Introduction: With the shift from a categorical to a dimensional model, ICD-11 has made substantial changes to the diagnosis of personality disorders (PDs), including obsessive-compulsive (anankastic) personality disorder (OCPD). The ICD-11 PD model proposes a single diagnosis of PD with specifications regarding severity and domains. However, a systematic overview of ICD-11 anankastia is lacking. In this review we address the reformulation of the OCPD diagnosis in the ICD-11, and draw comparisons with the DSM-5, with a particular focus on diagnostic validity and clinical utility. We hypothesized that the ICD-11 PD model provides a diagnostically valid and clinically useful approach to OCPD, with specific emphasis on the anankastia domain as the primary trait qualifier.Methods: Literature published from 2010 to 2020 was systematically searched using the PubMed/MEDLINE, PsychInfo, Cochrane, and Web of Sciences search engines, in order to find all articles that addressed ICD-11 anankastia. Relevant articles were collated, and themes of these articles subsequently extracted.Results: Out of the 264 publications identified, 19 articles were included in this review. Four themes were identified, namely (a) overlap of DSM-5 OCPD with the ICD-11 PD model, (b) the factorial structure of the ICD-11 PD model with respect to the anankastia domain, (c) the clinical utility of the ICD-11 PD model, and (d) comparison of the ICD-11 PD model of anankastia with the DSM-5 alternative model for OCPD.Conclusions: The ICD-11 anankastia domain overlaps with DSM-5 OCPD traits, and the factor analyses of the ICD-11 PD model further support the diagnostic validity of this domain. There is some support for the clinical utility of the ICD-11 PD model of anankastia but further studies are needed, including of its relationship to obsessive-compulsive and related disorders. Diagnostic and Statistical Manual of Mental Disorders    in the , DSM-5)  or anank ICD-10) , is char ICD-10) –6 and 23 ICD-10) , 8, OCPD ICD-10) , 10.The operationalization of PDs, including OCPD, in both the DSM and the ICD taxonomies, has been a subject of debate , 11, 12.A proposal to move to a dimensional conceptualization of PD, including OCPD, was put forward by the DSM-5 Personality and Personality Disorders Work Group , which oIn contrast, ICD-11 has moved away from a categorical framework of PDs to an entirely dimensional system  without A number of publications have addressed the ICD-11 conceptualization of PDs in general, and a number of studies have focused on the ICD-11 domain of anankastia in particular. However, we are not aware of any review that has synthesized the literature on ICD-11 anankastia. Given the recency of this conceptualization, we chose to conduct a scoping review to assess the existing body of literature and identify knowledge gaps .The current scoping review aims to provide a comprehensive overview and synthesis of empirical research on ICD-11 anankastia to date, with a particular focus on diagnostic validity and clinical utility. Due to the limited number of studies, the final pool of selected literature was not subjected to restrictions in terms of study population, intervention type, comparators or outcomes of interest (PICO). Our hypothesis was that the ICD-11 PD model is a diagnostically valid and clinically useful approach to OCPD.* AND ; (2) ICD-11 AND “personality disorder*” AND trait qualifier*; (3) ICD-11 AND “personality disorder*” AND domain*  . We used domain* .Studies were considered for inclusion if they addressed the classification of OCPD within the ICD-11 PD model. This included studies of the ICD-11 anankastia domain. There were no restrictions to inclusion criteria regarding country of origin, sample size, or PICO due to the relative scarcity of publications relevant to the study hypothesis. Due to the nature of the hypothesis, selection of studies was limited to those with a solely empirical research design . Publications that were not available in English –32 were The search yielded 19 studies for review , all of n =11) were published in 2020. Eight (n = 18) addressed the question of diagnostic validity, while a single study examined the clinical utility of the ICD-11 PD model.Most of the studies  that examined associations between ICD-11 anankastia and DSM-5 OCPD, the ICD-11 anankastia domain was more predictive of the presence of the DSM-5 OCPD than of other PDs . In a sample of 1,541 individuals comprised of the general population and psychiatric outpatients, the additional trait of distractibility (found in the low disinhibition domain) was also associated with the ICD-11 anankastia domain  overlap of DSM-5 OCPD with the ICD-11 PD model, (b) the factorial structure of the ICD-11 anankastia domain, (c) the clinical utility of the ICD-11 anankastia domain, and (d) comparison of the ICD-11 PD model of anankastia with the DSM-5 alternative model for OCPD.avoidance of, or reluctance to take risks (from the DSM-5 avoidant PD) was also associated with the anankastia domain  were found to be predictors of the ICD-11 anankastia domain  and a domain , 37, 40,a domain . These fa domain . Nevertha domain .Limitations of this scoping review deserve acknowledgment. In particular, the review was limited to articles written in English, so excluding a number of potentially relevant studies. In addition, key limitations of the literature itself deserve emphasis. First, the methods and instruments to assess the ICD-11 PD domains varied significantly, making it challenging to compare results across studies. Second, studies of the ICD-11 model are limited to only a small number of countries. Third, we found no longitudinal studies of the diagnostic reliability of the ICD-11 over time. Fourth, we found no papers exploring ICD-11 PD and the anankastia domain in individuals with obsessive-compulsive and related disorders. Thus, further longitudinal studies in more diverse cultural cohorts and in both community and clinical populations, using consensus instruments, are warranted.OCPD is a common mental health problem that is still relatively under-recognized and lacks empirical investigations. This scoping review suggests that the ICD-11 PD model is a diagnostically valid and clinically useful approach to OCPD. Specifically, the ICD-11 anankastia domain overlaps with DSM-5 OCPD traits, with factor analyses of the ICD-11 PD model further supporting the diagnostic validity of this domain. There is some support for the clinical utility of the ICD-11 PD model with regards to anankastia. Future studies investigating the clinical utility of ICD-11 PD in more diverse clinical and cultural samples are warranted. Finally, further work exploring the overlap of the ICD-11 anankastia domain with DSM-5 obsessive compulsive and related disorders  is needeThe original contributions presented in the study are included in the article further inquiries can be directed to the corresponding author.DS conceptualized the idea, supervised during the writing process, and provided critical revisions. JG-S conducted the systematic search, prepared the first draft of the manuscript, and revised the manuscript. CL, NF, and CM provided critical revision to its further development. All authors contributed to the article and approved the submitted version.JG-S serves as a consultant at FACITtrans. NF declares that in the past 3 years she has held research or networking grants from the ECNP, UK NIHR, EU H2020, MRC, University of Hertfordshire; she has accepted travel and/or hospitality expenses from the BAP, ECNP, RCPsych, CINP, International Forum of Mood and Anxiety Disorders, World Psychiatric Association, Indian Association for Biological Psychiatry, Sun; she has received payment from Taylor and Francis and Elsevier for editorial duties. In the past 3 years, she has accepted a paid speaking engagement in a webinar sponsored by Abbott. Previously, she has accepted paid speaking engagements in various industry supported symposia and has recruited patients for various industry-sponsored studies in the field of OCD treatment. She leads an NHS treatment service for OCD. She holds Board membership for various registered charities linked to OCD. She gives expert advice on psychopharmacology to the UK MHRA. DS has received research grants and/or consultancy honoraria from Johnson & Johnson, Lundbeck, Servier, and Takeda. The handling editor declared a past collaboration with one of the authors DS. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Both COFs containing a photoredox triphenylamine moiety are efficient photocatalysts for the cross-dehydrogenative coupling reactions and asymmetric α-alkylation of aldehydes integrated with a chiral imidazolidinone catalyst. Under visible-light irradiation, the targeted chiral products are produced in satisfactory yields with up to 94% enantiomeric excess, which are comparable to those of reported reactions using molecular metal complexes or organic dyes as photosensitizers. Whereas the COFs became amorphous after catalysis, they can be recrystallized through solvent-assisted linker exchange and reused without performance loss. This is the first report utilizing COFs as photocatalysts to promote enantioselective photochemical reactions.Covalent organic frameworks (COFs) show great promise as heterogeneous photocatalysts, but they have not yet been explored for asymmetric photocatalysis, which is important for the sustainable production of pharmaceuticals and fine chemicals. We report here a pair of twofold interpenetrated 3D COFs adopting a rare -connected  Covalent organic frameworks (COFs) show great promise as heterogeneous photocatalysts, but they have not yet been explored for asymmetric photocatalysis, which is important for the sustainable production of pharmaceuticals and fine chemicals. In this work, we demonstrated a new metal-free photocatalytic system for asymmetric catalysis based on 3D covalent organic frameworks (COFs) combined with a chiral imidazolidinone catalyst.Photocatalytic methods have shown great promise for bulk production and are widely accepted as convenient strategies in the field of asymmetric catalysis.14,15 By judicious choices of constituent building blocks, COFs have provided a powerful platform for engineering functional materials and hold promise for many applications such as molecule storage and separation,16 