, ϕH17-8b, and ϕH17-9b from V. harveyi; phages VV1, VV2, VV3, and VV4 from V. vulnificus; and φSt2 and φGrn1 of V. alginolyticus of A. salina. However development of specific cocktails of phage therapy for aquaculture of crabs, prawns, lobsters, and crabs is desirable. The phage therapy is an ideal method for the control of microbial infections that occur in dissimilar organisms at various stages from eggs to brood stock as well as in laboratory, tanks, or field applications, and extending such research to other organisms could be one of the valuable strategies, to provide evidences and validate the usefulness and therapeutic potential of the phage therapy. An understanding of the infection processes, phage resistance, the efficacy of phage therapy on the targeted pathogens, and their impact on the normal microbiome can help to improve biocontrol strategies. The potential applications of the phages for various purposes outside aquaculture and further research on the specificity and phage-host interactions are needed to establish their usefulness and exploitation. Future work may be carried out on the intricacies of phage lifestyles and their dynamics in natural systems, genome and viriomes, proteome analysis, genes coding for their proteins, and DNA polymerase phylogeny, which can help us in identifying novel methods of phage-host interaction and in understanding the way in which phages control their hosts. The use of probiotic organisms and phage therapy in crustacean aquaculture is found to be safe for the consumer of shrimp, and therefore a scientific rational approach is needed to evolve guidelines for phage therapy/probiotic applications in aquaculture and for their use and safety.
The author (PR) is thankful to Ms. Keerthana and Mrs. Rekha for their assistance and help in the work. The author acknowledges support from Sree Balaji Medical College and Hospital for providing research facilities and encouragement for technical assistance to complete the work.
The author declares that there is no financial or conflict of interests.
The author (Dr P. Ramasamy) has made a significant contribution to the conception, design, and execution of the reported study and drafted the manuscript writing and discussion.
The concept of Corporate Social Responsibility (CSR) stems from the need for companies to interconnect the needs of the community with the various sources of profit. The growing interest in CSR issues, especially in banks, is the result of a cultural journey that sees the company react to market changes and to be the protagonist of an increasingly sustainable future.
Corporate social responsibility is understood: “companies integrate social and environmental concerns in their business operations and in their interaction with their stakeholders on a voluntary basis” [1]. In other words, the company integrates social and environmental interest among its strategic objectives. Together with the financial and environmental aspects, the ethical value of banks is more important for the development of both productive and marketing strategies, representing a new tool of competitiveness [2]. At the beginning of the 1970s, the first CSR studies were born to analyze the correlation between social issues and economic performance. However, it was in the 1990s that there was a real explosion of the CSR issue [3, 4].
The prevailing approach up to this period was that there was a negative correlation between the ethical and social orientation of the investor and the economic performance. It was believed that investing in good behavior practices would reduce the number of available investment alternatives and possibly damage economic performance.
The spread of sustainable investments in financial markets, the development of ethical stock market indices and ethical rating methodologies, has helped to affirm the belief that there are economic benefits related to the assumption of corporate social responsibility. In fact, investing in socially responsible behaviors can also bring economic benefits.
In line with these considerations, CSR is not a follow-up to profit, but sees it as a profit-making option. In banking, CSR is an important aspect of the company’s strategy and it must have a substantial value in its business. In other words, it is necessary to integrate CSR into strategies, processes, operations as well as daily relationships with stakeholders. If sustainability enters these areas, then it can effectively contribute to the resilience of the economic and social fabric, foster confidence in the market and the acceleration of the recovery from the crisis [5].
Since the last economic crisis, the deteriorating economy along with numerous banking scandals has provided a new and challenging environment for the banking sector. At the beginning of the crisis, scholars discussed its impact on social investment [6, 7]. Some predicted a sharp reduction in CSR budget costs if they were perceived as non-core assets, while others believed that companies strategically engaged in CSR would continue to spend in this area, despite the challenging economic environment. Banks are blamed primarily for the financial crisis that caused economic turmoil [8, 9].
