Phage Endolysins as Potential Antimicrobials against Multidrug Resistant Vibrio alginolyticus and Vibrio parahaemolyticus: Current Status of Research and Challenges Ahead
Phage Endolysins as Potential Antimicrobials against Multidrug Resistant Vibrio alginolyticus and Vibrio parahaemolyticus: Current Status of Research and Challenges Ahead
Nandita Matamp and Sarita G. Bhat *
Department of Biotechnology, Cochin University of Science and Technology, Kochi-682022, Kerala, India; nandita.matamp@gmail.com
* Correspondence: saritagbhat@gmail.com; Tel.: 91-984-603-3486. Fax: 0484-257-7595
Received: 25 February 2019; Accepted: 15 March 2019; Published: 18 March 2019
Salmonid aquaculture has suffered from a number of different ailments. Infectious Salmon Anaemia, caused by the Isavirus from the orthomixoviridae family, led to a major crisis of the Atlantic salmon aquaculture industry in Chile, dramatically reducing production numbers.
Mycotoxins are recognized as toxic compounds of great concern in the context of human health and economy. Mycotoxins are toxic chemical products formed as secondary metabolites by some fungi that readily colonise crops in the field or after harvest. The toxicity syndrome resulting from the intake of such contaminated material by animal and manis termed Mycotoxicosis.These compounds pose a potential threat to human and animal health through ingestion of food products prepared from these commodities.Mycotoxicoses affect various systems of the body according to the target organs of the mycotoxin. This review revealed the major mycotoxins of fungal origin and their mycotoxicoses. The study also reviewed the history of mycotoxin, methods of mycotoxin detection, analysis and the health implications of consuming mycotoxin-contaminated foods/products. In most developing countries, majority are ignorant of the inherent dangers of consuming mouldy produce or food contaminated with fungi and moulds with possible contamination by mycotoxigenic fungi. In view of this, there is need for general and public education to sensitise the people on the health hazards posed by mycotoxins. Proper washing and cooking practices of food commodities, good agricultural practices, fast and effective analyses and detection, good produce handling and storage are some of the control/regulatory measures that should be encouraged, as to assist in mitigating the side effects of mycotoxins in food and health particularly in the tropical and sub-tropical countries and in African where there is enabling environment that promotes fungal growth.
Presentation by Charles Tyler on 'Sustainable Aquaculture Future (SAF) Aquaculture Projects in Bangladesh: Focus on the Environment' at the One Health Approach workshop on Tuesday, 23 March 2021.
The Edge of Tomorrow — Plant Health in the 21st CenturySophien Kamoun
Presented at the ICPP2018 International Congress of Plant Pathology Plenary Session - Plant Health is Earth’s Wealth, Monday, July 30, 2018. See notes and acknowledgments at http://kamounlab.tumblr.com/post/176385835530/the-edge-of-tomorrow-plant-health-in-the-21st
Salmonid aquaculture has suffered from a number of different ailments. Infectious Salmon Anaemia, caused by the Isavirus from the orthomixoviridae family, led to a major crisis of the Atlantic salmon aquaculture industry in Chile, dramatically reducing production numbers.
Mycotoxins are recognized as toxic compounds of great concern in the context of human health and economy. Mycotoxins are toxic chemical products formed as secondary metabolites by some fungi that readily colonise crops in the field or after harvest. The toxicity syndrome resulting from the intake of such contaminated material by animal and manis termed Mycotoxicosis.These compounds pose a potential threat to human and animal health through ingestion of food products prepared from these commodities.Mycotoxicoses affect various systems of the body according to the target organs of the mycotoxin. This review revealed the major mycotoxins of fungal origin and their mycotoxicoses. The study also reviewed the history of mycotoxin, methods of mycotoxin detection, analysis and the health implications of consuming mycotoxin-contaminated foods/products. In most developing countries, majority are ignorant of the inherent dangers of consuming mouldy produce or food contaminated with fungi and moulds with possible contamination by mycotoxigenic fungi. In view of this, there is need for general and public education to sensitise the people on the health hazards posed by mycotoxins. Proper washing and cooking practices of food commodities, good agricultural practices, fast and effective analyses and detection, good produce handling and storage are some of the control/regulatory measures that should be encouraged, as to assist in mitigating the side effects of mycotoxins in food and health particularly in the tropical and sub-tropical countries and in African where there is enabling environment that promotes fungal growth.
Presentation by Charles Tyler on 'Sustainable Aquaculture Future (SAF) Aquaculture Projects in Bangladesh: Focus on the Environment' at the One Health Approach workshop on Tuesday, 23 March 2021.
The Edge of Tomorrow — Plant Health in the 21st CenturySophien Kamoun
Presented at the ICPP2018 International Congress of Plant Pathology Plenary Session - Plant Health is Earth’s Wealth, Monday, July 30, 2018. See notes and acknowledgments at http://kamounlab.tumblr.com/post/176385835530/the-edge-of-tomorrow-plant-health-in-the-21st
Identification of Race/strain of Phytopathogenic Fungi through Conventional A...Sarda Konjengbam
Detection and identification of fungi has relied on a combination of microscopy and culture based techniques. Conventional methods often rely on identification of disease symptoms, isolation and culturing of environmental organisms, and laboratory identification by morphology and biochemical tests. These methods, although the cornerstone of fungal diagnostics, can lead to problems in identification, resulting in incorrect interpretation, diagnosis and ultimately treatment. The methods rely on experienced, skilled laboratory staff, the ability of the organism to be cultured, are time consuming, non quantitative, prone to contamination and error and often delay management (Atkins and Clark, 2004). During the last decades, the advent of molecular biology promised to offer radical alternatives in the detection and enumeration of fungal pathogens. Molecular technology increases understanding of the biology and population structures of plant pathogens, provides quick and accurate answers to epidemiological questions about plant diseases, and supports disease management decisions. New, rapid screening methods are being developed and increasingly used in all aspects of fungal diagnostics.
The quest for better food quality has invariably increased cases of food-borne infections which in turn contribute to the problem of antibiotic resistance as a result of drug abuse. This study is aimed at characterizing bacterial isolates from some seafood sold in Nembe, Bayelsa State, Nigeria. A total of 200 fresh seafood samples (crab, shrimp, oyster and periwinkle) were collected randomly from Nembe, Bayelsa State. Isolates were obtained using the conventional microbiological methods and the pure cultures were screened by gram staining and biochemical test for preliminary identification. Isolates were further characterized for 16SrRNA using Polymerase Chain Reaction and Sequencing. The most dominant species isolated were Staphylococcus gallinarum 27(22.5%), Vibrio rotiferanus 17(14.2%), Vibrio parahaemolyticus 48(40%), Klebsiella aerogenes 10(8.3%) and Klebsiella quasipneumoniae 18(15%). Analysis of variance (ANOVA) by single factor was done to determine the variation in colony counts of isolates from the different seafood samples and P value was > 0.05 indicating that there is no significant difference in colony counts among the different sea foods. The presence of these bacterial species in these seafood samples renders the food unsafe for consumption. Adequate handling as well as proper cooking of seafood before consumption is highly recommended so as to reduce the incidence of food-borne infections.
