Hereditary autoinflammatory syndromes with identified gene loci (adapted from Lachmann and Hawkins, 2009: 36).
\\n\\n
More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
\n'}],latestNews:[{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"},{slug:"all-intechopen-books-available-on-perlego-20201215",title:"All IntechOpen Books Available on Perlego"}]},book:{item:{type:"book",id:"117",leadTitle:null,fullTitle:"Artificial Neural Networks - Methodological Advances and Biomedical Applications",title:"Artificial Neural Networks",subtitle:"Methodological Advances and Biomedical Applications",reviewType:"peer-reviewed",abstract:"Artificial neural networks may probably be the single most successful technology in the last two decades which has been widely used in a large variety of applications in various areas. 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\r\n\tHysteresis phenomena manifest themselves with highly intriguing features that have been continuously attracting the interest of scientists and physicists who have attempted to unravel the origins and the underlying mechanisms. Hysteretic behaviour is ubiquitous to different physical systems, and despite displaying diversified characteristic elements depending on the specific context, hysteresis originates from the presence of several metastable energy minima, which relate to path-dependent and rate-dependent responses to various external stimuli. This is translated in a considerable number of experimentally observable events, such as hysteretic loops in ferroic media, Bauschinger effect in elastoplastic materials, characteristic magnetic hysteresis in superconductors, current-voltage hysteresis in various electronic devices, contact angle hysteresis in liquid-solid interfaces and bistability in cell biological processes, among others. Significant progress on understanding the mechanisms underpinning hysteresis in different systems has been made over the years and improved knowledge on how to tailor hysteretic materials behaviour for application purposes has been gained.
\r\n\r\n\tThe present book aims at compiling the main achievements and novel discoveries in the field of hysteretic materials, with particular focus on the control of materials hysteresis characteristics for engineering applications. Experimental and theoretical investigations from macro- to nanoscale on ferromagnetic, ferroelectric, ferroelastic, multiferroic materials and electronic devices, including solar cells, energy storage devices, memristors and tunnelling junctions are particularly attractive for the present compilation.
",isbn:"978-1-83881-972-9",printIsbn:"978-1-83881-971-2",pdfIsbn:"978-1-83962-472-8",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,hash:"6482387993b3cebffafe856a916c44ce",bookSignature:"Dr. Giuseppe Viola",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10568.jpg",keywords:"Thermodynamics of Hysteresis, Irreversibility, Modelling, Presaich Approach, Ferroic Materials, Hysteresis Loops, Resistive Switching, Non-Equilibrium Systems, Electronic Devices, Energy Conversion Devices, Energy Storage Devices, Actuators, Sensors",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"January 25th 2021",dateEndSecondStepPublish:"February 22nd 2021",dateEndThirdStepPublish:"April 23rd 2021",dateEndFourthStepPublish:"July 12th 2021",dateEndFifthStepPublish:"September 10th 2021",remainingDaysToSecondStep:"2 months",secondStepPassed:!0,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,biosketch:"Dr. Viola was awarded an individual Marie Curie Fellowship in 2014. His current research interests are mainly focused on the relationships between microstructure and properties of piezoelectric, ferroelectric/ferroelastic, and antiferroelectric ceramics for sensing and energy storage applications.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"173586",title:"Dr.",name:"Giuseppe",middleName:null,surname:"Viola",slug:"giuseppe-viola",fullName:"Giuseppe Viola",profilePictureURL:"https://mts.intechopen.com/storage/users/173586/images/system/173586.jpg",biography:"Giuseppe Viola (GV) graduated in Materials Engineering from Federico II, University of Naples in 2004. In 2009 he obtained his PhD from Queen Mary University of London which was focused on the study of the domain switching mechanisms in ferroelectric/ferroelastic ceramics. Afterwards, GV was employed as a Post-doctoral Reasearch Assistant at Queen Mary University of London (QMUL) and as a Materials Scientist at Nanoforce Technology Limited (a spin out company owned by QMUL). In 2014, GV was awarded an individual Marie Curie Fellowship and moved to Politecnico di Torino. In 2017, GV returned to the UK and joined UCL where he is currently based.\n\nGV's current research interests are mainly focused on the relationships between microstructure and properties of piezoelectric, ferroelectric/ferroelastic and antiferroelectric ceramics for sensing and energy storage applications. Previous research projects included the development of piezo-acoustic biosensors, investigation of domains switching mechanisms in ferroic materials and the exploitation of rapid sintering technologies for ceramics, metals and composites.",institutionString:"Independent Scientist",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University College London",institutionURL:null,country:{name:"United Kingdom"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"11",title:"Engineering",slug:"engineering"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"297737",firstName:"Mateo",lastName:"Pulko",middleName:null,title:"Mr.",imageUrl:"https://mts.intechopen.com/storage/users/297737/images/8492_n.png",email:"mateo.p@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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Unlike autoimmune disorders, autoinflammatory disorders lack the production of high-titer autoantibodies or antigen-specific T cells. These diseases primarily include hereditary syndromes (Table 1); Familial Mediterrenean fever (FMF), TNF receptor-associated periodic fever syndrome (TRAPS), hyperimmunoglobulinaemia D and periodic fever syndrome (HIDS), and the cryopyrin-associated periodic syndrome (CAPS) which involves familial cold autoinflammatory syndrome (FCAS), Muckle–Wells syndrome (MWS) and neonatal onset multi-system inflammatory disease (NOMID)/chronic infantile neurological cutaneous and articular syndrome (CINCA). Familial mediterrenean fever has been considered as the most prevalent of innate immune system disorders involving systemic autoinflammatory reaction effecting joints, skin, bones and the kidney. Systemic amyloidosis is the most severe manifestation of the disease, commonly effecting the kidneys (11% of cases), and sometimes the adrenals, intestine, spleen, lung, and testis [1]. As an innate immune system disorder, FMF is characterized by recurrent episodes of unseemingly unprovoked inflammation and fever with lasting 1- to 3-day attacks accompanied by sterile peritonitis, pleurisy, rash, arthritis, and in some cases amyloidosis leading to renal failure. this (Sohar et al., 1967). Apart from the typical implications of the disease, there is increasing evidence about the expanding clinical spectrum of FMF that embraces unusual clinical characters [2-4]. These are the rare presentations of the disease and therefore undescores the role of molecular analysis in particular for the suspicious and probable cases.
FMF is classically transmitted with autosomal recessive inheritance, and has been common among Mediterranean populations; however, previous reports have confirmed its presence worldwide. It has been describedin Mediterranean populations, including Italian, Spanish [5], Portuguese, French, and Greek, as well as in patients from Northern Europe and Japan. Nevertheless, only rare occurrences have been reported throughout the general population because of the low frequency of the causative alleles [6]. Among susceptible ethnic groups, FMF prevalence is between 1/500-1/2000, and the carrier rate is between 16-22%. Contrary to the traditionally known monogenic inheritance of the disease, it has been previously evidenced that there have been a number of patientswho have the typical FMF phenotype or FMF related symptoms with only one MEFV heterozygous mutation and/or even without any MEFV mutations [5-7], indicating the presence of clinical phenotype not only in homozygous patients, but also similarly in the heterozygous patients with mild disease.