catalysis,17 energy storage,18 and optoelectronics.19 From a structural perspective, this area is dominated by 2D COFs,20,21 which generally have eclipsed stacking structures with unidirectional channels. In contrast, 3D COFs are far less explored21 and, with few exceptions,22 they have only been reported for nets based on building blocks with the tetrahedral geometry,23 presumably because of their limited availability of building blocks and the difficulty of their crystallization. Compared to 2D COFs, 3D COFs can characteristically possess high surface areas and numerous open sites and fascinating confinement effects,23 which provide many opportunities for expanding COFs' potential applications. Therefore, the targeted synthesis of 3D COFs with novel topologies and functions is highly desirable. Here we reported imine condensations of rectangular and trigonal monomers to lead to 3D porous COFs with a rare twofold-interpenetrated ffc topology. We selected triphenylamine as the functional molecule, since it represents a type of important hole-conducting molecule with unique photophysical and redox properties.24 This led to the conclusion that the as-prepared 3D COFs can be used as photocatalysts for the cross-dehydrogenative coupling (CDC) reaction and the asymmetric α-alkylation of aldehydes integrated with a chiral imidazolidinone as the organocatalyst.3COFs are a new class of highly tunable, porous crystalline organic polymers with 2D or 3D network topologies.1 was prepared through the imine condensation of tetraamine ETTA and trialdehyde NBC in a mixture of o-DBC/n-butanol/6 M acetic acid  at 120 °C for 3 days, which afforded yellow crystalline powders in 78% yield. COF-2 was synthesized from the imine condensation of 4′,4′′′,4′′′′′,4′′′′′′-tetraaldehyde ETBC and triamine BADA in a mixture of o-DBC/n-butanol/9 M acetic acid  at 120 °C for 3 days, which produced yellow powders in 70% yield.The targeted 3D COFs were designed based on triangular and rectangular precursors connected by [3 + 4] condensation reactions. As shown in  PBM data was replaced with SVG by xgml2pxml:<glyph-data id="z.dbd" format="PBM" resolution="300" x-size="8" y-size="10" xml:space="preserve">00000000000000000000000000000000111111110000000011111111000000000000000000000000</glyph-data><svg xmlns="http://www.w3.org/2000/svg" version="1.0" width="13.200000pt" height="16.000000pt" viewBox="0 0 13.200000 16.000000" preserveAspectRatio="xMidYMid meet"><metadata>Created by potrace 1.16, written by Peter Selinger 2001-2019</metadata><g transform="translate scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg>N linkages were observed at 1621 and 1623 cm−1, respectively , (001), (111), (041) and (002) facets, respectively. For COF-2, the first and most intense peak corresponding to the (020) reflection plane appears at 1.88°, with other minor peaks at 3.37°, 3.68°, 4.89° and 6.29°, which can be attributed to the (001), (040), (041) and (311) facets, respectively  are reasonable for COF-1/2. After considering these possible nets with different space groups, the detailed simulations .The crystalline structures of the two COFs were determined by powder X-ray diffraction (PXRD) analysis with Cu Kα radiation . As reveectively . The cry ffc net . Full pr2 for 1 and 15.0 × 34.8 Å2 for 2. It should be noted that the networks reported for other 3D COFs are generally based on organic building blocks with the tetrahedral geometry.22,23 There are only two examples of -connected COFs with square planar four-connected motifs reported thus far, in which framework interpenetration was suppressed by using short organic linkers.23 It is likely that the interpenetration of theses COFs is controlled by the lengths of the building blocks. The ability to manipulate framework interpenetration is key to the future synthesis of new porous 3D COFs, which hold great promise in heterogeneous catalysis and molecule storage and separation.15,22,23 It is noted that a -connected 3D metal–organic framework with a two-fold interpenetrated ffc topology has been reported in the literature.25As shown in −5 Torr for 12 h. As shown in P/P0 < 0.01), which is characteristics of microporous materials. The Brunauer–Emmett–Teller (BET) surface areas were calculated to be 624 m2 g−1 for COF-1 and 570 m2 g−1 for COF-2, respectively , respectively  was only attached to the surfaces of the COFs and cannot enter the pores from the open channels, probably due to its larger size (3.68 × 3.18 nm2). The dye absorption amount was determined by measuring the UV-vis spectra in THF. As shown in A–C (9.4–19.2% of the COF weight), but had only negligible uptake of dye D. By carefully adjusting the CFM, we obtained the cross sectional fluorescence images across the crystals in different Z wide positions, which reflects the spatial arrangement of incubated dye molecules within the COF. Dye A and COF-1 after spectral separation can be assigned to red and green fluorescence by CFM, respectively  showed that both COFs have excellent thermal stabilities up to 350 °C under a nitrogen atmosphere Fig. S3. The che Fig. S10, further1 and 2 can absorb light in the UV and visible regions, with absorption edges at about 554 and 579 nm, respectively. These values are red-shifted by 46–147 nm in comparison to the solid state absorption spectra of the parent monomers, which might be attributed to a higher degree of conjugation in the extended structures. Based on the Kubelka–Munk formula, the optical band gaps of COFs 1 and 2 were calculated to be 2.24 and 2.14 eV, respectively, smaller than those of the building blocks such as NBC (2.44 eV) and BADA (2.87 eV) -1,2,3,4-tetrahydro isoquinoline (3a) was reacted with nitromethane in CH3CN at 40 °C for 40 h, the product (4a) was obtained in good yield. Under the optimized reaction conditions, a series of substituted tetrahydroisoquinoline derivatives can react with nitromethane, affording the products in 53–85% yield. Notably, substituted tetrahydroisoquinoline derivatives with electron-rich groups gave higher yields than substrates with electron-withdrawing substituents. For instance, when 1,2,3,4-tetrahydroisoquinoline with a methyl group (3a–c) was reacted under standard conditions, the desired products were isolated in 85%, 80% and 80% yields, respectively. However, substrates with electron-withdrawing substituents (3g and 3h) only gave the targeted products in low yield. In addition, COF-2 was also capable of promoting the CDC reactions, generating the products in 50–83% yield, close to those obtained with COF-1.The CDC reaction is one of the most efficient synthetic strategies for the construction of carbon–carbon bonds by oxidative coupling of two distinct C–H bonds.28 Compared with α-carbonyl substitution, direct β-activation of saturated carbonyls has demonstrated to be a more cumbersome and challenging task owing to the typically unreactive β-C(sp3)–H bonds and other competitive reactions. Recent studies indicated that the combination of organocatalysis and photoredox methods may provide a catalytic solution to this problem.3,7 Therefore, the enantioselective α-alkylation of aldehydes was selected as a test reaction to apply COF-based heterogeneous photocatalysts.The carbonyls as powerful building blocks play an important role in broad areas of organic synthesis.9a was obtained in 73% yield with 36% ee in this reaction under a white LED  and ee (73% or 69%). Fluorescence quenching titration showed that COF-1 and Macmillan catalyst 5 can form a stable host–guest adduct with association constants (Ka) of 3700 M−1 piperidine-1-carboxylate can also proceed smoothly in this reaction, and the product 9c was isolated in 55% yield with 85% ee. When 1-bromo-2,4-dinitrobenzene was subjected to the transformation, the expected products 9f were isolated in 80% yield with 86% ee. Besides, COF-2 can also act as a photosensitizer for these catalytic reactions, affording the products in 51–85% yield and 85–94% ee   in a mixture of o-DBC/n-butanol/6 M HOAc  in the presence of NBC or ETBC .After CDC and α-alkylation of aldehydes, both COFs  or ETBC , the cry or ETBC . The BET3a and CH3NO2 (or 7a and 8a). The careful exclusion of light completely suppressed the reaction process, confirming the photochemical nature of the reaction. The inhibition of the reactivity was also observed under an aerobic atmosphere for the CDC reaction or in the presence of the radical scavenger DMPO or TEMP (1 equiv.), with the latter experiment being indicative of a radical mechanism. An EPR spin-trapping technique was employed to confirm the general radical and superoxide anion process  between the ground state and the vibrationally related excited state of 2.40 eV  = −0.75]32 to generate COF-1+, which is rapidly reduced by the amine substrate 3 (Eox = 0.83 V)27 affording the amine radical cation THIQ+. The nucleophile attacks the intermediate 10 to obtain the target product 4 3.4 The excited state of triphenylamine could initiate a photoinduced single electron transfer from the COF to diethyl 2-bromomalonate (E1/2 = −0.49 V),3 rendering a radical anion that undergoes σ-bond cleavage to give an electrophilic radical. Meantime, the chiral organocatalyst serves as cooperative active sites, where a π-nucleophilic enamine combines with the electrophilic radical to forge a crucial reaction center that drives the reaction in an asymmetric manner. The electron-rich amino radical 13 is rapidly oxidized by COF-1+, which closes the redox cycle while regenerating the photocatalyst COF-1. Furthermore, subsequent hydrolysis of iminium 14 would regenerate the organocatalyst 5 while offering the enantioenriched product 9 . Similarly, the COPs promoted the reaction of 7a with 8a or 8b generating the products in 10% lower yields than COF-1 (9a and 9b). The improved catalytic performance of the COFs is probably a result of their crystallinity and permanent porosity, which may optimize substrate adsorption/activation and facilitate electron transfer for efficient reduction of the α-bromocarbonyl substrate and α-amino radical.The crystallinity and porosity of COFs play a vital role in determining their catalytic performances. In catalyzing CDC and α-alkylation of aldehyde, COFs ffc topology through condensation of rectangular and trigonal building blocks by imine linkages. The structure assignment was supported by PXRD analyses, modeling study, pore size distribution and dye-uptake experimental data. The 3D COFs were shown to be efficient photocatalysts for the CDC reaction and the asymmetric α-alkylation of aldehydes integrated with a MacMillan imidazolidinone as the chiral catalyst under visible light. The observed enantioselectivities are comparable to those of reported reactions using molecular metal complexes or organic dyes as photosensitizers. The COF materials that lost crystallinity after catalysis can be readily recrystallized and reused without performance loss. This work thus paves the way for future applications of COFs in visible-light-driven photoredox asymmetric catalysis and will promote the design of more 3D COFs with novel topologies and functions.We have designed and synthesized a pair of twofold interpenetrated 3D COFs with the There are no conflicts to declare.SC-011-C9SC04882K-s001