Prevalence, occurrence and biochemical characterization of Xanthomonas campes...INNS PUBNET
Xanthomonas campestris pv. vesicatoria the causal organism of bacterial spot in tomato results in heavy losses both in the form of quality and. In this study a survey was carried out to report the incidence of bacterial spot disease of tomato in district Swat. We reported maximum disease incidence in tehsil Kabal (71.66%), followed by Charbagh (61.66%) and Barikot (58.33%). For resistant screening a total of 13 tomato germplasms were screened against the disease. The foliar severity ranged from 3.33% to 73.33%, while severity for fruits was ranged from 18.33% to 30.66%. In case of phenotypic data the highest numbers of fruits obtained were 34, plant height 79.5cm and fruit weight was 470 grams/ten tomatoes. While the lowest average numbers of fruits were 6.67, plant height 45.7cm and fruit weight recorded was 215.67 grams/ten tomatoes. Line 1288 showed highest level of resistance followed by Red-stone. However, line 9708 showed highest susceptibility when exposed to artificial inoculation. Our study showed that bacterial spot is a major issue in some part of Pakistan and germplasm screening are linked to increased host resistance and could offer an important contribution to future integrated bacterial spot management programs.
Immunology and Microbiology,Host-Microbe Interactionsvarinder kumar
Immunology and Microbiology
Host-Microbe Interactions
Cellular Immunity
Principles of Immunization
Vaccines
Examples of bacterial exotoxins
Genetics of Pathogenicity
Mechanisms of Pathogenicity
Future developments & information
Applications of Principles of Immunity
Effects of Antigen-Antibody Interactions-2
This includes the detailed explanation on mycoviruses, its history, evolution, taxonomy, classification, hypovirulence, transmission, movement within the fungi and its use as a bio-control agent or its positive and negative impact on phenotypes. this is elaborately explained with recent case studies with special reference to dsRNA mycoviruses.
Kumaryasava is an alcoholic Ayurvedic formulation prepared by the fermentation of Aloe vera. Flowers of Woodfordia fruticosa are added as inoculums for fermentation process.
Host-pathogen Interactions, Molecular Basis and Host Defense: Pathogen Detect...QIAGEN
Host–pathogen interactions are strikingly complex during infection. This slidedeck provides an overview of the molecular basis of these intricate interactions: the impact of microbiota on innate and adaptive immunity, metabolism, and insulin resistance and host defense mechanisms. Various research tools will be introduced to simplify and streamline each step of studying the host response, enabling detection of pathogens, analysis of gene expression and regulation, epigenetic modification, genotyping and signal transduction pathway activation.
Structure-Activity Relationship Study of Synthetic Variants Derived from the ...CrimsonpublishersCJMI
Structure-Activity Relationship Study of Synthetic Variants Derived from the Highly Potent Human Antimicrobial Peptide hLF(1-11) by Carlo PJM Brouwer in Cohesive Journal of microbiology & infectious disease
Host microbial interaction in periodontal diseaseAnushri Gupta
Bacterial Evasion of Host Defense Mechanisms
Immunological Aspects of Microbial Host Interaction
Connective Tissue Alterations:Tissue Destruction in Periodontitis
BONE RESORPTION
Atlanta Botanical Garden Science Cafe: Medicines from Nature - 2014Cassandra Quave
In 2014, Dr. Quave presented a Science Cafe talk at Atlanta Botanical Gardens: "Medicines From Nature: Adventures of a Medical Ethnobotanist"
Abstract:
Ethnobotany is the study of human interactions with plants. It has perhaps been best defined as the science of survival as it involves the study of plants used in our daily lives for food, medicine, shelter, art, and much more. In this science café, Dr. Quave will discuss her research on medicinal plants for the treatment of infectious disease. From remote regions of the Amazon to volcanic islands in the Mediterranean and isolated mountains in the Balkans, Quave has conducted fieldwork in some of the most biologically diverse regions of the planet. She will share some of her adventures as a medical ethnobotanist and explain how traditional knowledge may be the key to the discovery of new life-saving medicines.
the presentation explains the most important disadvantages of genetically modified organisms and the risk they present against our life and the environment, the presentation has been presented in a debate about GMO's in the Mediterranean Agronomic Institute of Chania-Crete-Greece. 08/11/2019
Work is done by:
-BOUBLAT Hatem
-Ebid Mina
-GIAKOUMAKI Stella
-MAROUNI Zainab
-TUL Safiye
Engineering of Phage-Derived Lytic Enzymes: Improving Their Potential as Antimicrobials
Carlos São-José
ID
Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa,
Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal; csaojose@ff.ul.pt; Tel.: +351-217-946-420
Microbiome: The genes and genomes of the microbiota, as well as the products of the microbiota and the host environment” [the collective genomes of the micro-organisms in a particular environment. Although the composition of the gut microbiota varies between individuals, the community in each individual is relatively stable over time.
Identification of Race/strain of Phytopathogenic Fungi through Conventional A...Sarda Konjengbam
Detection and identification of fungi has relied on a combination of microscopy and culture based techniques. Conventional methods often rely on identification of disease symptoms, isolation and culturing of environmental organisms, and laboratory identification by morphology and biochemical tests. These methods, although the cornerstone of fungal diagnostics, can lead to problems in identification, resulting in incorrect interpretation, diagnosis and ultimately treatment. The methods rely on experienced, skilled laboratory staff, the ability of the organism to be cultured, are time consuming, non quantitative, prone to contamination and error and often delay management (Atkins and Clark, 2004). During the last decades, the advent of molecular biology promised to offer radical alternatives in the detection and enumeration of fungal pathogens. Molecular technology increases understanding of the biology and population structures of plant pathogens, provides quick and accurate answers to epidemiological questions about plant diseases, and supports disease management decisions. New, rapid screening methods are being developed and increasingly used in all aspects of fungal diagnostics.
