Aspergillus and Penicillium producing selected mycotoxins. During the 2021 grape harvest, grapes were collected from the Small Carpathians wine region. Eleven grape samples were analyzed by the plating method and plating method with surface disinfection. Emphasis was placed on Aspergillus and Penicillium species because of their importance in mycotoxin production. Of the 605 fungal strains detected, 11 genera were identified in the exogenous mycobiota. The most common and abundant genera were Alternaria and Botrytis. In the genus Aspergillus, A. section Nigri is the most abundant, while in the genus Penicillium, P. raistrickii reached the highest frequency and abundance. Of the 379 strains detected and identified from the endogenous mycobiota, the most common genera were again Alternaria and Botrytis and the most abundant genus was Botrytis. Penicillium species were detected in 17% of all fungi found, with P. raistrickii dominating. The A. section Nigri reached only 4% of the relative density of all isolates. Potentially toxigenic Aspergillus and Penicillium species were tested for toxinogenity by thin layer chromatography. The most important mycotoxin-producing species found were A. section Nigri but without ochratoxin A production.The microbiological characteristics of the grapes are made up of a wide variety of microorganisms, including filamentous fungi. Their presence in grapes is traditionally associated with deterioration in quality. The health of the grapes is very important for obtaining quality wine. The objective of this study was to investigate the diversity of mycobiota on the surface and inside of different grapevine varieties at harvest time in the temperate climate of Slovakia and to identify potentially pathogenic isolates of  The area under vine in the Slovak Republic is 14,642 ha, of which 3,947 ha is in the Malokarpatska wine-growing region, 4,171 ha is in the South Slovak wine-growing region, 2,993 ha is in the Nitra wine-growing region, 1,713 ha is in the Central Slovak wine-growing region, 726 ha is in the East Slovak wine-growing region, and 1,092 ha is in the Tokaj wine-growing region [Aspergillus and Penicillium produced 3-methylbutanol, 3-octanol, 1-octen-3-ol, 1-octanol, and 2-octen-1-ol along with a number of higher alcohols, aldehydes, and ketones [Grapes, like other foods, can be contaminated by various fungi. Fungal diseases in the vineyard affect the quality of grapes by influencing the volatile profile and consequently affecting the aroma, flavor, and color of juice and wine. Molds synthesize numerous compounds e.g.,  ketones .However, food and feed may be contaminated by several fungal species simultaneously or in rapid succession. Therefore, humans and animals are generally exposed to a cocktail of mycotoxins. Even very low doses of mycotoxins can synergistically impair the intestinal health, and the consequences of combined exposure to environmental and food contaminants are specific to the target organ . The preAspergillus and Penicillium are of particular concern [2, ochratoxin A (OTA), patulin, citrinin, and others [Aspergillus carbonarius and Aspergillus niger are the main fungi responsible for the accumulation of OTA in grapes [A. carbonarius is less frequently isolated than other black aspergilli, it is considered the most important producer of OTA because almost all isolates  produce the toxin, which occurs at high concentrations [Aspergillus and Penicillium mycotoxins such as citrinin and Penicillic acid, which also have nephrotoxic and carcinogenic effects, with AFs, or with the Fusarium mycotoxin fumonisin B1 [Among the contaminating microorganisms, the mycotoxin-producing fungi  concern ,8. Amongd others . OTA is d others ,11. Thisd others . Aspergin grapes ,11,12,13trations . More oftrations . OTA occnisin B1 . CompareBotrytis infections are the most common causes of severe crop loss in grapes. Sour rot usually affects dense grapes just before harvest and is typically characterized by vinegar odor and brown berries [Botrytis infections  often occur in vineyards exposed to cold and humid conditions during the ripening period and has a detrimental effect on the organoleptic properties [Sour rot and  berries . Botrytioperties . Good agoperties .22.1In the 2021 vintage, the grape samples were collected from a small family winery in the village Vrbove (48°37′12″S 17°43′25″V), which is situated in Vrbovsky subregion, the part of the Small Carpathians wine region [2.2Aspergillus strains were incubated on Czapek Yeast Agar  [Penicillium strains were incubated on CYA, MEA, Creatine Sucrose Agar [Aspergillus strains was according to Klich [Penicillium strains was according to Pitt and Hocking [We used two methods to cultivate and isolate the fungi: (a) direct plating without surface disinfection which corresponds to exogenous contamination and (b) direct plating with surface disinfection which corresponds to endogenous contamination. Fifty grape berries from each sample were randomly selected and then plated in Petri plates  with Dichloran Rose Bengal Chloramphenicol Agar . Petri plates were incubated at 25°C for 7–10 days. Fifty additional grape berries were surface disinfected for 1 min in 1% NaClO according to the methods of Magnoli et al.  and rinsGermany) , Malt ExGermany) , and CzaGermany) . Penicilose Agar , and Yeaose Agar  incubateto Klich , Pitt anto Klich , and ide Hocking , Samson  Hocking .2.3The obtained results were evaluated and expressed in terms of relative density (RD) and isolation frequency (IF). The RD (%) is defined as the percentage of isolates of the species or genus present in the analyzed sample . These vIF (%) is defined as the percentage of samples in which the species or genus occurred at least once. These values were calculated following González et al.  as follo2.41, AF G1, citrinin, and patulin – were inoculated on YES agar and potentially toxigenic species producing intracellular metabolites – roquefortine C, penitrem A (PA), and cyclopiazonic acid (CPA) – were inoculated on CYA agar. From the colonies (after 14 days of incubation), some mycelial pieces of about 5 mm × 5 mm were cut and placed in an Eppendorf tube containing 500 μL chloroform:methanol in the ratio of 2:1 . The contents of the tubes were shaken for 5 min with Vortex Genie® 2 . Mycotoxins extracted from cultures of fungal isolates were determined by TLC technique on a precoated silica gel plate . 30 μL of the liquid phase of the extracts was applied to the TLC plate together with 10 μL of standards . The plate was placed in TEF solvent . After elution, the plate was air dried. Mycotoxins were identified by comparison with appropriate reference standards for mycotoxins. Roquefortine C (RC) was visible as an orange spot after spraying with Ce(SO4)2 × 4H2O. PA was visible as a dark blue spot after spraying with 20% AlCl3 in 60% ethanol and heating at 130°C for 8 min. CPA was visible as a purple-tailed spot after spraying with Ehrlich reagent directly in daylight. Patulin was detected by spraying with 0.5% methylbenzothiazolone hydrochloride in methanol and heating at 130°C for 8 min and was then visible as a yellow-orange spot. Directly under UV light with a wavelength of 365 nm, OTA was visualized as a blue-green spot, citrinin as a yellow-green-tailed spot, AF B1 as a blue spot, and AF G1 as a green-blue spot.To evaluate the toxigenic potential we used the agar plug method on thin layer chromatography (TLC) according to Samson et al. , modifie33.1Alternaria and Botrytis , followed by Aspergillus (82% IF), Penicillium (73% IF), Cladosporium and Mucor , and Rhizopus (45% IF). The most abundant genus from exogenous mycobiota was Botrytis  and Aspergillus (6% RD) of all fungi found. The other genera were detected in less than 5% of all isolates. Sclerotiniaceae and genus Botrytis (43%), and Pleosporaceae and genus Alternaria (32%).The filamentous fungi of the surface mycobiota identified in eight white and three red grape varieties are shown in  43% RD, , followe3.2Alternaria and Botrytis , followed by Aspergillus, Penicillium , and Rhizopus (45% IF). Botrytis reached the highest number of isolates in most samples, except in two varieties of Chardonnay (4) and Feteasca Regala (10), where the genus Alternaria dominated, and Alternaria was not detected at all in 2 samples of Müller-Thurgau (6) and Pinot Blanc (8). The most abundant genus of endogenous mycobiota was Botrytis  and Penicillium (17%) of all fungi found. The remaining genera were detected in less than 5% of all isolates. The most isolated family of mycobiota by the direct plating method with surface disinfection is shown in Sclerotiniaceae and genus Botrytis (51.2%), and Pleosporaceae and genus Alternaria (21.1%).The filamentous fungi of the endogenous mycobiota identified in 11 grape varieties are shown in tis 51%, , followe3.3Aspergillus isolates and 52 Penicillium isolates representing A. flavus, A. section Nigri, P. crustosum, P. expansum, P. chrysogenum, and P. raistrickii. From