The major types of vaccines in the clinical application represent different strategies used to reduce the risk of illness, while retaining the ability to induce beneficial immune responses.
\\n\\n
Dr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\\n\\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\\n\\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\\n\\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\\n\\nThank you all for being part of the journey. 5,000 times thank you!
\\n\\nNow with 5,000 titles available Open Access, which one will you read next?
\\n\\nRead, share and download for free: https://www.intechopen.com/books
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Preparation of Space Experiments edited by international leading expert Dr. Vladimir Pletser, Director of Space Training Operations at Blue Abyss is the 5,000th Open Access book published by IntechOpen and our milestone publication!
\n\n"This book presents some of the current trends in space microgravity research. The eleven chapters introduce various facets of space research in physical sciences, human physiology and technology developed using the microgravity environment not only to improve our fundamental understanding in these domains but also to adapt this new knowledge for application on earth." says the editor. Listen what else Dr. Pletser has to say...
\n\n\n\nDr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\n\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\n\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\n\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\n\nThank you all for being part of the journey. 5,000 times thank you!
\n\nNow with 5,000 titles available Open Access, which one will you read next?
\n\nRead, share and download for free: https://www.intechopen.com/books
\n\n\n\n
\n'}],latestNews:[{slug:"intechopen-partners-with-ehs-for-digital-advertising-representation-20210416",title:"IntechOpen Partners with EHS for Digital Advertising Representation"},{slug:"intechopen-signs-new-contract-with-cepiec-china-for-distribution-of-open-access-books-20210319",title:"IntechOpen Signs New Contract with CEPIEC, China for Distribution of Open Access Books"},{slug:"150-million-downloads-and-counting-20210316",title:"150 Million Downloads and Counting"},{slug:"intechopen-secures-indefinite-content-preservation-with-clockss-20210309",title:"IntechOpen Secures Indefinite Content Preservation with CLOCKSS"},{slug:"intechopen-expands-to-all-global-amazon-channels-with-full-catalog-of-books-20210308",title:"IntechOpen Expands to All Global Amazon Channels with Full Catalog of Books"},{slug:"stanford-university-identifies-top-2-scientists-over-1-000-are-intechopen-authors-and-editors-20210122",title:"Stanford University Identifies Top 2% Scientists, Over 1,000 are IntechOpen Authors and Editors"},{slug:"intechopen-authors-included-in-the-highly-cited-researchers-list-for-2020-20210121",title:"IntechOpen Authors Included in the Highly Cited Researchers List for 2020"},{slug:"intechopen-maintains-position-as-the-world-s-largest-oa-book-publisher-20201218",title:"IntechOpen Maintains Position as the World’s Largest OA Book Publisher"}]},book:{item:{type:"book",id:"2676",leadTitle:null,fullTitle:"Current Concepts in Kidney Transplantation",title:"Current Concepts in Kidney Transplantation",subtitle:null,reviewType:"peer-reviewed",abstract:"Despite significant accomplishments to date, kidney transplantation is a relatively young field in medicine. Due to the armamentarium of agents available to effectively suppress the immune system, the past decade has seen a shift in focus from prevention of rejection to a focus on extending the life of the allograft and novel strategies to increase the organ donor pool. This book covers basic concepts in kidney transplantation while also addressing ways to manage kidney transplant recipients in order to maximize patient and graft survival. In addition, novel concepts to increase organ availability are addressed, including kidney paired donation and single site laparoendoscopic donor nephrectomy for living donor kidney transplantation, and utilization of marginal, hepatitis C positive, and older donor organs to increase deceased donor transplant opportunities.",isbn:null,printIsbn:"978-953-51-0900-6",pdfIsbn:"978-953-51-7052-5",doi:"10.5772/3048",price:139,priceEur:155,priceUsd:179,slug:"current-concepts-in-kidney-transplantation",numberOfPages:330,isOpenForSubmission:!1,isInWos:1,hash:"9513ddae671c1c1d920820209c22ca4c",bookSignature:"Sandip Kapur, Cheguevara Afaneh and Meredith J. Aull",publishedDate:"December 19th 2012",coverURL:"https://cdn.intechopen.com/books/images_new/2676.jpg",numberOfDownloads:39585,numberOfWosCitations:2,numberOfCrossrefCitations:5,numberOfDimensionsCitations:11,hasAltmetrics:0,numberOfTotalCitations:18,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"December 5th 2011",dateEndSecondStepPublish:"January 9th 2012",dateEndThirdStepPublish:"July 31st 2012",dateEndFourthStepPublish:"August 31st 2012",dateEndFifthStepPublish:"September 30th 2012",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,editors:[{id:"78020",title:"Dr.",name:"Sandip",middleName:null,surname:"Kapur",slug:"sandip-kapur",fullName:"Sandip Kapur",profilePictureURL:"https://mts.intechopen.com/storage/users/78020/images/3515_n.jpg",biography:"Dr. Sandip Kapur is Chief of Transplantation and Director of the Kidney and Pancreas Transplant Programs at NewYork-Presbyterian Hospital/Weill Cornell Medical Center. Dr. Kapur is an internationally recognized pioneer in advancing innovative strategies that allow more recipients to receive successful transplants, including high risk kidney transplantation and expanding opportunities for living donor kidney transplantation. Dr. Kapur led Cornell in performing one of the first living donor kidney transplant chains, a novel concept that is revolutionizing transplantation and dramatically improves the opportunity for kidney transplant candidates to find a compatible living donor. 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In 2009, he pursued a two-year research fellowship in the Department of Transplantation Medicine, where his worked focused on renal allograft recipients and urinary cell gene expression patterns, predicting and prognosticating outcomes. He has won several awards including the Genzyme’s Clinical Science Award, TTS/Astellas \nYoung Investigator Award, and the ERA-EDTA: Best Abstract by a Young Investigator Award. 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\r\n\tThe book covers the latest developments in the constantly emerging research associated with fungal endophytic microorganisms. The fungal endophytes are distinct group of organisms forming associations nearly ubiquitously throughout the plant kingdom. In view of the great availability of these microorganisms, significant attention is being paid towards their exploitation with regard to novel and beneficial effects. Thus, the biotechnological applications of fungal endophytes in the fields of discovery of high value metabolites as natural drugs, endophytic fungal derived biocatalysts and flavor in food processing industries, enhanced in planta production of bioactive compounds through regulation key genes of biosynthetic pathway, fungal endophyte assisted phytoremediation of contaminated environments, mycendophyte mediated synthesis of metallic nanoparticles and endophytic fungal biocontol agents for plant diseases are discussed in detail. This book intends to cover recent advances in fungal endophytic research in environment, medicine, agriculture and industry.