The quest for better food quality has invariably increased cases of food-borne infections which in turn contribute to the problem of antibiotic resistance as a result of drug abuse. This study is aimed at characterizing bacterial isolates from some seafood sold in Nembe, Bayelsa State, Nigeria. A total of 200 fresh seafood samples (crab, shrimp, oyster and periwinkle) were collected randomly from Nembe, Bayelsa State. Isolates were obtained using the conventional microbiological methods and the pure cultures were screened by gram staining and biochemical test for preliminary identification. Isolates were further characterized for 16SrRNA using Polymerase Chain Reaction and Sequencing. The most dominant species isolated were Staphylococcus gallinarum 27(22.5%), Vibrio rotiferanus 17(14.2%), Vibrio parahaemolyticus 48(40%), Klebsiella aerogenes 10(8.3%) and Klebsiella quasipneumoniae 18(15%). Analysis of variance (ANOVA) by single factor was done to determine the variation in colony counts of isolates from the different seafood samples and P value was > 0.05 indicating that there is no significant difference in colony counts among the different sea foods. The presence of these bacterial species in these seafood samples renders the food unsafe for consumption. Adequate handling as well as proper cooking of seafood before consumption is highly recommended so as to reduce the incidence of food-borne infections.
Prevalence, occurrence and biochemical characterization of Xanthomonas campes...INNS PUBNET
Xanthomonas campestris pv. vesicatoria the causal organism of bacterial spot in tomato results in heavy losses both in the form of quality and. In this study a survey was carried out to report the incidence of bacterial spot disease of tomato in district Swat. We reported maximum disease incidence in tehsil Kabal (71.66%), followed by Charbagh (61.66%) and Barikot (58.33%). For resistant screening a total of 13 tomato germplasms were screened against the disease. The foliar severity ranged from 3.33% to 73.33%, while severity for fruits was ranged from 18.33% to 30.66%. In case of phenotypic data the highest numbers of fruits obtained were 34, plant height 79.5cm and fruit weight was 470 grams/ten tomatoes. While the lowest average numbers of fruits were 6.67, plant height 45.7cm and fruit weight recorded was 215.67 grams/ten tomatoes. Line 1288 showed highest level of resistance followed by Red-stone. However, line 9708 showed highest susceptibility when exposed to artificial inoculation. Our study showed that bacterial spot is a major issue in some part of Pakistan and germplasm screening are linked to increased host resistance and could offer an important contribution to future integrated bacterial spot management programs.
Immunology and Microbiology,Host-Microbe Interactionsvarinder kumar
Immunology and Microbiology
Host-Microbe Interactions
Cellular Immunity
Principles of Immunization
Vaccines
Examples of bacterial exotoxins
Genetics of Pathogenicity
Mechanisms of Pathogenicity
Future developments & information
Applications of Principles of Immunity
Effects of Antigen-Antibody Interactions-2
This includes the detailed explanation on mycoviruses, its history, evolution, taxonomy, classification, hypovirulence, transmission, movement within the fungi and its use as a bio-control agent or its positive and negative impact on phenotypes. this is elaborately explained with recent case studies with special reference to dsRNA mycoviruses.
Kumaryasava is an alcoholic Ayurvedic formulation prepared by the fermentation of Aloe vera. Flowers of Woodfordia fruticosa are added as inoculums for fermentation process.
Host-pathogen Interactions, Molecular Basis and Host Defense: Pathogen Detect...QIAGEN
Host–pathogen interactions are strikingly complex during infection. This slidedeck provides an overview of the molecular basis of these intricate interactions: the impact of microbiota on innate and adaptive immunity, metabolism, and insulin resistance and host defense mechanisms. Various research tools will be introduced to simplify and streamline each step of studying the host response, enabling detection of pathogens, analysis of gene expression and regulation, epigenetic modification, genotyping and signal transduction pathway activation.
Structure-Activity Relationship Study of Synthetic Variants Derived from the ...CrimsonpublishersCJMI
Structure-Activity Relationship Study of Synthetic Variants Derived from the Highly Potent Human Antimicrobial Peptide hLF(1-11) by Carlo PJM Brouwer in Cohesive Journal of microbiology & infectious disease
Host microbial interaction in periodontal diseaseAnushri Gupta
Bacterial Evasion of Host Defense Mechanisms
Immunological Aspects of Microbial Host Interaction
Connective Tissue Alterations:Tissue Destruction in Periodontitis
BONE RESORPTION
Atlanta Botanical Garden Science Cafe: Medicines from Nature - 2014Cassandra Quave
In 2014, Dr. Quave presented a Science Cafe talk at Atlanta Botanical Gardens: "Medicines From Nature: Adventures of a Medical Ethnobotanist"
Abstract:
Ethnobotany is the study of human interactions with plants. It has perhaps been best defined as the science of survival as it involves the study of plants used in our daily lives for food, medicine, shelter, art, and much more. In this science café, Dr. Quave will discuss her research on medicinal plants for the treatment of infectious disease. From remote regions of the Amazon to volcanic islands in the Mediterranean and isolated mountains in the Balkans, Quave has conducted fieldwork in some of the most biologically diverse regions of the planet. She will share some of her adventures as a medical ethnobotanist and explain how traditional knowledge may be the key to the discovery of new life-saving medicines.
the presentation explains the most important disadvantages of genetically modified organisms and the risk they present against our life and the environment, the presentation has been presented in a debate about GMO's in the Mediterranean Agronomic Institute of Chania-Crete-Greece. 08/11/2019
Work is done by:
-BOUBLAT Hatem
-Ebid Mina
-GIAKOUMAKI Stella
-MAROUNI Zainab
-TUL Safiye
Similar to Phage Endolysins as Potential Antimicrobials against Multidrug Resistant Vibrio alginolyticus and Vibrio parahaemolyticus: Current Status of Research and Challenges Ahead
Engineering of Phage-Derived Lytic Enzymes: Improving Their Potential as Antimicrobials
Carlos São-José
ID
Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa,
Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal; csaojose@ff.ul.pt; Tel.: +351-217-946-420
Microbiome: The genes and genomes of the microbiota, as well as the products of the microbiota and the host environment” [the collective genomes of the micro-organisms in a particular environment. Although the composition of the gut microbiota varies between individuals, the community in each individual is relatively stable over time.
Background: The widespread use of antibiotics has resulted in emergence of community-acquired antibiotic resistance among uropathogens in outpatient’s population. This constitutes an impediment in the management of urinary tract infection (UTI) in both community and hospital settings. Objective: The aim of this study was to determine the current antibiotic resistance trends, extended spectrum beta-lactamase (ESBL) production and plasmid profile of uropathogens from outpatients. Methods: A total of 370 mid-stream urine samples were collected and cultured by standard methods. Isolated uropathogens were identified using appropriate biochemical methods. The modified Kirby Bauer disk method was used for antibiotic susceptibility test. The ESBL-producing uropathogens were identified and their plasmid DNA extraction and curing were carried out by standard methods. Results: About 35.7% and 32.7% of uropathogens were multi-drug resistant and ESBL-producing respectively. There was higher prevalence of ESBL-production among isolates from female patients (62.5%) when compared to that from male patients (37.5%). The isolated uropathogens were most resistant to Cefotaxime, and most sensitive to Imipenem. Resistance to antibiotics by ESBL-producing uropathogens was found to be plasmid-mediated. Conclusion: Community acquired Uropathogens from outpatients were multidrug resistant due to ESBL production localized on plasmids, a probable cause of treatment failures experienced in Uyo.
