Evaluation of Antimicrobial Potency on Skin Mucus of Three Major Indian Carp Species

 

 

 

 

 

 

 

 

 

 

       Evaluation of Antimicrobial Potency on Skin Mucus of Three Major Indian Carp Species

Student's Name

Institutional Affiliation

 

 

 

 

 

 

 

 

 

 

ACKNOWLEDGEMENT

I   thank the Almighty God for his guidance and protection throughout this study. I extend my gratitude to my family and friends for the financial and moral support. I would also like to acknowledge my supervisor for the guidance and support throughout the study.

 

 

 

 

 

  

 

 

 

 

 

 

 

 

 

 

 

 

 

ABSTRACT

Zoonotic infections resulted from bacterial pathogens threatens both humans and the aquaculture industry. This outbreak probed search for different epidermal mucus as natural products from three Indian Major Carp species to evaluate the potency of the fish skin mucus as an effective antimicrobial against human and fish pathogenic bacteria. The skin mucus of the three indigenous Indian Major Carp species got collected against pathogenic fish bacteria and human viz. Catla catla, Labeo rohita, and Cirrinus mrigala. The activities of the microbes were measured based on Zone of Inhibition (ZOI) in millimeters to assess the MICs of various mucus extracts against a variety of bacteria. The results were compared with standard antibiotics amikacin and chloramphenicol to determine fish skin mucus antimicrobial potency.

Moreover, the three selected fish species were also challenged with A. Hydrophila to evaluate the presence of any change in their mucus and antibacterial influence after exposure of bacterial. Afterward, the agar plate dilution test was used to analyze the minimum inhibitory concentration of HFM and CFM from every fish species against the selected microbial strains. Therefore the results of the study were useful in identifying the possibility of using the fish skin mucus as an alternative to antibiotics in fish and human health associated complications.

 

Keywords: Zone of Inhibition (ZOI), fish, antimicrobial activity, skin mucus, major Indian carps (IMCs), Healthy Fish Mucus (HFM), Challenged Fish Mucus (CFM), Colony Forming Units (CFU)

 

                  

    

 

Evaluation of antimicrobial potency on Skin Mucus of Three Major Indian Carp Species

INTRODUCTION

Fishes play a significant role in the aquatic environment. Their diversity forms the largest class of water habitat, especially cold-blooded vertebrate species.  They provide interconnection between food webs of the water ecosystems, which harbors a variety of pathogenic micro-organisms (Ellis 2001). Moreover, According to Alvarez-Pellitero (2008), infectious diseases are widely spread when fish get reared in intensive and high densities aquaculture. Increased population density and limited resources provide a favorable environment for the thriving of pathogens.

Aquatic organisms like fish get stressed in a crowded and unnatural environment characterized by a limited flow of water hence making the fish family more susceptible to diseases. Consequently, through the consumption of fish, the transmission of the bacterial pathogens from infected fish poses a severe danger to the aquaculture industry, and human health. Increased density of bacteria, viruses, and parasites are, therefore, critical for the development of aquaculture (Cooper and Shlaes 2011). Infectious diseases cause complications such as edwardsiellosis, columnaris, and microbial hemorrhagic ascites in fish. Interestingly, fishes pose mucus substances on their surfaces, which aides them in locomotion, osmoregulation, body lubrication that wades off pathogens and also immunological functions.

            According to Lirio et al. (2018), the mucus layer defends the fish against the microbe invasion. Moreover, Angeles Esteban (2012) asserts that aquatic life, such as fish gets covered with an innate immunological subsystem. This subsystem offers the primary defense line in epidermal mucus form consisting of components of immune through goblet cells secretion. The three types of fish species are the leading Indian Carps (IMCs) which are essential for aquaculture and mostly used in polyculture. According to Mukherjee et al. (2016), all three species have distinct food habitats, thereby making it easier to get grown together. Subsequently, the difference in food habitats among the three species presents an ideal discovery of different mucus components, hence difference in action against pathogenic bacteria. For example, Catla catla is a surface feeder, Labeo rohita get adapted as a mid-water feeder, and Cirrinus mrigala thrives best as a bottom feeder. 

Recent related studies have also indicated that skin mucus of fish plays essential roles in fish immune defense. For example, according to Lirio, De Leon, and Villafuerte (2019), fish skin mucus lies at the interface between aqueous surrounding and fish. It is a vital component of the innate immune system of fish that fights pathogens immediately they get in contact with fish skin. Angeles Esteban (2012) established that the skin mucus of fish is continuously replaced and secreted. Thus, the microbes and other substances get washed when entrapped, this aid in the prevention of the stable colonization of probable infectious bacteria. Alvarez-Pellitero (2008), reported that mucus act as a storeroom for components of innate immune such as proteolytic enzymes, pentraxins, alkaline phosphatase, and immunoglobulin. The parts offer strong antimicrobial point to skin mucus.

Al-Arifa, Batool, and Hanif (2013) asserted that the fish skin mucus layer consists of a complex fluid with a varying composition throughout the fish epithelial surface. However, extensive evidence on the epidermal mucus antimicrobial activity in freshwaters of various habitats and habits still lacks. Every fish species possess its habit and habitat, hence, surviving in the different aquatic surrounding, consumes different food types. This distinct adaptation of each species may influence the quantity of secreted mucus and its components between or within species and can be significant in offering different immune parts and response.

           The use of antibiotics to treat and prevent dynamic illness caused by the microbes in the aquaculture has intensified over the years. However, its effectiveness is compromised. This nature gets primarily attributed to the increased resistance of bacteria. Therefore uncontrolled use of the antimicrobial agents has immensely contributed to antimicrobial resistance of the fish microbes. Spread of the opposition from the aquatic environment to the terrestrial leads to massive treatment failure and more diseases.

 Bacterial infections proliferation among fishes can, therefore, get acquired unintentionally by a human being through ingestion (Mukherjee et al., 2016). The Indians occasionally experience emerging outbreaks of bacterial infections, for example, salmonellosis and typhoid. Hence, resulting in individuals becoming victims of nosocomial diseases contracted from medical equipment and health facilities. Other consequences of the disorders may include a prolonged usage of antibiotics, increased hospital bills, as well as economic losses. According to Mukherjee et al. (2016), drug resistance pervasiveness is currently a threat to the efficacy of the antibiotics used in the treatment of infections. Increased antibiotics dosage functions faster on pathogenic viruses, thus, leasing to an increased risk for neurotoxicity, hepatoxicity plus, several adverse effects to humans.