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Chandra Mouli was born and grew up in Andhra Pradesh, India. He completed M.Sc. and Ph.D degree at Sri Venkateswara University, Tirupati, India. Dr Chandra is working as Postdoctoral Scientist (Fellow) in the Department of Botany of Sri Venkateswara University. He has more than six years of teaching and research experience. His research interests have focused on biology, chemistry of medicinal plants and their associated fungal endophytes, protective effects of bioactive compounds derived from medicinal plants and endophytic fungi against obesity, diabetes and caner, enhanced in planta production of bioactive compounds by fungal endophytes through regulation of key genes involved in the biosynthetic pathway, fungal endophyte assisted phytoremediation of heavy metal contaminated environments. He has completed externally funded research project on 'In search of leptin inhibitors from fungal endophytes (LIFEs) for the development of novel immunemodulating lead compounds”. He has received awards like Fast Track Young Scientist Award (DBT, India), Young Scientist Award (Biotech Helpline Foundation , India), Rajiv Gandhi National Fellowship Award and APCOST Project Award (Andhra Pradesh, India). He has received fellow award from the Society for Applied Biotechnology, India. Dr. Chandra is serving as editorial board member and reviewer of several international reputed peer reviewed scientific journals. 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Type | \nDefinition | \nExample | \n
---|---|---|
Inactivated | \nVirulent microbes are previously destroyed with chemicals, heat, radiation, or antibiotics | \nInfluenza vaccine, whole cell pertussis vaccine, cholera vaccine, hepatitis A vaccine, rabies vaccine, plague vaccine, inactivated polio vaccine (IPV) | \n
Attenuated | \nLive microbes are cultivated under conditions to disable their virulence, or closely related but less dangerous microbes are used to induce broad immune responses | \nYellow fever vaccine, shingles vaccine, measles, mumps and rubella (MMR) vaccine, (1)typhoid (Ty21a) vaccine, BCG vaccine, rotavirus vaccine, live attenuated influenza vaccine (LAIV), oral polio vaccine (OPV) | \n
Toxoid | \nVaccines are made from inactivated toxic compounds that cause illness rather than microbes | \nTetanus toxoid, diphtheria toxoid | \n
Subunit | \nProtein-based vaccine: A fragment of protein creates an immune response, rather than introducing inactivated or attenuated microbes to an immune system. | \n(2)Hepatitis B vaccine, (2)human papillomavirus (HPV) vaccine | \n
Polysaccharide vaccine: A capsular polysaccharide antigen is covalently attached to a carrier protein and creates an immune response. | \n(1)typhoid (Vi) vaccine, (3)pneumococcus vaccine, (4)meningococcus vaccine | \n|
\n | Conjugate vaccine: Certain pathogens have poorly immunogenic polysaccharide outer coats. The immune system recognizes the polysaccharide as if it were a protein antigen by linking these outer coats to proteins | \ntype b (Hib) vaccine, (3)pneumococcus vaccine, (4)meningococcus vaccine | \n
The major types of vaccines in the clinical application represent different strategies used to reduce the risk of illness, while retaining the ability to induce beneficial immune responses.
A vaccine, an antigenic material, is a biological preparation that provides adaptive immunity to a specific disease. The agent triggers immune responses to recognize the disease-causing agent to destroy and remember it, so that the immune system can immediately recognize and destroy the foreign agent once it invades into the body later. Inactivated microbes live attenuated microbes, toxoids, subunits, or conjugates have been manufactured as vaccines and employed to stimulate adaptive immune responses (Table 1). A monovalent (univalent) vaccine is designed to immunize against a single antigen or microorganism such as measles vaccine, while a multivalent (polyvalent) vaccine is designed to immunize against more than one strain of the same microorganism or against more than one microorganism such as oral polio vaccine (OPV), three in one (e.g., diphtheria-tetanus-pertussis, DTP) vaccine, etc. Heterologous vaccines are pathogens of other animals that do not cause diseases or only cause mild diseases in the organism being treated, such as the cowpox vaccine and the Bacillus Calmette-Guérin (BCG) vaccine.
\n\n(1) Two types of typhoid vaccines are available: typhoid (Ty21a) vaccine (a live, attenuated vaccine given by mouth) and typhoid (Vi) vaccine (a subunit vaccine given by injection). (2) Current hepatitis B vaccine and human papillomavirus (HPV) vaccine may be produced by recombinant DNA technology; therefore, they are sometimes called recombinant vaccines. (3) Two types of pneumococcus vaccines given by injection are available: pneumococcal polysaccharide vaccine (PPSV) and pneumococcal conjugate vaccine (PCV). (4) Two types of meningococcus vaccines given by injection are available: meningococcal polysaccharide vaccine (MPSV) and meningococcal conjugate vaccine (MCV).
\nIt is well-known that prevention is better than cure for diseases control. Vaccination is the best approach to prevent infectious diseases ahead of the incidence of diseases. Vaccination can prevent or ameliorate morbidity from infection and result in herd immunity when a high percentage of population has been vaccinated. For the past centuries, some diseases are globally eradicated such as smallpox; some diseases are significantly controlled in much of the world such as polio, measles, and tetanus through vaccines distribution. The effectiveness of vaccination has been widely studied and verified such as the influenza vaccine [6], HPV (Human papillomavirus) vaccine [7], and the shingles (chicken pox) vaccine [8], etc. In 2012, the World Health Organization (WHO) reports that licensed vaccines are currently available to contribute to the prevention and control of 25 infections. From the discovery of the related pathogens, 25 infection have been effectively prevented using licensed vaccines; however, vaccines remain elusive for many important diseases, such as the infections of Zika viruses, cytomegaloviruses (CMV), hepatitis C viruses (HCV), Human immunodeficiency viruses (HIV), severe acute respiratory syndrome (SARS) viruses, Ebola viruses, respiratory syncytial viruses (RSV), and malaria parasites
Time line for the development of vaccines: 25 infections (
Vaccines have not only successfully reduced the incidence rate of many devastating diseases to be always low but also prevented many infectious diseases from persistently bursting out. At present, only few people have ever experienced the detrimental effects of many illnesses. The two main advantages of vaccination are noted as follows:
\nPrevention in advance
Vaccination can result in lower morbidity and mortality rate as well as faster recovery even though a partial, late, or weak immunity may only alleviate an infection. The early protective efficacy of vaccination is primarily conferred by the induction of antigen-specific antibodies. Vaccination can trigger adaptive immunity before contacting with the pathogen. Should a pathogen infect the body, there is no illness in that the immune responses stimulated by vaccines continuously guard against the disease. The quality of such antibody responses (avidity: a measure of the overall strength of an antigen–antibody complex) has been identified as a determining factor of vaccines efficacy. However, there is more to antibody-mediated protection than the peak of vaccine-induced antibody titers. Vaccination stimulates both humoral and cell-mediated immunity of one’s own throughout the body. This kind of defensive response is usually safer, more effective and with fewer side effects (e.g., inflammation, muscle aches, allergy, etc.) due to the active, acquired and specific immunity is established. Through vaccination, humans can eliminate or alleviate the attack of pathogens beforehand to avoid the suffering of diseases.
Immunity for long term
Vaccination stimulates adaptive immunity to create immunological memory that can maintain in the body for a long period of time. Therefore, people will be protected from the attack of diseases as long as these specific memory cells are still in the body. The disease control or elimination requires the induction of protective immunity in a sufficient proportion of the population. This is best achieved by vaccination programs capable of inducing the long term protection (adaptive immunity) that contrasts to the quick but short-lasting immune responses (innate immunity). Long-term immunity is conferred by the maintenance of antigen-specific immune effectors and/or by the induction of immune memory cells that may be efficiently and rapidly reactivated upon subsequent microbial exposure.
In spite of vaccines’ advantages for diseases control and prevention, three major limitations related to vaccines are noted as follows:
\nComplex vaccination schedules
The potency of vaccines may not be high enough in receiving immunization only once. Though the body develops antibodies, protection sometimes is not adequate due to the development of immunity might be too slow to be effective in time. Furthermore, the antibodies might not disable the pathogen completely, or there might be multiple strains of the pathogen, not all of which are equally susceptible to the immune responses. In order to provide the best protection, additional “booster” shots are often required to achieve full immunity. Vaccine schedules are usually regulated by the competent authority or physician groups to achieve maximum effectiveness. A vaccination schedule is a series of vaccinations, including the timing of all doses, which may be either recommended or compulsory. Many vaccines require multiple doses for maximum effectiveness, either to produce sufficient initial immune responses or to boost responses that fade over time. Over the past two decades, the recommended vaccination schedule has grown rapidly and become more complex as many new vaccines have been developed.