The Irish Johne's Control Programme (IJCP) provides a long-term approach to the voluntary control of Johne's disease (JD) in Ireland, strongly supported by Irish cattle industry leadership. It leverages the establishment of Animal Health Ireland for control of animal diseases not regulated by the European Union. The IJCP has four objectives: facilitate protection against spread of JD to uninfected farms; reduce the level of infection when present; assure markets of JD control in Ireland; and improve calf health and farm biosecurity. Key IJCP elements are an annual veterinary risk assessment and management plan (VRAMP), annual whole herd test (WHT) by ELISA on blood or milk samples with ancillary faecal PCR testing of ELISA reactors, and Targeted Advisory Service on Animal Health (TASAH) investigations of infected herds. There are pathways for assurance of herds with continuing negative tests and for management of test-positive herds. Herdowners are responsible for on-farm activities, and specifically-trained (approved) veterinary practitioners have a pivotal role as technical advisors and service providers. The programme is supported by training of veterinarians, performance of testing in designated laboratories, documentation of policies and procedures, innovative data management for herd and test activities and for programme administration, training, and broad communication and awareness activities. Tools and systems are refined to address emerging issues and enhance the value of the programme. An Implementation Group comprising industry, government and technical leaders sets strategic direction and policy, advised by a Technical Working Group. Shared funding responsibilities are agreed by key stakeholders until 2022 to support herds in the programme to complete requirements. Herd registrations have increased steadily to exceed 1,800. National bulk tank milk surveillance is also being deployed to identify and recruit test-positive herds with the expectation that they have a relatively high proportion of seropositive animals. The programme will continue to innovate and improve to meet farmer and industry needs. This case study describes the implementation of a control programme for Johne's disease  in Ireland based on recommendations of Jordan et al. , some isAgriculture is Ireland's oldest and largest industry. Agricultural exports, particularly dairy products, are economically and strategically important for the expanding Irish economy .Ireland has a moist temperate climate. Irish dairy production is highly seasonal, aligned with seasonal pasture growth, to maximise production from natural inputs and the time spent by cows on pasture in a moist temperate climate. During winter, cows are generally housed to preserve soil health and are fed forage principally derived from harvesting surplus summer pasture growth. Milk production in January (winter) is <10% of peak production in May (spring). Ireland's dairy industry is structured on many small co-operatives as well as multinational agri-businesses, each of which independently determines farm milk prices.Following abolition of the European Union quota system in 2015, the dairy industry has increased milk production by 40%. Ninety percent of dairy output is exported and the value of exports has grown from 2 to 5.2 billion in value since 2015 , furtherIreland has established an international reputation for reliable supply of agricultural and food products from pasture-based farm systems that are safe, of consistently high quality, and environmentally sustainable . HoweverStakeholders in the Irish dairy industry seek to enhance the reputation of existing products and to reassure customers through ongoing assessment of potential risks to which Irish food products could be exposed with an emphasis on preventative animal health measures .Mycobacterium avium subspecies paratuberculosis  is ever demonstrated to be a disease of zoonotic significance, trading success for Irish dairy products may be protected and enhanced by a demonstrable, effective and scientifically based programme to reduce the risk of presence of MAP. There is ongoing scientific investigation and research to determine the extent, if any, of an association between MAP and Crohn's disease in humans . Using a process involving participatory consultation surveys and expert opinion, stakeholders ranked JD consistently as an important biosecurity risk disease requiring future management, even though the prevalence and production impacts of the disease were considered low at the time . CollectIreland commenced a pilot voluntary Irish Johne's Control Programme in late 2013 to determine the feasibility of transitioning to a national programme. The pilot programme was based on the findings of a review , AHI commissioned an evaluation of testing strategies to determine the most appropriate approach  and a reIn Ireland, JD is notifiable to enable the Department of Agriculture, Food and the Marine (DAFM) to monitor the incidence, but there is no formal regulatory approach to control or eradication.Vaccination against JD is not permitted, due to potential interference with testing for bovine tuberculosis (bTB).The IJCP is a significant collaboration between the Irish dairy industry and DAFM, and is managed by AHI. AHI is an innovative not-for-profit partnership between farmers, agri-food businesses, private sector organisations and DAFM, that delivers programmes for non-regulated diseases of livestock . AHI proThe IJCP has the support of all stakeholders involved in the programme, recognising it as having the capability to deliver a sustainable and internationally credible programme for the Irish dairy industry. Costs are shared by DAFM, the Rural Development Programme, individual milk processors and farmers. DAFM and milk processors have committed to maintain financial supports until at least the end of 2022.The programme is advised by a Technical Working Group (TWG) comprising veterinary and technical personnel from private and government fields with interest or expertise in JD. This group ensures that the programme is evidence based and reflects contemporary scientific knowledge about JD control.The programme is directed by an Implementation Group (IG) comprising AHI, DAFM, milk processors, farmer and veterinary representative organisations, milk recording organisations, breed societies, the Chair of the TWG and Animal Health and Welfare Northern Ireland (a sister not-for-profit organisation operating in Northern Ireland). This wide-ranging representation ensures that AHI stakeholders have a voice in the direction, design and implementation of the IJCP.AHI takes advice from both the TWG and IG and has responsibility for the day-to-day management of the programme.Prior to establishment of the IJCP, the herd-level true prevalence of JD on Irish dairy farms was estimated at 20% in 2005, based on the results of a serological survey , and wasThe four objectives established by the IG for the IJCP are to:Enhance the ability of participating farmers to keep their herds clear of JD.Assist participating farmers to reduce the level of infection in their herds, where present.Provide additional reassurance to the marketplace in relation to Ireland's efforts to control JD.Improve calf health and farm biosecurity in participating farms.To achieve these programme objectives, the following activities are required of participating herds:Annual herd level veterinary risk assessment and management plan . The VRAMP is undertaken collaboratively by an approved veterinary practitioner (defined below) and farmer, to systematically review the bioexclusion and biocontainment risks of JD for the herd and agree on up to three management changes to reduce the likelihood of introduction and spread of MAP.Annual whole herd test (WHT) comprising ELISA screening tests with ancillary faecal culture or PCR testing of animals with positive or inconclusive ELISA results. The purposes of the whole herd test are either to increase herd-level assurance for test-negative herds or early detection and monitoring of progress towards infection control for test-positive herds. A WHT requires all bovine animals on the farm aged two years or more  to be tested by ELISA, using milk or blood samples.Ancillary testing is required for all animals with positive or inconclusive ELISA results unless the herd has a previous positive result for a faecal test. The purpose of the ancillary test is to confirm the presence of MAP in the herd.An epidemiological investigation follows the first confirmation of infection in a herd under a Targeted Advisory Service on Animal Health (TASAH) programme. The purpose is to identify the likely source and spread of infection and to inform VRAMP refinements.The IJCP provides standardised protocols for testing and risk management, underpinned by training of veterinary practitioners and standards of laboratory testing to provide quality and consistency across the programme.Private veterinary practitioners who provide essential support to herdowners under the programme must undertake specific training presented by AHI in the basic epidemiology of JD and programme operations. After completing training, an “approved veterinary practitioner” (AVP) may carry out VRAMPs, animal sampling and test interpretation. AVPs may also undertake a second tier of training to provide the TASAH epidemiological investigations.All testing is conducted in designated laboratories, which are