the exogenous mycobiota of grapes, 34 isolates belonging to 2 Aspergillus and 4 Penicillium species were screened. All strains of A. section Nigri tested were negative for OTA, and one strain of A. flavus was negative for AF B1, G1, and CPA. Three strains of P. crustosum were positive for RC and PA, and one strain of P. expansum was positive for RC, citrinin, and patulin. A positive toxinogenity was detected for RC by P. chrysogenum (three out of five strains) and for griseofulvin by P. raistrickii (five out of six).Food safety is a priority for humanity. Mycotoxins pose a major threat to food safety . The datAspergillus strains and 37 Penicillium strains isolated from the endogenous mycobiota of grapes were evaluated for their ability to produce mycotoxins. Negative toxinogenity for OTA by Aspergillus section Nigri was detected not only from exogenous but also from endogenous mycobiota. The ability to produce RC by P. chrysogenum in in vitro conditions was detected in 12 out of 14 strains and griseofulvin by P. raistrickii in 18 out of 23 strains.Ten 4Alternaria was also the most frequently occurring genus with the highest RD in Slovak grapes from the Small Carpathians wine region, the South Slovak wine region [Botrytis was a less frequently occurring fungal genus with a low RD in grape samples [Botrytis cinerea is considered to be one of the most destructive fungi in viticulture in cool climates, causing Botrytis bunch rot or grape grey mold and dramatically changing the physiochemical state of grape berries [Aspergillus, Alternaria, Botrytis, Penicillium, and Cladosporium were five of the most frequent genera in all four winemaking regions in Portugal [Aspergillus was higher than that by Penicillium in most samples, except for Alibernet, Chardonnay, and Pinot Blanc, in which Penicillium was more represented. However, the diversity of Penicillium species was higher than that of Aspergillus. Kizis et al. [Aspergillus was the most frequently isolated genus in all regions except Macedonia, where the genus Alternaria was most common. EL Khoury et al. [Aspergillus, while only 4.5% belonged to the genus Penicillium. The Aspergillus species found in our grapes from the Vrbovsky subregion were A. flavus and A. section Nigri. The species of section Nigri accounted for 97% of the Aspergillus isolates. These results are consistent with the reports by Battilani et al. [A. section Nigri was the most commonly isolated species in hot and dry areas. Among them, A. carbonarius and A. niger were the most abundant species in grapes [A. section Nigri were predominant in the mycobiota of grapes from the Small Carpathians area in Slovakia during 2011–2013 harvest [Aspergillus species is present worldwide, in all wine grape products, and under all environmental conditions, most frequently in warmer regions and on heat-producing substrates [A. clavatus and A. flavus were the other main species recorded with a high IF. The identified species of the genus Penicillium were P. raistrickii, P. chrysogenum, P. crustosum, P. brevicompactum, P. expansum, and P. polonicum. Most cultivars were colonized by P. raistrickii, which dominated in 27% of the samples examined and accounted for 41% of Penicillium isolates, especially in the Alibernet cultivar. Penicillium raistrickii is a soil fungus and has a widespread though sparse distribution in food [Penicillium species were P. sclerotiorum, P. citrinum, P. glabrum, P. decumbens, P. implicatum, and P. solitum [Penicillium brevicompactum, P. thomii, and P. glabrum were found in Portuguese vineyards [Penicillium chrysogenum was the most frequently isolated species in Argentina [P. brevicompactum was the species with the highest occurrence [P. expansum as the most common species in Portuguese [In previous studies ,31, Altee region , and thee region . Botryti samples ,32,33. T berries  or noble berries . AspergiPortugal . In our s et al.  studied y et al.  found sii et al. , who foun grapes  found si harvest ,32. Certbstrates . The tox in food . These r solitum . Penicilineyards . Penicilrgentina  and Slovrgentina . In Franrtuguese , French rtuguese , and Slortuguese ,32,40 viAlternaria and Botrytis in the endogenous mycobiota of grapes. Alternaria was the predominant genus in the endogenous mycobiota of grapes, and also in the Small Carpathians region [Penicillium is a common component of grapes and soils in temperate regions [Penicillium species were isolated, namely P. brevicompactum, P. crustosum, P. chrysogenum, P. polonicum, and P. raistrickii. Six samples were colonized by Penicillium species. The grape variety Cabernet Sauvignon was the most colonized by the species P. raistrickii. On the other hand, Penicillium was less frequently detected in grapes from the Small Carpathians area in 2018 with these four Penicillium species: P. citrinum, P. griseofulvum, P. hordei, and P. chrysogenum [Aspergillus was as frequently occurring genus as Penicillium, but its RD was lower. All 15 isolates were identified as A. section Nigri. A similar conclusion was also reached by other authors [In our study, almost all samples were colonized by the genera s region , in the s region , and in s region . Penicil regions . From thysogenum . In our  authors ,44,45,46Vitis vinifera grapes [P. crustosum from our samples were positive for RC and PA, and one strain of P. expansum was positive for RC, citrinin, and patulin. Penicillium was a common component of grapes from the Small Carpathians area from 2011 to 2013 [P. crustosum on PA and RC . Penicillium expansum produced RC (5 out of 5), patulin (3 out of 5), and citrinin (2 out of 5). Patulin is considered less risky because production of significant amounts of patulin is associated with visible rotting of fruit, so almost all of the toxin can be removed by rejecting rotten fruit through selection and sorting. Nevertheless, patulin concentrations in food are subject to regulatory control in some countries [Production of OTA was not confirmed in grapes from some Slovak wine-growing zones such as the Central Slovak wine region , the Easa grapes ,50,51,52 to 2013  where po5Botrytis and Alternaria as the most frequent and abundant genera that can affect the sensory profile of must and wine. OTA, griseofulvin, patulin, citrinin, RC, PA, AF B1, AF G1, and CPA production were tested by TLC method in in vitro condition. Although several grape samples analyzed were contaminated with A. section Nigri, the abundance of Aspergillus was minimal and none of the isolates produced OTA. This is of crucial importance for understanding the fungal hazards for grapes and subsequently for wine and for the knowledge of field ecology of the fungal species. We recommend regular monitoring of mycological colonization of grapes, especially during harvest, with a focus on toxinogenic genera. Attention should also be focused on research aimed at mitigating mycotoxins in grapes or wine.The present study provides a detailed description of the filamentous fungi found on the berry surface and inside of Slovak grapes collected from the wine region largest in size and the most important over the centuries, the Small Carpathians wine region in 2021. The results show


Crop production is becoming an increasing challenge as the global population grows and the climate changes. Modern cultivated crop species are selected for productivity under optimal growth environments and have often lost genetic variants that could allow them to adapt to diverse, and now rapidly changing, environments. These genetic variants are often present in their closest wild relatives, but so are less desirable traits. How to preserve and effectively utilize the rich genetic resources that crop wild relatives offer while avoiding detrimental variants and maladaptive genetic contributions is a central challenge for ongoing crop improvement. This Essay explores this challenge and potential paths that could lead to a solution. The wild relatives of domesticated plants harbor tremendous genetic diversity that has the potential to help adapt crops to changing climates and future challenges. This Essay looks at historical admixture between crops and their wild relatives, and how these traditional varieties provide an evolutionary filter that has selected for useful diversity. Plant domestication is a process that began with cultures around the world experimenting with alternative means of food production. These experiments have expanded, undoubtedly, beyond the imaginations of their originators. Much of the planet’s surface is now involved, and most of human sustenance derives from experiments that originated among fewer than 1,000,000 humans starting 10 to 12,000 years before present. Domestication has involved a mix of intentional  selection and unintentional (sometimes “unconscious”) selection—the simple result of differential survival and reproduction among individuals in a novel environment . Evo. Evo2]. Plants that have coevolved with humans. Most domesticates rely on humans for survival and