Gene (GenBank no) | ||||
FMF (249100) | Autosomal Recessive (dominant forms are rarely presented) | MEFV (NM_000243) Ch-16 | Pyrin (marenostrin) | Childhood |
HIDS (260920; 251170) | Autosomal Recessive | MVK (M88468) Ch-12 | Mevalonate kinase | Infancy |
TRAPS (142680; 191190) | Autosomal Dominant | TNFRSF1A (NM_001065) Ch-12 | TNF-receptor type I (p55) | Childhood |
CAPS (606416) - MWS - FCAS - CINCA/ NOMID | Autosomal Dominant | NLRP3 Ch-1 | Cryopyrin | -Childhood -Infancy -Neonatal |
PAPA Syndrome | Autosomal Dominant | PSTPIP1 Ch-15 | PSTPIP1 | Childhood |
Blau Syndrome | Autosomal Dominant | NOD2/CARD15 Ch-16 | NOD2/CARD15 | Childhood |
Hereditary autoinflammatory syndromes with identified gene loci (adapted from Lachmann and Hawkins, 2009: 36).
16p13.3 chromosomally located MEFV (Mediterranean Fever) gene has been found responsible for FMF disease, and the protein product, Pyrin, is a 781-amino-acid protein [8-11]. Evolutionary conserved domains of pyrin protein involves N-terminal pyrin domain, a B-box zinc-finger, a coiled coil and a C-terminal B30.2 PrySpry domains. Pyrin protein has been reported as a component of the inflammasome complex with both pro-inflammatory and anti-inflammatory role in the cytokine regulation [10-13]. Thus, a proapoptotic or antiapoptotic role have been still not precise for the pyrin protein in NF-kB activation and apoptosis [11-16]. By means of its PYD and B30.2 interacting domains, pyrin has been shown to bind different proteins of autoinflammatory disease genes. Each interacting protein that binds through the pyrin domains (PYD) consists of PSTPIP1 [17], 14-3-3 [18], Caspase-1 [19], ASC [20], and Siva [21].
In 1997, The International FMF Consortium and The French FMF Consortium reported four missense disease associated mutations in the MEFV gene involving M694V, M680I, V726A, and M694I.Major and minor mutations of MEFV gene are well documented in INFEVERS, the database of hereditary autoinflammatory disorder mutations, and exons 2 and 10 comprises the hot-spots [22]. To date, mutations have been mostly identified in exons 2, 3, 5, and 10 of the MEFV gene. According to previous reports by Touitou I. (2001), and by the Turkish FMF study group (2005), the most common MEFV mutation in Turkey is M694V (57.0 and 51.4%, respectively), followed by M680I (16.5 and 14.4%, respectively), and V726A (13.9 and 8.6%, respectively). Moreover, no correlation has been reported between various MEFV gene mutations and the severity of the phenotype in various populations supporting the genotypic and phenotypic heterogeneity present for FMF [5, 23-25]).
According to INFEVERS [22], the database of hereditary autoinflammatory disorder mutations, To date, approximately 222 sequence variants including both missense mutations (only one nonsense mutation; Y688X) and polymorphisms have been defined in the FMF gene (MEFV), INFEVERS, 100 of them was clinically associated with the phenotype, 33 of them was not associated with the disease and the remaining was of uncertain pathogenicity. The remarkably wide clinical variability of the disease, as indicated by previous reports, has been linked to the MEFV allelic heterogeneity that underlies genotypic and phenotypic heterogeneity [23, 26, 27], and this has made detailed mutation screening critically important. In particular, Turkish FMF patients are characterized by an increased genetic heterogeneity due to various mutation frequencies from different regions, explained by the intrapopulation differentiation.
With respect to our mutation screenings, a previous comprehensive study was performed with 3430 Turkish individuals from all regions of Turkey (ages range from 2 months to 67 years; 2101 females and 1329 males) including first and second-degree relatives of individuals with FMF clinical diagnoses (including suspicious, possible, and definitive cases) who referred to the Molecular Medicine Laboratory for genetic diagnosis between years May, 2005 and December, 2010. The Tel-Hashomer and Livneh criteria were used for the clinical diagnosis of FMF based on the model of major, minor, and supportive criteria, which stipulates the presence of either 1 major or 2 minor criteria or 1 minor and 5 supportive criteria for a diagnosis. A simple set of criteria for the diagnosis of FMF required 1 or more major and/or 2 or more minor criteria [28]. None of the patients with FMF had an immunological disorder or another rheumatic disease. Active clinical presentations (fever, abdominal pain, arthritis, and myalgia) and laboratory parameters (high levels of serum amyloid A [SAA], C-reactive protein [CRP], fibrinogen, white blood cell [WBC] counts and erythrocyte sedimentation rates [ESR]) were determined for each patient. For the detection of all coding and non-coding sequence variations along the MEFV gene, we performed bidirectional DNA sequencing analysis in all 10 coding exons and exon-intron boundaries of the respective gene, and reported frequencies of common and rare nucleotide substitutions and synonymous and non-synonimous single nucleotide polymorphisms obtained in the Turkish population [7].
2 ml peripheral blood was collected into ethylenediaminetetraacetic acid (EDTA)-anticoagulated tubes by the standard venipuncture method and DNA was extracted using the QIAamp DNA Blood Isolation kit (Qiagen GmbH, Hilden, Germany) following the manufacturer’s instructions. The extracted DNA concentration was determined using a Thermo Scientific NanoDrop spectrophotometer (Wilmington, USA). The quality assessment of the extracted DNA was determined by 2% agarose gel electrophoresis.
Reverse hybridization assay (FMF StripAssay, Viennalab Labordiagnostika GmbH) was used to investigate the mutations. According to the manufacturer’s instructions, in a first step multiplex PCR was performed using biotinylated primers for exons 2, 3, 5, 10 amplification. PCR products were selectively hybridized to a test strip presenting a paralel array of allele-specific oligonucleotide probes which includes 12 MEFV mutations [E148Q, P369S, F479L, M680I (G/C), M680I (G/A), I692del, M694V, M694I, K695R, V726A, A744S, R761H]. Hybridizations were illuminated by the reaction of streptavidin-alkaline phosphatase and color substrate.