AMR & Alternative Stratergies - MicrobiologySijo A
Antibiotic resistance poses one of the most important health challenges of the 21st century.
The rise of multidrug-resistant bacteria has already led to a significant increase in human disease and death.
The U.S. Centers for Disease Control and Prevention estimates that approximately 2.8 million people worldwide are infected with antibiotic-resistant bacteria, accounting for 35,000 deaths each year in the U.S. and 700,000 deaths around the globe.
Evaluation of resistance profile of pseudomonas aeruginosa with reference to ...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Background: Infectious diseases cause significant production losses in aquaculture every year. Since the gut
microbiota plays an essential role in regulating the host immune system, health and physiology, altered gut
microbiota compositions are often associated with a diseased status. However, few studies have examined the
association between disease severity and degree of gut dysbiosis, especially when the gut is not the site of the
primary infection. Moreover, there is a lack of knowledge on whether bath treatment with formalin, a disinfectant
commonly used in aquaculture to treat external infections, might affect the gut microbiome as a consequence of
formalin ingestion. Here we investigate, through 16S rRNA gene metabarcoding, changes in the distal gut
microbiota composition of a captive-reared cohort of 80 Atlantic salmon (Salmo salar L.), in consequence of an
external bacterial skin infection due to a natural outbreak and subsequent formalin treatment.
Results: We identified Tenacibaculum dicentrarchi as the causative disease pathogen and we show that the distal
gut of diseased salmon presented a different composition from that of healthy individuals. A new, yet undescribed,
Mycoplasma genus characterized the gut of healthy salmon, while in the sick fish we observed an increase in terms
of relative abundance of Aliivibrio sp., a strain regarded as opportunistic. We also noticed a positive correlation
between fish weight and Mycoplasma sp. relative abundance, potentially indicating a beneficial effect for its host.
Moreover, we observed that the gut microbiota of fish treated with formalin was more similar to those of sick fish
than healthy ones.
Conclusions: We conclude that external Tenacibaculum infections have the potential of indirectly affecting the host
gut microbiota. As such, treatment optimization procedures should account for that. Formalin treatment is not an
optimal solution from a holistic perspective, since we observe an altered gut microbiota in the treated fish. We
suggest its coupling with a probiotic treatment aimed at re-establishing a healthy community. Lastly, we have
observed a positive correlation of Mycoplasma sp. with salmon health and weight, therefore we encourage further
investigations towards its potential utilization as a biomarker for monitoring health in salmon and potentially other
farmed fish species.
Keywords: Microbiota, Atlantic salmon, Infectious diseases, Dysbiosis, Tenacibaculosis, Aliivibrio, Mycoplasma,
Biomarkers, Fish growth
Bacteriophages come in different sizes and shapes but most of them.docxrock73
Bacteriophages come in different sizes and shapes but most of them have the same basic features: a head or capsid and a tail. A bacteriophage’s head structure, regardless of its size or shape, is made up of one or more proteins which protectively coats the nucleic acid. Though there are some phages that don’t have a tail, most of them do have one attached to its head structure.
How Bacteriophages Work
n oder to infect a host cell, the bacteriophage attaches itself to the bacteria’s cell wall, specifically on a receptor found on the bacteria’s surface. Once it becomes tightly bound to the cell, the bacterial virus injects its genetic material (its nucleic acid) into the host cell. Depending on the type of phage, one of two cycles will occur – the lytic or the lysogenic cycle. During a lytic cycle, the phage will make use of the host cell’s chemical energy as well as its biosynthetic machinery in order to produce phage nucleic acids (phage DNA and phage mRNA) and phage proteins. Once the production phase is finished, the phage nucleic acids and structural proteins are then assembled. After a while, certain proteins produced within the cell will cause the cell wall to lyse, allowing the assembled phages within to be released and to infect other bacterial cells.
Viral reproduction can also occur through the lysogenic cycle. The main difference between the two types of cycles is that during lysogeny, the host cell is not destroyed or does not undergo lysis. Once the host cell is infected, the phage DNA integrates or combines with the bacterial chromosome, creating the prophage. When the bacterium reproduces, the prophage is replicated along with the host chromosomes. Thus, the daughter cells also contain the prophage which carries the potential of producing phages. The lysogenic cycle can continue indefinitely (daughter cells with prophage present within continuing to replicate) unless exposed to adverse conditions which can trigger the termination of the lysogenic state and cause the expression of the phage DNA and the start of the lytic cycle. These adverse conditions include exposure to UV or mutagenic chemicals and desiccation.
http://phages.org/bacteriophage/
Patients in hospitals, especially those on breathing machines, those with devices such as catheters, and patients with wounds from surgery or from burns are potentially at risk for serious, life-threatening infections.
n hospitals, where the most serious infections occur, Pseudomonas can be spread on the hands of healthcare workers or by equipment that gets contaminated and is not properly cleaned.
https://www.cdc.gov/hai/organisms/pseudomonas.html
P. aeruginosa can develop resistance to antibacterials either through the acquisition of resistance genes on mobile genetic elements (i.e., plasmids) or through mutational processes that alter the expression and/or function of chromosomally encoded mechanisms. Both strategies for developing drug resistance can severely limit the therapeutic ...
AMR in Animal Origin Products A ChallengeSarzamin Khan
The AMR and its origin from the products of animal based products has been discussed. The AMR as challenge has been described and recommendation to minimize the risk of AMR
Anti-Virulence Potential And In Vivo Toxicity Of Persicaria Maculosa And Bist...Jose Katab
Custom Writing Service
http://StudyHub.vip/Anti-Virulence-Potential-And-In-Vivo-To 👈
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[13386905 nova biotechnologica et chimica] phage endolysin a way to unders...Pig Farm Solution
PHAGE ENDOLYSIN: A WAY TO UNDERSTAND A BINDING FUNCTION OF C-TERMINAL DOMAINS
A MINI REVIEW
Department of Biology, Faculty of Natural Sciences, University of SS. Cyril and
Methodius, Trnava, SK-917 01, Slovak Republic (lenka.tisakova@ucm.sk)
식품 업계에서의 박테리오파지 요법
Endersen et.al/Phage Therapy in the Food Industry/Annu.Rev. Food Sci. Technol./January 9, 2014
1. 분석자 서문
최신 기술들의 발전에도 불구하고 식품 업계는 여전이 미생물 오염이라는 큰 위협에 계속 직면하고 있다. 항생제의 과도한 이용이 이 문제를 더욱 악화시켜서 항생제 저항성 식품 매개 병원체가 확산되고 있다. 때문에 식품과 식품 가공 환경에서 미생물 오염을 통제하는 새로운 방법의 개발이 매우 중요하게 되었다. 이런 상황에서 박테리오파지와 그 유도체들은 다양한 식품과 식품 가공 환경에서 이들 오염의 예방, 처리, 박멸을 가능하게 하는 새롭고 실행 가능하며 안전한 대안으로 대두되고 있다. 이번 리뷰에서는 박테리오파지, 변형된 박테리오파지, 이들의 유도체들의 현재의 활용과 미래의 가능성에 대하여 다루려고 한다.