accredited to ISO 17025 for relevant tests, use only test kits approved by the Frederich-Loeffler-Institut with sensitivities and specificities indicated by the kit manufacturer, and participate in proficiency testing. Laboratories report results by electronic transfer to the programme database. DAFM provides National Reference Laboratory services.The programme provides funded supports for activities. For all herds, costs of required ancillary PCR testing  are fully funded by DAFM, and TASAH veterinary investigations are also fully funded. Ancillary faecal culture results are recognised by the programme, but this test is rarely used as it is not funded and requires a relatively long incubation period. For dairy herds that complete both the VRAMP and WHT annual requirements, DAFM funds the VRAMP and the milk processors fund herd testing assistance under agreed cost-sharing. Testing assistance is provided at the rate of EUR 2.75 per tested eligible animal for all herds in their first year. This rate is approximately the cost of ELISA testing using milk samples collected for milk quality and volume testing (milk recording). These supports are valued at EUR 550 for an average participating herd of 130 eligible animals. For testing of blood samples, there are additional costs for veterinary attendance, sampling and laboratory submission, borne by the herdowner. For test-negative herds, herd testing assistance declines over 3 years; for test-positive herds, assistance is maintained at the rate of EUR 2.75.There is no compensation for culling test-positive animals as provided under some eradication programmes, since the IJCP only advises rather than requires such removals.Completion of the WHT requires all animals to be sampled; a “sweeper” test of animals missed in a herd test  is commonly required, usually using blood samples. Non-breeding animals which are held in an epidemiologically separate unit to the breeding herd may be exempted from testing.The IJCP advises against ELISA testing within 90 days after tuberculosis skin testing, or within 7 days after calving (milk sample only), due to increased likelihood of false positive results.Farmers and their advisers have access to a range of resources including the IJCP technical manual, user guides and standard operating procedures, forms and templates, monthly information bulletins and technical leaflets. The IJCP publishes an annual business plan with clearly articulated targets. While these elements that underpin the programme are not unique to the IJCP, and form the basis of control programmes in other developed countries, notably Canada , GermanyComplementing the IJCP, DAFM undertakes animal disease surveillance including bulk tank milk testing (BTM) for a range of diseases, including JD. BTM may detect high-risk herds, so herds with positive BTM results are advised by a DAFM veterinary officer to join the IJCP, to avail of the funded tools to confirm infection and to control the spread and impacts of JD. However, BTM is considered a poor indicator of herd prevalence .Farmer participation in the IJCP is supported and encouraged by key stakeholders including milk processors, milk recording organisations (MROs), DAFM and the veterinary profession.At the end of 2020, there were 1,750 dairy herds registered in the IJCP, representing 11% of dairy herds and 18% of dairy cows in Ireland.Six hundred thirty one herds continued from the pilot programme to Phase 1 of the IJCP. There were 301 new registrations in Phase 1 (late 2017–2018), 729 in 2019 and 139 in 2020 . In consThe numbers of herds that completed both annual requirements of VRAMP and WHT were 1,376 (82%) in 2019 and 1,325 herds (75%) in 2020. In 2020, 326 registered herds (13%) were inactive and the remaining 99 herds either part-completed the WHT and/or did not complete the VRAMP.There has been a substantial improvement in the number and proportion of animals and herds conducting the required ancillary PCR tests, from 30% in 2018 to 67% in 2020. Many animals requiring outstanding PCR tests are no longer available for testing. If required ancillary testing of an animal with a positive or inconclusive ELISA result is not conducted, the animal and its herd are considered by the programme to be infected despite the presence of JD not being confirmed, which may have adverse consequences for the herd's future assurance standing.Registered herds self-selected into the programme and thus do not constitute a random sample of Irish herds, therefore extrapolation of programme prevalence data to the national herd is inappropriate. Additionally, due to the large number of herds (691) that have had at least one animal with a positive or inconclusive ELISA test result without an ancillary PCR test, the number or proportion of herds in the programme that are truly infected cannot be accurately calculated.For the 2020 programme year (including an extension for completion of requirements until 31 January 2021-discussed later), there were 224,364 ELISA tests conducted, 105,642 (47%) on milk samples and 118,722 (53%) on blood. Of these ELISA tests, 8,466 (3.8%) results were positive or inconclusive .ELISA tests of milk samples have consistently higher rates of positive and inconclusive results than tests of blood samples ; howeverThere were 5,419 ancillary tests in 2020, with positive results for 281 (5.1%) samples (not herds). Since 2018, the proportions of herds undertaking the required ancillary testing of animals following positive or inconclusive ELISA results have increased  but the Diminishing participation, for both recruiting new herds and completion of annual requirements by registered herds, is a current challenge. Multiple IG members, particularly milk processors, PVPs and MROs, reported that Brexit and COVID-19 concerns disrupted efforts to promote registration to their suppliers and clients for most of 2020. Farm access for sampling (both blood and milk) and VRAMPs was constrained by government restrictions, and later by aversion of farmers and their veterinary advisors to the risk of spreading COVID-19. The leadership of the milk processors in promoting the IJCP was re-directed towards ensuring the safety of suppliers and staff, continuity of supply and managing additional market risks created by Brexit and the COVID-19 pandemic.The costs of the programme for participating farmers are low (or a minor cost if blood-sampling), compared to the benefits. However, the majority of dairy farmers in Ireland have yet to register in the programme.This reticence is consistent with findings that “farmers are not solely influenced by economic consequences of management changes: . Smith aSimilarly for mastitis research, Regan et al.  found thELISA testing of milk samples during lactation typically extends from April until October, whereas ELISA testing of blood samples collected by AVPs (for either the whole-herd or only sweeper tests) is concentrated towards the end of the calendar year, and due to logistical and seasonal consideration collection may be delayed, on occasion occurring in the first month of the following calendar year.For efficiency, AVPs often carry out blood and faecal testing at the same time as conducting the annual bTB test and/or annual VRAMP. The blood and faecal sampling and VRAMP activities may be purposely delayed until the winter housing period, to minimise inconvenience and animal time-off-pasture for sampling and to coincide VRAMPs with the pre-calving period when the most effective interventions can be deployed.Despite the financial incentive of testing assistance, many herds have been unable to complete the annual WHT and VRAMP requirements before the end of the calendar year, necessitating one-month extensions of the programme years. Thirty percent of herds utilised this extension to meet the programme requirements for the 2020 programme year. It is proposed to integrate this flexibility to the programme in future years, notwithstanding the financial and administrative complexities.Testing and VRAMP data are held in the Irish Cattle Breeding Federation (ICBF) database. Designated laboratories upload test results for the IJCP, and AVPs upload VRAMP and TASAH investigation reports. The database also contains genetic and production information for all registered cattle in Ireland, including animal birth dates, pedigrees, livestock movements, disposal of animals, and bTB test dates (but not results) to assist interpretation of JD ELISA test results.Herdowners and AVPs can readily access the database via computer or mobile devices. The primary ICBF screen  did not find ready acceptance. Industry advisors reported “on-line fatigue,” especially for herdowners and AVPs with urgent and operational demands on their time.The programme has recently been broadened to apply to beef herds. It is expected to appeal to herdowners of either pedigree herds or commercial beef breeding herds with confirmed or suspected clinical disease. Pedigree herds may benefit by using a standardised protocol for market assurance and by seeing an increased demand for low-risk bulls to the dairy sector.Cost sharing for herd testing assistance and VRAMP funding for beef herds has not yet been determined.Development and early implementation of the IJCP holds learnings that may assist others who are planning JD control programmes.Strategic engagement of key stakeholders in agreeing objectives and sharing responsibility for decision-making and funding was critical. Policies and procedures must balance technical precision and pragmatism in the context of commercial dairy production.Farmers value simple, consistent, relevant and timely communications. Veterinary practitioners operating busy private practises may benefit from training beyond the technical elements of Johne's control, to equip them for the role of herd health advisor and for managing conflicts of interest. The inclusion of behavioural science perspectives from the outset may have foreseen and resolved barriers to participation.Sustaining Johne's control programmes can be challenging . At the Future recruitment is likely to require a different approach to messages and communication channels to convince more conservative farmers—the “late majority” and “laggard” groups for the diffusion of innovations [, cited iThe context of Johne's control is changing, as the Irish government promotes a