Strict requirements for storage and shipment
Vaccines are temperature sensitive and their storage and shipment are usually restricted to cold temperature [11]. Potency can be negatively affected by exposing to extended or multiple temperature excursions (out-of-range temperatures). It is essential to have a label on the storage unit to indicate that vaccines should be stored in the freezer (−20 or −80°C) or in the refrigerator (2–8°C). Correct cold-chain (a temperature-controlled supply chain) procedures must be followed during production, warehousing, and transportation, that is, vaccines have to be stored properly from the time that they are manufactured, distributed until administered to the people. Most live, attenuated vaccines tolerate freezing temperatures, but deteriorate rapidly after being removed from cold temperature. Inactivated vaccines will be damaged by exposing to temperature fluctuations. Cold-chain failure has to be evaluated in addition to routine tests such as potency, sterility, toxicity, safety, chemical, visual, and pyrogen test.
Restricted routes of administration
Most vaccine products are administered by injection such as intramuscular (IM), subcutaneous (SC), and intradermal (ID) injection, though some can be given by oral administration or intranasal spray. IM injection administers the vaccine into the muscle mass. Vaccines containing adjuvants should be injected into muscle to reduce adverse local effects. SC injection administers the vaccine into the subcutaneous layer above the muscle and below the skin. ID injection administers the vaccine in the topmost (dermis) layer of the skin. BCG is the known vaccine with ID injection; this route reduces the risk of neurovascular injury. The professional person’s assistance is usually required for the injection administration. Oral administration vaccines (e.g., OPV and rotavirus vaccine) make immunization easier by self-administration and eliminating the need of an injector. The intranasal spray vaccine (e.g., influenza vaccine live) also offers a needle-free approach through the nasal mucosa. Percutaneous administration, such as a multiple puncture device (4–5 cm length, with 9 short needles) used for BCG vaccination in Japan, indicates that any medical procedure is done via needle puncture of the skin, rather than using an “open” approach where inner organs or tissues are exposed.
In the new era of subunit vaccines, to against both infectious diseases and cancers effectively, subunits made of proteins, peptides, DNA antigens, or potent adjuvants may provide a good choice to enhance immunogenicity. Subunit vaccines are not generally immunogenic in humans because of size, degradation, destruction, nonspecific targeting, lack of cross-presentation, and others [12]. This reveals that it is necessary to develop a new approach to package and present antigens to the immune system. Nanomedicine is an area of research that combines nanotechnology and medicine and provides new hopes for therapeutic strategies and diseases control.
\nAdvances in nanotechnology have shown to be beneficial for therapeutic strategies such as drug discovery, drug delivery, and gene/protein delivery. Through nanotechnology, drug consumption and side effects can be significantly decreased by locating the active agent at the desired site. Nanotechnology is the study and application of extremely small things and capable of being used in many science fields, such as biology, chemistry, physics, materials science, and engineering. It is referred to the particular technology with a goal to precisely manipulate atoms and molecules for fabrication of nanoscale products, which size below 100 nm (or sometimes up to 200–300 nm) [12] (Figure 2). Advances in nanotechnology in biomedicines have proved to be beneficial in vaccines development. The efficacy of vaccines can be improved and side effects also can be significantly decreased. For the past decades, a nanovaccine has become a novel approach for vaccination.
\nAdjuvants may be added to vaccines to modify the immune response by boosting it to give more antibodies and longer-lasting protection. Adjuvants have been an important component of vaccine formulations since aluminum salt (Alum) was approved in the 1920s [13] and adjuvant system 04 (AS04) was approved in 2009 in the United States, respectively. AS04 is a trade name for combination of adjuvants used in vaccine products by GlaxoSmithKline (GSK), which consists of aluminum hydroxide (Al (OH) 3) and 3-
Size comparison of biological substances: The size range of nanomaterials used in nanotechnology relative to the size of various biological system. Abbreviation: virus-like particle (VLP).
In accordance with the guidelines of WHO, US Food and Drug Administration (FDA) and EMA, vaccines belong to a heterogeneous class of agent; each has suitable, individual, and specific standards of evaluation. The major quality concerns in a vaccine human clinical trial are as follows:
\nTheoretical values and expected values of the trial: In preclinical studies stage, potency, and immunogenicity should be tested in addition to toxicity and safety. A suitable animal challenging model should be established; otherwise, an alternative method must be proposed. The contribution and clinical significance of vaccine should be explained, and a declaration of interest is usually needed.
Protection of the testers: The right and interest of testers should be emphasized either in the placebo control group (e.g., use insert placebos) or the active control group (e.g., use already licensed vaccines).
Selection of testing population group: Random health volunteers are tested in phase I, but the representation of testers’ population should be considered in phase II and III. The related factors include testers’ population, social economics, and the epidemiology of target disease, etc.
Condition of inclusion and exclusion for testing population group: Tester should be in good health condition and meet the criteria of examination, some people would be excluded such as allergy, cardiovascular disease, immunodeficiency, and neuropathy sicks.
Dosage, administration route, and schedule: Dose-response data should be achieved in the dose-finding study. The schedule of vaccination depends on the traits of antigens, the characteristics of testing population, and the pharmacokinetics induced by vaccines. Tolerance, safety, dosage, and administration routes should be evaluated in phase I. The optimal dose and schedule of vaccination should be defined in phase II and would be a reference in phase III.
Safety evaluation of testers: The adverse events should be recorded and evaluated in all phases of the clinical trial. Post-marketing surveillance for side effects in phase IV is usually required.
Limitation of concomitant administration: The interaction between drugs and vaccines is rare, but it is necessary to record the consequence while concomitant administering them.
Efficacy and effectiveness of vaccines: Vaccine efficacy is the measure of the reduced disease incidence after vaccination, compared with the unvaccinated group. Vaccine effectiveness is the measure of vaccine induced protection (not population related protection), which is influenced with many factors such as the protection coverage of vaccination, the immunological state of testers, and the epidemiology of target disease.
Lot-to-lot consistency study: It is necessary to evaluate the consistency of vaccine production and performance by comparing the different lots of vaccine products by the predefined standard (usually various parameters in immunology).
Vaccines, as all medicinal products regulated by national regulatory authorities (NRA) and national control laboratories (NCL), undergo a rigorous review of laboratory and clinical data to ensure their safety, efficacy, purity, and potency. In the stages of discovery and development of vaccines, the manufacturer has to comply with the related regulation, including nonclinical work, consultation, preclinical trials, and human clinical trials. Furthermore, vaccines approved for marketing may also be required to perform additional studies to give further evaluation and often address specific questions such as safety, efficacy or possible side effects and contraindications for post-marketing surveillance (Figure 3).
\nLot release is an independent assessment of each lot of a licensed vaccine or biologic prior to it is released onto the market. General practices for lot release involve the review of manufacturers’ production data and quality control test results by NRA and NCL. These practices can be performed by an agency or a contracted laboratory for the NRA. It may be supplemented by other documents such as the release certificate from the responsible NRA/NCL and in some circumstances, by tests in the other laboratory which is independent of the manufacturers’ quality control testing.
\nFrame work of vaccine research and development: The research, development, surveillance, and application of vaccines involve several practices performance and regulation requirements. (A) Stage: Vaccines discovery, development, and postmarked surveillance are three stages for the application of new vaccines. (B) Studies: There is much research work to do at every stage. (C) Phase: Each stage can be divided into different phases. (D) Regulation: Various regulations or guidelines are required for every stage. (E) Special procedure: fast track and communication. Abbreviations: Good Laboratory Practice (GLP); Good Clinical Practice (GCP); Good Distribution Practice (GDP); Investigational New Drug (IND), Pharmaceutical Inspection Convention and Pharmaceutical Inspection Co-operation Scheme/Good Manufacturing Practice (PIC/S GMP).