2. 목 차
1. 서론2. 식품 매개성 병원체3. 약물로서 박테리오파지4. 식품 업계에서 바이오요법제로서 박테리오파지와 이들의 유도체5. 안전성에 대한 우려6. 박테리오파지 제품의 현재 허가 현황
이 자료의 분석은 일양약품 중앙연구소의 배우철님께서 수고해 주셨습니다.
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H. Lee, S. J. Ohh, I. K. Kwon and B. J. Chae J Anim Sci
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A comparative study of the preventive use of tilmicosin phosphate (pulmotil premix) and mycoplasma hyopneumoniae vaccination in a pig herd with chronic respiratory disease
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Phage Endolysins as Potential Antimicrobials against Multidrug Resistant Vibrio alginolyticus and Vibrio parahaemolyticus: Current Status of Research and Challenges Ahead
1. microorganisms
Review
Phage Endolysins as Potential Antimicrobials against
Multidrug Resistant Vibrio alginolyticus and Vibrio
parahaemolyticus: Current Status of Research and
Challenges Ahead
Nandita Matamp and Sarita G. Bhat *
Department of Biotechnology, Cochin University of Science and Technology, Kochi-682022, Kerala, India;
nandita.matamp@gmail.com
* Correspondence: saritagbhat@gmail.com; Tel.: 91-984-603-3486. Fax: 0484-257-7595
Received: 25 February 2019; Accepted: 15 March 2019; Published: 18 March 2019
Abstract: Vibrio alginolyticus and V. parahaemolyticus, the causative agents of Vibriosis in marine
vertebrates and invertebrates, are also responsible for fatal illnesses such as gastroenteritis, septicemia,
and necrotizing fasciitis in humans via the ingestion of contaminated seafood. Aquaculture farmers
often rely on extensive prophylactic use of antibiotics in farmed fish to mitigate Vibrios and their
biofilms. This has been postulated as being of serious concern in the escalation of antibiotic
resistant Vibrios. For this reason, alternative strategies to combat aquaculture pathogens are in
high demand. Bacteriophage-derived lytic enzymes and proteins are of interest to the scientific
community as promising tools with which to diminish our dependency on antibiotics. Lysqdvp001
is the best-characterized endolysin with lytic activity against multiple species of Vibrios. Various
homologues of Vibrio phage endolysins have also been studied for their antibacterial potential. These
novel endolysins are the major focus of this mini review.
Keywords: antibiotic resistance; endolysin; Vibrio alginolyticus; V. parahaemolyticus
1. Introduction
Vibrio alginolyticus and V. parahaemolyticus, the normal inhabitants of estuarine and marine
environments, are notable human enteropathogens associated with seafood-borne mortality and
illness worldwide. V. alginolyticus is reported as the etiological agent of wound and ear infections
(both otitis media and otitis externa), intracranial infection, peritonitis and osteomyelitis among many
others [1–3], while V. parahaemolyticus causes bacterial gastroenteritis associated with the consumption
of raw or undercooked seafood [4,5].
In aquaculture, the hazard of infectious diseases has led to significant stock losses and problems
with animal welfare. Hence, intensive aquaculture promotes the indiscriminate use of anti-microbials,
thereby causing the dissemination of antimicrobial-resistant (AMR) bacteria and resistance genes in
aquaculture products and the environment [6]. This global concern has necessitated the exploration of
alternative therapies for bacterial pathogens in animal production, especially in aquaculture. Amongst
the several substitutes, that include probiotics, essential oils and anti-microbial peptides, phage therapy
has gained much attention for preventing and controlling pathogenic infections in aquaculture facilities.
Recent advances in phage genome sequencing have kindled the application of phage encoded enzymes,
especially endolysins, as biocontrol and therapeutic agents against major food-borne pathogens.
There have been numerous reviews on endolysins as antimicrobials against Gram-positive bacteria.
However, in this review, we concentrate on phage lysin biology against Gram-negative pathogens
V. alginolyticus and V. parahaemolyticus. The endolysin characteristics that are important to combatting
Microorganisms 2019, 7, 84; doi:10.3390/microorganisms7030084 www.mdpi.com/journal/microorganisms
2. Microorganisms 2019, 7, 84 2 of 11
multidrug resistant Vibrios are summarized, thereby outlining the remarkable potency of these enzymes
in the mitigation of similar pathogens in aquaculture.
2. Antibiotic Resistance in Vibrios alginolyticus and V. parahaemolyticus
Antimicrobial/chemotherapeutic agents against V. alginolyticus and V. parahaemolyticus are
used either as feed additives and/or as immersion bath solutions in fish farms. The
recommended antibiotics against Vibrios are fluoroquinolones (ciprofloxacin, levofloxacin),
tetracyclines (doxycycline, tetracycline), third-generation cephalosporins (cefotaxime, ceftazidime,
ceftriaxone), aminoglycosides (amikacin, apramycin, gentamicin, streptomycin) and folate pathway
inhibitors (trimethoprim-sulfamethoxazole) [7]. The excessive use of antibiotics has led to the evolution
of numerous strains that exhibit resistance to a single or a combination of antibiotics. However, as
reported by the United States Centers for Disease Control and Prevention (CDC), the occurrence of
Vibrio-related infections has increased dramatically since 2001 [8] (CDC, 2016).
Gram-negative bacteria have developed divergent mechanisms to bypass the inhibitory effects of
antibiotics such as (1) drug inactivation/destruction (2) decreased antibiotic penetration and efflux
(3) target site modification and (4) global cell adaptations [9–11]. Genes and associated insertion
elements, which confer antibiotic resistance, are usually found localized in plasmids as multi-resistance
regions (MRR) in these organisms [12]. Studies of antibiotic resistance in many pathogens such as
V. cholera, Staphylococcus aureus and Salmonella have been reported, but the mechanism of the same in
V. alginolyticus and V. parahaemolyticus is poorly documented [13].
Fluoroquinolones, a promising class of broad-spectrum antibiotics, are direct inhibitors of DNA
synthesis. They bind to the enzyme-DNA complex and stabilize DNA strand breaks created by the
enzymes DNA gyrase and topoisomerase IV. [14]. Kitaoka and his co-workers have reported the
presence of spontaneous chromosomal mutations in gyrA and parC genes in Vibrios that encode
subunits of DNA gyrase and topoisomerase IV, respectively. These mutations could alter the affinity
of theses enzymes, thus protecting Vibrios from quinolones. [15]. Self-transmissible plasmids that
confer resistance by plasmid-mediated quinolone resistance (PMQR) mechanisms were also being
investigated in V. parahaemolyticus [16]. Tetracycline has been recommended as the antimicrobial of
choice for the treatment of severe Vibrio infections, thanks to its ability to inhibit the synthesis of
pathogenic extracellular enzymes [17]. The emergence of V. alginolyticus possessing tet plasmids for
tetracycline resistance [18] and pVAS3-1 plasmids for β-lactamase resistance is alarming [19].