reproduction.Alleles that decrease the survival or reproductive capacity of an organism.One of the fundamental evolutionary processes, genetic drift refers to stochastic changes in allele frequencies unrelated to an allele’s impact on fitness.Natural selection that removes deleterious alleles from a population.Genetic resources related to the species being studied, including wild relatives, unimproved populations such as landraces or heirlooms, and improved varieties. These genetic resources—usually seeds but possibly including living plants or tissue culture—are collected and maintained for long-term preservation and are commonly used in genetic studies or breeding programs.Introduction of genetic material from one taxa or species into another. Introgression may occur naturally via hybridization or via inbreeding by traditional crossing and repeated backcrossing to a recurrent parent. The size of the introgressed region depends on the local recombination rate and how many backcrosses have occurred; each backcross with the recurrent parent results in reduction of the donor genome by approximately 50%.The recipient parent of an introgression, typically an improved variety with high yield or superior quality, that is otherwise lacking a particular trait to be introduced from a donor variety.In species that exhibit heterosis or hybrid vigor, there are often specific germplasm combinations that result in higher or lower levels of heterosis. Low levels of heterosis may result when crossing individuals from the same heterotic group, while crosses between individuals from 2 different heterotic groups result in higher levels of heterosis.The evolutionary path from wild species to domesticate is different for each crop ; thus, tWhether domestication entailed thousands of generations of gradual selection or the extraction of a single clonal genotype from wild populations, it nearly always results in the loss of genetic diversity in traditional varieties and modern cultivars compared with crop wild relatives. An understanding of the extent of loss in genetic diversity in domesticates is relatively new , as earlIn this Essay, we argue that allelic diversity from crop wild relatives likely already exists in cultivated populations conserved in germplasm  repositoCrop wild relatives contain a wealth of alleles that were lost during domestication and improvement –29. ThesCrop wild relatives and traditional varieties share much of their genetic makeup with modern cultivars. In the initial stages of cultivation and in incipient domesticates, long segments of the genome will be shared with wild relatives. These relationships reflect identity by descent (IBD) from parents to progeny. The size of IBD regions is reduced each generation by recombination and is dependent on factors such as the outcrossing rate and diversity within populations. Individuals from closely related populations can share large regions of IBD over hundreds of generations . SharingBecause domestication is a recent evolutionary process, the majority of gene-level variants in modern cultivars, including single-nucleotide changes and insertions and deletions, are a subset of those found in crop wild relatives c.f. ,34). Non. Non34])Direct introgression of crop wild relative alleles is a major strategy for increasing genetic diversity and genetic variation in commercial breeding programs. Breeders typically use introgression to respond to an emerging threat or existing deficiency in the commercial germplasm collection. Genetic variation is required for breeders to make genetic gains, and alleles that can confer a selective advantage for emerging threats such as a new disease or climatic extremes may not exist in modern cultivars. In this case, researchers and breeders look for the desired trait variation in crop wild relatives in hopes of finding individuals that contain alleles with large genetic effects capable of producing adapted progeny. This approach has been successful in many cases , includiIncorporating novel diversity from crop wild relatives into commercial breeding populations is the fundamental goal of “pre-breeding” for many crops. The pre-breeding process requires phenotyping crop wild relatives, identifying key lineages (donors) with beneficial or novel alleles, and introgressing the donor alleles. Introgression is most commonly done by crossing diverse germplasm with the relevant modern cultivar in conjunction with selection for the novel trait or allele using phenotype, genetic markers, or predicted breeding values . One sucBy definition, crop wild relatives are unimproved or less improved than modern cultivars and hence harbor alleles that are maladaptive under modern agronomic practices . These mOne solution to linkage drag is marker-assisted backcrossing, where flanking markers are used to track the desired allele in a breeding population and to make selections, and genome-wide markers are used to actively select against the remaining donor genome and for the recurrent parent, effectively prioritizing recombination events close to the target locus. The added expense and effort of implementing marker-assisted backcrossing in a breeding program is such that it essentially requires an allele with a large effect to recover the value. In addition, the result is unlikely to be a single gene introgression; larger introgressions can contain dozens to hundreds of genes depending on the genomic context. Each desirable allele will reside in a region of the genome that may contain numerous maladapted alleles, and the combination of recombination rate and haplotype structure  will determine the likelihood of breaking up linkage blocks . In the Overall, while the long-term advantages of increasing diversity and adding functional variation from crop wild relatives are well understood, the short-term challenges are often sufficient to prevent the effective utilization of such wild relatives in breeding programs. This is especially true in industry settings focused on short-term profits (though there are some notable exceptions). The commitment to existing heterotic patterns  makes wiHistorically, domestication has often been portrayed as the split between cultivated plants and their crop wild relatives. However, empirical studies from a variety of systems highlight that domestication was a complex process that unfolded across a diverse landscape and involved genetic exchange both with a crop’s direct progenitor, as well as with additional wild relatives ,56. HumaFar from being accidental or detrimental, gene flow with crop wild relatives has often been instrumental in the evolution of domesticated taxa. In maize, for example, a meaningful subset of recent selection in traditional varieties has been for alleles introgressed from a wild relative , and intIf introgression from crop wild relatives generally increases maladaptation due to linkage drag and deleterious alleles, why have these processes not prevented historical gene flow? In fact, it is likely that hybridization with crop wild relatives was constrained by maladaptation. For example, ongoing gene flow between traditional varieties of maize and one of its wild relatives is depleted around loci important for maize domestication . But thiHow can wild relatives and existing germplasm resources be best used to adapt crops to the novel environments and agronomic practices that will accompany changing climates? One approach that has garnered much public attention is the use of novel genome editing techniques to “domesticate” wild plants or introduce alleles from crop wild relatives into modern germplasm. In one recent study, researchers edited a number of key genes to dramatically change the architecture and agronomic suitability of a wild relative of tomatillo . In anotHowever, we would argue that such approaches are not likely to be the most fruitful avenue for using wild relative diversity to improve crops. Domestication invariably involves changes at hundreds or thousands of alleles , such thWhile genome editing is undoubtedly a useful tool, we argue that an effective and efficient avenue for incorporating crop wild relative diversity into modern germplasm is to use wild relative alleles already present in traditional varieties housed in germplasm banks. Germplasm repositories maintain a wealth of historical and modern genetic diversity. They increasingly include additional genetic and phenotypic data that can be used a priori to help narrow down useful material –84. IncrThe incorporation of crop wild relative and traditional variety alleles into elite breeding programs is dependent on a number of factors, including the complexity of the trait or traits of interest, the ease of intercrossing, generation time, and the timeline for trait development. At one extreme, some crops may require lengthy pre-breeding interventions involving multiple crosses to bridge between diverse germplasm and relevant modern cultivars. Such bridging crosses limit the potential for genetic