Renata Laks

Marinho Jerson

Aimee Mograbi

Elodie Bertrand

Renata Naylor
email: rpbnaylor@gmail.com

Valeska Marinho

Jerson Laks
email: jersonlaks@gmail.com

Aimee Spector
email: a.spector@ucl.ac.uk

Daniel Correa Mograbi

Cognitive Stimulation Therapy for Brazilian people with dementia: examination of implementation' issues and cultural adaptation

Keywords: dementia, Cognitive Stimulation Therapy, Brazilian population, cultural adaptation

Objectives: The prevalence of dementia has been increasing particularly in developing countries. However, people with dementia (PwD) in Brazil are currently offered no psychosocial treatment upon diagnosis. Cognitive Stimulation Therapy (CST) that originated in the UK has proven benefits on cognition and quality of life in PwD. We investigate the possible issues for the implementation of CST for the Brazilian population and its cultural appropriateness.

Method: Individual interviews and focus groups were conducted with PwD, their caregivers and health professionals (n=37). Data were recorded and transcribed, before being analyzed using Framework Analysis.

Results: Regarding the issues for implementation of CST in the Brazilian population, two main themes emerged, "Barriers" and "Facilitators", along with nine subthemes. Overall, the activities and materials were seen as being appropriate for use with the Brazilian population, some minor changes were suggested.

Conclusions:

The results indicate that CST is appropriate for use in the Brazilian population, only some cultural adaptations are necessary. In the stakeholders' opinions, CTS intervention is needed in Brazil, due to the lack of treatment options for PwD in developing countries.

Introduction

Increases in life expectancy worldwide have led to growing numbers of people with dementia (PwD) [START_REF] Ferri | Global prevalence of dementia: a Delphi consensus study[END_REF]. The prevalence of dementia has been increasing particularly in developing countries, with an estimated number of 1.6 million PwD in Brazil (ADI, 2005). In this context, interventions that can ameliorate the symptoms of dementia and improve quality of life are research priorities, but dementia care provision is still limited in lower and middle income countries (ADI, 2005). Indeed, PwD in Brazil are currently offered no psychosocial treatment upon diagnosis.

Cognitive Stimulation Therapy (CST) is a 14 session group intervention for PwD developed in the UK. It aims to mentally stimulate people through complex psychological techniques (including implicit leaning, multi-sensory stimulation) embedded in structured group activities, such as word association and current affairs. It has a robust evidence-base, significantly improving cognition and quality of life [START_REF] Orrell | Maintenance cognitive stimulation therapy for dementia: single-blind, multicentre, pragmatic randomised controlled trial[END_REF][START_REF] Spector | Efficacy of an evidence-based cognitive stimulation therapy programme for people with dementia: randomised controlled trial[END_REF] and is the only non-drug intervention recommended to treat cognition by UK NICE guidelines (National Institute for Health and Clinical Excellence, 2006). It is endorsed by Alzheimer's Disease International, used in over 24 counties globally and translated into at least eight languages.

To facilitate CST implementation in other cultures, adaptation guidelines have been developed [START_REF] Aguirre | Guidelines for adapting cognitive stimulation therapy to other cultures[END_REF] with an approach involving stakeholders in the cultural adaptation process. This 'bottom-up' approach consists of five phases, namely, (i) generating information and collaborating with stakeholders; (ii) integrating the generated information with clinical and theoretical knowledge; (iii) reviewing and revising; (iv) pilot testing; and (v) synthesizing stakeholder feedback and finalizing. This adaptation process should precede a feasibility study, which will use a randomized controlled trial design.

The present study was performed to understand the possible issues for the implementation of CST for the Brazilian population and its cultural appropriateness by interviewing stakeholders. Two complementary formats were used for the interviews: focus groups and individual interviews. As highlighted by previous studies [START_REF] Harmer | What is meaningful activity for people with dementia living in care homes? A comparison of the views of older people with dementia, staff and family carers[END_REF][START_REF] Thorgrimsen | Whose quality of life is it anyway? The validity and reliability of the Quality of Life-Alzheimer's Disease (QoL-AD) scale[END_REF], the use of individual interviews allows the participants to express freely their opinions regarding the program. Moreover, based on the fact that PwD frequently present deficits in short-term memory, individual interviews appeared more appropriate for this population (Murphy, Killick, & Allan, 2001). The aim of this study was to identify any practical issues that might affect the delivery of the program, and to gather data about the quality and appropriateness of the activities presented in the translated manual, which would inform the development of further drafts of the materials.