Treatment Costs for Vibriosis
According to CDC Outbreak Surveillance Data, around 6680 cases of V. parahaemolyticus and
165 cases of V. alginolyticus have been reported annually. The annual health costs of Vibrio infections are
estimated to be over $30 million. These data are quite imprecise due to limitations of surveillance data
and underreporting. Under such circumstances, the costs are likely higher, leading to considerable
uncertainty into the overall estimate. In addition to the hike in treatment costs, antimicrobial resistance
can lead to protracted hospital stays and escalations in morbidity and mortality rates [20].
3. Bacteriophage Endolysins-‘the Holy Grail’ to Control Food Borne Pathogens
Bacteriophages or phages are viruses that specifically infect and lyse bacteria. Following their
discovery by Twort and Felix D’Herelle, it became clear that they exhibit two kinds of life cycles:
lytic (used by both virulent and temperate phages) and lysogenic (used exclusively by temperate or
pro-phages) [21]. Lytic or virulent phages have evolved a lytic system to weaken the bacterial cell
wall, leading to bacterial lysis. This bacterial lysis is achieved by phage-encoded muralytic enzymes
called Endolysins (or lysins) that degrade the peptidoglycan (PG) layer present in the bacterial cell
wall during the final stage of the phage reproduction events. The lysis events of double-stranded DNA
bacteriophages can be elucidated by three different mechanisms. The most explicitly demonstrated
mechanism is canonical lysis, where lysins act on PG layer with the help of a second phage encoded
3. Microorganisms 2019, 7, 84 3 of 11
protein called holin, in a timely-controlled fashion [22]. Holins depolarize the cytoplasmic membrane
by allowing endolysins to diffuse through pores in the membrane and target the PG layer. The
second pathway is mediated by a special class of holins designated as pinholins which forms small,
heptametrical channels in the membrane instead of large holes as seen in canonical lytic pathway.
These pinholins work in association with Signal-arrest-release (SAR) endolysins which are inactive
tethered enzymes accumulated in the periplasm. Using proton motor force (PMF), pinholins trigger
the activation of these pro-enzymes, refolding their configuration leading to their release from the
bi-layer, thereby degrading PG. Pinholins act as timers for endolysin activation playing no lead role in
their export. In Gram-negative hosts, the lysis of OM is by a third functional class of lysis proteins
called the spanins [23]. Spanin complex consists of small outer membrane lipoprotein (o-spanin) and
an integral cytoplasmic membrane protein (i-spanin) which disrupts OM by 3 modes: (i) enzymatic
degradation of PG cross links [24] (ii) pore formation [25] and (iii) inner membrane-outer membrane
fusion [26]. Phage researchers have termed these enzymes as ‘enzybiotics’; they can be exploited for
their ability to kill variety of pathogens [27].
4. Gram-Negative Endolysins as Antimicrobials
Basic Structure and Function
The peptidoglycan layer is the major structural component of the bacterial cell wall responsible
for protection, physical integrity and shape. It is composed of chains of alternating residues of
N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc), connected by β-1,4 glycosidic
bonds, linked to a short stem of tetrapeptide [28]. The cell wall of Gram-negative organisms has an
outer membrane (OM) situated above a thin PG layer and the limited permeability of OM [29] poses a
major hurdle for development of novel antimicrobials against Gram-negative pathogens preventing
many compounds from reaching their intracellular targets. Since the endolysin susceptible layer (PG)
is found between an inner and outer membrane, effective strategies, like use of peptides, detergents,
and chelators, should be applied in combination with hydrolytic enzymes to improve the applicability
of phage lysins. As an example, 5 mM EDTA used in combination with E. coli phage endolysin PlyE146
400 µg/mL, decreased titers of E. coli K12 by ca. 2 log10 CFU/mL upon 2 hours of incubation. [30].
Moreover, the peptide moiety made of L- and D-amino acids is highly conserved (chemotype A1γ) in
Gram-negative organisms, whereas the carbohydrate backbone is conserved in both Gram-positive
and negative bacteria.
Phage endolysins are analogous in structure and function to bacterial lysins, and are closely
related to the small family of mammalian PG recognition proteins [31]. They can have either a
globular or modular structure. Endolysins from phages infecting Gram-negative hosts are mostly
small single-enzymatically active domain (EAD) globular proteins (molecular mass 15–20 kDa) without
a specific cell wall binding domain (CBD) module [32,33]. An EAD cleaves a specific bond in the PG
structure, whereas a CBD targets the EAD to its substrate by binding PG or another cell wall component.
Apart from these two domains, recent reports of some Gram-negative antibacterial endolysins have
revealed another domain CHAP (cysteine,histidine-dependent amidohydrolase/peptidase) belonging
to amidase family whose role is to facilitate hydrolysis of the PG layer [34,35]. This feature enables
them to enhance their catalytic skills by binding to multiple sites during cell lysis. An endolysin
isolated from a phage infecting a Gram-negative species is therefore enzymatically-active on the PG of
any other Gram-negative strain [36].
The first endolysins infecting Gram-negative bacteria were reported in the 1960s, and were mostly
encoded by T-phages infecting Escherichia coli. Earlier, they were simply referred to as ‘lysozymes’
based on their functional similarity to egg white lysozyme, a muralytic enzyme well noted for its
anti-bacterial activity. Later, based on the mechanism of action, PG hydrolases were classified into 4
groups: (a) glycosidases which cleave the glycan component of peptidoglycan, (b) amidohydrolase,
that cleaves amide bond between the glycan moiety (MurNAc) and the peptide moiety (L-alanine)
4. Microorganisms 2019, 7, 84 4 of 11
of the PG (c) endopeptidase which cleaves peptide bonds between two amino acids, and finally, (d)
lytic transglycosylases that cleave the β(1-4) linkages between NAM and NAG residues of the PG.
Transglycosylases are not true hydrolases, as they do not require water to catalyze PG cleavage. Most
of the endolysins reported so far are lytic transglycosylases. The complexity of endolysins can be
further illustrated by the fact that an elaborate motif search of approximately 723 putative endolysins
in database has revealed the presence of 24 types of catalytic domains, 13 binding domains, and 89
possible architectural organizations [37].