exchange and slow the process of introgression. But in species with genetic compatibility among wild relatives or traditional varieties, the development of multiparent populations can accelerate the identification of loci contributing to trait variation, facilitate recombination, and uncover multiple alleles at a locus that contribute to a trait . In someFor many perennial crops, including trees and clonally propagated species, conservation of wild relatives and traditional varieties can be complicated by the limited potential for reproduction by seed or the need to preserve particular strains. Species such as apple, citrus, grape, and avocado are preserved in living nurseries, where many plants may consist of distinct genetics aboveground grafted to rootstocks that are tolerant of local growing conditions and soil pests . While tTriticum tiompheevii, for example, is a wild relative of wheat that is most likely extinct in situ but still available from ex situ collections [We are losing genetic resources from farmers’ fields through the replacement of traditional varieties by modern cultivars  and fromlections .Germplasm repositories are increasingly pushing to make their resources available. They do so most effectively by sharing germplasm under agreed terms as described under the multilateral system of the International Plant Treaty  or simil


Each year an estimated 48 million people are bereaved by suicide internationally. Following traumatic events, experiencing intrusive mental imagery relating to the trauma is not uncommon. This phenomenological study aimed to explore the nature, experience and impact of intrusive mental imagery after suicide bereavement.Semi-structured interview transcripts with 18 adults bereaved by the suicide of a close contact were analysed using thematic analysis to explore patterns and themes within the data, with particular consideration of the content of images, how people experience and relate to the imagery, and the impact that the imagery has on the bereaved.Thematic analysis identified common characteristics in the experience of intrusive mental imagery following suicide loss, summarised under two main themes capturing: 1) the descriptive characteristics and 2) the emotional experience of intrusive mental imagery following suicide loss. The majority of participants found the experience of intrusive imagery distressing, but most also described positive aspects, including help in making sense of the death and retaining memories of the deceased.Findings inform our understanding of the distressing experience of intrusive imagery after suicide loss, also revealing perceived value in processing the death. Annually nearly 700,000 people die by suicide worldwide, exposing an estimated 48 million people to suicide bereavement , 2. SuicTrauma-related intrusive memories are an involuntary cognitive-perceptual phenomenon characterised by distress and interference with activities of daily living . ClinicaThe majority of studies describing intrusive mental imagery (and interventions to address this) are situated in the context of PSTD, and whilst there are valuable insights to be gained from this clinical research there is also a need for research specific to suicide loss. PTSD is a common reaction to traumatic events, characterised by repeated and unwanted reexperiencing of the traumatic event in the form of intrusive memories and distressing dreams . People Gaining a better understanding of the nature and impact of intrusive cognitions experienced after suicide bereavement is an essential first step in clarifying whether mental imagery is an appropriate target for intervention in people bereaved by suicide experiencing intrusive imagery-related distress. In this study we aimed to investigate the phenomenon of intrusive mental imagery after suicide bereavement in a community sample by analysing data from interviews exploring individual experiences of such imagery following the suicide of a close contact.This study was approved by the UCL Research Ethics Committee .We recruited a community sample of bereaved adults from the general population in the UK through an advert distributed via social media and contact lists of an umbrella group of UK suicide bereavement support charities (Support After Suicide Partnership). Participants responding to the advert were screened for eligibility via an online questionnaire. Inclusion criteria included: adults aged 18 years and over (following bereavement in adulthood or childhood), UK residence, bereavement by suicide of a close contact , experience of intrusive imagery relating to that loss, and proficient English. Exclusion criteria included: recent suicide bereavement (within six months), recent suicide attempt and apparent cognitive impairment. Participants were not screened for pre-existing mental health disorders. Our risk protocol set out clear steps for managing any risk issues identified during screening or interviews. All participants gave written informed consent which included seeking the participant’s general practitioner and gaining consent to contact them in the event of risks being identified. Participants were also provided with a list of support services.Eligible participants were contacted by the researcher to facilitate informed consent. Purposive sampling of those eligible was used to capture different experiences of suicide bereavement .The interview topic guide was developed in collaboration with the Support After Suicide Partnership see  and desiWe analysed interview transcripts using thematic analysis to identify themes within the data capturing the phenomenon of intrusive mental imagery: the content and form of the imagery, how the imagery was experienced, its emotional impact, and any relationship of the imagery to factors such as kinship to the deceased or characteristics of the relationship .Initial analysis of interview transcriptions involved detailed notation of units of meaning and impression, developing initial codes. Two transcripts were independently coded by two researchers (KQ and PJ), discussing any coding differences to agree an initial coding framework. One author (KQ) then coded all remaining transcripts, collating codes into potential themes and refining themes in the iterative development of the thematic framework through discussions with the wider research team. Such discussions also involved a consideration of the differing perspectives and experiences of the analytic team to address reflexivity.We presented our findings in line with COREQ guidelines on the reporting of qualitative research .Of those responding to the study advert, 21 potential participants were contacted for an initial eligibility screen. All were deemed suitable to participate, and 18 subsequently consented for interview. Demographic characteristics of the sample are described in Thematic analysis identified two overarching themes capturing 1) the descriptive characteristics of intrusive imagery following suicide loss and 2) the cognitive and emotional responses to this. The first theme comprised three subthemes: (1.1) nature of imagery; (1.2) degree of control over imagery; and (1.3) changes in imagery over time. The second theme comprised four subthemes: (2.1) intrusive imagery is unhelpful and distressing; (2.2) impact of imagery; (2.3) ways of coping; and (2.4) positive aspects of the experience of imagery.These themes and sub-themes are set out below with illustrative quotes. Participants’ individual characteristics are shown in 1.1. Nature of imagery. Generally, participants’ descriptions of intrusive imagery identified differences in the form and quantity of the imagery (imagery characteristics): these defined people who experienced the same image repeatedly, those who reported a limited number of images relating to the suicide, and those who perceived multiple images relating to the suicide. For some people these patterns changed across the course of bereavement. There was substantial variation in the perceived frequency of the intrusive images, which appeared to be influenced by factors such as mood, and participants often lacked a sense of the time spent experiencing imagery. Intrusive imagery was experienced as both static images and film-like images, often first-person perspective. A key distinction appeared to be between images reported as mostly based on real memories  and images reported as mostly based on simulating elements (experienced by both those who had and had not been present at the time of discovery). This distinction appeared to be influenced by the degree to which there was a perception of complete information about the suicide, and therefore confidence in the memories of the event. Beyond these differences in the form and quantity of the imagery, participants also described the content of the imagery as largely focused on describing the story of the deceased’s suicide. While there was not one distinguishing narrative of the deceased’s suicide, some key shared features emerged around the content of imagery. The following sub-themes distinguish key features of the content of the images described.Five participants described the discovery of the deceased as the core content of their intrusive imagery, providing images of their memories of discovering the deceased, or (if not present) how they imagined the discovery.I see my son when I found him lying there dead.” (P10).