National Farmed Animal Biosecurity Strategy that references JD  and milkA practical protocol for scoring herd risk, incorporating objective measures of risk from testing and histories of animal movements into each herd, and recognising VRAMP measures implemented to address individual farm mitigation priorities, is under development and expected to be released in 2021. This may provide additional incentives for farmers to register by providing tangible evidence of a herd's individual level of assurance. This could reward test-negative herds with voluntary marketing opportunities for low-risk breeding stock, encourage herdowners with infected herds towards effective biocontainment for their herds, and generally raise awareness of Johne's control.A proposed behavioural science study will take an inductive approach to examine the experiences of participating farmers. By identifying motivations of participants and barriers and facilitators to completing yearly requirements, the study will support effective recruitment strategies to increase farmers' intrinsic motivation to join the programme, identify who is best to communicate key messages and provide recommendations to improve timely completion rates of annual WHTs and VRAMPs. It will also explore farmers' experiences of receiving test results and involve a collaborative co-design exercise to identify how best to communicate the complexities of the programme and the benefits of risk management recommendation uptake to end-users. This study will include determining whether the dissociation of annual cycles, between the programme based on the calendar year and farming practises based on seasonal events, is a significant deterrent. A second study will collate the experiences of AVPs with the aim of improving IJCP support to them.AHI will continue to further incorporate JD control within broader biosecurity management, which was suggested by McAloon  as a meaWork on developing metrics to determine progress in achieving programme objectives is to continue with the support and input from stakeholders.The IJCP has adopted a number of proven activities and new technologies to address the strategic perspectives of Irish dairy herds, viz the protection against infection for the estimated 70% of low-risk herds, and controlling the spread and impacts of JD for the estimated 30% of herds that are infected.The most immediate challenges for sustaining and growing the IJCP are to maintain or improve the rate of recruitment of new herds, increase the completion rates for ancillary PCR testing and simplify the currently complex logistics of completing annual WHTs and VRAMPs within the seasonal cycles of milk production and bTB testing. The judgement of insufficient reward for the risk, inconvenience and expense of participation, especially for low-risk herds, remains a constant limitation and is recognised as an inhibitor to uptake of programmes internationally.Information, tools and processes will continue to be refined, based on global and local scientific knowledge, to address the four agreed objectives of the programme.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.LG and LC prepared the manuscript. DG and SM provided guidance and critical reviews. LG, DG, and SM are currently, and LC was formerly, members of the JD Implementation and the Technical Working Groups and have contributed to the development and implementation of the IJCP. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The purpose of this study was to investigate the effects of reconstituted whole wheat flour (WWF) particle size on flour characteristics and northern-type steamed bread (NTSB) quality. In this study, hard white (HW), hard red winter (HRW), and hard red spring (HRS) wheat classes, and four different bran particle size distributions  were blended at a ratio of 85% refined flour + 15% bran to create reconstituted WWF and make reconstituted WWF NTSB. Farinograph water absorption and water solvent retention capacity (SRC) increased as bran particle size decreased. Flour and dough strength tests such as lactic acid SRC and Farinograph and Mixolab development time and stability did not show any clear trends with bran particle size. HRW WWF tended to be the exception as Farinograph development time and stability generally increased as particle size increased. Resistance to extension increased as bran particle size decreased for HRW WWF and increased as particle size increased for HW and HRS. These differences in WWF dough rheology trends were likely due to differences in gluten characteristics between the classes. The results showed that larger particle sizes (105 μm and 125 μm) were more conducive to achieving desirable whole wheat NTSB specific volume, color, and texture. Chinese steamed bread (CSB) is a popular food in China and many Asian countries . CSB is Steamed bread accounts for approximately 40% of China’s wheat usage, and its market has spread to North America and Europe ,7. TaiwaWWF refers to flour that uses 100% of the wheat kernel, including all bran, germ, and endosperm materials ,12. The The presence of the bran and germ results in darker product color, reduced product volume, and firmer texture for products containing WWF . These eUsing WWF to replace refined flour is not always simple. Whole wheat flour comprised of coarse bran produced CSB with better texture and crumb structure and equivalent specific volume, skin color, smoothness, and shape compared to a fine bran WWF . Xu et aThe effects of bran particle size on water absorption and dough rheology are also varied. It has been reported that water absorption for fine bran WWF was greater ,14 and lThese effects on volume, rheology, and texture may be explained by studies that have indicated that the presence of bran in dough not only affects gluten structure ,17, but Another factor affecting the baking quality of WWF bread is the variability inherent in wheat bran. The addition of bran is challenging because it does not have a consistent chemical composition and quality . Wheat bIn this study, three different classes of wheat  and four different particle size distributions of bran  were blended at a fixed ratio (85% refined flour + 15% bran) to create reconstituted WWF and make reconstituted WWF NTSB. The purpose of this study was to investigate the effects of reconstituted WWF particle size on flour characteristics, dough rheology, and NTSB quality. The results show that larger particle sizes produce NTSB with more desirable quality attributes than smaller particle sizes.Wheat from three U.S. wheat classes  was used in this study. Their protein contents (dry basis) were 15.1% (HW), 13.8% (HRW) and 17.1% (HRS). The three wheat samples were tempered in plastic buckets at room temperature to a final moisture level of 14.5% and milled on a Miag Multomat experimental mill . The refined flours  were the controls in this study. Bran from each respective wheat sample was milled by Prater Industries  on a laboratory scale air classifier mill to four particle size distribution targets with D(50) values of 53 μm, 74 μm, 105 μm, and 125 μm. Other materials included instant dry yeast , sugar, salt, and shortening .w/w) to create reconstituted whole wheat flour (WWF). The final sample set included: HW , HRW , and HRS . This study utilized reconstituted whole wheat flours to control variability stemming from attempting to combine different ratios of bran and shorts from the wheat classes studied.After milling, control flours  were mixed with bran from each particle size distribution target at a ratio of 85% refined control flour to 15% bran  . BecauseWet gluten content (AACC 38-12.01) is commonly used to indicate quality differences, especially among flours with similar protein contents , and is To evaluate the flour quality characteristics , Solvent Retention Capacity (SRC) (AACC Method 56-11.02), Farinograph (AACC Method 54-21.01) , Mixolab (AACC Method 54-60.01)  and Kieffer rig extensibility  tests were conducted . The KieThe steamed-bread making process is outlined in L, a, b color was determined using a chroma meter . The texture profile analysis (TPA) of the steamed bread was determined using the TA.XTPlus Texture Analyzer equipped with a 35 mm acrylic cylindrical probe. Steamed bread was sliced horizontally, and a flat piece of 15 mm thickness was compressed to 50% of its original height in a two-compression cycle. The test conditions were pretest speed, 2 mm/s; test speed, 1 mm/s; post-test speed, 1 mm/s; and trigger force, 5 g.All steamed bread samples were evaluated for specific volume, exterior color, and texture. The specific volume of steamed bread was determined using a laser volume analyzer  and dividing the measured volume by the weight of the bread piece. The exterior p < 0.05 was used to determine significant differences between mean values. The results of the study were presented as the average ± standard deviation.All measurements were performed at least in duplicate. A one-way Analysis of Variance (ANOVA) was performed using SAS statistical analysis software. Duncan’s test was used for the comparison of means. The value Damaged starch content of the control flours varied depending on wheat class, likely due to differences in kernel texture and milling characteristics . Starch Damaged starch is one of the most important factors affecting quality characteristics of flour . There aA comparison of the starch damage results obtained from each method are presented in Whereas the Megazyme method is considered a direct method for damaged starch measurement, the amperometric method utilized by the SD-matic is indirect. Thus, a slight algorithm correction to the SD-matic may be necessary to pull these results into better alignment. Another factor to consider is that the SD-matic mechanism is based to some degree on the formation of an iodine-amylose complex. Therefore, the amylose content of the flours could be a factor that contributed to the shift towards lower starch damage values for the SD-matic . McAllisOne final observation is that the refined flours for HRW and HRS exhibited slightly greater starch damage than their WWF counterparts across all bran particle size distributions. This finding was unexpected and does not match the results for HW refined and WWFs. If this were an issue of dilution due to the presence of bran, we would expect