Corporate scandals, lack of transparency and subsequent government bailouts have undermined public confidence in the banking sector. Several authors argued that the positive results of the CSR be particularly remarkable in the banking sector, as banks have had a reputation tarnished in the wake of the financial crisis [8, 10]. Transparency is very relevant in restoring bank reputation, which may explain why financial companies report significantly more information about CSR than other industries [11]. CSR acts as a protection of the company’s market value in times of crisis [12, 13]. While general mistrust in the financial sector has had a negative effect on reputation and therefore performance, CSR strategies could mitigate these results. In this way, CSR can be considered preventative in times of non-crisis because it improves reputation. However, it is also interesting to consider the effect of CSR in a post-crisis situation as a tool to restore reputation and mitigate a reputational crisis following corporate scandals [14, 15].
Absent or incorrect CSR policies have a much greater negative effect on performance than the positive effects of correct policies. However, the recent recession in the world economy, particularly in Europe, has shed light on some management scandals and the lack of integrity in the European banking sector. This has had a negative impact not only on bank returns but also on bank reputation. Banking governance plays a crucial role in the implementation of CSR practices. It is believed that sustainable measures lead to reputation and performance improvement when management demonstrates strong ethical leadership [16, 17]. In the banking sector, some sustainable policies have not been able to improve reputations and returns since the start of the financial crisis [18]. Unethical practices and mismanagement in several European banks have caused anger, and distrust of the sector that has received public bailouts, while some bank executives have been paid exorbitant bonuses. As a result, the ethical leadership and credibility of the banks were called into question, resulting in a major loss of reputation, as the public perceived discrepancies between the CSR directives of bank executives and their effective behaviors [19]. In this scenario, investments in CSR have failed to improve reputation due to weak business leadership. After one of the deepest economic crises in history, banks perceive CSR as a means of restoring their image and credibility [20, 21, 22, 23]. The banking sector’s commitment to more sustainable practices has interesting implications. In fact, banks can play an important role in economic development [24] because they decide how to allocate financial resources to different companies and sectors. Non-responsible companies pay an additional cost on bank financial income than the companies responsible because investments in CSR reduce risk and are more attractive to lenders [25]. Therefore, the involvement of banks in CSR practices should benefit the bank itself and promote the adoption of sustainable practices by potential borrowers, thereby having a positive impact on sustainable growth [26]. This makes the financial sector unique when considering the effects of CSR practices. In the banking sector, CSR covers many activities such as lending, wealth management, the operation of payment systems and risk management [27]. All of these factors are able to significantly influence society and its surroundings. For this reason, banks should fully integrate CST into their business strategies and see it as a strategic tool that can improve relationships with stakeholders, resulting in positive impacts both in terms of consensus and confidence and performance. If a bank acts in a socially responsible way, it creates the basis for consolidating its long-term presence in the market, emphasizing its contribution to environmental quality and society. CSR’s business affects all stakeholders involved in the business with different capabilities and with different expectations [28]. The CSR is taking on a crucial role among academics and researchers, thanks to its ability to jointly consider all aspects of operations: economic, environmental and social [29]. This is the approach of the so-called triple bottom line [30], according to which the assessment of benefits must cover not only the economic aspects, but also the environmental and social aspects.
Undoubtedly, there is the need for integrated communication between the criteria for implementing CSR practices. Disclosure of CSR is regulated by national and international self-regulatory measures. It is a voluntary disclosure and this faculty is linked to the very essence of ethics, inevitably influenced by specific business activities and difficult to define without proper contextualization.
Among the most relevant CSR provisions are the OECD Guidelines [31], which suggest that integrated relationships should be adopted. In addition, the Global Reporting Initiative (GRI) guidelines for sustainable reporting include the principles needed to define report content (Materiality, stakeholder inclusion, sustainable context and comprehensiveness) and relationship quality. They also include standard disclosure: organizational strategy and profile, management approach and performance indicators (economic, environmental and social).
European banks are more concerned about environmental, social and governance issues than their competitors based in other parts of the world. This can be confirmed, for example, by the proportion of signatories to Equator Principles [32], with European institutions accounting for 42% of all adopters compared to North American, Latin American and Asian entities, representing 17%, 12% and 9% of all signatories respectively [33].