“Imagined narratives were also described as feeling very real.The images in relation to the finding of him come and go, maybe because I know I’m imagining them, [they’re] not necessarily real because I didn’t see them with my own eyes, however they feel real.” (P4).“For fifteen participants the method was a key feature in the intrusive imagery and was described as very graphic, even for those not present at discovery. Often, intrusive images featuring the method were located at the scene of the suicide. However sometimes imagined images featured the deceased in other contexts, for example holding items used in the suicide or appearing with the stigmata of the suicide.I was imaging [him] walking down the hallway into the bedroom, but there was sort of like [items relating to the suicide method present].” (P6).“Nine participants mentioned that only the deceased featured in their imagery, regardless of whether others had been present at the time.Never anyone, not even the jogger that found him. …Nothing else, just [the deceased]. …But no, no nothing, nobody else at all. It all centred on him.” (P9)“Fifteen participants described their loved one as deceased in the imagery they experienced. For some, imagery might include aspects of events leading up to the death, but the images would always come to focus on the deceased after death.weren’t always right from the beginning of the day, they would be sections of it, but … they would always concentrate eventually on what he looked like when he died.” (P9).“[The images] Twelve participants described their images as piecing together what had happened in the last moments for the deceased.one thing I really visualise is the very last moments, like I can see her now in my head [in the act of suicide].” (P17).“For some this included considering how the deceased must have experienced those moments.one thing I remember was sort of picturing [him] … when he was sort of thinking through what he was doing … what was going through his mind. And he, he must have been scared and frightened and a whole host of emotions.” (P11).“Eight participants explained that their imagery reflected unanswered questions about aspects of the suicide and their search for meaning.although it’s a very short video, in my mind video, sort of moving image… it’s lots of different variations of it. It’s various nuances. … I know how it ends but I don’t know which version is the right one. And I don’t know … why it’s important for me to know that.” (P6).“1.2. Degree of control over imagery. 1.2.1. Lack of control. Seventeen participants described the involuntary nature of the imagery. A lack of control over its occurrence was in itself a distressing feature of the experience, whilst imagery conjured up voluntarily was experienced as less distressing.it was almost like I couldn’t stop it. And I would even go back to the beginning and start again, because I got no satisfaction from it … it was just completely out of my control.” (P9).“when… it’s just me by myself and I’m reflecting it’s different because, yeah, I think I’m more in control of it, and it’s a conscious thing that I’m doing, and I think there is that healing side of it which I’ve never really thought about before. And I think it’s different because it’s not like… an attack, … it’s me, myself in my personal space.” (P18).“Although many participants described the onset of intrusive imagery in the absence of a known trigger, eleven participants also mentioned common triggers. These included objects referencing the setting for the suicide, reminders of the deceased as an individual, or references to suicide more generally.the word ‘body’, like ‘oh the Police found a body’. If I hear anything, I see my son when I found him lying there dead.” (P10).“1.3. Changes in imagery over time. For most participants the experience of intrusive imagery appeared to persist for some years following suicide bereavement. Eleven participants described a reduction in the emotional impact or frequency over time, whether spontaneously or through gaining mastery over them.I think you become deadened. … your emotions become depleted. So that actually you become, you know, a mixture of exhausted and accepting. … so obviously my emotional response to visualisations and memories and events and so on, is less dramatic now …and more resignation I suppose. Whereas obviously in the early days it was incredibly violent and very, very difficult. … I couldn’t breathe. I couldn’t move.” (P12).“For some, the attenuation of the distress of imagery over time appeared to be characterised by the degree to which they felt in control, expressing their ability to manage their response to the experience, reducing associated distress.at the beginning it felt quite like, erm like life-debilitating because it meant that I would really struggle to go into particular bathrooms. … I’ve kind of learnt to live with the image when it comes up.” (P16).“it was about being able to [experience imagery] but in a safe way and a controlled way. And at the beginning I had no control and it just didn’t feel safe because it could come out of nowhere.” (P6).“All participants described a range of cognitive and emotional responses to the experience of intrusive mental imagery and ways of coping. Some also described positive aspects of the experience.2.1. Intrusive imagery as unhelpful and distressing. Fourteen participants described intrusive imagery as distressing or interfering with positive memories.Thirteen participants described their experience of intrusive imagery as distressing at various stages of the bereavement.In the earlier days, you know I just wanted it to stop, you know, just very, very, very, very distressed and wanted it to stop. … they’re still very distressing.” (P15).“Seven participants described how intrusive imagery relating to the suicide could interfere with their recollection of happier memories of the deceased’s life.I would at some stage be grateful for, for more of the images of the positive things in [the deceased]’s life, you know, the memories… . I think the negative thing with these [intrusive images] being there, whether they’re distressing or comforting, is the fact that they’re taking the space … and preventing other images of when [the deceased] was young … from being there. … if I didn’t so desperately need that connection, maybe it would be better, maybe the images would slightly fade, possibly. And allow some of those other, more positive images to come back…” (P15).“Participants spoke of active attempts to recall the more pleasant memories of the deceased, but reported that this was often difficult because intrusive imagery relating to the suicide was more dominant.I think I probably have more negative images than positive images. …I suppose one thing that I tried to kind of do … I would look through photos that we had from holidays, … his Facebook profile and all the images and posts on there. …I kind of force myself to review those to try and remember the happy times … I’d say generally the images that come to mind are associated with his death rather than his life.” (P11).“Five participants described their intrusive imagery simulating the suicide as unhelpful, preventing closure because it impeded an acceptance of not knowing what really happened.I think it would just give some closure if I knew the images were real. So, if I knew exactly what she would look like, it would just stop my mind from having so many other images. I think having real images, which I know will never happen … I guess that would help because then it stops my, it stops your imagination.” (P17).“2.2. Emotional and physical impacts of intrusive imagery. 2.2.1. Negative emotional impacts. Eleven participants spoke about the negative impacts that intrusive imagery had had on their everyday lives, with the distress causing them to feel agitated, distracted, or unable to sleep, and hampering functioning.I had them for months and months and months, until I was back at work, I realised that these imaginings were destroying me. I was getting quite careless at work.” (P9).“Twelve participants described a range of intense, conflicting emotional responses to the intrusive imagery, which could sometimes change quickly.Bereaved and just sad [tearful] and feeling like we somehow failed him. And also, really angry with why he didn’t ask us to help. So, loads of conflicting stuff.” (P7).