Fast-track procedures can be implemented under special circumstances, such as disease outbreaks or an acute shortage in global vaccine supply. It is a process designed to facilitate the development and expedite the review of drugs that show promise in treating a serious or life-threatening disease and address an unmet medical need as follows:
\nSerious condition: To determine if the drug will have an impact on such factors as survival, day-to-day functioning or the disease will progress more serious if left untreated such as acquired immune deficiency syndrome (AIDS), Alzheimer’s disease, heart failure, cancer, epilepsy, depression, and diabetes.
Unmet medical need: The drug must be developed to treat or prevent a disease that does not have a current therapy. If there are available therapies, a fast-track drug must show some advantage over available therapy such as showing of superior effectiveness, avoidance of serious side effects, improvement of the diagnosis, decrease of clinically significant toxicity, and the need of emerging or anticipated public health.
The purpose of a fast-track designation is to get important new drugs to the patient earlier through the consultation system. Once a drug receives fast-track designation, early and frequent communication between the NRA and a drug manufacturer is encouraged throughout the entire development and review process of drugs. The communication ensures that all questions and issues can be solved quickly, often leading to earlier drug approval and access by patients. For example, US FDA is able to offer accelerated approval or priority review if the requisite criteria are met. Priority review rapids the review process for a new drug from 6 to 10 months even can have US FDA make a final decision within 60 days.
\nIn the application of nanoparticles as vaccines, some obstacles need to be overcome such as stability during manufacturing and storage, sterilization by nonthermal methods, and reproducibility of formulation during large-scale production [16]. Reproducibility of formulation during manufacturing is one of the major hurdles [17]; for instance, size-dependent immunogenicity has been reported [18] and it is observed that nanomaterials may change toxicity only by changing size and shape but not composition [19]. If the nanoparticles are cleared slowly over a long period of time, they may induce toxic effect. In addition, large counterparts may accumulate in vital organs to cause toxicity, even though small nanoparticles can be removed quickly from the body. Because of the toxic concerns of nanomaterials, the potential effects on the human body, public health, and environment pollution should be taken into account. It is necessary to evaluate the effects on DNA damage, gene expression, cell death, and growth in the body when different types of cells are exposed to nanoparticles. The safety surveillance and quality control of nanovaccines are as central as the evaluation of their efficacy. Regulatory policies are based mainly on the purity and safety of the vaccine. For risk management, some special regulations or guidelines for nanotechnology application in vaccines are as follows.
\nThe approach to nanotechnology products\n
Product-focused and science-based regulatory policy
Attention to nanomaterials being incorporated into standing procedures
In legal standards for different product-classes
The regulation of pharmacopoeia (European Pharmacopeia 7.0 and 8.0; EP7.0, 8.0)\n
General provisions of products: The production method is validated to demonstrate that the product would comply with the test for abnormal toxicity for immunosera and vaccine for human use. Additionally, this method has to yield consistent vaccines comparable with the vaccine of proven clinical efficacy and safety in humans.
Preparation of nanoparticles (virosomes): Inactivated influenza virions are solubilized using a suitable detergent and purified by high-speed centrifugation to obtain supernatants containing mainly influenza antigens. After the addition of suitable phospholipids, virosomes are formed by removal of the detergent either by adsorption chromatography or another suitable technique.
Tests of final products: residual infectious virus, pH, phospholipids, ratio of hemagglutinin to phospholipids, antimicrobial preservative, free formaldehyde, ovalbumin, total protein, sterility, virosome size, and bacterial endotoxin
Preclinical efficacy and toxicity testing of nanoparticles\n
Physicochemical characterization: size measurement, topology, molecular weight, aggregation, purity, chemical composition, surface characteristics, functionality, zeta potential, stability, and solubility test
Inflexal® V, Epaxal®, GardasilTM, and CervarixTM, marketed in many countries for over 10 years, are four common vaccine products based on micro/nanotechnologies. They use nanoparticles-virosomes (special liposomes) or virus-like particles (VLPs) as a delivery and adjuvant system, and all should be administered by IM and stored at 2–8°C (Table 2).
\nProduct | \nInflexal® V | \nEpaxal® | \n(1)GardasilTM and (2)CervarixTM | \n
---|---|---|---|
Indication | \nInfluenza | \nHepatitis A | \nCervical cancer | \n
Formulation | \na mixture of three monovalent virosome pools | \na vaccine adjuvanted with IRIV | \n(1)Aluminum adjuvant composed of AAHS (2)AS04 adjuvant composed of Al(OH)3 and MPLA | \n
Nanoparticle | \nVirosome | \nVirosome | \nVLP | \n
Size in diameter | \n150 nm (100–300 nm) | \n150 nm (100–300 nm) | \n20–60 nm | \n
Comparison of different vaccine products based on nanotechnology.
Abbreviations: adjuvant system 04 (AS04); amorphous aluminum hydroxyphosphate sulfate (AAHS); aluminum hydroxide (Al(OH)3); immunopotentiating reconstituted influenza virosomes (IRIV); 3-
A virosome is a kind of vaccine delivery tool with a typical size in diameter about 150 nm. It consists of a phospholipid membrane vesicle incorporating viral-derived proteins and tends to form a pure fusion-active vesicle. Virosomes are only able to fuse with target cells without replicating inside their hosts. In contrast to liposomes, virosomes contain functional viral envelope glycoproteins: influenza virus hemagglutinin (HA) and neuraminidase (NA) intercalated in the phospholipid bilayer membrane. Virosomes represent an innovative, extensively applicable adjuvant and carrier system and are used in areas beyond conventional vaccines.
\nVirus-like particles (VLPs), derived from viruses and having no genetic materials, are self-assembling nanoparticles (20–60 nm in diameter) that expose multiple epitopes on their surface and resemble native virions [20, 21]. VLPs can differentiate themselves from soluble recombinant antigens, and their structure is capable of inducing stronger immune responses. They also can express one or several viral structural proteins in a recombinant heterologous system [22], for example, the recombinant baculovirus system results in the expression of massive and various virus proteins. It is possible to engineer vaccines with multiple viral epitopes to elicit protective immunity due to the availability of viral three-dimensional structures [23]. VLPs co-administered with oil, mutant
Influenza is an important respiratory infection of humans, responsible for many deaths worldwide every year. The influenza viruses are genetic variable due to continuous and gradual mutation as well as rearranged genome segments between viruses [26]. Frequent mutations of the surface glycoproteins HA and NA result in the gradual evolution of viral strains (antigenic drift). Consequently, it is required to develop a version of new influenza vaccine almost every year in that infectious viruses can no longer be neutralized effectively by the antibodies raised against previously circulating strains.
\nInflexal® V, an influenza vaccine, is composed of a mixture of three monovalent virosome pools, each formed with specific glycoproteins HA and NA of one influenza virus strain. The vaccine is completely biodegradable and contains no thiomersal, formaldehyde, and very low levels of ovalbumin. Virosomes, reconstituted by nanotechnology, can deliver influenza antigens to stimulate a strong immune response; thus, this product is highly efficacious by mimicking natural viral infection [22]. The product has been approved by many countries and is the only adjuvanted influenza vaccine licensed for all age groups.
\nHepatitis A is an acute infectious disease of the liver caused by the hepatitis A virus (HAV), which symptoms include nausea, vomiting, diarrhea, jaundice, fever, and abdominal pain. Around 10–15% of people experience a recurrence of symptoms during the six months after the initial infection. Acute liver failure may rarely occur with this being more common in the elderly.