As a way out, the development and discovery of new-fangled alternatives for available antibiotics are presently commercially promoted worldwide. New options of drugs from compounds extracted from animals, plants, and micro-organisms get considered as promising products. Substances produced naturally by organisms are relatively reasonable, cost-efficient, and environmentally friendly and have less serious side effects as compared to the commercially present antibiotics (Dash, Samal, and Thatoi, 2014). As a result, natural products antibacterial activities of organisms, particularly of wide-range of fish species has drawn interests as a promising source of compounds of antibiotics against fish and humans pathogens.

            India currently is the second-largest fish producer worldwide consisting of 6.3 percent of the total world fish production. It is a vital production mainly in its coastal states, making employment to over 14 million individuals. For example, during the 2017and 2018 season, the total output of fish got projected to be over 12.61 million metric tons and, inland fishing constituting 65 percent. On the other hand, culture fisheries represented about 50 percent of the total production (Jayanthi et al., 2018). Therefore, there might be a potency of antibiotic properties in the diverse fish species produced in India aquatic ecosystem.

           The primary goal of this research was to contribute to the general awareness of the search for novel antimicrobials. Therefore, this research explored extensively on an assessment of the potency of three major Indian fish of indigenous origin. The study examined the dominant skin mucus of Carp species, including Catla catla, Labeo rohita, and Cirrinus mrigala as an effective antimicrobial against human and fish pathogenic bacteria. The study particularly sought to test the effects of antimicrobial of the mucous against human and fish pathogenic bacteria to expound on possible agents in the mucus of fish for drug discovery.

This study presents the variation in fish mucus secretion by the three Indian carp species concerning their habits and habitats. The objectives of the study were, therefore, guide in the evaluation of the effectiveness of fish skin mucus that could constitute compounds of antibacterial. Hence, the findings could get utilized as an alternative to antibiotics that can get employed in both aquaculture and for humans. The study, therefore, presents the following research objectives; to evaluate the effect of antibacterial of skin mucus among the three IMCs viz. Catla catla, Labeo rohita, and Cirrinus mrigala, secondly, to determine any changes in secretion of mucus and its antibacterial influence after bacterial challenge, and to examine both challenged and healthy fish skin mucus among the three species under study and assess the MICs against all chosen microbial strain for investigations.          

Materials and Methods

Fish collection

Experimental fish constituted healthy yearlings from the three species which were obtained from the fish farms and transported to the Department of Zoology, University of Kurukshetra. Hygienic environment and the water quality were observed to ensure sustainability.

The daily recording of the fish health got conducted, and observations on dead fish noted.

Maintenance

Maintenance of fish involved checking of the health of fish, the quality of water as well as feeding the fish with proper food during acclimation.

Collection of Mucus: 

The fish species then acclimated for seven days. Mucus was then collected at regular intervals by careful scraping from the dorsal body surface using a sterile spatula. Mucus collection in the ventral area was avoided to eliminate urogenital and intestinal contamination (Al-Arifa, Batool, and Hanif, 2013). After that, the samples of mucus got frozen in ice at 0 degrees Celsius. This low temperature prevents bacterial growth (Al-Arifa, Batool, and Hanif, 2013).

 

 

 

Test of Micro-organisms

Both gram-negative and positive bacterial strains included human pathogens such as Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, which are gram-negative. The gram-positive strains of bacteria consisted of Bacillus cereus, Staphylococcus epidermidis, Staphylococcus aureus plus one Gram-negative bacterial strain of the fish pathogen, Aeromonas hydrophila. The bacterial strains got procured from a recognized institution of microbial technology in India, Institute of Microbial Technology (IMTECH), Chandigarh. The microbial strains were cultured regarding microbiological safety conditions and procedures (Eder et al., 2009).  The growth of all microbial strains took place at 37°C in nutrient broth. Afterward, CFU of every strain of bacteria got calculated by the method of colony-forming units (CFU).

Challenge Test

Ten fishes from every species then challenged with pathogenic bacteria of fish using A. Hydrophila maintained and cultured in the media of agar. For the first immersion, the mucus got collected on third and seventh days in a suspension of approximated107 CFU ml-1, and significant changes recorded regarding the amount of mucus secreted both by challenged and healthy fish. After that, second immersion performed by immersion of about109 CFU ml-1 for fifteen days and mucus collection conducted after a one-week interval through the method of a skin scraping. 

 

Preparations of Mucus Extracts

Extract of thick mucus prepared for both challenged and healthy fish species through centrifuging raw mucus at a revolution of 5000 per minute for 300 seconds. On the other hand, aqueous mucus, a similar amount of sterilized physiological saline was mixed with thick mucus and centrifuged for the same revolutions and time. The levels of protein in mucus got determined by the use of Lowry et al. (1951) for different fish species.  

Agar Well Diffusion Assay  

Agar well diffusion method was used to assay extracts of every tested fish species on the chosen bacterial strains of both HFM and CFM. Seeding involved 15ml nutrient agar medium contained in Petri-plates with a one day culture of averagely 107 CFU ml-1   strains of bacteria.  The diameter of six to seven (6 to 7) mm of well was then pierced aseptically with a cork borer. The cork borer got sterilized at first, and extracts of antibiotics and mucus added. Amikacin and chloramphenicol (positive extracts) got used for extracts of aqueous and crude in a concentration of 20µgml-1 and 40µgml-1, respectively. Incubation period took one day (24 hours) at a temperature of 37°C. The bacterial effect evaluation got measured by using the ZOI diameter (in mm) established around the well (Eder et al., 2015). The results of the antibacterial activity got compared with the two standard positive control drugs. The negative control comprised of 0.85% NaCl, which got applied in the assay of aqueous mucus antibacterial.

Agar Dilutions Assay 

The CFM and MIC of HFM of each species of fish got determined using agar plate dilution assay against every chosen microbial strain. All skin mucus of fish made by dilution of the mucous extract in sterile distilled water. The extract of desired dilutions of fish mucus got prepared and transferred in various wells. The different fish MICs get determined by the measurement of ZOIs to varying levels against different bacteria.   