“it’s sort of nostalgic and then sorrowful and then angry and the images just provoke rapid emotional changes like instantly and … there’s no kind of control over that, because the triggers are so strong.” (P4).Nine participants described unpleasant visceral impacts of the intrusive imagery, which arose as sudden, traumatic and uncontrollable sensations, including nausea or being frozen in whatever they were doing at the time.it’s so unexpected … It’s like being punched in the stomach … it takes my breath away.” (P2).““That image can be so powerful, and the immediate kind of emotional response means that somehow or another you’re, you know, quasi-paralysed. … It’s so traumatic as to sort of freeze you on the spot. … I have been known to stand still in the street for no particular reason.” (P12).2.3. Ways of coping. Ten participants described coping approaches that we categorized as three types of cognitive strategies they had developed.Five participants used distraction as a strategy to cope with the experience of intrusive imagery, but felt its effectiveness was reduced when low in mood or when prone to negative thinking styles.it will depend on my mood,… my state of mind on a particular day… And if I’m not able to distract with other things then I think it will be … harder to pull away from the [image].” (P15).“Six participants described overwriting the intrusive imagery by attempting to swap intrusive images with more pleasant memories of the deceased, or more helpful thought processes.Whenever I start imagining these things about [him] being dead in his car, and what position he was in, what did he look like and this sort of thing, I would tell myself, ‘No, I’m going to think about that day when we bought those coffee tables’.” (P9).“Eight participants reported attempting to “shake off” or block out the imagery when it arose, although this was not always successful.…it depends on what the image is, usually with the ones of him you know and the actual suicide, I’m actually quite good at shaking off or having that mental block.” (P3).“2.4. Positive aspects of the experience of intrusive imagery. Despite the distressing experiences of intrusive imagery described, seventeen participants mentioned some positive aspects.Six participants felt that intrusive imagery helped them to reflect on the enormity of a life-changing life event, and a reminder of their healing through the process of grief.it reminds me of what I’ve been through and that I’ve survived … and that I’ve got this far.” (P11).“it’s taught me resilience, it’s helped me compartmentalise my thoughts and… it actually brings something tangible to work with, and measurable, … if I can feel it getting less and less frequently, I know I’m healing.” (P1).“Twelve participants spoke of the role that intrusive imagery had in helping them process their suicide loss, facilitate meaning-making, and understand aspects of the suicide.rather than specific images, it’s more like the story … I’m trying to make sense of kind of what happened.” (P11).“You realise actually it’s your brain trying to, starting to process the whole thing that you’ve been through.” (P6).“For seven participants imagery provided the opportunity to connect with or feel close to the deceased, providing comfort. The vivid nature of the images meant they captured a sense of having the deceased with them again.it’s a way to be connected to the person. … I still am not able really to connect, to focus on all the good memories of [her] and the life she had before. … But … because they’re so strong, the images … it’s almost as though she’s still there. … Even though it’s horrible, it’s all you’ve got left. … I wonder whether the brain’s trying to keep connected in that way because they’re such powerful images … because the imagery is powerful it may be a way of connecting to her.” (P15).“Participants expressed a reluctance for the intrusive imagery to attenuate, as this would mean losing a valued connection with the deceased.[images] are … the last tie I have to [him], and if I didn’t have those images, I wouldn’t really have much to go on. … [the images] have kept me company for the best part of ten years and have allowed me to have kind of some kind of tie or relationship. So, I don’t think I’d want to lose them. …it would probably feel like really letting go of [him].” (P18).“Six participants described a reluctance to lose the imagery for fear of eroding their stock of memories of the deceased.I think I might miss [the images] because it’s [his] last moments… and anything related to him is still important. Like I would be forgetting him, or part of him.” (P6).“Four participants explicitly recognised a contradiction in the value they placed on imagery despite the intense distress it could bring.Should I miss [the image]… should I be relieved that I’d not have [it]?… At the moment I don’t know … maybe I would be relieved not to have pain anymore, but I wouldn’t like to not have [the image] because of the love.” (P10).“We did not identify any patterning of themes by kinship group, but we did identify a pattern defined by the quality of the relationship with the deceased. Where relationships were relatively disenfranchised, intrusive imagery was experienced as more unhelpful or with less potential to provide comfort, creating a greater motivation for the bereaved to eliminate the imagery. However, for those who described close relationships with the deceased, imagery was perceived as more important in maintaining a connection with the deceased or providing important memories of the deceased despite the associated distress. This suggests that for people bereaved by suicide, imagery can encapsulate the individual narrative of the loss, shaped particularly by the nature of the relationship with the deceased.Our analysis of qualitative interview data from 18 suicide-bereaved adults described clearly the phenomenon of intrusive mental imagery following suicide loss, setting out its characteristics, as well as positive and negative aspects. The range of emotional and cognitive responses to intrusive imagery described included striking impacts on social and occupational functioning but also opportunities for meaning-making and post-traumatic growth. Individuals had developed coping strategies to deal with the negative aspects of the imagery, although with limited success. Whilst all participants found the experience upsetting, nearly all identified some positive aspects. Despite a distressing lack of control over the occurrence, frequency or intensity of the images, some participants explained that the imagery helped them make sense of the death, hold onto their memories of the deceased, and retain a sense of connection. This is an important finding given that meaning-making is viewed as a key tool in the formation of continuing bonds) . ParticiFrom a phenomenological perspective our findings illustrate some parallels and contrasts with the experiences of intrusive imagery in individuals with psychological disorders, including PTSD , 12. In Similarly to intrusive imagery reported across specific psychological disorders, the occurrence of intrusive imagery following suicide loss was experienced with a distressing sense of lack of control. Participants were sometimes aware of triggers for their intrusive imagery, comparable to the triggers that are common for PTSD flashbacks , 12. At Many participants in our study identified positive aspects of their experience, contrasting with experiences of intrusive imagery described in the context of psychological disorders, where they appear largely devoid of positive aspects . This fiOverall, findings highlight similarities in the experience of intrusive mental imagery following suicide bereavement with intrusive imagery experienced in the context of PTSD, complicated grief and other psychological disorders . HoweverTo our knowledge, this is the first qualitative study of the phenomenon of intrusive mental imagery experienced by people bereaved by suicide, addressing a gap in the literature and providing valuable information that can be applied to intervention development. Our use of remote video-linked interviews is likely to have enhanced disclosure and expanded the reach of the study nationally. Discussions within a multi-disciplinary research team enhanced reflexivity and improved the validity of findings. However, limitations included our sampling methods, which meant we were unable to estimate the prevalence of intrusive mental imagery following suicide loss, nor compare it to that in other bereaved groups. Despite efforts to recruit a diverse sample nationally, the sample over-represented white females, and our findings are therefore not generalizable to males or non-white individuals experiencing suicide loss. However, given cultural dimensions to the experience of intrusive mental imagery and of bereavement, we hope that this study prompts further phenomenological work sampling in specific cultural groups.These findings provide valuable insights for anyone supporting people bereaved by suicide by describing a distressing aspect of their grief. We also shed light on the potential mechanisms underlying the observed adverse