Hot-spots, exons 10, and 2; with 3 and 5, and when necessary exons 1, 4, 6, 7, 8 and 9 of the MEFV gene were analyzed for MEFV mutations by PCR amplification followed by automated DNA sequence analysis. One microliter (100 ng) of genomic DNA was added to Polymerase Chain Reaction (PCR) amplification buffer containing 20 mM Tris (pH 8.3); 50 mM KCl; 1.5 mM MgCl2; 0.2 mM each of dATP, 2’-deoxycytidine 5’-triphosphate, dGTP, and 2’-deoxythymidine 5’-triphosphate; 10 pmol each of reverse and forward primers provided by Invitrogen; and 1.0 U of PlatiniumTaq DNA Polymerase (Invitrogen, Carlsbad, CA) in a total volume of 25 µl. The cycling conditions included a hot-start denaturation step at 95°C for 10 min, followed by 35 amplification cycles of denaturation at 95°C for 30 s, annealing at 61°C for exon 10, 58°C for exons 2 and 3, or 57°C for exon 5 for 40 s, and elongation at 72°C for 45 s; a final extension was performed at 72°C for 7 min (the oligonucleotide sequences are available upon request). Prior to sequencing, PCR products were purified using an ExoSAP-IT PCR Product Clean-Up kit. BigDye Terminatorv3.1 Cycle Sequencing Kit(Applied Biosystems, San Diego, CA, USA) was used in cycle sequencing reactions. Cycle sequencing PCR products followed purification with the BigDyeXT kit(Applied Biosystems,) and the data were analyzed using an ABI3130xl Genetic Analyzer (Applied Biosystems). DNA sequencing was performed in both directions, initiated from the forward and reverse primers that were used in the initial PCR reaction. SeqScape 2.0 sequence analysis software (Applied Biosystems, San Diego, CA, USA) was employed for sequence evaluation.
A RFLP was identified in the mutation siteand was utilized for mutation detection. Amplicons encompassing exon 5 were digested with the restriction enzyme Tsp509I, and electrophoresed on a 1% agarose gel.
We found that M694V accounted for the majority of FMF chromosomes (44%), followed by E148Q (19%), V726A (10%), M680I (10%), P369S (4%), R408Q (3%), K695R (2%), M694I and R761H (1.6%), A744S (1.4%), and F479L (0.09%) (Tables 2, 3). Missense disease-causing mutations and synonymous polymorphisms accounted for 38% and 54% of MEFV chromosomes, respectively. Among the Turkish general population, the most frequent healthy heterozygous carrier mutation was found E148Q (6.9%), and the carrier rate was found 16%, with a mutation frequency of 8% (Berdeli et al., 2011). Except for the known major FMF mutations, by DNA sequencing, we frequently detect additional rare and novel mutations and critical SNPs about which we have only limited information in Turkish FMF patients. Remarkable consequences of sequencing analysis have been found relative to mutation-SNP combination underlying the combined existence of nucleotide variations in the same haplotype.
For patients whose MEFV gene does not contain mutations of exons 2, 3, 5, and 10, we performed bidirectional DNA sequencing also in exons 1, 4, 6, 7, 8, and 9. However, we could not find any disease related mutation except for an exon 9 homozygous SNP, P588P, which is thought to be symptomatic with disease relation. This SNP was always in homozygous state and was not seen in combination with any of the major and minor mutations or any of the SNPs in the entire coding and non-coding regions of the gene. Relative to our experiences, this SNP has a disease relation to a minor degree, however possible validation of other autoinflammatory disease gene mutations should need to be considered. Single P588P SNP was associated with continuously high SAA levels and musculoskeletal complications which has a good response to colchicine in a three-member family who did not have any sequence variations along other coding and non-coding regions of the MEFV gene.
Number of Patients | ||||||||
M680IG-C/Wt | 69 | 5.24 | ||||||
M680IG-A/Wt | 5 | 0.37 | ||||||
M680IG-C/M680I | 12 | 0.91 | ||||||
M680IG-C/V726A | 23 | 1.74 | ||||||
M680IG-C/ M694V | 42 | 3.19 | ||||||
E230K | M680IG-C/ M694V | 2 | 0.15 | |||||
M680IG-C/A744S | 1 | 0.12 | ||||||
M680IG-C/ R761H | 2 | 0.15 | ||||||
E148Q | M680IG-C | 4 | 0.3 | |||||
E167D | F479L | M680IG-C | 1 | 0.07 | ||||
E167D | F479L | 2 | 0.15 | |||||
F479L/Wt | 1 | 0.12 | ||||||
M694V/Wt | 322 | 24.4 | ||||||
M694V/M694V | 91 | 6.91 | ||||||
M694V/V722M* | 1 | 0.12 | ||||||
M694V/V726A | 42 | 3.19 | ||||||
M694V/R761H | 5 | 0.37 | ||||||
M694V/K695R | 2 | 0.15 | ||||||
M694V/A744S | 1 | 0.12 | ||||||
R241K | M694V/ | 1 | 0.12 | |||||
E230K | M694V/ | 3 | 0.22 | |||||
E148Q | P369S | M694V | 1 | 0.12 | ||||
E148Q/S179N* | M694V | 1 | 0.12 | |||||
E148Q/Wt | 237 | 18 | ||||||
E148Q/E148Q | 19 | 1.44 | ||||||
E148Q | M694V | 42 | 3.19 | |||||
E148Q | L709R | 1 | 0.12 | |||||
E148Q | P369S | 4 | 0.3 | |||||
E148Q | V726A | 7 | 0.53 | |||||
E148Q | A744S | 1 | 0.12 | |||||
E148Q | M694I | 7 | 0.53 | |||||
E148Q | K695N | 1 | 0.12 | |||||
E148Q | R761H | 4 | 0.3 | |||||
E148Q | I72OM | 2 | 0.15 | |||||
E148Q/L110P | 6 | 0.45 | ||||||
E148Q/R151S | 1 | 0.12 | ||||||
E148Q | K695R | 2 | 0.15 | |||||
E148Q/T267M | 1 | 0.12 | ||||||
E148Q/E230K | 4 | 0.3 | ||||||
E148Q/T267I | 1 | 0.12 | ||||||
E148Q/L110P | M694I | 1 | 0.12 | |||||
E148Q | P369S/R408Q | 18 | 1.36 | |||||
E148Q | P369S/R408Q | M680I | 1 | 0.12 | ||||
M694I/A744S | 1 | 0.12 | ||||||
V726A/Wt | 111 | 8.43 | ||||||
V726A/V726A | 4 | 0.3 | ||||||
E167D | V726A | 3 | 0.22 | |||||
V726A/M694I | 2 | 0.15 | ||||||
V726A/R761H | 3 | 0.22 | ||||||
V726A/R761H/ M680IG-C | 1 | 0.12 | ||||||
V726A/K695R | 1 | 0.12 | ||||||
F479L | V726A | 3 | 0.22 | |||||
K695R/Wt | 38 | 2.88 | ||||||
A744S/Wt | 19 | 1.44 | ||||||
P369S/Wt | 7 | 0.53 | ||||||
P369S/R408Q | 40 | 3.03 | ||||||
P369S/R408Q | M694V | 4 | 0.3 | |||||
M694I/Wt | 10 | 0.75 | ||||||
R761H/Wt | 31 | 2.35 | ||||||
R761H/ A744S | 1 | 0.12 | ||||||
R653H/Wt | 1 | 0.12 | ||||||
E685K/E685K | 1 | 0.12 | ||||||
L110P/L1010P | 1 | 0.12 | ||||||
E230K/E230K | 1 | 0.12 | ||||||
E230K/ Wt | 1 | 0.12 | ||||||
T267M/Wt | 3 | 0.22 | ||||||
R241K/R241K | 1 | 0.12 | ||||||
E148V/Wt | 5 | 0.37 | ||||||
E148L/Wt | 2 | 0.15 | ||||||
E167D/Wt | 2 | 0.15 | ||||||
P350R/Wt | 1 | 0.12 | ||||||
P350R | A744S | 2 | 0.15 | |||||
G456A/Wt | 1 | 0.12 | ||||||
S503C/Wt | 2 | 0.15 | ||||||
I506V/Wt | 1 | 0.12 | ||||||
Y471X/Wt | 1 | 0.12 | ||||||
G340R/Wt | 1 | 0.12 | ||||||
S141I/Wt | 3 | 0.22 | ||||||
S166L/Wt | 2 | 0.15 | ||||||
A511V/Wt | 1 | 0.12 | ||||||
R354W/Wt | 1 | 0.12 | ||||||
S339F/Wt | 4 | 0.3 | ||||||
R329H/Wt | 3 | 0.22 | ||||||
R329H/ | M694V | 1 | 0.12 | |||||
E148Q | R329H/ | 1 | 0.12 | |||||
Heterozygotes | 885 | 67.2 | ||||||
Compound heterozygotes | 271 | 20.5 | ||||||
Homozygotes | 130 | 9.87 | ||||||
Complex genotypes | 30 | 2.27 | ||||||
Total number of patients with mutations | 1316 | 38.36 | ||||||
No mutation or SNPs identified | 231 | 6.7 | ||||||
Total number of patients with only SNPs ( | 1883 | 54.8 | ||||||
Total number of patients | 3430 | 100 |
DNA sequencing results ofMEFV genotyping among 3430 Turkish patients.