to see it manifest across all three wheat classes. As the bran fraction used for reconstitution contained only the outermost bran layers, our observations may be more a function of bran differences among wheat classes and how they affect fractionation into different mill streams. Indeed, Hemery et al.  reportedThe characteristic dense and firm texture of NTSB is achieved primarily by using a 10–12% protein hard wheat flour exhibiting medium to high dough strength ,12. SRC Water SRC (W-SRC) is influenced by the swelling of gluten, damaged starch, and AX , and it Lactic acid SRC (LA-SRC) is specific for swelling of gluten polymers, specifically glutenins, with greater values potentially indicating stronger gluten and greater protein functionality. The protein contents of HWf, HRWf, and HRSf were 13.77%, 12.36%, and 15.66%, respectively . The LA-Sucrose SRC (Su-SRC) evaluates the functional contribution of AX to flour and baking performance. The bran contains more AX than the endosperm , and henSodium carbonate SRC (SC-SRC) provides information on the functional contribution of damaged starch to the flours. The SC-SRC values of the control groups  showed that the contribution of damaged starch from low to high was HWf < HRSf < HRWf . This alThe refined flour development time (DT) and stability (ST) were on the order of HRSf > HWf > HRWf . This orThe effect of bran on Farinograph DT and ST compared to a refined flour has been varied. Farinograph DT has been reported to be reduced for coarse WWF compared to refined flour ,35,42 whThe HRW WWF DT and ST tended to increase as bran particle size increased. It may be that HRW bran provided structural reinforcement, thereby increasing dough resistance against the mixing blades and delaying curve departure from 500 BU as observed by other researchers ,45. The Milling technique and mill flow, whole wheat and bran particle size reduction strategy , wheat genetics, and growing conditions are several factors that influence Farinograph WA, DT, and ST results. The lack of agreement in the literature is likely a combination of these factors. Establishing a standardized method for whole wheat flour milling would resolve the extrinsic factors contributing to conflicting flour quality results, leaving the remaining variability to intrinsic factors such as genetics and environment.Mixolab results for the refined and reconstituted WWF are shown in Bran particle size distribution generally did not appear to affect C1 and ST . AlternaC2 is a low torque point in the curve that occurs when heating is initiated A–C. It tPoints C3, C4, and C5 are related to starch pasting properties A–C. C3 iC4 is the lowest torque point after the C3 peak, and C3–C4 is regarded as an indication of amylolytic activity and hot gel stability related to the shear thinning of the dough after starch gelatinization A–C. The Retrogradation potential is indicated by the C5 value, also called setback in starch pasting terminology A–C. ThisWithin the WWFs, HRS had the greatest extensibility . In the The resistance to extension is generally used as a proxy for gluten strength in flour + water doughs and an indicator of gas-holding capacity, although it is not as straightforward in full formula doughs. Resistance to extension has been reported to decrease with the addition of wheat bran compared to a refined flour dough ,15,50. TInterestingly, HRW WWF samples still stand out from HRS and HW samples in terms of bran particle size trends. A reduction in bran particle size resulted in an increase in HRW resistance to extension. The HRW WWF samples aligned with previous findings that the resistance to extension increased as WWF particle size decreased ,15,16,42The ratio of resistance to extensibility (R/E) value shows the balance of elastic to viscous properties of the dough after relaxation. The larger the R/E value, the stronger the gluten and/or less the extensibility . The HRW3/g, respectively). These differences were not statistically significant within this data set. The addition of bran universally resulted in decreased specific volume across all classes for Chinese steamed bread, which is consistent with previous findings [findings ,12,49,50findings , dilute findings  and inhifindings ,53.A clear trend was also observed for bran particle size, with smaller bran particle sizes exhibiting a smaller specific volume . This trWhen compared with the specific volume of their respective control flours, HWb125 decreased by 16.4%, HRWb125 decreased by 6.0%, and HRSb125 decreased by 11.9%, respectively . SteamedAnother factor that could have influenced specific volume is related to the Su-SRC results. The Su-SRC values increased as bran particle size decreased, indicating increased accessibility of AX by the solvent . This coL, a, b, and whiteness index (WI) for the steamed breads. In all cases, the reconstituted WWF L and WI were lower than those of the control flours. Smaller bran particle sizes resulted in lower L and WI values than those from larger bran particle sizes. This may be related to differences in specific volume as larger volumes tend to result in brighter, whiter colors. The effect of specific volume is the most likely explanation for the differences in steamed bread color as WWF with a fine particle size has been reported to have a lighter color than coarse ground WWF [ound WWF .The textural characteristics of steamed breads as determined by TPA are shown in The greatest firmness was observed in HW and HRW samples. The protein content of HRSf was 15.66%. Adding bran or other materials in refined flour affects the formation of the gluten network during dough mixing. For refined flour with lower protein content, bran can have a more negative effect on the end product. It can therefore be speculated that HRS, with its naturally greater protein content and strong, extensible gluten characteristics, is more capable of overcoming the deleterious impact of bran in whole wheat steamed bread than either HW or HRW.Other research has shown that smaller bran particle sizes can adversely affect bread quality. This study presents similar findings, with larger bran particle size distributions (105 μm and 125 μm) being more suitable for producing whole-wheat steamed bread with a larger specific volume, brighter color, and softer texture. HRS appeared to be the most suitable wheat class for whole wheat steamed bread across the majority of the instrumentally measured product characteristics, although this would need to be confirmed with a sensory study.From a flour quality and dough rheology standpoint, none of the tests performed in this study offered sufficient predictive potential related to final product quality. The SRC GPI offered some promise in predicting steamed bread volume, with values approaching ~0.80 leading to better specific volumes. The most promising rheological method appeared to be dough extensibility and resistance to extension, with R/E values approaching ~1.50 being more conducive to optimal specific volume and texture (hardness) of whole wheat steamed bread for a given wheat class. However, this needs to be investigated with a larger sample set encompassing a wider range of flour quality characteristics.Future work should focus on investigating large bran particle sizes across a wider range of samples to identify optimal wheat quality, flour quality, and dough rheology parameters for CSB, as well as new methodology that is optimized for WWF.


In this study, a hydrothermal approach known as heat-moisture treatment (HMT) was identified as a simple and effective method in reinforcing the nutritional benefits of sbeIIb/Lgc1 rice. As a result of HMT treatment at 120 °C for 2 h, significant reductions in in vitro digestibility and enhancements in RS content were observed in sbeIIb/Lgc1 rice flour when the rice flour mass fraction was 80% and 90%. The low-glutelin feature of sbeIIb/Lgc1 rice was not compromised by HMT. The potential impacts of HMT on a range of physicochemical properties of sbeIIb/Lgc1 rice flour have also been analyzed. HMT resulted in a darker color of rice flour, alteration in the semi-crystalline structure, an increase in gelatinization temperatures, and reductions in the pasting viscosities as the moisture content increased. This study provides vital data for the food industry to facilitate the application of this dual-functional rice flour as a health food ingredient.A novel rice germplasm  Rice is one of the most important cereal crops worldwide and the processed rice grain, known as white rice, contains mainly starch as its nutritional component ,2,3. StaProtein is second only to starch as an important nutritional component in a rice grain ,17, 90% japonica and indica by disrupting the expression of the rice starch branching enzyme IIb (SBEIIb) gene that controls the synthesis of amylopectin through chemical mutagenesis /[Absorbance (1 mg/mL D-glucose solution) − Absorbance (reagent blank)] × 5 × 12.5.The digestibility of native or HMT flour was evaluated by measuring the released glucose as described previously  with somThe RS contents of native and HMT flours were measured using a resistant starch assay kit , referring to the Association of Official Analytical Chemists (AOAC) official method 2002.02. To determine the RS content of gelatinized flour, a sample of 100 mg native or HMT flour was mixed with 1 mL distilled water and then boiled at 100 °C for 20 min before measurement using the K-RSTAR kit. When the gelatinized samples were cooled down to room temperature, the sample was used for the measurement using the K-RSTAR kit immediately.A 24-well plate containing individual samples of native or HMT flour of 500 mg in weight was photographed using a Nikon D90 digital single-lens reflex (DSLR) camera . The chromaticity of each sample was determined by using a spectrocolorimeter CM-700d  following the manufacturer’s instructions. The total color difference (ΔE) and whiteness index (WI) were calculated as following formulas :ΔE=ΔL∗2+g for 15 min, the yellow supernatant was removed. The pellet was washed with distilled water three times by centrifugation at 3000× g for 15 min. Following the removal of the supernatant, the pellet was dried at 55 °C for 24 h to obtain coarse starch. The dried coarse starch was then dissolved into 4 mL of a 5 g/L alkaline protease solution , the pH value of which