health and social impacts of suicide bereavement, informing the development of evidence-based psychological interventions to promote recovery, in line with government suicide prevention strategy , 27.Despite the distressing nature of intrusive mental imagery after suicide loss, some aspects of such imagery may benefit the bereaved individual, and interventions designed to target imagery should acknowledge such positive aspects. Approaches such as imagery rescripting, whereby individuals construct an alternative, vivid and more helpful image, which can include communicating with the deceased, may be appropriate . ImageryTo inform future interventional work, further research should investigate the phenomenological experience of intrusive mental imagery in groups under-represented in this study, and how this might vary by kinship closeness and quality of the relationship with the deceased. Researching this phenomenon in a range of countries would provide an understanding of variations in the experience cross-culturally.In conclusion, this qualitative analysis of subjective experience of intrusive mental imagery following bereavement by suicide of a close contact described clear characteristics of the phenomenon and its emotional impact. Whilst intrusive mental imagery after suicide loss was universally experienced as distressing, with characteristics similar to imagery observed in common mental disorders, many described valued features. This work provides a greater understanding of the experience of suicide loss, helping professionals provide more appropriate support, and informs the development of imagery-based interventions.S1 Appendix(DOCX)Click here for additional data file.S2 Appendix(DOCX)Click here for additional data file.S3 Appendix(DOCX)Click here for additional data file. 1 Sep 2022 <div> PONE-D-22-08780 </div>  <div> Exploring the phenomenon of intrusive mental imagery after suicide bereavement: a qualitative interview study in a British sample </div>  <div> PLOS ONEDear Dr. Pitman,Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. 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Reviewer #1: Thank you for an opportunity to comment on a qualitative study exploring the phenomenon of intrusive mental imagery after suicide bereavement. The study is of interest as it explores in detail an underresearched phenomenon. I have some questions and suggestions regarding the mss.The starting point (Introduction) is intrusive imagery in PTSD. This seems to be a quite narrow point of view to approach intrusive mental imagery in suicide bereavement. Can the authors provide information on outcomes of previous studies on intrusive mental imagery in bereavement more generally, and suicide bereavement more specifically (if such data exist)?Further, intrusive imagery may also be related to the phenomenon of continuing bonds in suicide bereavement . Continuing bonds after loss by suicide: a systematic review. International journal of environmental research and public health, 19(5), 2963). This should be considered in the design of the study and discussion of results.Further, the study sample has not been assessed for PTSD. Again, the discussion seems to be centred around this diagnosis.Although 2.4.1 Post-traumatic growth is one of the subthemes in the analysis, it has been relatively overlooked in the the Discussion . “Turning personal tragedy into triumph”: A systematic review and meta-analysis of studies on posttraumatic growth among suicide-loss survivors. Psychological trauma: theory, research, practice, and policy, 13(3), 322).Reviewer #2: Thank you for the opportunity to review this manuscript. It is well-written and provides a clear rationale for the importance of this research. A rigorous qualitative analysis has been conducted and an insightful interpretation of results provided. In my opinion, the manuscript is of a high standard and provides an important contribution to understanding the experiences and needs of people bereaved by suicide to help inform future interventions. I recommend publication.Reviewer #3: This paper aims at investigating the phenomenon of intrusive mental imagery after suicide bereavement. Utilising qualitative approach, the authors explored individual experiences of mental imagery following the suicide of close contacts of the bereaved. Semi-structured interviews of 18 adults who experienced suicide of a close contact were conducted. Thematic analysis (TA) yield insights surrounding the characteristics of intrusive imagery, and the cognitive and emotional response to the imagery. The authors highlighted that there are some similarities with intrusive imagery experienced in PTSD, complicated grief and other psychological disorders.The thematic analysis is clear and straightforward, but the sub-themes on the characteristics of intrusive imagery could be explored further to create a better and a more coherent link to the participants’ narrative of the deceased’s suicide. The present version is rather disjointed.My key concern lies on the contribution of the paper. Based on the finding, the impact of intrusive mental imagery after suicide bereavement appears to be no different from other form of bereavement, and the clinical implication (from an intervention point of view) is similar. As discussed in the limitation section, the scope for generalization to other culture is very limited given the under-representation of the sample. Bereavement is a social and cultural phenomenon, and I strongly believe recognizing intrusive mental imagery cross-culturally is important.Minor comments: Further details regarding participants such as whether they have underlying mental health conditions should be outlined. It is not clear from the exclusion criteria. I am curious whether certain pre-existing conditions might give rise to emotional complications following loss, and hence affect the interpretation of the results.********** <font color="black"> 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.If you choose “no”, your identity will remain anonymous but your review may still be made public.Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. </font> Reviewer #1: NoYes: Victoria RossReviewer #2: Reviewer #3: No**********https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool,  21 Oct 2022Reviewer #1: Thank you for an opportunity to comment on a qualitative study exploring the phenomenon of intrusive mental imagery after suicide bereavement. The study is of interest as it explores in detail an under-researched phenomenon. I have some questions and suggestions regarding the mss.The starting point (Introduction) is intrusive imagery in PTSD. This seems to be a quite narrow point of view to approach intrusive mental imagery in suicide bereavement. Can the authors provide information on outcomes of previous studies on intrusive mental imagery in bereavement more generally, and suicide bereavement more specifically (if such data exist)?We agree that the introduction cites few studies in relation to intrusive mental imagery after suicide bereavement, but we have edited this to be clear that none exist (that we are aware of) and to report the findings of the one study we were aware of in relation to intrusive mental imagery after any bereavement. We also agree that the early focus on PTSD in the Introduction disrupted the narrative arc in terms of setting out the aims of the study and implied that this study sampled those with a diagnosis of PTSD. We have edited the text (page 3-4) to make it clearer how the PTSD literature fits into this topic, explaining that this represents the majority of the literature in this area, and that whilst we can extrapolate some understanding of the phenomenon from this, we need to investigate this in a suicide-bereaved sample. We thereby justify why it is important to conduct a phenomenological study specifically in a non-clinical sample of people bereaved by suicide. Further, intrusive imagery may also be related to the phenomenon of continuing bonds in suicide bereavement . Continuing bonds after loss by suicide: a systematic review. International journal of environmental research and public health, 19(5), 2963). This should be considered in the design of the study and discussion of results.Thank you for suggesting this additional discussion point and citation. Regarding the design of the study, our methodological approach was to ensure that our questioning did not only assume negative aspects of imagery, recognising the potential role of imagery as part of maintaining bonds with the deceased. Our topic guide had included questions such as - Do you find these images helpful or comforting in any way? - Is there any other way in which these images impact upon you positively or negatively?We have now added the topic guide as an Appendix, as we felt it was important for readers to see the open questioning style, and the focus on the form of the imagery rather than the narrative of the suicide. Thus:“The interview topic guide