 

 

 

Data Analysis

All the samples were analyzed in triplicate using statistical analysis performed by T-test as well as one-way ANOVA at significant level 0.05. The test used SPSS Version 11.5 for Windows among all the selected species and antibiotics. Besides, the student's test was used to evaluate the considerable difference between ZOIs gotten from extracts of CFM and HFM of each selected species of fish against various pathogenic strains performed under research. Finally, Statistical significance got settled at P\0.05 probability value. 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

References

Al-Arifa, N., Batool, A., & Hanif, A. (2013). THE JOURNAL OF ANIMAL & PLANT SCIENCES-Effects of Alkaline PH on Protein and Fatty Acid Profiles of epidermal Mucus From Labeo rohita. Retrieved August 3, 2019, from http://www.thejaps.org.pk/docs/v-23-4/16.pdf

Alvarez-Pellitero, P. (2008). Fish immunity and parasite infections: From innate immunity to immunoprophylactic prospects. Veterinarian Immunology and Immunopathology. Retrieved from https://www.researchgate.net/publication/23249497_Alvarez-Pellitero_P_Fish_immunity_and_parasite_infections_From_innate_immunity_to_immunoprophylactic_prospects_Veterinarian_Immunology_and_Immunopathology

Ángeles Esteban, M. (2012). An overview of the immunological defences in fish skin. ISRN Immunology, 2012.

Balasubramanian, S., Baby Rani, P., Arul Prakash, A., & Prakash, M. (2012). (PDF) Antimicrobial properties of skin mucus from four freshwater cultivable Fishes (Catla Catla, Hypophthalmichthys molitrix, Labeo rohita and Ctenopharyngodon idella). Retrieved from https://www.researchgate.net/publication/268509222_Antimicrobial_properties_of_skin_mucus_from_four_freshwater_cultivable_Fishes_Catla_catla_Hypophthalmichthys_molitrix_Labeo_rohita_and_Ctenopharyngodon_idella

Dash, S., Samal, J., & Thatoi, H. (2014). A comparative study on innate immunity parameters in the epidermal mucus of Indian major carps. Aquaculture international, 22(2), 411-421.

Eder, A. F., Kennedy, J. M., Dy, B. A., Notari, E. P., Skeate, R., Bachowski, G., ... & Benjamin, R. J. (2009). Limiting and detecting bacterial contamination of apheresis platelets: inlet‐line diversion and increased culture volume improve component safety. Transfusion, 49(8), 1554-1563.

Jayanthi, M., Thirumurthy, S., Muralidhar, M., & Ravichandran, P. (2018). Impact of shrimp aquaculture development on important ecosystems in India. Global environmental change, 52, 10-21.

Lirio, G., De Leon, J., & Villafuerte, A. (2019). Antimicrobial Activity of Epidermal Mucus from Top Aquaculture Fish Species against Medically-Important Pathogens | LIRIO. Retrieved August 3, 2019, from https://wjst.wu.ac.th/index.php/wjst/article/view/6287

Mukherjee, A., Dutta, D., Banerjee, S., Ringø, E., Breines, E. M., Hareide, E., ... & Ghosh, K. (2016). Potential probiotics from Indian major carp, Cirrhinus mrigala. Characterization, pathogen inhibitory activity, partial characterization of bacteriocin and production of exoenzymes. Research in veterinary science, 108, 76-84.

Nigam, A. K., Kumari, U., Mittal, S., & Mittal, A. K. (2017). Evaluation of antibacterial activity and innate immune components in skin mucus of Indian major carp, Cirrhinus mrigala. Aquaculture Research, 48(2), 407-418.

Music in a film

 

 

 

 

 

 

Music in films

Name

Institution

 

Music in films

Music is employed for making the ideal environment in a specific film scene by setting the correct tone and temperament. The way the music is portrayed in the movie, one can appropriately get the degree of show or air or the state of mind expected of such a film or scene. Music got a respectful significance in films and is very respected for controlling the feelings audience has and it helps in inundating into the story. Music is amongst the prime components in the film. Its absence makes the film sound not exciting and dull. There exist various components in a film: the first is acting, followed by an image, and lastly the music. It is a sacred trinity; if deficient, there occurs lack of fervor and sensation. Combination of picture and acting are independent from each other, yet the music entirely triggers the film to be vital. There are a large number of movies musicians worldwide in the music business, a rare sort of people who triumph and whose work is outstanding to practically a half.

Whenever the actor puts activity music in an exciting scene; by rationale, it would not be right. The group of spectators may miss feeling the punch of the stage. That is the reason there ought to be a feeling connected to the song with the goal that this music arrives at the most significant sensations. While the well-made sound structure and audio cues are so fundamental in the film making, music contacts the feeling, the mind, and the aspects you cannot see. Sound plan and impacts do not do this so proficiently and frequently. In absence of music, it could be substantially more hard to pursue the passionate good and difficulties of a film. Clearly, this clarifies the noteworthiness of the music and the jobs within the films. Music needs to coordinate the feelings that the entertainers are appearing and the nature in it. In case it neglects to do as such, the film may be a repulsive thing to watch. An additional feature of motion picture music is leitmotifs. "Wagner's main specialized advancement was the improvement of the leitmotif framework where short, unmistakable melodic expressions become related with account and topical thoughts and grow symphonically to reflect the advancement of the show". Composers utilize this to distinguish characters, situations, feelings or thoughts. One case of that could be the score for Lord of the Rings. Each character has a focal melodic topic. Amongst the unmistakable could be the leitmotifs subjects of the character Gollum.

Moreover, among the delights of having a group of individuals' part in a cinema is the capacity of having an encounter outside the movie. The responses of the audience increases through an assortment of ways — regardless of whether stun, pity or delight — yet maybe among the surefire approaches to accomplishing some reaction exists in a straightforward part of pretty much involvement of every audience.

With the previous end of the week's chief and film industry achievement of Guardians of the Galaxy Vol. 2, audiences not just demonstrate that Marvel is still on a series of wins in the hero advertise however that innovation rewards movie producers with fans' love (Gunn et al., 2017). The continuation additionally got much acclaim for its soundtrack and unique score, even though the soundtrack has been a friendly exchange amongst different news sources and fans, for the most part on account of its retro choices.