Methods

Semi-structured interviews were conducted to assess the cultural appropriateness of the CST program in Rio de Janeiro (Brazil), including possible issues for its implementation.

The quality of the first adaptation of the CST material was also discussed. Two complementary formats were used: focus groups and individual interviews.

Recruitment

Health professionals taking part in the focus groups were recruited from invitation, including staff from the hospital where a CST feasibility RCT is being conducted and other clinical services. Caregivers were recruited from the same hospital or from a caregivers' association.

Individual interviews were conducted with PwD and caregivers who consented to take part in the feasibility study and were randomly allocated to the control group, not being exposed to the intervention.

Procedure

Both focus groups and individual interviews were audio-recorded and were then transcribed verbatim.

Focus groups

Two focus groups with multidisciplinary health professionals, including psychiatrists, geriatricians, neuropsychologists and nurses (n=9) and two focus groups with caregivers of PwD (n=15) were conducted. Each group was attended by two members of the research team, one of whom took on the role as facilitator and led the group discussion, while the other observed the group and made notes to supplement the audio data collected. The interview schedule was developed in a semi-structured style, guided by a series of predetermined focus points (Table 1). A description of the general principles of CST was given, with each session of the program being presented in more detail. The participants discussed the feasibility of the activities suggested and the need of adapting the material for the Brazilian elderly population. Each group lasted approximately 90 minutes in total.

PLEASE INSERT TABLE 1 HERE

Individual interviews

PwD and their caregivers (n=13) were invited to participate to individual interviews, which were conducted separately by one member of the research team. Each interview lasted approximately 20 minutes. After being provided with a description of the CST program, both PwD and their caregivers were invited to discuss the possible facilitators and barriers that could affect the implementation of the program.

Analyses

The transcripts of the focus groups and the individual interviews were analysed using Framework Analysis [START_REF] Ritchie | Qualitative data analysis for applied policy research[END_REF]. Five key stages were followed: (i) Familiarization; (ii) Thematic framework identification; (iii) Coding; (iv) Charting; and (v) Mapping and Interpretation. First, two researchers read all transcripts in order to become familiar with the broad themes expressed. Then, identified themes were compiled and refined leading to a coding key, which included broad themes and sub-themes. Transcripts were then categorized and charted accordingly. Finally, mapping and interpretation was realized to map the factors which might affect the implementation of the CST program in Brazil. No specialist software was used to perform the data analyses.

Ethics

The research project was approved by the research ethics committee of the Institute of Psychiatry of the Federal University of Rio de Janeiro (IPUB-UFRJ; project ref: 57019616.5.0000.5263).

Results

Summing the focus groups and the individual interviews, a total of 37 persons were interviewed. Two themes and nine sub-themes were identified consistently across the interviews (Table 2). Illustrative quotes are provided for each sub-themes.

PLEASE INSERT TABLE 2 HERE Theme 1: Barriers

Sub-theme: More work for the caregivers

Both health professionals and caregivers of PwD said that one of the barriers for the implementation of the program is that it will entail more work for the caregivers. "It's essential to have collaboration from the family because we would be giving more work to the caregivers" (health professional) "What can be difficult for the family is to find somebody who has (the) time to bring the patient to the intervention site." (caregiver)

Sub-theme: Patients' motivation to go to CST

Participants identified the lack of patients' motivation to participate in the program as one barrier for the implementation of CST in Brazil. This is linked to the barrier "More work for the caregiver" considering that, if the patient is not willing to go, the caregiver will need to convince the PwD to participate in CST.

"(…) to take them away from home, to make them get up from the couch (…) he does not want to leave the house" (caregiver)

"As the principal barrier, I identify the fact that the patient does not have the initiative to participate (…) in relation to the patient's motivation." (health professional)

Sub-theme: Frequency of sessions

There was a consensus among participants that the frequency of the sessions would be a barrier for the implementation of the CST in Brazil. This barrier is also link to the first barrier identified ("More work for the caregiver"), due to the fact that, in Brazil, PwD will almost always have to go to the intervention site with the help of a caregiver.

" [about] the frequency of this program, I believe we would have some problems to get people with dementia to come" (health professional)

Sub-theme: Distance from patient's neighborhood

Participants, especially caregivers, reported that PwD generally live far from the intervention site and considered this a potential barrier to the implementation of CST in Brazil.