European banks, as the first to adopt sustainability practices, can be a benchmark for their peers in other regions. In addition, in Community area the banking sector is known for the relevance of bank income in overall financial intermediation compared to other regions, such as the United States, where capital markets are the main source of financing. In fact, in the European banking-based financial system [34], banking is three times the EU’s total GDP [35], unlike other advanced economies, namely the United States, where a market-based system prevails that derives in a lower percentage of banking intermediation in the economy, where bank assets roughly correspond to GDP.
Today, banks pay attention to corporate social responsibility as an additional lever of innovation and development to better compete in the market in the medium and long term. Taking a CSR path is an opportunity for the bank to: (i) improve proactive risk governance by integrating social, environmental and government variables into their corporate governance system; (ii) listen to the needs of your stakeholders and innovate the development of products, services and business models; (iii) make explicit the implications that the role of money brokerage has on the company and maximize the creation of a shared value.
This chapter presents the results of a survey of a sample of banks belonging to the Global Systematically Important Institutions (G-SII) universe, as defined by the EBA. The list of banks included in this section follows the EBA’s guidelines on the dissemination of indicators of global systemic importance in order not only to increase the transparency of the G-SII identification process, but also to achieve a level playing field in terms of disclosure requirements between systemically important institutions and other large institutions. The EBA guidelines directly follow the Recommendations of the Basel Committee to identify global systemically important banks (G-SIBs) and provide data that help assess the systemic riskiness of EU banks.
In line with the EBA’s guidelines, all European institutions with a leverage ratio of more than 200 billion euros are required to participate in this disclosure. Our sample includes 25 G-SII operating on European territory in 2018. The following table (Table 1) shows the banks included in the sample. Of the 25 banks, 5 are from the United Kingdom, 4 in Spain and Sweden respectively, 3 in France, 2 in Germany and Italy and 1 for Austria, Belgium, Denmark and the Netherlands respectively.
Financial institution | Country |
---|---|
E | Austria |
K | Belgium |
D | Denmark |
B | France |
C | France |
S | France |
C | Germany |
D | Germany |
I | Italy |
U | Italy |
A | Netherlands |
D | Norway |
S | Spain |
B | Spain |
L | Spain |
S | Spain |
N | Sweden |
S | Sweden |
H | Sweden |
S | Sweden |
B | UK |
H | UK |
L | UK |
R | UK |
S | UK |
Sample.
In terms of assets managed in December 2018 (Figure 1), UK banks are at the top of the ranking (36% of assets attributable to the entire sample). In second place are Spanish banks with 18% of assets managed, followed by German banks with 12% and Italian banks with 11%. Overall, French and Swedish banks manage 18% of the assets. Netherlands ranks seventh with only 3%, followed by Austria and Norway with a total of 4% of assets managed. Finally, the Belgian and Danish banks are included in the final part of the rankings, with a total of 0.02% and 0.01% respectively.
Total asset by country.
In order to ascertain the degree of integration of CSR practices by the selected banks, several areas of investigation were analyzed, selected because they were considered relevant according to an analysis of the studies on the subject.
The research focused on four areas of investigation, relating to the composition, size and configuration of the Boards of Directors of the 25 banks examined. In particular, they were examined for each company:
The size of the board of directors
The presence of independent directors on the board
The presence of women on the board
The presence of endo-council committees specifically dedicated to sustainability issues.
In order to achieve our goal, we analyzed all the bank’s official documents on governance and sustainability policies, as well as we used Datastream database with regard to some qualitative aspects.
The importance and efficiency of CSR practices in banks depends almost exclusively on the board of directors and the information provided to stakeholders. The CSR disclosure helps to increase the well-being of stakeholders and communicate information on the bank’s economic, social and environmental performance [36]. This reporting also reduces the information asymmetry between shareholders and bank executives [37]. In line with these considerations, CSR is a valuable tool to increase shareholder confidence and improve the bank’s ethical behavior. It is therefore one of the key factors in influencing the bank’s competitiveness and long-term success [38].
The growing interest in CSR has led many countries to introduce their respective regulatory frameworks. CSR regulations have been imposed for banks in different countries over the years (e.g. 2003 in Austria, 2007 in Malaysia, 2009 in Sweden, 2010 in China, 2012 in Spain, 2016 in Belgium and 2017 in Hungary and Singapore). Other countries, such as Australia, Canada and Cyprus, have soft regulations in the form of recommendations to encourage the disclosure of CSR [39]. Banks should follow standards (e.g. GRI, designed for the financial services sector) or employ independent external auditors to ensure the quality and reliability of the information disclosed.