"In the middle of the thuddingly evident music prompts of Suicide Squad, and the Sturm und Drang scores of most other comic-book flicks," composed the editors of Vulture, "the Guardians movies have cut out a quite certain melodic specialty, utilizing various affectionately (however not over-) recalled radio works of art from the 1970s."

Vulture's utilization of "niche" is actually what Guardians of the Galaxy Vol. 2 figured out how to fit into. Director James Gunn's utilization of Fleetwood Mac's "The Chain" during a scene when Peter, Gamora and Drax fly away on Ego's spaceship isn't just suitable yet the ideal melody decision.

Nevertheless, that cannot be a new event, as a large number of clique works of art have utilized outstanding arranged soundtracks that emitted elegant vibes tuned to the tone of a specific moment.

Quentin Tarantino is a prime case of a movie producer who consolidates unique scores with soundtracks based on his personal preference. It is tough to overlook the bone-chilling yet hypnotizing tune that Elle Driver whistles as she advances toward a debilitated Beatrix's medical clinic bed to kill her in Kill Bill Vol. 1 (Bill, 2003), or when Vince and Mia do the curve to Chuck Berry's "You Never Can Tell" in Pulp Fiction.

Producers have additionally begun coordinating orchestrated scores that have fiercely compared soundtracks, for example, director Barry Jenkins his awesome Academy Award-winning Moonlight, where symphonic violin strings were heard in a single scene and afterwards a cleaved and-screwed variant of Jidenna's "Classic Man" could be heard in the following. While "Classic Man" is amongst the most peculiar melody decisions ever heard in the movie despite everything, it figures out how to fit pleasantly through the scene it's utilized.

However, exemplary film movies, for example, Star Wars, Jurassic Park and The Lord of the Rings never utilize soundtracks, quitting for the absolute famous movie themes always seen in movies. There exist exceptional cases to the soundtrack, yet that demonstrates how many parts of experience music can be. Never shall there be uncertainty that without writer John Williams, pivotal moments within Star Wars —, for example, Luke Skywalker's enthusiastic gaze at the binary sunset of the desert world Tatooine — could not have been as significant or wholly figured it out.

Music, at large, dictates the feelings audiences feel through what might be the most exceptional significant snapshots of the movie. It contributes to the life of the story, and it can at last break or make the experience of the audience.

 

References

Gunn, J., Bates, T., & Feige, K. (2017). Guardians of the Galaxy: Vol. 2. Blackstone.

Bill, K. (2003). Vol. 1. Directed by Quentin Tarantino. USA.

Media analysis essay

 

 

 

 

 

 

Music in films

Name

Institution

 

Music in films

Music is employed for making the ideal environment in a specific film scene by setting the correct tone and temperament. The way the music is portrayed in the movie, one can appropriately get the degree of show or air or the state of mind expected of such a film or scene. Music got a respectful significance in films and is very respected for controlling the feelings audience has and it helps in inundating into the story. Music is amongst the prime components in the film. Its absence makes the film sound not exciting and dull. There exist various components in a film: the first is acting, followed by an image, and lastly the music. It is a sacred trinity; if deficient, there occurs lack of fervor and sensation. Combination of picture and acting are independent from each other, yet the music entirely triggers the film to be vital. There are a large number of movies musicians worldwide in the music business, a rare sort of people who triumph and whose work is outstanding to practically a half.

Whenever the actor puts activity music in an exciting scene; by rationale, it would not be right. The group of spectators may miss feeling the punch of the stage. That is the reason there ought to be a feeling connected to the song with the goal that this music arrives at the most significant sensations. While the well-made sound structure and audio cues are so fundamental in the film making, music contacts the feeling, the mind, and the aspects you cannot see. Sound plan and impacts do not do this so proficiently and frequently. In absence of music, it could be substantially more hard to pursue the passionate good and difficulties of a film. Clearly, this clarifies the noteworthiness of the music and the jobs within the films. Music needs to coordinate the feelings that the entertainers are appearing and the nature in it. In case it neglects to do as such, the film may be a repulsive thing to watch. An additional feature of motion picture music is leitmotifs. "Wagner's main specialized advancement was the improvement of the leitmotif framework where short, unmistakable melodic expressions become related with account and topical thoughts and grow symphonically to reflect the advancement of the show". Composers utilize this to distinguish characters, situations, feelings or thoughts. One case of that could be the score for Lord of the Rings. Each character has a focal melodic topic. Amongst the unmistakable could be the leitmotifs subjects of the character Gollum.

Moreover, among the delights of having a group of individuals' part in a cinema is the capacity of having an encounter outside the movie. The responses of the audience increases through an assortment of ways — regardless of whether stun, pity or delight — yet maybe among the surefire approaches to accomplishing some reaction exists in a straightforward part of pretty much involvement of every audience.

With the previous end of the week's chief and film industry achievement of Guardians of the Galaxy Vol. 2, audiences not just demonstrate that Marvel is still on a series of wins in the hero advertise however that innovation rewards movie producers with fans' love (Gunn et al., 2017). The continuation additionally got much acclaim for its soundtrack and unique score, even though the soundtrack has been a friendly exchange amongst different news sources and fans, for the most part on account of its retro choices.

"In the middle of the thuddingly evident music prompts of Suicide Squad, and the Sturm und Drang scores of most other comic-book flicks," composed the editors of Vulture, "the Guardians movies have cut out a quite certain melodic specialty, utilizing various affectionately (however not over-) recalled radio works of art from the 1970s."

Vulture's utilization of "niche" is actually what Guardians of the Galaxy Vol. 2 figured out how to fit into. Director James Gunn's utilization of Fleetwood Mac's "The Chain" during a scene when Peter, Gamora and Drax fly away on Ego's spaceship isn't just suitable yet the ideal melody decision.

Nevertheless, that cannot be a new event, as a large number of clique works of art have utilized outstanding arranged soundtracks that emitted elegant vibes tuned to the tone of a specific moment.

Quentin Tarantino is a prime case of a movie producer who consolidates unique scores with soundtracks based on his personal preference. It is tough to overlook the bone-chilling yet hypnotizing tune that Elle Driver whistles as she advances toward a debilitated Beatrix's medical clinic bed to kill her in Kill Bill Vol. 1 (Bill, 2003), or when Vince and Mia do the curve to Chuck Berry's "You Never Can Tell" in Pulp Fiction.