\nEpaxal®, a hepatitis A vaccine, is adjuvanted with immunopotentiating reconstituted influenza virosomes (IRIV). This vaccine is based on formalin inactivated HAV adsorbed to the surface of virosomes that replace the adjuvant-aluminum hydroxide with phospholipids. A single injection of virosomal HAV vaccine is well tolerated and highly immunogenic, with 88–97% of seroprotection 2 weeks after the first dose (reinforcing immunization: one dose at 6–12 months after the first dose).
\nHPV is a sexually transmitted infection and results in serious illness including genital warts and cervical cancers. Cervical cancer is the second most common cause of cancer deaths in females worldwide, only next to breast cancer [27]. HPV causes many cervical cancer cases every year, of all the cases, type 16 and 18 are responsible for about 70%.
\nGardasilTM and CervarixTM are subunit vaccines containing VLPs assembled from the major capsid protein (L1 protein) of HPV type 6, 11, 16, and 18 (GardasilTM) and type 16 and18 (CervarixTM) [21]. Vaccination with these two products has been shown to protect women against a high proportion of precursor lesions of cervical cancer caused by HPV.
\nGardasilTM is a sterile liquid suspension prepared from the highly purified VLPs composed of L1 proteins of HPV types 6, 11, 16, and 18. The L1 proteins of these HPV types (6, 11, 16, and 18) are produced by separate fermentations in the recombinant
CervarixTM is a preparation of purified VLPs composed of L1 proteins of HPV type 16 and 18, which is formulated in AS04 adjuvant-containing aluminum hydroxide and 3-
The advantages that nanotechnology-based vaccines may over conventional vaccines are as follows:
\nSubstitution for traditional adjuvants: Nanovaccines demonstrated immunogenicity similar to conventional vaccines without the presence of traditional adjuvant (Alum) which is found to cause irritation.
Extended history of safety and shelf life: Nanoparticles as vaccine delivery systems avoid the use of toxic substances such as thimerosal and formaldehyde and potentially extend the stability of vaccines on the shelf.
Less frequency of administration for boost: Nanovaccines can be administered for reinforcement immunization less frequently than conventional vaccines, which are usually multi-injection or multi-dose delivery systems.
Nanotechnology may provide us a new choice for safe and effective vaccines. However, frequency of boost, refrigeration of vaccines, and routes of administration are still challenges for vaccines distribution in small areas or villages of some countries. Nanovaccines are expected to be more effective, convenient, and economic than conventional vaccines (Figure 4). We will have a great future ahead in nanovaccines if the following three aims are achieved.
\nControlled release at specific location
Biodegradable polymers are superior to nondegradable polymers because the former may not need additional removal procedures. For reliability and reproducibility, synthetic biodegradable polymers are the best choice for antigen encapsulation in single-dose vaccine production. Biodegradable polymer microsphere such as poly-1-lactic acid (PLA) and poly lactic-co-glycolic acid (PLGA) has been used to control the time and rate of vaccine antigen release due to their safety and ability to provide long-term controlled vaccine antigen release [29]. Sustained release and pulsatile release are two possible types of vaccine antigen release with biodegradable PLGA microspheres. Sustained release, continued vaccine antigen diffusion after the initial release, mimics the administration of several small boosters. Pulsatile release, second vaccine antigen diffusion distinct from the first release, mimics the current vaccination schedule [30]. PLA and PLGA allow encapsulation within a hydrophobic core or adsorption to the hydrophilic shell, which can manipulate to encapsulate vaccine within the interior [31, 32]. For example, tetanus toxoid (TT) can be used to prevent tetanus which is caused by the bacterium
Nanoscale carriers can enhance the potency of vaccines to develop safer and more effective vaccine formulations. A nanoparticle-based strategy was demonstrated to safely deliver and detain intact toxins to mount a potent anti-toxin immune response. A biomimetic nanoparticle cloaked in biological membranes was used to sequester membrane-active toxins. As the event of contacting with the nanoparticles, the toxins will retrained and lose toxicity in that they are prevented from interacting with cellular targets. The resulting particles and toxin complex adopts a nanoparticulate morphology that facilitates the intracellular delivery of toxins. This technique assists to design a novel toxoid vaccine that promises more effective anti-toxin immune responses. The potent nanotoxoid formulations provide a viable anti-virulence approach to combat with microorganisms that contain membrane damaging toxins such as
Stable at room temperature
The common adjuvant -alum is known to cause irritation if it is used in the conventional hepatitis B vaccine. However, the use of needle-free nasal immunization with a combination of nanoemulsion and hepatitis B antigen was found to be tolerable, effective, and safe without side effects [36]. The solvent extraction or evaporation from a water-in-oil-in water (W/O/W) emulsion is usually used to prepare the antigen-encapsulated nanoparticles [37]. Interestingly, refrigeration was not required for this nanoemulsion, as it was effective at 25°C for one month and at 40°C for 6 weeks, respectively [16].
To make an effort to develop nanoparticles from biodegradable and biocompatible polymers (e.g., PLA, PLGA) as vaccine delivery systems can induce both humoral and cell-mediated immunity. These biodegradable polymers may make nanovaccines have extended history of safety and proven biocompatibility at room temperature. The additional advantage is their property to control the time and rate of polymer degradation and antigen release [38].
Replaceable routes of administration
In addition to injection (IM, SC or ID), mouths and nostrils are two successful alternative routes for administration. The vaccine can be given by these two routes without the help of trained health workers and the equipment of sterile injection. These two methods are relatively effective, inexpensive, and convenient, compared with injection. For example, Sabin vaccine (OPV) and FluMist® are administered by oral administration and intranasal spray, respectively, in contrast to Salk vaccine (IPV) and influenza vaccine (flu shots), which are both given by injection.
A. Sabin vaccine: An OPV is indicated for the prevention of poliomyelitis, which trivalent contains a mixture of attenuated strains of three poliovirus types. The live polioviruses vaccine produces antibodies to all types of poliovirus and triggers a local immune response in the mucous membrane of the intestines. Once infected with wild polioviruses, these antibodies protect against paralysis by restricting the viral replication inside the intestine to avoid invading to the nervous system. Later, the live attenuated vaccine virus would be excreted in the faces and spreaded to others in close contact. This means that OPV may result in the passive immunization of unvaccinated people, so that OPV can rapidly stop person-to-person transmission of wild poliovirus in areas where there is poor public health.
\nB. FluMist®: An intranasal spray attenuated vaccine is indicated for the prevention of influenza which quadrivalent contains four virus strains (a type A/H1N1, a type A/H3N2, and two type B strains). The live, attenuated influenza vaccine (LAIV) contains neither thimerosal nor preservatives and is sprayed into the nostrils. LAIV produces a significantly stronger immune response than inactivated vaccines, because it is delivered via the natural site of entry of the influenza virus [39–41]. In addition, this intranasal spray vaccine can prevent more 50% cases than the flu shots given by IM in younger children [42].
\nNanotechnologies applied in biomedicines’ vaccine (a cartoon figure for the perspective): Nanoparticles used in the vaccine production potentially overcome the disadvantages of conventional vaccines. (A) Vaccines encapsulated in nanoparticles which are polymers in solid form may be assistance in the stabilization at room temperature, elongation of storage time, exploration of alternative administration routes, and facilitation of controlled release. (B) Nanovaccines may only need to be boosted once; however, conventional vaccines usually need to be boosted many times. (C) Soluble antigens released from nanovaccines may induce both humoral immunity (B cell responses) and cell-mediated immunity (helper and cytotoxic T cell responses). *Ab (antibody): serum immunoglobulin G (IgG), mucosal IgG and mucosal IgA. Abbreviations: antigen (Ag); antibody (Ab); antigen presenting cell (APC); antibody forming cell (AFC); antibody-dependent cytotoxic cell (ADCC); cytotoxic T cell (Tc); granulocyte (GC); helper T cell (Th); macrophage (MAC); memory cell (MC); natural killer cell (NKC).