M694V | 908 | 44.7 |
E148Q | 386 | 19 |
V726A | 204 | 10 |
M680IG-C | 170 | 8.3 |
P369S | 75 | 3.69 |
R408Q | 63 | 3.1 |
K695R | 43 | 2.11 |
M694I | 21 | 1 |
R761H | 47 | 2.31 |
A744S | 26 | 1.28 |
E148V | 5 | 0.24 |
E167D | 8 | 0,39 |
T267M | 4 | 0.19 |
L110P | 8 | 0.39 |
R241K | 3 | 0.14 |
I720M | 2 | 0.09 |
E230K | 12 | 0.59 |
M680IG-A | 5 | 0.24 |
E148L | 2 | 0.09 |
F479L | 7 | 0.34 |
E685K | 2 | 0.09 |
R653H | 1 | 0.04 |
T267I | 1 | 0.04 |
V722M | 1 | 0.04 |
S141I | 3 | 0.14 |
S339F | 4 | 0.19 |
R151S | 1 | 0.04 |
I506V | 1 | 0.04 |
S503C | 2 | 0.09 |
L709R | 1 | 0.04 |
K695N | 1 | 0.04 |
P350R | 3 | 0.14 |
G340R | 1 | 0.04 |
G456A | 1 | 0.04 |
Y471X R329H S166L S179N A511V R354W | 1 5 2 1 1 1 | 0.04 0.24 0.09 0.04 0.04 0.04 |
Allelic frequencies of totally 40 MEFV mutations involving major, rare and, novel sequence changes among the detected mutations in 1316 mutation positive patients group (mutation frequency for the studied mutations; complex alleles excluded).
Additionally, sequence analysis revealed that there was a single FMF-associated mutation in the MEFV coding region of 76% of the Turkish individuals studied, and 80% of these individuals initiated colchicine treatment following molecular diagnosis. The prevalence of a single mutation in patients experiencing a pathogenic effect in Turkey (76%) is contrary to the expected pattern of autosomal recessive inheritance and does not support the ‘’heterozygous advantage’’ selection theory. However, the expression of the FMF phenotype may be influenced by other candidate modifier gene loci, autoinflammatory pathway genes or FMF-like diseases [29-31]. For this reason, genome-wide association studies involving more patients should be performed and the data included in future investigations covering critical coding and noncoding gene SNPs for Turkish FMF patients.
As an ancestral population of FMF, Turkey was one of the regions which involves most of the rare and novel mutations. As referenced in INVEFERS, most of the rare mutations in view of the ethnic origins were found to be symptomatic. Novel Y471X mutation found in the present study was the second nonsense mutation in FMF era. Among the newly identified mutations, involving R151S, S166N, S179N, and G340R; P350R, G456A, Y471X, S503C, I506V, L709R and K695N; Y471X, R151S, L709R, and K695N were observed as pathogenic reflecting the typical FMF character. The main clinical characteristics of the patients were as follows: abdominal pain (92.1%), fever (93.9%), thoracic pain (59%), myalgia (67.8%), arthritis (55.1%), erysipelas like erythema (ELE) (21.8%). None of the patients developed amyloidosis.This finding verifies the importance of molecular diagnosis and detailed sequencing which is recommended to perform in particular for the ancestral populations of FMF.
In this report, from a large scaled heterogeneous group of patients, we describe a 44-year-old Turkish patient from Western Turkey with clinical diagnosis of periodic fever. The case presented here is a 44-year-old Turkish woman, from western Turkey. The course of the patient includes short and rare episodes of fever, ongoing abdominal pain, temporary myalgia and arthralgia since her childhood. Physical examination revealed no pathology except for arthritis on the right knee. Her weight, height, and blood pressure were normal. Primarily, she had diagnosed as having conditions secondary to FMF. Although family and relatives screening are of great importance, her family (parents are dead in an accident) and past history were noncontributory and unhappy. She had undergone antibiotherapy, steroid treatment and appendectomy. Laboratory tests revealed the acute phase reactants as follows; ESR 81 mm/h, SAA 76 mg/dl, CRP 3.46 mg/dl, and fibrinogen 526 mg/dl. Renal function tests and other biochemical parameters were normal. No molecular genetic diagnosis was done except for Strip Assay in other centers. The clinical figure associated with her was not much contributed to the start of colchicine not fulfilling most of the clinical criteria, so in our laboratory, FMF strip assay was used as the first stage of mutation detection method involving 12 common mutations. However, no particular mutation was identified. Thereafter, DNA sequence analysis revealed the responsible nonsense mutation, p.Y471X, in MEFV gene (Figure 1). By means of the molecular diagnosis, colchicine therapy (1.5 mg/day) was started properly. She had no symptoms after the colchicine therapy and had a good response to 1,5 mg/d, and the acute phase reactants were completely normal in the last 3 years. So, other autoinflammatory genes, MVK, TNFRSF1A, CIAS1, were not considered to evaluate as the suspicious genes in this case and were not evaluated as molecular diagnostics.