Producers have additionally begun coordinating orchestrated scores that have fiercely compared soundtracks, for example, director Barry Jenkins his awesome Academy Award-winning Moonlight, where symphonic violin strings were heard in a single scene and afterwards a cleaved and-screwed variant of Jidenna's "Classic Man" could be heard in the following. While "Classic Man" is amongst the most peculiar melody decisions ever heard in the movie despite everything, it figures out how to fit pleasantly through the scene it's utilized.

However, exemplary film movies, for example, Star Wars, Jurassic Park and The Lord of the Rings never utilize soundtracks, quitting for the absolute famous movie themes always seen in movies. There exist exceptional cases to the soundtrack, yet that demonstrates how many parts of experience music can be. Never shall there be uncertainty that without writer John Williams, pivotal moments within Star Wars —, for example, Luke Skywalker's enthusiastic gaze at the binary sunset of the desert world Tatooine — could not have been as significant or wholly figured it out.

Music, at large, dictates the feelings audiences feel through what might be the most exceptional significant snapshots of the movie. It contributes to the life of the story, and it can at last break or make the experience of the audience.

 

References

Gunn, J., Bates, T., & Feige, K. (2017). Guardians of the Galaxy: Vol. 2. Blackstone.

Bill, K. (2003). Vol. 1. Directed by Quentin Tarantino. USA.

INDIAN CARP FISH SPECIES

 

 

 

 

 

 

 

 

 

 

       Evaluation of Antimicrobial Potency on Skin Mucus of Three Major Indian Carp Species

Student's Name

Institutional Affiliation

 

 

 

 

 

 

 

 

 

 

ACKNOWLEDGEMENT

I   thank the Almighty God for his guidance and protection throughout this study. I extend my gratitude to my family and friends for the financial and moral support. I would also like to acknowledge my supervisor for the guidance and support throughout the study.

 

 

 

 

 

  

 

 

 

 

 

 

 

 

 

 

 

 

 

ABSTRACT

Zoonotic infections resulted from bacterial pathogens threatens both humans and the aquaculture industry. This outbreak probed search for different epidermal mucus as natural products from three Indian Major Carp species to evaluate the potency of the fish skin mucus as an effective antimicrobial against human and fish pathogenic bacteria. The skin mucus of the three indigenous Indian Major Carp species got collected against pathogenic fish bacteria and human viz. Catla catla, Labeo rohita, and Cirrinus mrigala. The activities of the microbes were measured based on Zone of Inhibition (ZOI) in millimeters to assess the MICs of various mucus extracts against a variety of bacteria. The results were compared with standard antibiotics amikacin and chloramphenicol to determine fish skin mucus antimicrobial potency.

Moreover, the three selected fish species were also challenged with A. Hydrophila to evaluate the presence of any change in their mucus and antibacterial influence after exposure of bacterial. Afterward, the agar plate dilution test was used to analyze the minimum inhibitory concentration of HFM and CFM from every fish species against the selected microbial strains. Therefore the results of the study were useful in identifying the possibility of using the fish skin mucus as an alternative to antibiotics in fish and human health associated complications.

 

Keywords: Zone of Inhibition (ZOI), fish, antimicrobial activity, skin mucus, major Indian carps (IMCs), Healthy Fish Mucus (HFM), Challenged Fish Mucus (CFM), Colony Forming Units (CFU)

 

                  

    

 

Evaluation of antimicrobial potency on Skin Mucus of Three Major Indian Carp Species

INTRODUCTION

Fishes play a significant role in the aquatic environment. Their diversity forms the largest class of water habitat, especially cold-blooded vertebrate species.  They provide interconnection between food webs of the water ecosystems, which harbors a variety of pathogenic micro-organisms (Ellis 2001). Moreover, According to Alvarez-Pellitero (2008), infectious diseases are widely spread when fish get reared in intensive and high densities aquaculture. Increased population density and limited resources provide a favorable environment for the thriving of pathogens.

Aquatic organisms like fish get stressed in a crowded and unnatural environment characterized by a limited flow of water hence making the fish family more susceptible to diseases. Consequently, through the consumption of fish, the transmission of the bacterial pathogens from infected fish poses a severe danger to the aquaculture industry, and human health. Increased density of bacteria, viruses, and parasites are, therefore, critical for the development of aquaculture (Cooper and Shlaes 2011). Infectious diseases cause complications such as edwardsiellosis, columnaris, and microbial hemorrhagic ascites in fish. Interestingly, fishes pose mucus substances on their surfaces, which aides them in locomotion, osmoregulation, body lubrication that wades off pathogens and also immunological functions.

            According to Lirio et al. (2018), the mucus layer defends the fish against the microbe invasion. Moreover, Angeles Esteban (2012) asserts that aquatic life, such as fish gets covered with an innate immunological subsystem. This subsystem offers the primary defense line in epidermal mucus form consisting of components of immune through goblet cells secretion. The three types of fish species are the leading Indian Carps (IMCs) which are essential for aquaculture and mostly used in polyculture. According to Mukherjee et al. (2016), all three species have distinct food habitats, thereby making it easier to get grown together. Subsequently, the difference in food habitats among the three species presents an ideal discovery of different mucus components, hence difference in action against pathogenic bacteria. For example, Catla catla is a surface feeder, Labeo rohita get adapted as a mid-water feeder, and Cirrinus mrigala thrives best as a bottom feeder. 

Recent related studies have also indicated that skin mucus of fish plays essential roles in fish immune defense. For example, according to Lirio, De Leon, and Villafuerte (2019), fish skin mucus lies at the interface between aqueous surrounding and fish. It is a vital component of the innate immune system of fish that fights pathogens immediately they get in contact with fish skin. Angeles Esteban (2012) established that the skin mucus of fish is continuously replaced and secreted. Thus, the microbes and other substances get washed when entrapped, this aid in the prevention of the stable colonization of probable infectious bacteria. Alvarez-Pellitero (2008), reported that mucus act as a storeroom for components of innate immune such as proteolytic enzymes, pentraxins, alkaline phosphatase, and immunoglobulin. The parts offer strong antimicrobial point to skin mucus.