The skin is also an attractive organ for vaccine delivery because of its large surface area that is easily accessible for administration, and dermal delivery is almost pain-free [43, 44]. Furthermore, the skin contains many antigen presenting cells (e.g., Langerhans cells and dermal dendritic cells) and is available for the induction of immune responses [44–46]. Besides, the delivery via the skin circumvents the problem of swallowing, the first-pass effect of the liver, and the absorption and stability of vaccines in the gastrointestinal tract. However, the stratum corneum (the outermost layer of the epidermis) barrier may form significant resistance for vaccines to permeate into the body. To overcome the stratum corneum barrier, percutaneous administration such as microneedle arrays (needle-like structures with a length of less than 1 mm) [43–46] is one of the most promising delivery systems.
\nMicroparticles or nanoparticles (e.g., PLA, PLGA) have been proved to be capable of making an inactive
Nanotechnology has extensively been utilized in the drug preparation and delivery for clinical applications. Nanomedicine has already been an attractive research area and the development of nanovaccines is one of the most promising enterprises. A variety of nanoparticles in vaccines have been employed as a delivery tool or immunity booster, inducing not only the highest efficacy but also the least side effects. The significance of nanovaccine development is to make it practical to have a perfect vaccination system including refrigeration-free, single-dose, and painless administration. Indeed, we might have effective, safe, convenient, and cheap vaccines through nanotechnology in the near future.
\nMycotoxins are secondary toxic metabolites with a wide variety of chemical structures synthesized by fungi (mold) [1]. Mycotoxins are thought to be a kind of “chemical defense system” to protect mold from insects, microorganisms, nematodes, grazing animals, and humans [2]. Molds reproduce by means of spores, and their small molecular weight spores are easily disseminated to environment by wind. They cannot be affected by the adverse environmental conditions and can be present in the latent state for long periods. Moreover, when the environmental conditions are appropriate, spores return to vegetative form and can form into new mold colonies. Agricultural products can be contaminated with mold in pre-harvest via insect and bird damage and harsh weather condition damage such as hail damage. In addition, selected harvesting method is one of the most important reasons in contamination of the mold to the products. Improper storage, transport, and marketing can also cause the mold growth and synthesis of mycotoxins [3].
Mycotoxin can occur in food and agricultural products via many contamination pathways, at any stage of production, processing, transport, and storage (Figure 1) [4]. Factors that affect mold growth and mycotoxin production are temperature, relative humidity, fungicides and/or fertilizers, interaction between the colonizing toxigenic fungal species, type of subtract and nutritional factors, geographical location, genetic requirements, and insect infestation [5, 6].
Factors affecting mycotoxin occurrence in the food and feed chain [
Approximately 400 fungal secondary metabolites are known to be toxic, and one quarter of agricultural products have been reported to be contaminated with mycotoxins in the world [5, 6, 7, 8, 9]. While a type of mold may form more than one mycotoxin, a mycotoxin can be synthesized by many molds. The most common types of mold which are known to produce mycotoxins are
According to the result of many studies in poultry and mammals, mycotoxins can be carcinogenic, mutagenic, teratogenic, hepatotoxic, nephrotoxic, immunosuppressive, and embryotoxic [11]. The phenomenon of toxicity is called mycotoxicosis occurring after consumption of mycotoxin-contaminated product by human and animal [12].
Especially cereals, grains, nuts, oilseeds, fruits, dried fruits, vegetables, cocoa and coffee beans, wine, beer, herbs, and spices are major mycotoxin vectors since they are consumed by a large mass of people and animals [4]. Mycotoxins cause different degrees of toxicity according to exposure time, mycotoxin amount, physiological state, and sensitivity of the organism in humans and animals.
In addition to risk of public health, mycotoxins generate high level of economical loses for food industry due to reduced crop yields, lost trade revenues (local and international), and livestock illnesses [13, 14]. Elimination of mycotoxin is quite though due to resistant to physical, chemical, and biological methods; however, some of the measures described in the following sections may help to prevent mycotoxin. The methods used for mycotoxin determination are chromatography such as high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC), gas chromatography-mass spectrometry (GC-MS), and also enzyme-linked immunosorbent assay (ELISA) technique and biosensor-based screening methods [15]. Detection is complicated due to limitations in analytical methodology [16]. Therefore, prevention of mold contamination and mycotoxin synthesis is essential for food safety in food industry.
According to the Food and Agricultural Organization (FAO), 77 countries have established guidance and regulations on mycotoxin in food and feed to control the level of mycotoxin. On the other hand, 13 countries including African countries still do not have specific regulation for food safety [4].
Ergotism is one of the oldest determined mycotoxicoses (disease) in human and results from consumption of the ergot body in rye or other grains infected by a parasitic fungus of the genus
Mycotoxins can occur in the food in several ways (Figure 1), but technically divided into two groups; first is mold growth as a pathogen plant in field, another one is grow on stored. After plant materials are contaminated with mold spores from soil and air, they easily contaminate other food source, production area, laboratory, and even kitchen of our homes. Certain species of mold are capable of mycotoxin synthesis; therefore, each food contaminated with mold always may not contain mycotoxins. Nevertheless, moldy products are considered to be risky products in terms of mycotoxin.
Mycotoxins appear in almost all kinds of animal feed and products such as wheat bran, noug cake, pea hulls, maize grain, milk and meat, and also human food such as cereal, fruit and vegetables, spice, etc. [5]. Consuming these foods creates serious health risks in human and all animal species. Mycotoxin intake by feed or food causes chronic intoxication rather than acute symptoms. Acute toxicity is observed in high-dose mycotoxin exposure, and symptoms show a rapid effect such as borborygmy, abdominal pain, diarrhea, etc. On the other hand, low-level mycotoxin exposure in long period causes serious impairments in the liver, kidney, and immune system organs and tissues. Therefore, mycotoxin plays a significant role in cancer in these organs [2]. Some important mycotoxin health effects are shown in Figure 2. Toxic effects on humans and animals of important mycotoxins are shown in Table 1 [19].
Aflatoxin (AFL), ochratoxin A (OTA), patulin (PAT), fumonisin (FUM), trichothecenes (TCT), and zearalenone (ZEA) mycotoxin health effects [
Mycotoxins | Genus/species | Major food | Toxic effects and diseases |
---|---|---|---|
Aflatoxin | Cereals, feeds, oilseeds and pulp, coconut | Carcinogenic, hepatotoxicity, teratogenicity, decreasing immune systems, affecting the structure of DNA, hepatitis, bleeding, kidney lesions | |
Fumonisin | Cereals, corn | Encephalomalacia, pulmonary edema, carcinogenic, neurotoxicity, liver damage, heart failure, esophageal cancer in humans | |
Ochratoxin OTA | Cereals, herbs, oil seeds, figs, beef jerky, fruits, and wine | Kidney and liver damage, loss of appetite, nausea, vomiting, suppression of immune system, carcinogenic | |
Patulin | Silage, wheat, feeds, apples, grapes, peaches, pears, apricots, olives, cereals | Neural syndromes, brain hemorrhage, skin lesions, skin cancer, lung, mutagenicity, antibacterial effect | |
Trichothecenes (T2, DON, DAS, HT2) | Cereals, feeds, silage, legumes, fruits, and vegetables | Immune suppression, cytotoxic, skin necrosis, hemorrhage, anemia, granulocytopenia, oral epithelial lesions, GIS lesions, hematopoietic, alimentary toxic aleukia (ATA), hypotension, coagulopathy | |
Zearalenone | Cereals, corn, silage, timothy grass, fodder | Carcinogenic, hormonal imbalance estrogenic effect, reproductive problems, teratogenic |
Name of some important mycotoxin-producing fungi, susceptible foods, and mycotoxin effects on humans and animals [19].