was developed in collaboration with the Support After Suicide Partnership (see Appendix 1) and designed to use open questioning, avoiding assuming only negative aspects of imagery.”We have edited the discussion early on (page 22 and 23) to address this point about continuing bonds, highlighting the opportunities that positive aspects of the imagery presented participants with, in terms of making sense of the death and post-traumatic growth. We have therefore expanded our Discussion to relate our findings under theme 2.4 (positive aspects of the experience of intrusive imagery) to the literature on continuing bonds and post-traumatic growth, including this study and related ones. This relates the findings from theme 2.4 to the wider qualitative and quantitative literature in relation to suicide loss, and we believe is now much improved. Further, the study sample has not been assessed for PTSD. Again, the discussion seems to be centred around this diagnosis.As we recruited a community sample, we did not use screening or diagnostic instruments to characterise our participants, but instead sought to describe the phenomenon of intrusive imagery after suicide loss, regardless of whether an individual also had the core symptoms of PTSD . We have edited our introduction to be clear about the role of the PTSD literature in relation to what we know about intrusive imagery in the context of suicide loss. This avoids suggesting that this study focusses on a population of those with PTSD. In the discussion we relate our findings to those reported in the literature covering a range of psychological disorders, including PTSD, other anxiety disorders, depression, and addiction, and also self-harm. As the features of intrusive imagery are best described in the context PTSD, this does feature in more detail, but we do also relate our findings to those for these other psychological disorders. Although 2.4.1 Post-traumatic growth is one of the subthemes in the analysis, it has been relatively overlooked in the Discussion . “Turning personal tragedy into triumph”: A systematic review and meta-analysis of studies on posttraumatic growth among suicide-loss survivors. Psychological trauma: theory, research, practice, and policy, 13(3), 322).We agree that although we had identified this explicitly as a sub-theme, we had neglected this in the discussion. We have now edited our discussion (page 23) to relate our findings to the quantitative summarised in Levi-Belz et al.’s meta-analysis, thus making an important link between the qualitative and the quantitative literature. Reviewer #2: Victoria Ross Thank you for the opportunity to review this manuscript. It is well-written and provides a clear rationale for the importance of this research. A rigorous qualitative analysis has been conducted and an insightful interpretation of results provided. In my opinion, the manuscript is of a high standard and provides an important contribution to understanding the experiences and needs of people bereaved by suicide to help inform future interventions. I recommend publication.Thank you very much for your comments. Reviewer #3: This paper aims at investigating the phenomenon of intrusive mental imagery after suicide bereavement. Utilising qualitative approach, the authors explored individual experiences of mental imagery following the suicide of close contacts of the bereaved. Semi-structured interviews of 18 adults who experienced suicide of a close contact were conducted. Thematic analysis (TA) yield insights surrounding the characteristics of intrusive imagery, and the cognitive and emotional response to the imagery. The authors highlighted that there are some similarities with intrusive imagery experienced in PTSD, complicated grief and other psychological disorders.The thematic analysis is clear and straightforward, but the sub-themes on the characteristics of intrusive imagery could be explored further to create a better and a more coherent link to the participants’ narrative of the deceased’s suicide. The present version is rather disjointed.We were quite careful when writing up the findings of our thematic analysis not to provide too many details of the narrative relating to the deceased’s suicide, both to avoid upsetting participants and readers, and to protect participants’ anonymity. It was also because our aim was not to look for a unifying narrative emerging from the imagery of bereaved individuals (which is a different research question) but to identify the characteristics of the imagery. The themes may come across as disjointed because not all participants had a coherent narrative of the suicides and instead focussed on describing the form of the images. This is function of our topic guide, where only the first three questions probe the background to the suicide, before moving on to characteristics of the imagery. We had used our sub-themes within Theme 1 to capture broad features of the content of the images . However, we have now edited the opening paragraph of theme 1 (page 9-10) to distinguish between differences in the form and quantity of the images  as an overarching feature of this theme, and specific features of the content of the images. To clarify this, we have added the following text to theme one:“While there was not one distinguishing narrative of the deceased’s suicide, some key shared features emerged around the content of imagery”. We have also added the following text to the end of the results:“This suggests that for people bereaved by suicide, imagery can encapsulate the individual narrative of the loss, shaped particularly by the nature of the relationship with the deceased.”My key concern lies on the contribution of the paper. Based on the finding, the impact of intrusive mental imagery after suicide bereavement appears to be no different from other form of bereavement, and the clinical implication (from an intervention point of view) is similar. As discussed in the limitation section, the scope for generalization to other culture is very limited given the under-representation of the sample. Bereavement is a social and cultural phenomenon, and I strongly believe recognizing intrusive mental imagery cross-culturally is important.https://pubmed.ncbi.nlm.nih.gov/27792974/), and it is likely that the same would apply to imagery in those bereaved by suicide.Members of our team have previously conducted quantitative studies comparing mental health outcomes in groups bereaved by suicide and those bereaved by other causes, but our intention in this study was not to compare these groups’ experiences of intrusive imagery but to conduct a phenomenological study focussed on understanding how bereavement by suicide might influence cognitions and how intrusive imagery might be experienced in this very specific (and high risk) group. As no previous phenomenological studies have been conducted specific to suicide bereavement, we aimed to carry out the first of its kind. As we noted in our original submission, although we were able to recruit nationally, and tried hard to balance our sample across demographic characteristics, our sample was nonetheless over-represented by white females typical of studies of this kind. We agree that our findings may only have resonance to those with similar socio-demographic characteristics to those in our sample. We had acknowledged this in the limitations but have edited this section (page 24) to make this point more clearly, and to recommend that this study should prompt future studies exploring the experience of intrusive imagery in specific cultural groups. Previous research on mental imagery of suicide in clinical populations has identified both shared characteristics across cultures and more culturally-specific elements  psychiatric symptoms on the nature and course of intrusive mental imagery after suicide loss and are planning quantitative longitudinal work to address this.AttachmentResponse to Reviewers 2022 10 12.docxSubmitted filename: Click here for additional data file. 12 Apr 2023Exploring the phenomenon of intrusive mental imagery after suicide bereavement: a qualitative interview study in a British samplePONE-D-22-08780R1Dear Dr. Pitman,We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.Within one week, you’ll receive an e-mail detailing the required amendments. 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For more information, please contact Kind regards,Barbara Dritschel, PhDAcademic EditorPLOS ONEAdditional Editor Comments :Reviewers' comments: 17 Apr 2023PONE-D-22-08780R1 Exploring the phenomenon of intrusive mental imagery after suicide bereavement: a qualitative interview study in a British sample Dear Dr. Pitman:I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. onepress@plos.org.If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact plosone@plos.org. If we can help with anything else, please email us at Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staffon behalf ofDr. Barbara Dritschel Academic EditorPLOS ONE