Al-Arifa, Batool, and Hanif (2013) asserted that the fish skin mucus layer consists of a complex fluid with a varying composition throughout the fish epithelial surface. However, extensive evidence on the epidermal mucus antimicrobial activity in freshwaters of various habitats and habits still lacks. Every fish species possess its habit and habitat, hence, surviving in the different aquatic surrounding, consumes different food types. This distinct adaptation of each species may influence the quantity of secreted mucus and its components between or within species and can be significant in offering different immune parts and response.

           The use of antibiotics to treat and prevent dynamic illness caused by the microbes in the aquaculture has intensified over the years. However, its effectiveness is compromised. This nature gets primarily attributed to the increased resistance of bacteria. Therefore uncontrolled use of the antimicrobial agents has immensely contributed to antimicrobial resistance of the fish microbes. Spread of the opposition from the aquatic environment to the terrestrial leads to massive treatment failure and more diseases.

 Bacterial infections proliferation among fishes can, therefore, get acquired unintentionally by a human being through ingestion (Mukherjee et al., 2016). The Indians occasionally experience emerging outbreaks of bacterial infections, for example, salmonellosis and typhoid. Hence, resulting in individuals becoming victims of nosocomial diseases contracted from medical equipment and health facilities. Other consequences of the disorders may include a prolonged usage of antibiotics, increased hospital bills, as well as economic losses. According to Mukherjee et al. (2016), drug resistance pervasiveness is currently a threat to the efficacy of the antibiotics used in the treatment of infections. Increased antibiotics dosage functions faster on pathogenic viruses, thus, leasing to an increased risk for neurotoxicity, hepatoxicity plus, several adverse effects to humans.

As a way out, the development and discovery of new-fangled alternatives for available antibiotics are presently commercially promoted worldwide. New options of drugs from compounds extracted from animals, plants, and micro-organisms get considered as promising products. Substances produced naturally by organisms are relatively reasonable, cost-efficient, and environmentally friendly and have less serious side effects as compared to the commercially present antibiotics (Dash, Samal, and Thatoi, 2014). As a result, natural products antibacterial activities of organisms, particularly of wide-range of fish species has drawn interests as a promising source of compounds of antibiotics against fish and humans pathogens.

            India currently is the second-largest fish producer worldwide consisting of 6.3 percent of the total world fish production. It is a vital production mainly in its coastal states, making employment to over 14 million individuals. For example, during the 2017and 2018 season, the total output of fish got projected to be over 12.61 million metric tons and, inland fishing constituting 65 percent. On the other hand, culture fisheries represented about 50 percent of the total production (Jayanthi et al., 2018). Therefore, there might be a potency of antibiotic properties in the diverse fish species produced in India aquatic ecosystem.

           The primary goal of this research was to contribute to the general awareness of the search for novel antimicrobials. Therefore, this research explored extensively on an assessment of the potency of three major Indian fish of indigenous origin. The study examined the dominant skin mucus of Carp species, including Catla catla, Labeo rohita, and Cirrinus mrigala as an effective antimicrobial against human and fish pathogenic bacteria. The study particularly sought to test the effects of antimicrobial of the mucous against human and fish pathogenic bacteria to expound on possible agents in the mucus of fish for drug discovery.

This study presents the variation in fish mucus secretion by the three Indian carp species concerning their habits and habitats. The objectives of the study were, therefore, guide in the evaluation of the effectiveness of fish skin mucus that could constitute compounds of antibacterial. Hence, the findings could get utilized as an alternative to antibiotics that can get employed in both aquaculture and for humans. The study, therefore, presents the following research objectives; to evaluate the effect of antibacterial of skin mucus among the three IMCs viz. Catla catla, Labeo rohita, and Cirrinus mrigala, secondly, to determine any changes in secretion of mucus and its antibacterial influence after bacterial challenge, and to examine both challenged and healthy fish skin mucus among the three species under study and assess the MICs against all chosen microbial strain for investigations.          

Materials and Methods

Fish collection

Experimental fish constituted healthy yearlings from the three species which were obtained from the fish farms and transported to the Department of Zoology, University of Kurukshetra. Hygienic environment and the water quality were observed to ensure sustainability.

The daily recording of the fish health got conducted, and observations on dead fish noted.

Maintenance

Maintenance of fish involved checking of the health of fish, the quality of water as well as feeding the fish with proper food during acclimation.

Collection of Mucus: 

The fish species then acclimated for seven days. Mucus was then collected at regular intervals by careful scraping from the dorsal body surface using a sterile spatula. Mucus collection in the ventral area was avoided to eliminate urogenital and intestinal contamination (Al-Arifa, Batool, and Hanif, 2013). After that, the samples of mucus got frozen in ice at 0 degrees Celsius. This low temperature prevents bacterial growth (Al-Arifa, Batool, and Hanif, 2013).

 

 

 

Test of Micro-organisms

Both gram-negative and positive bacterial strains included human pathogens such as Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, which are gram-negative. The gram-positive strains of bacteria consisted of Bacillus cereus, Staphylococcus epidermidis, Staphylococcus aureus plus one Gram-negative bacterial strain of the fish pathogen, Aeromonas hydrophila. The bacterial strains got procured from a recognized institution of microbial technology in India, Institute of Microbial Technology (IMTECH), Chandigarh. The microbial strains were cultured regarding microbiological safety conditions and procedures (Eder et al., 2009).  The growth of all microbial strains took place at 37°C in nutrient broth. Afterward, CFU of every strain of bacteria got calculated by the method of colony-forming units (CFU).

Challenge Test

Ten fishes from every species then challenged with pathogenic bacteria of fish using A. Hydrophila maintained and cultured in the media of agar. For the first immersion, the mucus got collected on third and seventh days in a suspension of approximated107 CFU ml-1, and significant changes recorded regarding the amount of mucus secreted both by challenged and healthy fish. After that, second immersion performed by immersion of about109 CFU ml-1 for fifteen days and mucus collection conducted after a one-week interval through the method of a skin scraping. 

 

Preparations of Mucus Extracts

Extract of thick mucus prepared for both challenged and healthy fish species through centrifuging raw mucus at a revolution of 5000 per minute for 300 seconds. On the other hand, aqueous mucus, a similar amount of sterilized physiological saline was mixed with thick mucus and centrifuged for the same revolutions and time. The levels of protein in mucus got determined by the use of Lowry et al. (1951) for different fish species.  