Electropherogram of the p.Y471X nonsense mutation in the MEFV gene revealed by DNA Sequencing analysis in the Turkish patient.
The case presented here was one of the patients who had misdiagnosis in particularly during the childhood losing time by unnecessary processes and treatments. Therefore, certain diagnosis determined by detailed DNA sequence analysis is essential for suspicious and undefined cases, and for cases disestablished by other limited screening methods. In the molecular analysis ofMediterranean fever gene, c.1413C>A nucleotide change in exon 5 resulting in p.Tyr471X nonsense mutation was determined (Figure 1). We also exploited the fact that the p.Y471X creates a novel recognition site for the Tsp509I restriction enzyme to develop a PCR-RFLP assay in order to screen the affected families and healthy controls for the mutation.
Y471X nonsense mutation in MEFV gene is the first noted in Turkish FMF patients [7], and the second nonsense mutation of FMF mutation database worldwide. Inherited missens mutations reported in the 5th exon of MEFV gene in FMF patients are very rare. Though the fifth exon of the gene could not called as a critical region carrying the mutational hotspots, the result could demonstrate there is still way to walk on the road through the hidden side of FMF. Novel Y471Xmutation in exon 5 of the MEFV gene located in the coiled coil domain of pyrin protein is implicated in association with actin binding interacting selectively with monomeric or multimeric forms of actin. Since effects of nonsense mutations in the amino acids are known damaging and pathogenic, we did not use the PolyPhen software [32] in order to evaluate the potential pathogenicity of this newly found amino acid substitution which we carry out regularly in our laboratory. Nevertheless, expression studies will be required.
Due to the abundance of mutations in exon 10 and clinical heterogeneity of the disease, different screening methods have been developed. As long been known the majority of FMF patients in classically affected populations were screened by routine methods for only common mutations, which primarily targets only the most prevalent MEFV mutations in a specific population; thus, rare or novel mutations can be overlooked. The first nonsense mutation in FMF era, Y688X, was evaluated by Touitou I. [5], and was suggested to have a location between two well-known hotspots for FMF mutations (codons 680 and 694) in exon 10. This finding contributed to the critical role of exon 10 for the MEFV function as an hotspot. Here, it is discussed that, the newly found Y471X nonsense mutation has a great significance in screening asymptomatic individuals since it was not found in one of the hot-spots of MEFV gene.
Autoinflammatory diseases are heterogeneous group of disorders, thus FMF like phenotypes and related genes most likely exists [33-36]. In some cases, the causal genes may not only be the unique causes of the diseases. It is well known that Mendelian disorders caused by the dysfunction of a single gene have a wide heterogeneity of disease phenotypes [37]. FMF has both genetic and phenotypic heterogeneity and mutations within a single gene are known to cause different clinical phenotypes in Turkey. Thus, all MEFV gene sequence variations found in symptomatic cases should not be considered as causative pathogenic disease mutations. In particular, FMF related Turkish patients with no MEFV mutation or with only single MEFV mutations may not actually reflect the phenotype seen in FMF.
Another point is subclinical inflammation concerning asymptomatic heterozygous patients without a second mutation mostly continues with the typical disease characteristics possibly due to the presence of other modifier genes and/or environmental factors. Therefore, factors other than casual MEFV gene and other pyrin-dependent effects should be contributing to the sustainable systemic inflammation that is sufficient for the occurrence of the symptomatic FMF related phenotype. Previously, MICA, TLR2 and SAA loci were shown as modifying alleles in FMF [5, 38]. Synonimous or non-synonimous sequence variations of MEFV relevant genes involving SAA and TLR2 were previously considered as critical factors for the course of the disease. Both SAA1 locus and Arg753Gln TLR2 polymorphism were implied as genetic susceptible loci for a risk factor of developing secondary amyloidosis in different ethnic populations of FMF patients [26, 27, 30]. Against the traditionally considered monogenic inheritance pattern, compound heterozygotes of 2 autoinflammatory disease genes were also reported describing patients who were found to have 2 or more reduced penetrance mutations, involving E148Q in MEFV, R92Q or P46L in TNFRSF1A, V377I in MVK, and V198M in CIAS1 (29, 34, 35). For the purpose of screening mutations in other known autoinflammatorygenes for typical FMF patients carrying 1 single heterozygous MEFV mutation, Booty et al screened 6 candidate genes that encode proteins known to interact with pyrin or genes functioning in IL-1B pathway involving ASC/PYCARD, SIVA, CASP1, PSTPIP1, POP1, and POP2 (6). A novel PSTPIP1 nucleotide mutation, two novel substitutions in ASC/PYCARD and SIVA genes were identified while Casp1, POP1, and POP2 were mutation negative. In a Jewish patient with FMF, novel W171X (513G>A) mutation was identified which is presumed as a stop codon, to remove the last 2 of the 6 helices in the CARD domain of ASC/PYCARD. In FMF patients with only 1 MEFV mutation, including milder FMF-associated mutations, 1 Turkish patient was identified as a carrier of W171X (6). To date, SNPs in ASC/PYCARD gene were identified in 5’/3’ region, exon 1, intron 1, exon 3 coding region involving rs79351176, rs8056505, rs11648861, rs79464842, rs73532217, rs75471387, rs11867108, rs61086377, rs76878620, and rs75216100. In the ASC/PYCARD protein, the conserved PyD domain is 91 aa in lenght (1-91) and CARD domain is 89 aa in lenght (107-195). The previously reported W171X (513G>A) mutation (31) corresponds to the exon 3 coding region of the ASC/PYCARD gene and results with a stop codon. Thus, in our sequencing analysis, we also searched the presence of mutations in the ASC/PYCARD gene in our entire patients group. However, this sequence was not mutated, and we have neither identified the above substitutions along the entire coding regions and flanking segments of ASC/PYCARD gene (unpublished data).
For investigating of mutations in other periodic fever disease genes, in a study of our group, a total of 75 Turkish patients and 25 ethnically matched healthy control individuals diagnosed with periodic fever was molecularly diagnosed for having mutations in causative disease genes (apart from the present patients group; unpublished data). Mutation screening of coding and noncoding regions of MVK, TNFRSF1A, and NLRP3/CIAS1 genes were carried out for different group of patients according to their clinical implications.
MVK gene transcript variant 1 (12q24; NM_000431.2→NP_000422.1) was fully sequenced in 25 periodic fever patients. Molecular diagnosis revealed the following results: p.Ser52Asn missense mutation was identified in 6 patients. In addition, p.Asp170Asp and p.Ser135Ser synonimous aminoacid mutations and IVS6-18 A>G, homozygous IVS9+24 G>A, and IVS 4+8 C/T intronic nucleotide substitutions were observed in the remaining patients group.