The efficiency of the banks’ board of directors is important to ensure their stability, compliance with regulations, the protection of stakeholders as well as to form long-term strategies that also include sustainability issues [37, 40, 41]. Diversity in the composition of the Board of Directors is considered one of the key elements to resolve complex issues and satisfy the interests of different actors. Diversity on company boards should improve good corporate governance. The diversity of the Board of Directors is examined in terms of the composition of the board of directors with a focus on the size of the board, the independence of the board of directors and gender diversity.
The size of the board of directors in banks is much larger than the boards of directors of non-financial corporations [42]. These differences in the size of the board of directors may depend on the complexity of banking activities and regulatory recommendations. Several studies examine the relationship between the size of the board of directors and the various performance measures of banks. The size of a bank’s board of directors has positive effects on performance; this is probably due to the fact that banks are complex businesses and the advantages of larger boards outweigh costs, improving monitoring functions and mitigating risks.
In order for the Board of Directors to carry out its functions efficiently, it is necessary to diversify the skills and experience of its members [43]. More board members are associated with better monitoring mechanisms for performing their functions as well as an improvement in CSR practices [44]. As more directors provide a more diverse and broader variety of skills and opinions, larger boards of directors are expected to focus more on the CSR [45, 46]. The banking sector, being subject to strict information disclosure requirements, is more transparent than non-financial companies.
Also the independence of the Board of Directors is considered one of the most efficient governance mechanisms [47]. Independence is linked to the presence of non-executive directors who ensure the correct behavior of the company [37, 48]. Independent directors therefore act as guardians of the company’s legitimacy by ensuring compliance with regulations and meeting the expectations of the external environment, including social and environmental concerns [49]. Non-executive directors can be guided by personal interests and consequently pursue goals that are misaligned with the company’s strategy. Since CSR information is obtained by management, there is a risk of spreading misleading information [50]. In that case, independent directors may reduce that risk. Much of the existing literature is agreed that non-executive board members are positively associated with the disclosure of the CSR of banks or its quality [49].
Nowadays a large part of CSR studies believe that a key success factor is represented by the diversity of the board in terms of gender, ethnicity or background. Diversity on boards, expressed in terms of the number of women on the board, should increase the independence of the board and focus on the interests of different stakeholders [40]. Leadership styles based on gender diversity suggest that women tend to be more democratic, showing more empathy for diversity [39, 51]. This indicates that women should have a positive influence on the functioning of the board of directors as they should promote collaboration and integration of more complex issues in discussions and decision-making. Much of the literature on the subject is in agreement in affirming the positive association between the number of women on the board of directors and the information on the CSR of the banks [43, 45, 48].
Finally, it is worth noting that in recent year companies, in order to achieve sustainability goals, more frequently choose to set up a committee. The CSR or Sustainability Committee assists the Board of Directors in overseeing the company’s liability practices, but they can also play a key role in monitoring and evaluating the company’s CSR performance by ensuring compliance with regulations that manage sustainability risks. In other words, the CSR Committee helps to improve the ethical culture of the company by ensuring that the potentially dangerous risks to the company’s reputation are properly assessed [52, 53].
The CSR advisory committee periodically reports to the board on sustainability issues affecting the company, while managing public disclosure on sustainability issues. The existence of a CSR committee is evidence of the company’s commitment to CSR and therefore to the pursuit of ethical and sustainable objectives [54, 55].
In line with these considerations, a company that decides to set up a CSR committee demonstrates not only its CSR commitment to stakeholders, but also its intention to make sustainability a key strategy to improve the extent or quality of sustainability disclosure [56, 57, 58].
The size of the board of directors as a lever to make the function of the bank’s board of directors efficient is analyzed by several academics and scholars. In line with the introductory considerations, a greater number of members of the board of directors is associated with better monitoring mechanisms for carrying out the functions of the board as well as an improvement in CSR practices. In line with these considerations, the analysis carried out revealed that the average size of Board of Directors is 13 members within a range that varies from a minimum of 6 to a maximum of 21 members. Although a positive correlation between the number of members of the Board of Directors and size - measured in terms of assets managed - can be detected, it does not however assume particularly significant values (correlation coefficient: 0,14) (Figure 2).