Agar Well Diffusion Assay  

Agar well diffusion method was used to assay extracts of every tested fish species on the chosen bacterial strains of both HFM and CFM. Seeding involved 15ml nutrient agar medium contained in Petri-plates with a one day culture of averagely 107 CFU ml-1   strains of bacteria.  The diameter of six to seven (6 to 7) mm of well was then pierced aseptically with a cork borer. The cork borer got sterilized at first, and extracts of antibiotics and mucus added. Amikacin and chloramphenicol (positive extracts) got used for extracts of aqueous and crude in a concentration of 20µgml-1 and 40µgml-1, respectively. Incubation period took one day (24 hours) at a temperature of 37°C. The bacterial effect evaluation got measured by using the ZOI diameter (in mm) established around the well (Eder et al., 2015). The results of the antibacterial activity got compared with the two standard positive control drugs. The negative control comprised of 0.85% NaCl, which got applied in the assay of aqueous mucus antibacterial.

Agar Dilutions Assay 

The CFM and MIC of HFM of each species of fish got determined using agar plate dilution assay against every chosen microbial strain. All skin mucus of fish made by dilution of the mucous extract in sterile distilled water. The extract of desired dilutions of fish mucus got prepared and transferred in various wells. The different fish MICs get determined by the measurement of ZOIs to varying levels against different bacteria.   

 

 

 

Data Analysis

All the samples were analyzed in triplicate using statistical analysis performed by T-test as well as one-way ANOVA at significant level 0.05. The test used SPSS Version 11.5 for Windows among all the selected species and antibiotics. Besides, the student's test was used to evaluate the considerable difference between ZOIs gotten from extracts of CFM and HFM of each selected species of fish against various pathogenic strains performed under research. Finally, Statistical significance got settled at P\0.05 probability value. 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

References

Al-Arifa, N., Batool, A., & Hanif, A. (2013). THE JOURNAL OF ANIMAL & PLANT SCIENCES-Effects of Alkaline PH on Protein and Fatty Acid Profiles of epidermal Mucus From Labeo rohita. Retrieved August 3, 2019, from http://www.thejaps.org.pk/docs/v-23-4/16.pdf

Alvarez-Pellitero, P. (2008). Fish immunity and parasite infections: From innate immunity to immunoprophylactic prospects. Veterinarian Immunology and Immunopathology. Retrieved from https://www.researchgate.net/publication/23249497_Alvarez-Pellitero_P_Fish_immunity_and_parasite_infections_From_innate_immunity_to_immunoprophylactic_prospects_Veterinarian_Immunology_and_Immunopathology

Ángeles Esteban, M. (2012). An overview of the immunological defences in fish skin. ISRN Immunology, 2012.

Balasubramanian, S., Baby Rani, P., Arul Prakash, A., & Prakash, M. (2012). (PDF) Antimicrobial properties of skin mucus from four freshwater cultivable Fishes (Catla Catla, Hypophthalmichthys molitrix, Labeo rohita and Ctenopharyngodon idella). Retrieved from https://www.researchgate.net/publication/268509222_Antimicrobial_properties_of_skin_mucus_from_four_freshwater_cultivable_Fishes_Catla_catla_Hypophthalmichthys_molitrix_Labeo_rohita_and_Ctenopharyngodon_idella

Dash, S., Samal, J., & Thatoi, H. (2014). A comparative study on innate immunity parameters in the epidermal mucus of Indian major carps. Aquaculture international, 22(2), 411-421.

Eder, A. F., Kennedy, J. M., Dy, B. A., Notari, E. P., Skeate, R., Bachowski, G., ... & Benjamin, R. J. (2009). Limiting and detecting bacterial contamination of apheresis platelets: inlet‐line diversion and increased culture volume improve component safety. Transfusion, 49(8), 1554-1563.

Jayanthi, M., Thirumurthy, S., Muralidhar, M., & Ravichandran, P. (2018). Impact of shrimp aquaculture development on important ecosystems in India. Global environmental change, 52, 10-21.

Lirio, G., De Leon, J., & Villafuerte, A. (2019). Antimicrobial Activity of Epidermal Mucus from Top Aquaculture Fish Species against Medically-Important Pathogens | LIRIO. Retrieved August 3, 2019, from https://wjst.wu.ac.th/index.php/wjst/article/view/6287

Mukherjee, A., Dutta, D., Banerjee, S., Ringø, E., Breines, E. M., Hareide, E., ... & Ghosh, K. (2016). Potential probiotics from Indian major carp, Cirrhinus mrigala. Characterization, pathogen inhibitory activity, partial characterization of bacteriocin and production of exoenzymes. Research in veterinary science, 108, 76-84.

Nigam, A. K., Kumari, U., Mittal, S., & Mittal, A. K. (2017). Evaluation of antibacterial activity and innate immune components in skin mucus of Indian major carp, Cirrhinus mrigala. Aquaculture Research, 48(2), 407-418.

MORAL DILEMMA

 

 

 

 

 

 

MORAL DILEMMA

 

Student's Name

Subject

Date

 

 

 

 

 

 

 

 

 

When a situation arises within the organization, the manager is responsible for undertaking appropriate action to resolve it. In this case, the problem is the ship is not following the ordinary course as targeted. If this is a police case, then the manager should not worry because the vessel is under the right hand. If the business is legal, then it means that the patrol officers will release it and the ship will return to its ordinary course[1]. However, if the vessel attacked, the manager is supposed to follow the necessary protocols as set by the company to minimize or curb the effects of the attack. However, the manager does not know what to do because there is no reference to this case; he only received a phone call[2].

Considering the problem can be said to be a moral dilemma. Here, the individual faced with a question of decision-making between two probable ethical imperatives, and none of them is unambiguously preferable or acceptable. In this case, the commanding officer faces the problem where he has to decide upon two cases that he has no reference. Neither of these situations is unequivocally acceptable. If he decides to take measures against an attack, he may find out it was a police case. If he chooses to relax and let the police do their work, he risks the matter to be an attack. In an ethical dilemma, the complication comes in out of the situational conflict where one transgressed if the individual decides to follow the other. If the commanding officer takes the information provided by the first group of engineers, then he will be forced to send the police or other security officials that protect the well-being of the company to the sea. On the other hand, they will be free if the ship is under police officers.