NLRP3 gene (CIAS1; 1q44; NM_004895.4→NP_004886.3) NACHT, LRR and PYD domains-containing protein 3 isoform a was fully sequenced in 25 periodic fever patients. Molecular diagnosis revealed the following nucleotide substitutions in the screened gene region: K608fsX611 frameshift mutation, p.Ser726Gly and p.Gln703Lys missense mutations, together with Ser34Ser, Ala242Ala, Arg260Arg, Thr219Thr ve Leu411Leu synonimous aminoacid mutations.
TNFRSF1A gene (12p13.2; NM_001065.3→NP_001056.1) tumor necrosis factor receptor superfamily member 1A precursor form was fully sequenced in 25 periodic fever patients. Molecular diagnosis revealed the following nucleotide substitutions in the screened gene region: p. Arg92Gln and p. Ala301Thr missense mutations with IVS6+10 A>G and IVS8-23 T>C intronic nucleotide substitutions.
Intronic nucleotide substitutions and synonimous aminoacid mutations of all the screened gene regions were also observed in the 25 ethnically matched healthy control individuals. Mutation frequency was 4% (1/25), 32% (8/25), and 40% (n:10/25) in TRAPS, HIDS, and CAPS patients.
Nonetheless, finding of symptomatic rare MEFV mutations in particular for at-risk populations and the individuals who have been asymptomatic and negative for common mutations makes detailed mutation screening critically important in FMF. It has been previously evidenced that there have been a number of patientswho have typical FMF phenotype or FMF related symptoms with only one MEFV heterozygous mutation and/or even without any MEFV mutations [6, 7].
The majority of FMF patients in classically affected populations are screened by routine methods that are limited to the detection of common mutations. These tests primarily target the most prevalent MEFV mutations to rule out asymptomatic cases in at-risk populations. Therefore, while searching for the common mutations that underlie typical FMF symptoms, we should primarily consider the entire coding sequence of the MEFV gene before analyzing other recurrent fever genes. Patients with no mutation or with only single pyrin mutations may not actually reflect the phenotype seen in FMF. Compound heterozygotes of 2 autoinflammatory disease genes involving MEFV,TNFRSF1A, CIAS1, andMVK were reported [29, 34, 35]. Thus, screening of other autoinflammatory disease genes, e.g. CIAS, were considered for the MEFV gene mutation/SNP negative FMF patients. In conclusion, by using sequencing analysis, we can prevent less common, population-restricted, novel sequence variants from being overlooked. This has implications for the characterization of typical and atypical FMF; screening for the most common mutations by routine methods is sufficient for the initial laboratory diagnosis of FMF in Turkish patients; however, the results should be confirmed by specific DNA sequencing of all coding exons and exon-intron flanking regions.
Among the newly identified mutations in this comprehensive study, Y471X, R151S, L709R, and K695N were observed as pathogenic reflecting the typical FMF character involving abdominal pain, fever, thoracic pain, myalgia, arthritis, and erysipelas like erythema. Rare mutations and SNPs have great importance for FMF pathogenesis. For this periodic fever disorder, heterogeneity is present in phases of allelic, frequency and critical locations of mutant alleles, and clinical appearance. Therefore, in particular for the suspicious cases; possible presence of other autoinflammatory disease gene mutations as we outlined above and rare mutations and SNP variations in the MEFV gene, molecular techniques, sample sizes, ethnic origins, and regions in the ancestral countries should be regarded as critical and determinative keys in FMF clinical and molecular diagnosis.
Sequencing analysis not only the common major mutations but also the detection of rare mutations can be carried out which have great importance in particular for at-risk populations. By means of sequencing analysis, we could prevent the missing of less common rare variants that might be restricted to the populations by routine techniques. The majority of FMF patients in classically affected populations are screened by routine methods that are limited to the detection of common mutations. These tests primarily target the most prevalent MEFV mutations to rule out asymptomatic cases in at-risk populations. Therefore, while searching for the common mutations that underlie typical FMF symptoms, we should primarily consider the entire coding sequence of the MEFV gene before analyzing other recurrent fever genes. In conclusion, by using sequencing analysis, we can prevent less common, population-restricted, novel sequence variants from being overlooked. This has implications for the characterization of typical and atypical FMF; screening for the most common mutations by routine methods is sufficient for the initial laboratory diagnosis of FMF in Turkish patients; however, the results should be confirmed by specific DNA sequencing of all coding exons and exon-intron flanking regions. We should consider gene mutation screening in early diagnosis and the follow-up of the clinical course in particular for the asymptomatic cases. Early determination of the disease causing mutation will be favorable in order to prevent abundant treatments in newly diagnosed patients.
We would like to thank patients and clinicians for their participation and contribution in our study.
Probiotic are the live microbial feed supplements which provide the beneficial impact on the host by producing the useful metabolites [1]. Many probiotics have been available in the market for improving animal and human health in safe and healthy way. The commercially available probiotic product contains mostly lactic acid bacteria (
Representative scheme of development of target-based probiotic (TBP): The right side covers the main steps involving in the preparation of the TBP, the internal part covers the legalistic evidence of the interrelationship between, host and microbes. The left side covers the mechanistic activity of the TBP; including the improve gut microbial balance which leads to the improve feed digestion resultantly improve host health and production in cost effective manner.
In the situation of high animal feed cost, we must identify the cost-effective probiotic by using the concept of ITP to improve poor quality feed into high quality milk and meat. We had already given the concept of indigenous probiotic yeast our previous book chapter [31]. A clear understanding regarding the proposes guidelines to develop the ITP to improve gut microbiota resultantly improve milk and meat production. This book chapter will discuss the identification of the microbial strain from local ecological breed and its mode of action for preparation of target based probiotic products. We will also support our concept of ITP with our lab conducted experiments.
Yeast is a very useful microorganism with broad range of industrial application, because of their unique genetics and physiology. Yeast cells have many useful metabolites (protein, carbohydrate, vitamins; vitamin B6, thiamin, biotin, riboflavin, nicotinic acid and pantothenic acid and minerals; zinc and magnesium) [10]. The utilization of the naturally prepared yeast would be accelerated in coming years due to the nature-oriented mind set of the consumers. Therefore, research on the isolation of the nutritious rich yeast strains for preparation of probiotic product has rapidly increased [11, 12]. Yeast is an important single cell microorganism, belongs to fungus family and it multiplies by cell division. The genetics and physiology of the yeast are very unique, and, therefore, a broad range of research work in biological sciences is being carried out on this microbe. The yeast cell size is composed of 5 × 10 μm and the size of the baker’s yeast genome is 12.1 Mb containing 16 chromosomes and 5400 coding genes approximately [13]. Members of the order Saccharomycetales are mainly used for the animal probiotics when serves as reliable and economical source of essential amino acids, vitamins, carbohydrates, and minerals from yeast cell. Thiamin, Riboflavin, Niacin and Biotin are present in yeast [14]. The antagonistic ability of the yeast to block bacterial pathogenicity is also makes its very useful [15]. Yeast cell has competition for nutrients, pH changes in the medium, high concentrations of ethanol production, secretion of antibacterial compounds and release of antimicrobial compounds are major antagonistic steps. Yeast cell has many useful fermentation metabolites (protein, vitamins, carbohydrates) which makes it important microbial feed supplement. Yeasts are naturally present (1.3 X 105 yeasts ml-1) inside the rumen fluid [16]. Literature showed that, yeasts (
The role of the probiotic yeast in dairy animal is well studied [25]. They have been extensively used for improve milk yeast and its composition in cost effective manners. The benefits to cost ratio of probiotic yeast is 4:1 in dairy animals. They have also used as preventer against digestive problems, and rumen acidosis.