was adjusted to 10 with 1 M NaOH, prior to being stirred at 45 °C for 2 h. After centrifugation at 3000× g for 15 min and removal of the supernatant, the pellet was washed with distilled water three times by centrifugation at 3000× g for 15 min. Following the removal of the supernatant, the pellet was washed with 1 mL methanol three times by free sedimentation for 1 h and centrifuged at 10,000× g for 10 min. Following the removal of the supernatant after the last wash, the pellet was dried at 55 °C for 24 h prior to grounding and sieving through a 100-mesh sieve to obtain pure starch.Approximately 1 g native or HMT flour was added into 4 mL of a 4 g/L NaOH solution prior to being stirred for 2 h. After centrifugation at 3000× The pure starch powder was mounted to an aluminum stub using double-sided adhesive tape and coated with gold prior to morphological observation of starch granules by using a scanning electron microscope (SEM) Zeiss EVO-LS10  following the manufacturer’s instructions.The native and HMT flours were filtered by a 100-mesh sieve prior to being exposed to the monochromatic Cu-Kα radiation (λ = 0.15406 nm) produced by an X-ray powder diffractometer Bruker D2 PHASER  at 40 kV and 40 mA. The scanning regions of the diffraction angle 2θ were 5–60°. The scan rate was 4°/min with a step interval of 0.02°. The relative crystallinity was calculated and the original XRD patterns were processed by the software EVA 4.1  referring to the method described previously . The ressbeIIb/Lgc1. The gelatinization properties were analyzed using a Differential Scanning Calorimeter (DSC) DSC-Q20 .Following filtration by a 100-mesh sieve, a sample of 100 mg native or HMT flour was equilibrated and dried at 37 °C for 24 h, 5 mg of which was mixed with 10 μL deionized water in a hermetical aluminum cup prior to being heated from 20 °C to 110 °C at a rate of 10 °C/min for the native and HMT rice flours of SXG100, and from 20 °C to 130 °C at a rate of 10 °C/min for the native and HMT rice flours of Distilled water was added to a series of samples of native and HMT flours which were filtered by a 100-mesh sieve, and then measured by a rapid visco analyzer RVA4800 . The standard moisture content was set as 14%, under which the amounts of flour and water added were 3 g and 25 g, respectively. For each native and HMT sample, the amounts of flour and water were calculated by the built-in calculator of the RVA4800 software , depending on the actual moisture content. The rotation speed of the paddle was set to 160 rpm, and the resulting flour slurries were then maintained at 50 °C for 1 min before being heated from 50 °C to 95 °C at a rate of ~12.2 °C/min, at which they were maintained for 3.5 min before cooling down to 50 °C at a rate of ~12.2 °C/min, and finally maintained at 50 °C for another 2 min.w/v SDS, 4 M Urea, 5% v/v β-mercaptoethanol, 125 mM Tris-HCl) [w/v coomassie brilliant blue R-250, 25% v/v isopropanol, 10% v/v glacial acetic acid) for about 3 h at room temperature. Following the treatment by a destaining solution  overnight, the protein gel was photographed by using a Nikon D90 DSLR camera .A sample of 100 mg native or HMT flour was dispersed in 800 μL extraction solution (4% ris-HCl) , and incsbeIIb/Lgc1 or SXG100 was soaked in 20 mL tap water for 30 min in a glass beaker and sealed by a piece of filter paper prior to being steamed on boiling water for 30 min. The freshly steamed rice was photographed by a Nikon D90 DSLR camera .A sample of 10 g of the polished rice grains of p < 0.01 was considered as being significantly different for all the comparisons.All experiments were conducted with three replications, except the analysis of chromaticity which was conducted with nine replications. Statistical analyses were conducted using IBM Statistics 22.0 . The significance of variation in RS content of gelatinized or ungelatinized flours was analyzed using F-test to detect the homogeneity of variances, followed by a two-tailed t test, while the other measured values were analyzed by ANOVA and compared by least significant difference (LSD) analysis. sbeIIb/Lgc1 rice flour was significantly lower than that of SXG100 at all the time points tested. This result is consistent with previous research about sbeIIb [sbeIIb/Lgc1, the amount of released glucose at a rice flour mass fraction of 100% was comparable to that of sbeIIb/Lgc1 native flour. When the rice flour mass fraction was 90% and 80%, the glucose release amounts of HMT sbeIIb/Lgc1 rice flour decreased gradually, and at the time point of 120 min, the amounts of released glucose were ~72% and ~64% of that of native sbeIIb/Lgc1 rice flour, respectively. When the rice flour mass fraction of sbeIIb/Lgc1 was 80%, the starch hydrolysis rate at 120 min of digestion was only about 30%, which was the lowest among all the tested samples. According to the above results, we demonstrated that when the rice flour mass fraction was 90% and 80%, HMT at 120 °C for 2 h could further lower the in vitro digestibility of sbeIIb/Lgc1 rice flour, and they were expected to have better performance than native rice flour when used as an ingredient of food for diabetic patients.Following treatment by α-amylase and amyloglucosidase in combination, the amount of released glucose at different time points  of digestion in the native or HMT rice flour was measured to predict the changes of digestibility . It is at sbeIIb . For thesbeIIb/Lgc1 was reduced down to 70% under which the moisture content was slightly higher than conventional HMT standard, the amount of released glucose was higher than that of native flour. Such an observation might be attributed to the severe impact on the starch granule structure by high intensity heat treatment and the occurrence of partial gelatinization [sbeIIb/Lgc1 at a rice flour mass fraction of 70% was still lower than that of native rice flour of SXG100. Considering the energy supply, the lowest digestibility may not be always desirable. Through in vitro digestion assay, this study established that the effects of HMT on digestibility vary by the moisture content of treated rice flour. Therefore, the moisture content of HMT flour could be adjusted to achieve the optimal digestion rate to suit specific dietary requirements.When the rice flour mass fraction of nization ,39, evensbeIIb/Lgc1. Under HMT of 100% rice flour mass fraction, the digestion rate of the SXG100 flour began to decrease, and the lowest amount of released glucose for SXG100 was observed when the rice flour mass fraction was 90%. In contrast, when the rice flour mass fraction was 100%, the digestibility of HMT sbeIIb/Lgc1 rice flour was comparable with that of native flour and decreased when the moisture content increased. This might well be attributed to the characteristics of sbeIIb/Lgc1 starch granules and starch molecules which require more water to alter the interactions of starch molecules [sbeIIb/Lgc1 rice flour was still recognizable but those of SXG100 had already begun to disintegrate and fuse together .The effect of HMT on the digestibility of SXG100 rice flour was also observed, which showed a similar trend of reduction in hydrolysis rate, but to a lesser extent compared to olecules . Such a sbeIIb/Lgc1 rice flour was about 10.24% which was in sharp contrast to that of SXG100 (0.09%) which is in line with a previous study [sbeIIb/Lgc1, the RS content of the ungelatinized rice flour of 100% mass fraction was significantly lower than that of the native flour, which might be partially attributed to the disruption in the crystalline structure as reflected by the decrease of relative crystallinity . As the rice flour mass fraction decreased, the RS content was significantly raised relative to native flour at both rice flour mass fractions of 90% and 80% as a result of heat treatment. It might be that as the moisture content of treated rice flour increased, more water molecules made entry into the starch granules, which might alter the interactions of starch molecules to a great extent and reinforce the entanglements between starch chains or starch with other molecules, e.g., lipid or protein [sbeIIb/Lgc1 samples, which might also be caused by the partial gelatinization. The above observations demonstrated that HMT played a substantial role in the further increase of RS content of sbeIIb/Lgc1 when the moisture content of the sample was selected appropriately.In addition to in vitro digestibility, the comparative analysis of the RS contents of native and HMT flours is shown in us study . For HMT protein ,33. At tsbeIIb/Lgc1 were drastically reduced relative to their ungelatinized counterparts. Since gelatinization will cause an irreversible collapse of the starch granules and crystalline structures [sbeIIb/Lgc1 rice flour might be mainly attributed to the crystalline structure of starch and the products formed during HMT with resistance to amylase but lower thermal stability [sbeIIb/Lgc1 rice flours were significantly higher than that of native flour with the highest content being 3.30% at the flour mass fraction of 80%. For SXG100, though a slight enhancement of RS content was also observed when the sample moisture content was higher, the enhancement was not statistically significant and the absolute RS amounts were still quite low compared with those of sbeIIb/Lgc1. These observations revealed that HMT had a greater effect on sbeIIb/Lgc1 than SXG100 on reinforcing the interactions between starch molecules and promoting the formation of the thermally stable amylose-lipid complex that contributes to RS5, which might be attributed to higher AAC content and lipid content in sbeIIb/Lgc1 compared to SXG100 [The RS contents of the gelatinized native and HMT rice flours were also determined to facilitate the investigation of the mechanisms by which RS formed. Following gelatinization at 100 °C for 20 min, the RS contents of both the native and HMT rice flours of ructures , this obtability . Nevertho SXG100 ,7,40.sbeIIb rice was about 5~9%, varying among independent lines [sbeIIb rice makes it quite difficult to cook. In contrast to SXG100, the rice grains of sbeIIb/Lgc1 largely maintained their original morphology following the cooking process, with the core of many grains remaining undercooked even after being steamed at 100 °C for 30 min  was calculated to describe the extent of color difference of two