Board size.
The second area of investigation concerned the examination of the number of independent directors. In line with existing literature, independent directors can reduce the risk of manipulation or distortion of CSR reporting. The boards of directors of the banks examined present an average of 64% of independent directors, in a range that varies from a minimum of 24% to a maximum of 64%. Only in one case is the board of directors made up exclusively of independent directors. However, it should be noted that most banks have at least 50% of independent directors (18 out of 25 banks), while in the remaining 7 banks the percentage of independent directors varies between 24% and 45% (Figure 3).
Number of independent directors.
Gender diversity on boards of directors, usually expressed in terms of the number of women on the board of directors, should have a positive influence on the functioning of the board of directors and information on banks’ CSR.
The empirical analysis shows that in 2018, the representation of women on the boards of directors of the banks analyzed was 35%. In three of the banks examined, the number of women on the board of directors is equal to the number of men. 18 of the banks examined have a percentage of women on the board of directors of more than 30%, while in the remaining 7 banks there is a percentage varying between 13 and 29% (Figure 4).
Number of women on the board.
Establishing a committee dedicated to CSR is a widespread practice (92% of the sample). The analysis showed a strong heterogeneity in the behavior of banks. On the one hand, some banks decide to set up coordination committees that control other units dedicated to specific CSR issues. On the other hand, in other cases there is cooperation between officials at group level or committees focusing on specific issues relating to the environment, society and governance. The range of activities carried out by CSR functions include: stimulating CSR initiatives and increasing internal awareness of CSR issues; formulation and monitoring of policy and accountability programmes; responsibility for coordinating and implementing the company’s sustainability strategy and action plan; measures to deliver the sustainability strategy and achieve agreed company-wide goals. In the cases examined, there is often a special committee for responsible investments in the asset management business area to ensure that banks’ responsible investment policy is respected (Figure 5).
CSR committee.
The concept of Corporate Social Responsibility stems from the need for companies to interconnect the needs of the community with the various sources of profit. The growing interest in CSR issues, especially in banks, is the result of a cultural journey that sees the company reacting to market changes and being the protagonist of an increasingly sustainable future.
Banks integrate social and environmental interest into their strategic objectives. Together with the financial and environmental aspects, the ethical value of the banks assumes greater importance for the development of both production and marketing strategies, representing a new tool for competitiveness.
Banks pay attention to corporate social responsibility as an additional lever of innovation and development to better compete on the market in the medium and long term. More precisely, CSR contributes to the improvement of proactive risk management, integrating it with social, environmental and government variables; improves the relationship with stakeholders, promoting an analysis of the needs of bank interlocutors and the development of products, services and commercial models. Finally, the CSR makes explicit the implications that the role of intermediation of money has on society and favors the creation of a shared value. In light of the above, this chapter has set itself the objective of exploring the level of integration of Corporate Social Responsibility in the banking system. To achieve this, we carried out an exploratory analysis on a sample of 25 banks, belonging to the universe of Global Systematically Important Institutions in 2018. All the bank’s official documents on governance and sustainability policies were analysed, and we used the Datastream database for some qualitative aspects. Our study focused on four areas of investigation relating to the composition, size and configuration of the boards of directors.
The main results show a favorable attitude of banks towards the integration of sustainable policies. More precisely, with regard to the first area ofinvestigation, a greater number of members of the board of directors (average of 13 directors) are associated with an improvement in CSR practices.
The examination of the number of independent directors (second area of investigation), as a tool to reduce the risk of manipulation or distortion of CSR relationships, showed positive trends. In fact, the boards of directors of the banks examined present an average of 64% of the independent directors.
A further crucial element for examining the implementation of CSR policies in banks concerns gender diversity on boards of directors. It is believed that more women on the board of directors positively influence the functioning of the board of directors and the disclosure on CSRs in banks. In 2018 the representation of women on the boards of banks of the banks analyzed was 35%.