The main target of the PY used in new born ruminate diet are; (a) improvement in the rumen maturation; (b) stop the pathogenic bacterial growth; (c) establishment of the normal growing animals like microbial flora [26, 27, 28]. Microbial based feed can improve the rumen development during the growing phase of the dairy animals. The new born gut is sterile and have no germ [29]. After 6 months of age the rumen is colonized with diverse microbial flora. PY provides beneficial metabolites and enzymes like thiamine for fast growth of the fungi. The poor fungal growth of the animal fed on PY might be due to the low production of thiamine [30]. At the same time, the animal plays an important role in the maximum colonization of the beneficial microbial population [31]. If there is any imbalance bacterial species, it would result in digestive problems and leads to the economic loss. The establishment of the useful bacterial strains results in the development of strong and balanced rumen which resultantly strong immunity and health condition [32, 33]. PY provide the improve the rumen maturation and its microbial flora is also in strong balance. PY provide the useful bacterial species for feed digestion, like cellulolytic bacterial species and ciliate protozoa [34]. The balance in rumen microbial flora plays a crucial role in feed utilization and could result in better animal productivity [35]. PY remove oxygen from rumen and provides a more anaerobic environment for its growth of key beneficial microbial groups [36]. The newborn gut can easily be modulating by PY. The new born key beneficial microbial
For clear understanding of the ruminal gut microbiota using latest genomic methods to get useful information for preparation of specific probiotics. The ruminants feed consists of concentrate, silage, seasonal fodders etc. There diet mostly contains cellulose, hemicellulose starch and water-soluble carbohydrate. The rumen microbes play an important role in feed digestion. The animal feed is digested inside rumen and then energy is released for animal use. Cow and its microbes are mutually benefiting each other (Figure 2). Rumen is the first and the largest anaerobic chamber of the cow GIT. The temperature inside the rumen chamber is between 38 to 41 oC, with 6-7 pH (depends on feed type). There are three different types of microbes present inside the rumen including, bacteria, fungi and ciliated protozoa [41, 42, 43, 44] . The location and size of the rumen microbes depends on the feed formulation and host genetic. Mostly, bacteria are associated with fibrous feed particles; fungi, protozoa [45, 46]. Some are freely living and some are bound with rumen mucous membrane. 1 ml of the rumen is composed of 109 to 1010 per ml bacteria with 200 different species, 104 to 106 per ml protozoa with 20 different species, and 103 per ml fungi with 20 different species [47]. The rumen bacteria are gram negative 1-2 micrometer in size and cocci, and rod shaped mostly. Rumen bacterial are mostly non-spore producing, facultative anaerobes. 1- 5 % of the bacterial cells in rumen are cellulose digesters [48]. The rumen fungi (gut fungi) also play an important role in fiber digestion by stimulating growth of fibrolytic bacteria [49]. The rumen microbial features are heritable; moreover, animals age, feed and genome plays an important role in the microbial colonization. The composition of the diet describes the type of gut microbial species [50]. Therefore, the rumen microbiota can be manipulated by using the yeast-based probiotic to obtained the useful products. The feed must be targeted for modulating the rumen microbiota (Figure 3).
Major factors effects on the mode of action of probiotic.
A scheme describing the mutually benefits between host microbes.
The modulation of the rumen microbiota is mostly for the enhanced colonization of the fiber digesting microbiota [35, 36]. Literature showed that, animal diet has an important role in the manipulation of the rumen microbiota. Low amount of fibrous feed builds up fast working microbes (fibre-degrading Butyrivibrio fibrisolvens and F. succinogenes) and high amount of fibrous build up slow working fiber degrading microbes
Potential mechanisms of microbial ruminal acidosis: This figure suggested that, the live yeast supply different growth factors (amino acid, peptides, vitamins and organic acids). These growth factors have the knock-on impact of increases the stimulation and metabolism of lactic acid utilizing anaerobic bacteria, such as M. Elsdenii or S. ruminantium (that control the acidosis). Yeast cells has a affinity for sugar which outcompete S.bovis for the utilization of sugar.
A proposed flowsheet to explain mechanistic pathway of IPY: Steps involved in the mode of action of PY and its impact on animal.
A simple scheme proposed to explain mode of action of probiotic yeast in gut: IPY improve carbohydrates, protein and lipid digestion rate by improving the production of cellulolytic, hemi- cellulolytic and proteolytic and lipolytic bacteria and fungi as compare to FPY and no yeast animal.
The gut microbiota can digest the animal feed and produce nutrients for improve host health and well beings. Animal feed and host genetics play important role in shaping and composition of gut microbiota [18]. Same is the case of the rumen microbiota, which is highly variable and is depended on various factors like animal breed, physiology, feed type and geographical location. It has been commonly accepted that commercially available probiotic yeast may not showed equal impact to all animal breeds [65, 66] The compatibility of PY could be variable among animals. The local prepared yeast probiotic isolated from same ecological niche may have more beneficial impact than any exotic probiotic yeast [3]. The local isolated probiotic yeast may have fast adaptability and colonization in the local rumen ecosystem [24]. The origin of the probiotic strain determines the best prepared probiotic product. The strain selection is the most important step for the development of right probiotic for animal. Being precise during the strain’s selection could yield positive outcomes from the probiotic. The probiotic yeast may use for the rumen microbial manipulation [67] Different types of PY have been used for improve animal health and production [7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68]. Some PY strains produced beneficial results in animals while others did not. The difference of that variable results of PY may be explained by different host and PY associated factors [69, 70, 71]. These factors are; animal age, breed, sex, feeding dose, PY strains isolation source and some unknown factors [3]. The major factors might be the low compatibility of the exotic probiotic yeast strain with animal having diverse biological inheritance and gut microbial composition. The right probiotic strain should be novel, so we must use latest molecular methods to isolate the target specific/local isolated microbial strains. The local isolated and molecular identified probiotic strains may have more impact on local animals in cost effective manners. The probiotic are species specific by targeting the indigenous strains and local dairy farms can get the cost-effective probiotic product for improve milk production and composition.
The main steps involved in the preparation of the breed specific probiotic yeast are as following [3].