"I live far from here, if I had to bring my wife I would depend on my two sons-inlaw, if either is free, to bring me here, but if they're not, I would have to rent a car to come??." (caregiver) "We are living far away (...) The distance is a big barrier for me. (..) So it is complicated for my sister to bring me here." (PwD) "The travel from the patient's house to the IPUB, they are relying on someone to bring them there, because nobody is going alone. There is also a financial problem, we are dealing with people from the IPUB, so we are dealing with persons… financially… they do not have money, so…" (health professional)

Sub-theme: Cognitive stimulation uncommon for PwD

The participants explained that one barrier to the implementation of the CST in Brazil is that cognitive stimulation therapy is not commonly offered to PwD in this country. There is a lack of information around treatment options, and health professionals, based on caregivers' demands, usually treat dementia using pharmacological interventions only.

"The caregiver's demand is not for cognitive stimulation, the caregiver's demand is for patient's silencing. Doctors meet the caregiver's demand and they also believe that once dementia begin there is nothing else to do other than giving a pharmacological "We have to count on the caregivers' help. They are the ones who will take the people with dementia there, give their support, stimulate the people with dementia to go to the clinical service" (health professional) "To facilitate for me to come here? The only thing is that my sister has to agree."

(PwD)

Sub-theme: Have two sessions per day (once a week).

There was a clear consensus amongst participants that having the two weekly sessions on the same day would facilitate the implementation of the program. Indeed, due to some Brazilian characteristics (e.g. PwD leaving far from the intervention site or lack of secure transportation for PwD or financial difficulties), limiting the number of travels to participate weekly to the CST was considered a facilitator.

"I think that for Brazil's reality, for the kind of people with dementia that come here (...) they live far from here and can't afford coming twice a week, (...) so I think it would be more productive to have the intervention once a week." (health professional) "I think once (a week) would be good because... less, less work... less time wasted...

I would come once a week" (caregiver)

Sub-theme: Give the caregivers something to do while they wait for the sessions to end.

Both health professionals and caregivers said that providing some kind of entertainment to the caregivers while they are waiting for the PwD would facilitate the implementation of the intervention. Indeed, due to the distance between the PwD's home and the intervention site, the caregivers would have to wait for the patients there.

"If it were me, I would have to be around, I could bring my knitting material and I would be completely entertained doing it, but, also, a movie would be good"

(caregiver)

"What are we going to do with the caregivers while the patients are in the intervention? (…) I think that we could do psychoeducational groups with them."

(health professional)

Sub-theme: Group activities

Participants, especially caregivers of PwD, said that one facilitator of the implementation of the CST is that it is a group activity.

"In my opinion, the fact that the activities are made in group facilitates the stimulation, because one can help the other, there are more interactions" (caregiver) " [about] the bonds they are creating with the other participants, they start to attend the group weekly, this also stimulates, like "you are going to see you friends there"." (caregiver)

Adaptation of the sessions and materials

Based on their opinions, modifications were made. Most of the activities suggested in the original CST manual were kept. However, some of the materials for these activities were adapted (e.g. faces of famous Brazilian people or typical Brazilian foods). See Table 3 for a detailed description of the cultural adaptations made in the CST for the Brazilian population.

PLEASE INSERT TABLE 3 HERE

Discussion

Summary of the results

The present study aimed to evaluate the feasibility of the CST program in the Brazilian context by understanding the possible barriers and facilitators for the implementation of the program. The results suggested that there was a consensus among health professionals, PwD and their caregivers regarding the factors that can facilitate or difficult the delivery of the CST in the Brazilian population. Indeed, two themes (i.e. Barriers and Facilitators) and nine sub-themes were identified consistently across all the interviews.

Interpretation of findings

Participants reported that the frequency of the sessions could be a barrier for the adherence to the treatment. Indeed, PwD would have to rely on their caregivers to bring them to the intervention site, in part because most of them are living far from there. This point was also highlighted by the participants as a principal barrier to the patients' participation. As pointed by [START_REF] Gratao | The demands of family caregivers of elderly individuals with dementia[END_REF], most of the caregivers in Brazil are daughters, within an age range in which they have several work and personal life commitments, or elderly spouses. Therefore, having the PwD taking part in the CST program would indeed lead to more work for the caregiver, who is frequently stressed and overwhelmed by the act of caring for a demented family member [START_REF] Etters | Caregiver burden among dementia patient caregivers: A review of the literature[END_REF]. To facilitate recruitment, it should be important to emphasize to the caregivers that studies showed an improvement of cognition in PwD after the CST intervention [START_REF] Aguirre | Cognitive stimulation for dementia: A systematic review of the evidence of effectiveness from randomised controlled trials[END_REF] and that this improvement could lead diminished caregiver burden. Based on these issues raised by the participants, we suggest that the two weekly sessions of the CST should be performed on the same day following the structure: first session of 45 minutes, break of 15 minutes, second session of 45 minutes.