The modular structure of endolysin has facilitated development of engineered lysins with desired
properties such as higher stability, solubility and broad killing spectrum. Because of the independent
functions of N-terminal catalytic domain (CD) and a C-terminal cell-wall binding domain (CBD), lysins
can be constructed by fusing them from different origins or with other molecules [38]. Among
the engineered lysins, chimeolysins and artilysins are worth mentioning. Several chimeolysins
have been constructed with extended broad spectrum activity against Gram-positive pathogens
like Staphylococcus, Streptococci and E. faecalis [39–43]. Recently, novel chimeolysin (ClyF) active against
planktonic and biofilm MRSA designed from a chimeolysin library with different combinations of CDs
and CBDs was expressed in E. coli [44]. Artilysins are outer membrane-penetrating lysins constructed
by fusing a fragment of natural lysin with peptides or other proteins with high anti-bacterial activity
against Gram-negative pathogens. The lipopolysaccharide destabilizing peptides of artilysins can
be effectively exploited against Pseudomonas, E. coli, Salmonella and Yersinia [45,46]. The concept of
endolysin delivery against Gram-negatives is further expanded by the development of Innolysins
which are constructed by combining receptor binding proteins (RBPs) of candidate phages. Zampara
and his co-workers constructed twelve Innolysins using phage T5 endolysin and receptor binding
protein Pb5, which bind irreversibly to the phage receptor FhuA involved in ferrichrome transport in
E coli. It was proved that they pass through the outer membrane and degrade the PG layer, thereby
killing the target bacteria [47].
5. Vibrio Phage Endolysins
Phage therapy experiments have shown promising results in the eradication of several pathogenic
Vibrios (V. harveyi, V. parahaemolyticus, V. alginolyticus, V. splendidus, V. anguillarum,) in aquaculture
settings since 1999 [48–52]. The extensive amount of genetic information assembled from phage whole
genome sequencing has opened up new horizons to design novel antimicrobial agents. In this respect,
timely exploration into utilization of Vibrio phage endolysins has sparkled interest among active
phage researchers. Table 1 shows the complete list (to date) of all endolysins/putative ORFs coded by
V. alginolyticus and V. parahaemolyticus. The three dimensional structures of Vibrio phage endolysins
have been predicted by homology modeling (Figure 1).
5. Microorganisms 2019, 7, 84 5 of 11
Table 1. Showing endolysins of Vibrio alginolyticus and V. parahaemolyticus phages.
Bacteria Phage
Putative
Endolysins/Predicted
Orfs
Features Reference
Vibrio alginolyticus
PVA1 gp60 Putative lysozyme family protein * [50]
phi-Grn1 phiGrn1_0012 SLT domain protein/endolysin * [53]
ValKK3 ORF304 Tail lysozyme [54]
Athena1 Cds006 Protein with lysozyme activity * [55]
VEN gp50 Cell wall hydrolase-like protein * [56]
Vp670 cwlQ endolysin [57]
vB_ValP_IME271 CDS64 endolysin [58]
V. parahaemolyticus
qdvp001 Lysqdvp001 modular endolysin [59]
VP06 PP_00050
membrane-bound lytic murein
transglycosylase *
[60]
VPp1 LysVPp1 endolysin [61]
VPMS1 LysVPMS1 endolysin [62]
VpKK5 ORF62
N-acetylmuramoyl-L-alanine
amidase *
[63]
pTD1 BAW98403.1 Endolysin * [64]
* indicates the newly identified features of sequenced endolysins as part of the present study.
Microorganisms 2019, 7, x FOR PEER REVIEW 5 of 11
VPMS1 LysVPMS1 endolysin [62]
VpKK5 ORF62
N-acetylmuramoyl-L-
alanine amidase *
[63]
pTD1 BAW98403.1 Endolysin * [64]
* indicates the newly identified features of sequenced endolysins as part of the present study.
Figure 1. Structural modeling of Vibrio phage endolysins.
5.1. Lysqdvp001 and Its Homologues
5.1.1. Structure, Function and Physiochemical Properties
The endolysin Lysqdvp001 is derived from V. parahaemolyticus bacteriophage qdvp001, a lytic
broad-spectrum phage belonging to Myoviridae family with genome length of 134,742-bp. [59]. The
endolysin gene (ORF 60) of qdvp001 has a close relationship to Vibrio (cholerae) phage ICP1, which
Figure 1. Structural modeling of Vibrio phage endolysins.
6. Microorganisms 2019, 7, 84 6 of 11
5.1. Lysqdvp001 and Its Homologues
5.1.1. Structure, Function and Physiochemical Properties
The endolysin Lysqdvp001 is derived from V. parahaemolyticus bacteriophage qdvp001, a lytic
broad-spectrum phage belonging to Myoviridae family with genome length of 134,742-bp. [59]. The
endolysin gene (ORF 60) of qdvp001 has a close relationship to Vibrio (cholerae) phage ICP1, which
shares the same modular structure with ORF 60. The PG_binding _1 domain of the Vibrio phage ICP1
endolysin gene is 58 % homologous to ORF 60, whereas the CHAP domain shares a 66 % amino acid
sequence identity. Interestingly, SMART analysis of endolysin has shown an unusual structure with
two domains: a PG binding (PF01471) domain and a CHAP (PF05257) domain. Lysins with dual
domains have been mostly reported in phages infecting Gram-positive pathogens and rare among
bacteriophages infecting Gram-negative bacteria [65]. Lysqdvp001 is a modular endolysin with no
transmembrane regions or signal peptide regions. Bioinformatic analysis also revealed the absence
of any holins to assist the function of Lysqdvp001. The estimated molecular weight is 25.9 kDa and
pI value is 5.97. The endolysin was cloned in competent E. coli BL21 StarTM (DE3) strain and the
recombinant endolysin has a good yield of 10.4 g from 300 mL of E.coli culture. Turbidity reduction
assay of the purified product demonstrated promising results as the endolysin reduced turbidity
of host bacteria by 0.6 log upon 5 min of incubation. This effective reduction was observed due to
pretreatment of bacterial culture with EDTA for 5 min. Furthermore, Lysqdvp001 was able to lyse
11/11 V. parahaemolyticus strains tested, whereas the parent bacteriophage qdvp001 had a shorter host
range of lysing 3/11 strains suggesting a broader anti-bacterial spectrum of the purified phage enzyme.
5.1.2. LysVPMS1
LysVPMS1 was obtained from V. parahaemolyticus bacteriophage VPMS1. The host used to
propagate the phage was isolated from shrimp farms in northwestern Mexico during an acute
hepatopancreatic necrosis disease (AHPND) outbreak in 2014. This endolysin is the first reported
phage lytic enzyme against V. parahaemolyticus AHPND strains. LysVPMS1 showed lytic activity
against 17 AHPND strains and 5 non- AHPND strains. The highest rate of muralytic activity was
observed in case of V. parahaemolyticus ATCC-17802 strain (96%). This information is quite significant
in terms of the ability of purified LysVPMS1 to lyse strains from different origins with different degrees
of pathogenicity. Moreover, this endolysin has the unique ability to lyse the cell wall of other Vibrio
species specifically V. alginolyticus, V. harveyi and V. campbellii. More information on biochemical and
bactericidal properties of the LysVPMS1 endolysin is presently unavailable [62].