Mycotoxins have caused many serious outbreaks worldwide. There was an outbreak that occurred in 1967, and 26 people were poisoned because of the consumption of moldy rice for up to 3 weeks in Taiwan [21]. An outbreak of aflatoxicosis affecting humans, reported in India, led to the death of 100 people in 1974 [22]. Another outbreak was reported in India in 1995, affecting 1424 people due to sorghum and maize contaminated with fumonisin [23]. During January–June 2004, an aflatoxicosis outbreak in eastern Kenya resulted in 317 cases and 125 deaths [24].
Mycotoxin contamination in foods and fodder has been becoming a global concern day by day. According to Food and Agricultural Organization (FAO) reports, it is estimated that mycotoxin affects nearly 25% of the world’s crop each year and is causing huge agricultural product and industrial losses in billions of dollars [25]. For example, estimated annual loss in the United States is approximately $ 0.5–1.5 billion [19]. The main effects of mycotoxins on national economies can be thought in five ways:
Product yield losses due to toxigenic mold diseases
Decrease in commercial value because of contaminated food and feed
Human and animal health losses due to harmful impacts associated with mycotoxin-contaminated food and fodder consumption
Cost of analysis of mycotoxin
Strategies to control mycotoxin contamination
Economic impacts are felt by agricultural chain such as manufacturer of plant and animal, especially cereal industry, consumers, and briefly all farm-to-fork steps.
Aflatoxins are a group of toxic secondary metabolites of filamentous fungi,
Ochratoxin A (OTA) is a natural mycotoxin produced mainly by fungal type of
Fumonisins are generated by various fungal species such as
The International Agency for Research on Cancer (IARC) identified FB1 as possibly carcinogenic to humans (group 2B). Recent studies reported that FB1 causes an increased prevalence of esophageal and liver cancer in humans [59]. Furthermore, this mycotoxin has been found to have toxic effects against several organs (nervous and cardiovascular systems, liver, lung, kidney) in animals [60]. Fumonisins are largely found in corn and corn-based foods and also FB1 in rice, beer, sorghum, cowpea seeds, triticale, beans, asparagus, and soybeans [61].
Zearalenone (ZEA), known as an estrogenic mycotoxin, is a secondary metabolite produced by
Several in vivo studies found that ZEA disrupts hormonal balance due to its similarity to naturally occurring estrogens [64]. The mycotoxin has high affinity for estrogen receptors, causing reproduction and fertility disorders in mammals [65]. In addition, it is known that progressive exposure to endocrine-modulatory compound has been linked with carcinogenesis in human [64]. According to the European Food Safety Authority (EFSA) report in 2014, the bioavailability of toxin is up to 80% in human and animals such as rats, rabbits, and pigs [66]. Moreover, recent works report ZEA is metabolized in the liver and has shown hepatotoxic, immunotoxic, carcinogenic, and nephrotoxic effect in animal tests [67, 68, 69]. As this mycotoxin possesses such consumer health risks, the European Union (EU) has prescribed the limits of ZEA (20–350 μg/kg) for various processed and unprocessed cereals [66].
Trichothecenes are a large group of mycotoxins produced predominantly by
The mechanism of action of trichothecenes is based on the inhibition of protein synthesis in eukaryotes. This mycotoxin affects peptidyl transferase enzyme binding the 60S ribosomal subunit, thus causing the inhibition of protein translation and ribotoxic stress [75]. Also, Pestka reported these groups of mycotoxins cause immunosuppression or immune stimulation by affecting the leucocytes [76].
The family of trichothecenes has a significant impact on cereal and grain production due to health risk for human consumption, livestock feed, or malting purposes [77, 78]. According to report from the FDA, economic losses associated with mycotoxin ranges from USD 0.5 million to over USD 1.5 billion from aflatoxin (corn and peanuts), fumonisin (corn), and deoxynivalenol (wheat) in the United States. [72]. Hence, control of these mycotoxins is essential for human and animal health and economic reasons.
Deoxynivalenol (DON), known as vomitoxin, is the most commonly detected trichothecenes in grains such as wheat, barley, oats, rye, and corn and less often in rice, sorghum, and triticale [79]. Even though NIV presence of cereals appears generally to be lower than DON [80], it has been reported that the occurrence of NIV in of wheat and barley is as prevalent as that of deoxynivalenol (DON) in Japan [81]. According to animal toxicity studies, NIV shows higher toxicity than DON. The LD50 values for DON and NIV in tests in mice were 78 and 39 mg/kg, respectively, and DON and NIV, similarly to other trichothecenes, show inhibitor effect on cell metabolism such as protein, DNA, and RNA synthesis [82]. In addition, these mycotoxins affect cell division and mitochondrial functions [83, 84, 70]. Both mycotoxins exhibit major symptoms such as abdominal discomfort, diarrhea, vomiting, and inflammation of the throat, weight loss, and anorexia [85].
The World Health Organization (WHO) reported that trichothecenes shows fatal and chronic intoxications on human and livestock and also DON shows teratogenic, neurotoxigenic, and immunosuppressant effects [86].
According to the conducted BIOMIN World Mycotoxin Survey, DON appeared in 81% of livestock feed from 81 countries worldwide followed by fumonisins that were detected in 71% of samples. Therefore, DON is reported as the most common mycotoxin worldwide (https://www.biomin.net/en/biomin-mycotoxin-survey/).
Food safety is a key component in public health issue, and a mycotoxin is a huge food safety risk in developing countries. Prevention is the most important and effective way in reducing fungal growth and mycotoxin production to ensure food safety. The following steps that explain prevention and control of mycotoxin occurrence include good agricultural practices (GAP) in field, control practices of harvesting and storage, physical methods (cleaning, milling, etc.), implementation of biotechnological application, biological control through the use of controlled atmosphere during storage, detoxification/degradation, and fermentation techniques.
Pre-harvesting is considered first and one of the most important stages to prevent mold growth and mycotoxin synthesis. Several strategies are available for the produce of healthy products and reduce the mold formation at pre-harvesting, including selection of plants according to the soil structure and production capacity, use of plant which is resistant to fungi and insects, irrigation time, make fertilization, use of insecticides to prevent insect damage [87].
Harvesting at the appropriate time periods (low moisture and full maturity) is essential for reducing the risk of a mycotoxin contamination since overmaturity creates sensitivity to mold growth. Additionally, suitable harvesting equipment and procedures should be used, and crops should be dried after maturity to both reduce grain moisture to safe levels [88].
The latest technological advances provided new paths in mycotoxin control strategies that include the use of a controlled atmosphere with inhibitory or a protective effect and use of naturally occurring compounds under different conditions and essential oils with antioxidant properties to decrease fungal growth and mycotoxin production in grains during storage [89]. Moreover, these strategies also include using regularly cleaned transport vehicles to prevent cross contamination of products; monitoring of temperature, humidity, aeration and pest infestation periodic during storage [90]; using mold inhibitors (propionic acid) to contaminated food and feed; and application of disinfectant such as sodium hypochlorite to storage area [91].