samples and the whiteness index (WI) was calculated to describe the closeness of a sample to white . It has sbeIIb/Lgc1 was slightly darker and browner than that of SXG100, possibly because of the different chemical compositions. As a result of HMT, the color of HMT rice flours turned darker progressively as reflected by the L*-value for both sbeIIb/Lgc1 and SXG100. Meanwhile, the increases in the a*-value and b*-value indicated that the degrees of redness and yellowness of the HMT flours of both sbeIIb/Lgc1 and SXG100 were also enhanced. Consequently, the WI of HMT rice flours was also gradually decreased as the moisture content increased. A similar observation was also made when rice flour was treated by HMT under some other conditions [sbeIIb/Lgc1 were always brown and darker than those of SXG100, but for specific treatment the color difference between sbeIIb/Lgc1 and SXG100 was not dramatically higher than that between two native flours as reflected by comparable ΔE values when the rice flour mass fraction was 100% and 90%, suggesting the similar extent of effects of HMT on these two rices in spite of their difference in chemical composition. It has been reported that the extent of the browning of rice flour was positively correlated to the intensity of HMT [The appearances of native and HMT rice flours are shown in nditions ,42. Acconditions ,42. Undey of HMT ,42. ThersbeIIb/Lgc1 and SXG100 was only 0.8 and the ΔE between HMT and native flours of SXG100 was higher than that of sbeIIb/Lgc1. Considering the great color changes of HMT rice flours when the mass fraction was 70%, this observation suggested a promoting effect of partial gelatinization on the browning of heat-treated rice flours.Interestingly, when the rice flour mass fraction was 80%, the ΔE between the HMT samples of sbeIIb/Lgc1 showed irregular shapes with variable sizes, which is consistent with a previous study [To determine the effects of HMT on the morphology of starch granules, the starch derived from native and HMT flours was observed by SEM as shown in us study .sbeIIb/Lgc1 was severely impacted as the intact starch granules could be recognized readily when the rice flour mass fraction was above 80%. As the moisture further increased, i.e., when the rice flour mass fraction was 70%, the integrity of starch granules of sbeIIb/Lgc1 was impaired to a large extent and the individual starch granules began to agglomerate and fuse into clumps. A similar tendency was observed for the starch of HMT flours of SXG100. However, when the rice flour mass fraction was decreased to 80%, the starch granules in SXG100 HMT flour already showed severe collapse and fusion. These observations demonstrated that when the moisture content reached a threshold level, the physical integrity of starch granules collapsed to a great extent as the result of partial gelatinization, which is in agreement with previous reports [After HMT, neither the morphology nor the integrity of starch granules of  reports ,33,39.sbeIIb/Lgc1 was maintained to a large degree. Taken together, these observations suggested that the starch granules of sbeIIb/Lgc1 were more resistant to gelatinization inflicted by HMT with higher moisture content, which might be attributed to the high ACC of sbeIIb/Lgc1 [sbeIIb/Lgc1 and SXG100 HMT flours when the flour mass fraction was 80% and 70%.It is worth noting that when the flour mass fraction was 80%, which is considered as HMT, the starch granules of SXG100 had already collapsed and fused together, while the integrity of the starch granules from IIb/Lgc1 . This obsbeIIb/Lgc1 flour resembled B-type as manifested by the diffraction peaks at 2θ angles of 15°, 17°, 22°, 24° and ~5.6°, whereas the diffraction pattern of native SXG100 flour was a typical A-type manifested by the diffraction peaks at 2θ angles of 15°, 17°, 18°, and 23°. It appears that the XRD patterns of the native rice flour of both sbeIIb/Lgc1 and SXG100 were comparable to those of pure starches [sbeIIb/Lgc1, when the rice flour mass fraction was 100%, the relative crystallinity decreased and the feature peak of B-type crystalline at ∼5.6° 2θ angle disappeared. Intriguingly, the diffraction peak at the 17° 2θ angle appeared to split while the shoulder peaks at 22° and 24° showed signs of fusion when the rice flour mass fraction was 90%, and the B-type polymorph crystallites in native sbeIIb/Lgc1 rice flour transitioned to A-type when the rice flour mass fraction was reduced to 70%, as manifested by the diffraction peaks at 15°, 17°, 18° and 23° 2θ angles , the peak gelatinization temperature (Tp) and the conclusion gelatinization temperature (Tc) of native sbeIIb/Lgc1 flour were all significantly higher than those of SXG100, which is consistent with the previous study suggesting the naturally higher thermal stability of sbeIIb/Lgc1 than common rice [DSC assays were conducted to investigate the changes of the gelatinization behaviors of rice flour after HMT as presented in mon rice ,14.sbeIIb/Lgc1, when the rice flour mass fraction was 100%, the pattern of the thermogram was similar to that of native flour (o, Tp and ΔH were slightly lower and Tc was comparable with that of native flour. This observation suggested that the lower intensity HMT did not strengthen the interactions of starch molecules dramatically, but even disrupted the interactions of double helices in starch fractions with a lower thermal stability of sbeIIb/Lgc1 which might be related to a slight reduction of relative crystallinity (p become apparent (p corresponding to starch fractions with higher thermal stability became dominant (p was apparent (For HMT rice flour of ve flour , while Tallinity  and the allinity . When thapparent , which wdominant , while Δapparent  and ΔH fapparent , caused o, Tp, Tc and ΔH were comparable with those of native flour while slight increases were observed when the rice flour mass fraction was 90%. The patterns of thermograms were also similar to that of native flour at both rice flour mass fractions of 100% and 90% (p were also observed, suggesting the formation of starch fractions with a higher thermal stability (For SXG100, when the rice flour mass fraction was 100%, the T and 90% . When thtability . Howevertability . This obsbeIIb/Lgc1 was affected more profoundly by HMT and a higher amount of ordered starch fractions with intensified double helices structures and enhanced thermal stability formed, which might be attributed to higher AAC and lipid content of sbeIIb/Lgc1 than SXG100.Taken together, compared with SXG100, the native rice flour of sbeIIb/Lgc1 rice flour were all significantly lower than those of SXG100, reflecting stronger resistance of sbeIIb/Lgc1 starch granules to swelling than common rice due to higher content of amylose, which is congruent with numerous previous studies [sbeIIb/Lgc1 rice flours decreased drastically, especially when the rice flour mass fraction was 80% and 70%, under which the consistency of starch might be a suspension of starch granules instead of a gel [RVA assay was conducted to analyze the changes in the pasting properties of HMT rice flour. The measured pasting parameters of the native and HMT flours were shown in  studies ,14,26,46of a gel .For the HMT rice flours of SXG100 with rice mass fractions of 100% to 80%, the PV and BD were also lower than those of native flour and decreased gradually as the rice flour mass fraction decreased . NeverthsbeIIb/Lgc1 rice flours were more severe than those of SXG100 (sbeIIb/Lgc1, including higher AAC and lipid content, might have enhanced the effects of HMT and resulted in more profound changes in PV and BD.The reductions of the PV and BD of HMT f SXG100 . The redsbeIIb/Lgc1 rice flour [sbeIIb/Lgc1 rice flour might serve as a desirable ingredient of heat-processed food that is expected to have lower digestibility.Combined with the results of DSC assays, HMT further enhanced the thermal stability of ce flour ,40, suggsbeIIb/Lgc1 rice. To analyze the changes of protein composition of rice flour after HMT, total protein of the native and HMT flours was extracted and separated by SDS-PAGE referring to the method described previously [sbeIIb/Lgc1 native and HMT flours was verified by the significantly lower intensity of the bands corresponding to two glutelin subunits and dramatically enhanced intensity in the prolamin and globulin bands when compared to SXG100 (Except for starch-related properties, the low glutelin content is another desired characteristic of eviously . The releviously ,38. The o SXG100 . This obIn the HMT flour samples, all the bands on the gel became weaker along with the reduction in the flour mass fraction. The fact that all the bands on the protein gel became weaker following HMT suggests a reduction in free proteins, which might be owing to the increased interactions between proteins and between starch and proteins, which reduced the solubility and digestibility of proteins during the HMT process as reported by previous studies ,39,48,49sbeIIb/Lgc1 rice. In this study, when the rice flour mass fraction of sbeIIb/Lgc1 was 80%, which is the highest moisture content of the HMT samples, the lowest digestibility and a higher RS content were observed. Compared with common rice flour, HMT had a greater effect on the reduction of digestibility and enhancement of RS content of sbeIIb/Lgc1 rice flour which might be attributed to the intrinsic features of sbeIIb/Lgc1, such as higher AAC, higher lipid content and B-type crystalline structure. HMT also caused the alterations of other physicochemical properties of sbeIIb/Lgc1 rice flour including darker and browner color, alteration in the semi-crystalline structure, enhanced gelatinization temperatures and reduced pasting viscosities.Our study has demonstrated the feasibility of HMT as a simple approach to enhance the desirable traits of low digestibility and high RS content in rice flour by taking advantage of the intrinsic properties of sbeIIb/Lgc1 rice flour on stabilizing postprandial blood sugar level when adopted as an ingredient of functional food such as meal replacement powders or yogurt, which are suitable for patients suffering diabetes.This research paved the way for potential commercial applications of this promising rice flour as a vital ingredient in functional foods. Further studies are warranted to investigate the potential improvements of the HMT