Finally, the last area of investigation relating to the presence of a committee dedicated to CSR reveals a strong heterogeneity in the behavior of banks. On the one hand, some banks decide to set up coordination committees that control other units dedicated to specific CSR issues. On the other hand, in other cases there is cooperation between group level officials or committees focused on specific issues relating to the environment, society and governance. In the cases examined, there is often a special committee for responsible investment in the commercial asset management sector to ensure compliance with the responsible investment policy of banks.
To sum up, the integration of CSR policies will allow banks to compete better on the market in the medium and long term, satisfy the requests of their stakeholders as well as protect the ethical and social values of the banks themselves.
This chapter represents an exploratory study on the level of integration of CSR practices in banks and in particular on the boards of directors of banks. The elements considered in this study may be further investigated, through future empirical analyzes. Future research could be oriented towards an in-depth examination of the sustainable investments put in place by banks over time.
The authors declare no conflict of interest.
This article is the result of the joint efforts of the authors, who equally contributed to the work.
The Internet has irrevocably changed the dynamics of scholarly communication and publishing. Consequently, we find it necessary to indicate, unambiguously, our definition of what we consider to be a published scientific work.
",metaTitle:"Prior Publication Policy",metaDescription:"Prior Publication Policy",metaKeywords:null,canonicalURL:"/page/prior-publication-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"A significant number of working papers, early drafts, and similar work in progress are openly shared online between members of the scientific community. It has become common to announce one’s own research on a personal website or a blog to gather comments and suggestions from other researchers. Such works and online postings are, indeed, published in the sense that they are made publicly available. However, this does not mean that if submitted for publication by IntechOpen they are not original works. We differentiate between reviewed and non-reviewed works when determining whether a work is original and has been published in a scholarly sense or not.
\\n\\nThe significance of Peer Review cannot be overstated when it comes to defining, in our terms, what constitutes a published scientific work. Peer Review is widely considered to be the cornerstone of modern publishing processes and the key value-adding contribution to a scholarly manuscript that a publisher can make.
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\\n\\nIn order to facilitate the tracking of a manuscript’s publishing history and its development from its earliest draft to the manuscript submitted, we encourage Authors to disclose any instances of a manuscript’s prior publication, whether it be through a conference presentation, a newspaper article, a working paper publicly available in a repository or a blog post.
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\\n\\nSome basic information about the editorial treatment of different varieties of prior publication is laid out below:
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\\n\\nGiven that conference papers and presentations generally pass through some sort of peer or editorial review, we consider them to be published in the accepted scholarly sense, particularly if they are published as a part of conference proceedings.
\\n\\nAll submitted manuscripts originating from a previously published conference paper must contain at least 50% of new original content to be accepted for review and considered for publication.
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\\n\\nSubmitted manuscripts stemming from a previous newspaper or magazine article will be accepted for review and considered for publication. However, Authors are strongly advised to report any such publication in an accompanying note to the External Editor.
\\n\\nAs with the conference papers and presentations, Authors should obtain any necessary permissions from the newspaper or magazine that published the work, and indicate that they have done so in a note to the External Editor.
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\\n\\nWhite papers, working papers, technical reports and all other forms of papers which fall within the scope of the ‘Luxembourg definition’ of grey literature do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense.
\\n\\nAlthough such papers are regularly made publicly available via personal websites and institutional repositories, their general purpose is to gather comments and feedback from Authors’ colleagues in order to further improve a manuscript intended for future publication.
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\\n\\n4. SOCIAL MEDIA, BLOG & MESSAGE BOARD POSTINGS
\\n\\nWe feel that social media, blogs and message boards are generally used with the same intention as grey literature, to formulate ideas for a manuscript and gather early feedback from like-minded researchers in order to improve a particular piece of work before submitting it for publication. Therefore, we do not consider such internet postings to be publication in the scholarly sense.
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\\n"}]'},components:[{type:"htmlEditorComponent",content:'A significant number of working papers, early drafts, and similar work in progress are openly shared online between members of the scientific community. It has become common to announce one’s own research on a personal website or a blog to gather comments and suggestions from other researchers. Such works and online postings are, indeed, published in the sense that they are made publicly available. However, this does not mean that if submitted for publication by IntechOpen they are not original works. We differentiate between reviewed and non-reviewed works when determining whether a work is original and has been published in a scholarly sense or not.