Some studies have shown that using physical methods (dehulling, washing, sorting, and cleaning of visible moldy seed) reduces different mycotoxin species in foods regardless of grain genre [70]. Scudamore and Pascale et al. [92] and Patel [93] observed a reduction of T-2 (62%) and HT-2 (53%) and DON (50%) in wheat seeds after cleaning. Scudamore and Patel also reported a 32% reduction in fumonisin levels in corn in an industrial enterprise [94]. Moreover, milling is an important effect in the reduction of
One of the best applicable strategies for the prevention of mycotoxin formation is the cultivation of fungal infestation-resistant plants and improvement of the genetic composition to suppress mycotoxin production [96]. The benefits of biotechnological applications were observed with Aflasafe. Aflasafe is a biocontrol product that includes a blend of four fungal species covered over grains which reduce aflatoxigenic fungi that produce AFs in maize and groundnuts (https://aflasafe.com/).
Mycotoxins are resistant to heat and cannot be completely destroyed under normal cooking process. On the other hand, mycotoxin reduction has been determined after heating, and this may be the result of reactions changing the chemical structure [70]. Ryu et al. reported heat treatment (at temperature 120–160°C) causes a reduction between 66 and 83% of ZEN [97]. Scott and Lawrence also reported a reduction of 60–100% of fumonisins with a heat treatment at 190°C (60 min) and 220°C (25 min).
Biological control of mycotoxins via detoxification/degradation offers a promising alternative method [98]. Recently the effectiveness of fermentation for the reduction and elimination of mycotoxins has also been proven. Studies documented in the literature generally show that mycotoxins are reduced by conversion, detoxification, binding, degradation, and decontamination after food fermentation [99]. Modification of the chemical structure of the mycotoxin molecule, removal or detoxification/inactivation, and adhesion to bacterial cell walls provide a reduced toxicity during fermentation [99]. Implementation of these preventive methods cannot solve the problem alone; also it must be an integral part of an integrated food safety management system based on the hazard analysis and critical control point (HACCP).
HACCP is a food management system where food safety is addressed through the analysis, control, and monitoring of physical, chemical, and biological hazards from raw material manufacturing, supply, and handling to production, distribution, and consumption of the finished product [100]. The National Advisory Committee on Microbiological Criteria for Foods (NACMCF) published a guideline about HACCP containing seven basic principles, decision tree, and all plans in 1992 [101]. Implementation of HACCP is an effective strategy for prevention, control, and periodic monitoring of mycotoxin in all stages from field to the consumer. There are 12 successive steps recommended to implementation of HACCP system. Previous HACCP studies can be researched to set up tasks from 1 to 5 that specify each food process, and tasks required for mycotoxin control begin at 6 (Principle 1).
Establish the HACCP team.
Describe the product.
Identify the product’s intended use.
Draw up the commodity flow diagram.
Confirm the flow diagram on-site.
Identify and analyze hazard(s) (Principle 1).
Determine the critical control points (CCPs) (Principle 2).
Establish critical limits for each (CCP) (Principle 3).
Establish a monitoring procedure (Principle 4).
Establish corrective action (Principle 5).
Verify the HACCP plan (Principle 6).
Keep record (Principle 7).
Crops and tolerated levels of mycotoxins (μgkg−1) | |||||
---|---|---|---|---|---|
Country | Mycotoxins | Rice | Maize | Spices | Fruit juices |
Brazil | AFB1/AFG1 | 30 | 30 | 30 | 30 |
China | AFB1 | 10 | 20 | — | — |
France | FB1 | 1000 | 1000 | — | — |
Hungary | Total AF OTA | 50 5 | 50 5 | - - | - - |
Japan | AFB1 Patulin | 10 | 10 | 10 | — |
The United States | Total AF Patulin | 20 - | 20 - | 20 - | - 50 |
Turkey | AFB1 Patulin | 2 - | 2 - | 5 - | - - |
Global regulation of mycotoxin contamination in agricultural products [103].
Microbiological and/or chemical tests can be used to confirm which product is meeting CCP.
Asking questions especially to CCP employees.
Internal or external audit by independent person to check whether HACCP system is being implemented.
Step/CCP | Hazard analysis | Monitoring | Corrective action | |||
---|---|---|---|---|---|---|
Hazard | Control | Critical limit | Monitoring | Frequency | ||
Pre-harvest/ growing | Low soil moisture leading to plant stress during kernel development | Irrigate | Lower limit of critical water activity (aw) (check with your agronomist/extension staff for an exact value) | Measure soil moisture and record | Weekly on Monday morning | Additional irrigation; record amounts |
Insufficient soil nutrients leading to plant stress during kernel development | Fertilize | N, P, and K applications as recommended for hybrid by local agronomists (insert the values) | Fertilizer applied (appropriate for soil type and hybrid); amounts and type recorded | As recommended for hybrid | Additional fertilizer; record amounts added | |
Insect attack leading to damaged kernels | Integrated pest management (IPM) plan | Insect population within acceptable limits as determined by control program | Visual inspection and sample, with results recorded | Weekly | Apply pesticide in accordance with IPM plan | |
Harvest | Damage to kernels from harvester | Harvest when kernels are dry | Moisture content ≤14% | Measure and record grain moisture | Prior to harvest | Delay harvest till kernels are dried enough |
Storage | Excessive moisture content of kernels | Do not store until kernels are dry | Moisture content ≤14% | Measure and record grain moisture | Immediately prior to storage | Dry mechanically |
Insect attack, allowing fungi to penetrate kernels | IPM plan | No evidence of insect or rodent infestation using inspection protocols specified in IPM plan | Visual inspection with results recorded | Weekly | Apply pest control methods in accordance with IPM plan | |
High ambient humidity and temperature | Aerate grain to control temperature and humidity | Temperature and humidity within limits recommended in industry literature | Measure and record humidity, ambient temperature, and airflow | Daily during storage | Adjust aeration time of day or airflow to achieve desired temperature and humidity |
HACCP plan of maize [102].
Mycotoxin is a well-known food safety risk, which is a threat to human and livestock health, and has high economic significance in food industry. Recently, the food industry has become aware of the new term modified mycotoxins introduced by Rychlik et al. (masked mycotoxin) [104]. Food safety risk has risen since masked mycotoxins which pose many difficulties including the unknown occurrence/co-occurrence of these compounds and their toxicological properties. In addition, Lorenz et al. reported that the European Food Safety Authority (EFSA) has taken into account efforts to address this emerging issue in food safety by developing strategies on how to evaluate potential added health risk due to the occurrence of modified mycotoxins [104].
Mycotoxigenic molds are difficult to prevent and control due to their widespread presence in nature. Prevention of mycotoxin synthesis in all stages of food processing is an essential point for public health and economic reasons. Many practices used for prevention of mycotoxin include good agricultural practices (GAP) in field, control practices of harvesting and storage, physical methods (cleaning, milling, etc.), implementation of biotechnological application, biological control through the use of controlled atmosphere during storage, detoxification/degradation, and fermentation techniques.
Meanwhile a number of techniques for mycotoxin control and management prove to be quite costly and/or unenforceable in some cases. On the other hand, using fermentation process for appropriate process has been recommended for mycotoxin reduction by Adebiyi et al. [99]. In the future, more emphasis should be given to nanotechnology and genetic engineering practices in the development of durable product types to ensure food safety.
In addition to these applications, food safety management systems such as HACCP, GAP, and good manufacturing practices (GMP) should be integrated at all stages of production, transport, and storage, in order to minimize contamination in food industry. Also fairly new food safety system including threat assessment critical control points (TACCP), vulnerability critical control points (VACCP), and hazard analysis and risk-based preventive controls (HARPC) should be investigated and implemented to ensure an effective control system.
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\n\nThe first step in obtaining funds for your Open Access publication begins with your institution or library. IntechOpen’s publishing standards align with most institutional funding programs. Our advice is to petition your institution for help in financing your Open Access publication.
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