5.1.3. LysVPp1
VPp1 is a double-stranded DNA phage capable of infecting V. parahaemolyticus strains belonging
to Myoviridae family. Its genome consists of 50,431 bp with a G+C content of 41.35%. The ability of
VPp1 to reduce bacterial load during depurination in oysters was reported back in 2014 [36]. Recently,
endolysin (LysVPp1) derived from VPp1 was purified and assessed for its anti-bacterial activities.
LysVPp1 is a soluble lytic transglycosylase related to hen egg white lysozyme with a molecular weight
of ~44 kDa and yield of 1 mg/mL. Peptidoglycan binding domain was not reported in LysVPp1.No
holins/antiholin were also annotated in the phage genome. The antibacterial spectrum of the lysin
was evaluated via two methods-(1) gel diffusion assay and (2) turbidity reduction assay. In gel
diffusion assay, V. parahaemolyticus ATCC 17802 was used as the substrate. The hydrolase activity was
determined by color changes around the holes in gels (0.01% potassium hydroxide + 0.1% methylene
blue) containing peptidoglycan. Gel holes treated with LysVPp1 showed a light blue color resulting
from the hydrolysis of peptidoglycan, thereby validating the hydrolytic activity of recombinant enzyme.
In addition, the turbidity of EDTA-pretreated V. parahaemolyticus cells was reduced by 0.4 log after
5 min of incubation. The lytic spectrum assay of parent strain VPp1 lysed only V. parahaemolyticus
7. Microorganisms 2019, 7, 84 7 of 11
isolates whereas the recombinant lysin LysVPp1 could hydrolyze 9 of 12 Vibrio strains tested, which
included closely related Vibrio strains such as V. parahaemolyticus, V. campbellii, and V. azureus [61].
5.2. cwlQ- First Recombinant Endolysin with Holin Assistance
Vp670 is lytic phage of Podoviridae family capable of infecting V. alginolyticus strains. This is the
first report of a V. alginolyticus phage whose lysis cassette was annotated, cloned and expressed. The
genome size of Vp670 is 43,121 bp which codes for 49 ORFs and contains a lysis module, composed
of two components- holA (holin) and cwlQ (endolysin). cwlQ is a relatively small protein(15–20 kDa)
belonging to hydrolase-2 domain superfamily (Pfam 07486). TM pred analysis showed holA has a
transmembrane helix with a hydrophilic C-terminal region inside the cytoplasmic membrane. Clones
were expressed in E. coli (LPN028 and LPN030) and V. alginolyticus (LPN041 and LPN043) strains and
these cells were able to survive under L-arabinose induction conditions. The clone expression was
further studied by TEM analysis. Clones with holA and cwlQ had their OM layer disrupted and their
cellular contents released from channels in the cell membrane. Cells without expressed genes had
intact cellular structures with no morphological difference. Coexpression of both genes has resulted in
severe cell damage compared to the expression of holA alone in the cells [57].
The therapeutic potential of the above reported endolysins been investigated in neither in vitro
nor in vivo models.
6. Challenges of Endolysin Engineering and Delivery
There are numerous reports supporting the antibacterial activity of endolysins in vivo, but only
a few of them have been proven by human clinical trials. There are numerous reports supporting
the antibacterial activity of endolysins in vivo, but little information has been published on human
clinical trials. SAL200 is the first endolysin based therapeutic formulation with a recombinant form of
phage endolysin SAL-1 (rSAL-1) derived from the bacteriophage SAP-1, as its active pharmaceutical
ingredient. SAP-1 infects Staphylococci, including MRSA and vancomycin-resistant S. aureus (VRSA)
strains [66,67]. The first in-human phase 1 study of SAL200 provided preliminary information on safety,
tolerability, pharmacokinetics, and pharmacodynamics of the product upon intravenous injection
among healthy adults [68]. No serious adverse effects were observed in volunteers except mild and
temporarily observed effects such as fatigue, headaches and myalgia. Similarly Staphefekt SA.100,
a recombinant phage endolysin formulated ointment against infections caused by MRSA strains is
available in a cetomacrogol-based cream/gel as over-the-counter treatment in Europe since 2017 [69].
Many challenges need to be addressed and overcome to deliver engineered chimeric endolysins.
Lysins are non-replicating proteinaeous molecules with short half-life in systemic circulation [21,70].
They also elicit immunological response when applied systemically leading to catalytic loss of
the enzyme [65]. Endolysins can be used in combination with other anti-bacterials, as they
have been proven to act synergistically with antibiotics [71]. Currently, studies on lysin dosage
are underdeveloped. Safe and successful therapeutic application of endolysin requires detailed
information on bioavailability, immunogenicity and lysin synergy. More human clinical trials are
anticipated to investigate phage endolysin treatments to combat several human pathogens. This is
extremely important due to the substantial increase of multi-drug resistant pathogens and the steady
decline in the discovery of new classes of antibiotics.
7. Conclusions
The extensive use of antibiotics has resulted in the emergence of multidrug resistant ‘superbugs’
worldwide. Bacteriophage encoded lytic enzymes ‘endolysins’ have enormous anti-microbial potential
to fight against food borne pathogens in this multi-drug-resistance era. Promising results have
encouraged active phage researchers to apply phage enzymes in various fields, such as food safety,
pathogen detection, surface decontamination and nanotechnology. All reported endolysins showed
a broad activity spectrum for the genus Vibrio. Research on Vibrio phage-encoded lytic enzymes has
8. Microorganisms 2019, 7, 84 8 of 11
intensified since 2016. Lysqdvp001 and its homologues are highly divergent enzymes which are capable
of superior lytic and antibacterial activity compared to their parent phages. Several attributes, such as
high catalytic activity, modular structure and dual catalytic domains, support the robust development
of them as novel alternatives to conventional antibiotic therapy. As the most abundant biological entity
on earth, Bacteriophages’ lytic proteins are also considered structurally and functionally divergent.
Bio-informatic and proteomic studies will allow researchers to expand endolysins as a powerful tool
with diverse applications.
Author Contributions: This review paper is a part of the Ph.D. work of author N.M. All the authors contributed
to the writing and the proofreading of all versions of this article.
Funding: This research received no external funding.
Acknowledgments: The authors acknowledge Department of Biotechnology, Cochin University of Science and
Technology for providing all facilities for research.
Conflicts of Interest: The authors declare no conflict of interest.
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