\n\nThe significance of Peer Review cannot be overstated when it comes to defining, in our terms, what constitutes a published scientific work. Peer Review is widely considered to be the cornerstone of modern publishing processes and the key value-adding contribution to a scholarly manuscript that a publisher can make.
\n\nOther than the issue of originality, research misconduct is another major issue that all publishers have to address. IntechOpen’s Retraction & Correction Policy and various publication ethics guidelines identify both redundant publication and (self)plagiarism to fall within the definition of research misconduct, thus constituting grounds for rejection or the issue of a Retraction if the work has already been published.
\n\nIn order to facilitate the tracking of a manuscript’s publishing history and its development from its earliest draft to the manuscript submitted, we encourage Authors to disclose any instances of a manuscript’s prior publication, whether it be through a conference presentation, a newspaper article, a working paper publicly available in a repository or a blog post.
\n\nA note to the Academic Editor containing detailed information about a submitted manuscript’s previous public availability is the preferred means of reporting prior publication. This helps us determine if there are any earlier versions of a manuscript that should be disclosed to our readers or if any of those earlier versions should be cited and listed in a manuscript’s references.
\n\nSome basic information about the editorial treatment of different varieties of prior publication is laid out below:
\n\n1. CONFERENCE PAPERS & PRESENTATIONS
\n\nGiven that conference papers and presentations generally pass through some sort of peer or editorial review, we consider them to be published in the accepted scholarly sense, particularly if they are published as a part of conference proceedings.
\n\nAll submitted manuscripts originating from a previously published conference paper must contain at least 50% of new original content to be accepted for review and considered for publication.
\n\nAuthors are required to report any links their manuscript might have with their earlier conference papers and presentations in a note to the Academic Editor, as well as in the manuscript itself. Additionally, Authors should obtain any necessary permissions from the publisher of their conference paper if copyright transfer occurred during the publishing process. Failure to do so may prevent Us from publishing an otherwise worthy work.
\n\n2. NEWSPAPER & MAGAZINE ARTICLES
\n\nNewspaper and magazine articles usually do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense. Articles appearing in newspapers and magazines rarely possess the depth and structure characteristic of scholarly articles.
\n\nSubmitted manuscripts stemming from a previous newspaper or magazine article will be accepted for review and considered for publication. However, Authors are strongly advised to report any such publication in an accompanying note to the External Editor.
\n\nAs with the conference papers and presentations, Authors should obtain any necessary permissions from the newspaper or magazine that published the work, and indicate that they have done so in a note to the External Editor.
\n\n3. GREY LITERATURE
\n\nWhite papers, working papers, technical reports and all other forms of papers which fall within the scope of the ‘Luxembourg definition’ of grey literature do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense.
\n\nAlthough such papers are regularly made publicly available via personal websites and institutional repositories, their general purpose is to gather comments and feedback from Authors’ colleagues in order to further improve a manuscript intended for future publication.
\n\nWhen submitting their work, Authors are required to disclose the existence of any publicly available earlier drafts in a note to the Academic Editor. In cases where earlier drafts of the submitted version of the manuscript are publicly available, any overlap between the versions will generally not be considered an instance of self-plagiarism.
\n\n4. SOCIAL MEDIA, BLOG & MESSAGE BOARD POSTINGS
\n\nWe feel that social media, blogs and message boards are generally used with the same intention as grey literature, to formulate ideas for a manuscript and gather early feedback from like-minded researchers in order to improve a particular piece of work before submitting it for publication. Therefore, we do not consider such internet postings to be publication in the scholarly sense.
\n\nNevertheless, Authors are encouraged to disclose the existence of any internet postings in which they outline and describe their research or posted passages of their manuscripts in a note to the Academic Editor. Please note that we will not strictly enforce this request in the same way that we would instructions we consider to be part of our conditions of acceptance for publication. We understand that it may be difficult to keep track of all one’s internet postings in which the researcher´s current work might be mentioned.
\n\nIn cases where there is any overlap between the Author´s submitted manuscript and related internet postings, we will generally not consider it to be an instance of self-plagiarism. This also holds true for any co-Author as well.
\n\nFor more information on this policy please contact permissions@intechopen.com.
\n\nPolicy last updated: 2017-03-20
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