For this moral dilemma, the best advice to the commanding officer to follow the correct protocol as set by the company to help in resolving this dilemma. Since he is not sure whether the ship is under the police or terrorists, then he should follow the correct standards that are set by the company in resolving such an issue. It recommended that the officer uses deontological theory of ethics. This theory believes that firms ought to follow some preset principles, no matter what their consequences are. As opposed to consequentialism, deontology does not consider the results of any action taken[3].  In this case, the commanding officer may opt to send security officials to attack the presumed attackers. He also has the option of letting the vessels alone if they belong to the police. A board meeting should immediately be organized to find a solution to the problem. Whether they are police or terrorists, the board needs to meet and come up with a defined course of action so that they may ensure the supply chain follows the right path. [4]The consequences of the activity undertaken might be functional, but the process may be against the conditions followed within the organization. Therefore, the best thing for the commanding officer to do before sending security officers into the sea is first receiving reliable information from relevant sources so that they may meet with the board and come up with the right course of action while following the appropriate procedures. If the effects considered, the commanding officer will do anything possible to ensure that the ship is protected so that the supplies reach the destination on time and satisfy the needs of the customer and maintain the company's right name. However, this may mean going against some codes of conduct, such as not following the outlined protocol while making the decision, only to save the company's reputation. However, the course of action is critical and what described within the code should be followed without exception. Thus, deontology is the right theory that should guide the correct course of action that the commanding officer will undertake[5].

Kant, a significant proponent of this theory, believed that a moral act follows standard ethical law, such as "do not lie." This theory requires that individuals should only follow preset conditions while carrying out their duties[6]. The approach does not require that an individual weighs between costs and benefits. It helps to avoid possible uncertainty and subjectivity since the individual will be needed to follow a set of rules.

Generally, leadership decision-making, alongside other features and roles that the leadership of the company has provided continuous creativity and crucial new insights into management and leadership activities in all the organizational undertakings. Therefore, any organizational leadership has a significant influence on how moral dilemmas solved. First, the command is responsible for recruiting and hiring new employees. Employees should be experts in their respective fields of study to ensure that they can follow the codes of ethics with ease and that they can deliver the best in case a case of moral dilemma needs resolution. Board members should be influential that they make the most rational decisions ever. Secondly, they educate and train workers on the meaning and importance of ethics and moral behaviors in all their undertakings. For example in the current case, it is unethical that members of the same team are providing conflicting reports of a similar situation. Thirdly, leadership in any organization is responsible for the way finances utilized[7].

 

Bibliography

 

 

Guevara, Daniel. Kant's theory of moral motivation. Routledge, 2019.

 

Nucci, Larry. "Recovering the role of reasoning in moral education to address inequity and social justice." Journal of Moral Education 45, no. 3 (2016): 291-307.

 

Pojman, Louis P., Katie McShane, and Paul Pojman. Environmental ethics: Readings in theory and application. Nelson Education, 2015.

 

Yoon, Cheolho. "Theory of planned behavior and ethics theory in digital piracy: An integrated model." Journal of business ethics 100, no. 3 (2011): 405-417.

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[1] CheolhoYoon, Theory of planned behavior and ethics theory in digital piracy, Journal of business ethics 100, no. 3 (2011): 407

[2] Louis P.Pojman,  McShane Katie, and Pojman Paul . Environmental ethics: (Nelson Education, 2015), 95

[3] Daniel Guevara, Kant's theory of moral motivation.( Routledge, 2019), 5.

[4] Larry Nucci, "Recovering the role of reasoning in moral education to address inequity and social justice." Journal of Moral Education 45, no. 3 (2016): 301.

[5] CheolhoYoon, Theory of planned behavior and ethics theory in digital piracy, Journal of business ethics 100, no. 3 (2011): 407

 

[7] Larry Nucci, "Recovering the role of reasoning in moral education to address inequity and social justice." Journal of Moral Education 45, no. 3 (2016): 301.

ux?

 It is what supports the internet and many scientific breakthrough open source and community-developed operating system for computers, servers, mainframes, mobile devices and embedded device l 

2. Describe the components of Linux operating system.

OS Kernel- it is the part of operating system which is situated near the computer hardware .It is the one that controls the central processing unit. It also controls the access to the memory and any other peripheral devices.

Graphics server- This is the one that provides a graphical sub-system which gives all the images and shapes found on the monitor of a computer.

Bootloader- This is the one which facilitates the process of starting of the a 

Desktop environment- This refers to the part that one to interact with the graphical server... There are various options of the desktop environment. They include the KDE, Unity and Cinnamon. It also includes other applications such as the files web browsers and games. And Cinnamon. 

Applications – This is the one that allows for installation of apes since the since the desktop environment can't cater for every application needed. 

OS Shell- it is also known as the command line. This is the one that that tells the operating system what the user what's to do.  

3. Important features of the Linux operating system.

Multi-user – This is an important feature of the Linux OS where by multiple users Are able to access the system resources such as the memory and to apply programs at the same time.

Multi-programming –This is a feature where by multiple application can run at the same time and the applications do not interrupt each other.

 Hierarchical File System- It is a   standard file provided by the Linux which allows the arranging of system and user files

Portable – This means that the software can work on different type's hardware in the same way. 

Security – Security is provided by allowing the user to use some characteristics such as passwords and controlled access to data and data encryption.

 Shell – This is the one that allows for interpretation of programs and send command to the operating system on what the user is intending to do...

 Open Source – This is a feature of the lune that its code is readily available bad it is based on the development of the community... 

4. Describe the architecture of the Linux So?

The architecture of the lune operating system comprises of the hardware layer, the system library, the shell, the system utility and the kernel.

The hardware layer it is the part of the Linux operating system that consists of peripheral devices.

The system library refers to special functions that are used to implement the operating system functionality.

The shells sends command to the operating system on what the user wants to do.

System utility this allows the performance of individual and specialized tasks.

The kernel is the one that it is responsible of all the activities of the operating system

5.  What are the benefits of multi-threading?

It allows for better communicant

It ensures that all program strictures are simplified

The responsive of the server improves

The resources which are used are minimized

There is improved throughput

6. Why use threads?

 They use very little resources of the operating system in which they are working. Thus they do not need new global data program codes or operating system resources

They allow faster switching of the contexts hence saving and restoring of registers

Thread are cheap to create since the only require a stack and storage for registers.