Pre-plan ruminate diet for isolation of probiotic yeast
Identification of yeast strain based on the molecular techniques
Probiotic potential of selected yeast strains
In vitro probiotic potential
Safety assessment/In-vivo animal model
The first mode of action of the probiotic yeast is competitive exclusion (CE) [27]. The CE is a probiotic mode of action that involves the colonization of the beneficial microbial strains to GIT tract to reduce the addition of disease-causing microbial flora [18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74]. The ability of probiotic yeast cell to fight with other useless microbial flora can improve growth and function of beneficial microbial flora. The IPY has the indigenous strain, which has the advantage that it drives from animal of interest (Cow). IPY has an environmental modification capability. The concept of co-evolution of host microbial has been seen in case of IPY mode of action. The local strain gains an advantage because of its ability to adjust/modify itself in new environment by producing the antimicrobials e.g (lactic acid) to make its less suitable for its competitors. The FPY has the foreign origin strain, which has the less environmental modification capability less, competition for available nutrients, and mucosal adhesion sites. Second mode of action of the PY is reported as a good pH stabilization. Rumen microbial flora can work under stable pH [75]. Rumen pH is highly affected by animal feed intake and its composition. Ruminants eat different types of feed, like high energy concentrate diet, fodder, and silage. These types of feed have a quick impact on rumen pH. If rumen pH is not stable, the animals may have different types of metabolic diseases [76] Literature showed that PY has a stabilizing effect on the rumen pH. [77, 78] Some studies reported a rise rumen pH when animal was fed on diet with high energy supplemented with PY. Sometimes, the increased pH might be due to the decreased VFAs inside the rumen [3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79]. The lower pH leads to the rumen acidosis, PY can prevent the acidosis condition of the dairy animals [7]. The third proposed mechanism is that yeast cell provides the anaerobic condition inside rumen by removing the oxygen thus facilitated the useful feed digestion microbes [35, 36]. The main microbial flora are bacteria fungi and protozoa. These microbial species have a fiber digestion role by secreting the cellulase and hemicellulase enzymes. Fiber is the main part of the ruminant diet. Therefore, fiber digestion, nature blessed them with unique fibrolytic digestion bacteria (
Based upon the above discussion, we have conducted two research experiments on dairy animals by using the IPY concept to improve the gut health. In experiment 1, eight dairy heifers (87 ± 5 kg and 6–7 months) were divided into two equal groups (control n = 4 and probiotic n = 4)[80]. Control group animals fed on NRC recommended diet and probiotic group animals fed control diet FPY (Yea-Sac1026; 5 g/animal). After 120 days results showed that the FPY significantly affected the serum glucose, and urea levels in dairy heifers [24].
That means, we had a proof of positive impact of PFY on animal health. We had isolated the yeast from dairy animals fed on yeast. After careful assessment of the probiotic potential, we conducted another experiment to determine the impact of FPY Vs IPY on the health of lactating dairy cattle. Mix breed (
Items | Feeding regime | p-Value | |
---|---|---|---|
Control2 | FPY3 | ||
Before treatment4 | 30.10 ± *0.711 | 31.14 ± 0.974 | 0.012 |
After treatment5 | 33.34 ± 0.432 | 29.23 ± 0.494 | 0.01 |
Before treatment | 62.67 ± 4.04 | 60.86 ± 2.80 | 0.605 |
After treatment | 63.31 ± 2.60 | 65.47 ± 2.84 | 0.600 |
Blood serum metabolites (Means ± SEM) in dairy heifers fed on control and foreign probiotic yeast.
n = 4 per treatment.
Control feed without yeast.
Probiotic feed compose of control feed supplemented with 2.5×10 07 cfu/g commercially available probiotic yeast (Yac-Sac1026) at the rate of 5 g per animal/day * ± Standard error of the mean.
Before treatment (day 0).
After treatment (day 120).
Parameters | Feeding regime | ||
---|---|---|---|
Control2 | IPY3 | FPY4 | |
Before treatment5 | 14.55 ± *0.57 | 14.18 ± 0.21 | 15.54 ± 0.32 |
After treatment6 | 14.18a ± 0.58 | 12.31b ± 0.22 | 13.68ab ± 0.90 |
Before treatment | 75.70 ± 1.24 | 73.99 ± 2.51 | 75.08 ± 2.30 |
After treatment | 73.84b ± 0.71 | 77.42a ± 1.28 | 78.97a ± 0.54 |
Effect of indigenous Vs foreign probiotic yeast on blood parameters (Means ± SEM) in lactating dairy cattle.
n = 3 per treatment.
Control feed without yeast.
LAB-Probiotic feed compose of control feed supplemented with 3.13 × 1007 cfu/g laboratory produces probiotic yeast (QAUSC03) at the rate of 8 g/day/animal.
COM-Probiotic feed compose of control feed supplemented with 2.5×10 07 cfu/g commercially probiotic yeast (Yac-Sac1026) at the rate of 10g/day/animal.
Before treatment (day 0).
After treatment (day 120) * ± SEM = standard error of the mean.
a,b Values on the same row with different superscripts differ significantly (P < 0.05).
We highlighted that improved animal health condition might be due to improved digestive enzymes produced from well propagated IPY. The VFAs have a capability to reduce the triglycerol and cholesterol in liver cells and might be change the animal lipid profile. Results of the ruminal gut microflora showed that the average, beneficial
Total Lactococcus count (CFU/g) in the ruminal gut of dairy heifers fed on control feed (control, ♦; no yeast) or commercial probiotic feed (COM-P, ■; control feed plus commercial yeast) (n = 4).
Total Enterococcus count (CFU/g) in the ruminal gut of dairy heifers fed on control feed (control, ♦; no yeast) or commercial probiotic feed (COM-P, ■; control feed plus commercial yeast) (n = 4).
It can be concluded IPY improves the, gut health, and wellbeing of lactating dairy cattle in cost effective manner. IPY strain may adopt well in the cattle gut than FPY [80].
Ruminants of developing and developed countries have different types of gut microbiota due to their living standard, feeding type, their managemental style. Although from above discussion we have a clear understanding that the interlink between gut microbiota and fiber digestion plays a key role for obtaining maximum profit from dairy animals. Therefore, the PY must be target specific which give maximum outcomes in cost effective manners. For animals of specific geographical region, a unique and precise YP must be designed by isolating the local yeast strains from that population, only then maximum beneficial outputs can be obtained. The reason beings, compactivity of the local strains with normal microbiota of the rumen ecosystem (Figure 9).
Target based Probiotic Preparation strategy: This figure showed probiotic preparation of by using the local animal GIT tract as preparation of local yeast probiotic. Interlinked factors involved in the application of probiotics in the ruminant’s nutrition.
The recommendations are outlined as follows;
Pre-plane feed formulation for the manipulation of the rumen microbiota to digest the fibrous feed
Identification of breed specific probiotic strains with same target.
Whole genome sequencing of the probiotic strains as well as animal for maximum outputs
Mode of action of the probiotic should studied well for understanding of the useful and useless probiotic.
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