As a facilitator, health professionals and caregivers of PwD reflected on the possible activities that should be offered to the caregivers while they are waiting for the session to end. Indeed, due to characteristics of the Brazilian context already discussed (e.g. patients living far from the intervention site and/or lack of secure transportation for PwD), the caregivers would have to bring the patients to the intervention site and wait for them until the end of the session. However, to avoid biasing the feasibility study, it was decided that no specific activity would be provided for caregivers. Nevertheless, for an eventual larger scale implementation of CST in Brazil, intervention sites should be aware that offering activities to caregivers while PwD receive the program could increase adherence to the treatment. Indeed, studies showed that combined intervention programs for both the caregiver and the PwD are effective, especially in improving the general mental health of both the caregiver and the PwD [START_REF] Smits | Effects of combined intervention programmes for people with dementia living at home and their caregivers: a systematic review[END_REF].

Finally, participants said that CST is not commonly offered to PwD in Brazil, in part because of the lack of information regarding treatment options for PwD and that could limit the implementation of CST in the Brazilian context. Therefore, before larger scale implementation of the program, an awareness and information campaign regarding dementia and its treatment options might be initiated, especially targeting caregivers of PwD. In fact, this campaign, by increasing awareness regarding the efficacy of cognitive stimulation for PwD, could also lead to stronger support from caregivers in order to deliver the program. Indeed, participants of the present study highlighted that it would be essential to be able to count on caregivers' support for the CST implementation in the Brazilian context.

Regarding the different activities and the material presented in the CST manual, health care professionals and caregivers of PwD responded positively and suggested some adaptations.

Only minor changes were made in order to address cultural issues, such as removing some activities which are uncommon in Brazil (e.g. gardening) and using material reflecting the Brazilian population reality (e.g. typical Brazilian sweets in the session "Childhood").

Limitations

Limitations of the current study include the difficulty for the interviewer/moderator to have PwD and their caregivers answering specifically to the topic during focus groups and individual interviews. This issue has also been reported in other studies using qualitative methods with service users [START_REF] Qazi | User, carer and staff perspectives on anxiety in dementia: A qualitative study[END_REF][START_REF] Yates | Service users' involvement in the development of individual Cognitive Stimulation Therapy (iCST) for dementia: a qualitative study[END_REF]. Regarding PwD, the difficulty to stay "on topic" can be explained by the deficits in short-term memory and verbal communication frequently experienced by this population (Murphy et al., 2001). The tendency of PwD's caregivers to talk about issues other than the particular topic of discussion could reflect the need to share experiences and to talk about the caring role in general [START_REF] Qazi | User, carer and staff perspectives on anxiety in dementia: A qualitative study[END_REF][START_REF] Yates | Service users' involvement in the development of individual Cognitive Stimulation Therapy (iCST) for dementia: a qualitative study[END_REF]. Despite this issue, overall collected data was informative and "off-topic" data was minimal.

Conclusion

During this study, health professionals and caregivers highlighted the need of the CST intervention in Brazil, expressing the lack of treatment options offered for PwD. Indeed, the prevalence of dementia is increasing and care provisions for PwD are still scarce, especially in developing countries [START_REF] Shaji | Dementia care in developing countries: The road ahead[END_REF]ADI, 2005). Based on the adaptation guidelines developed by [START_REF] Aguirre | Guidelines for adapting cognitive stimulation therapy to other cultures[END_REF], we used a bottom-up formative research method and found CST appropriate for the Brazilian population, with some minor amendment needed to address cultural issues.

Table 1.

Focus points of the semi-structured interviews

  treatment for the person to stay calm." (health professional) "Basically, in our culture it is a little… the Brazilian society is not yet open for this kind of things [about cognitive stimulation]" (caregiver) Theme 2: Facilitators Sub-theme: Have the caregivers' support PwD and health professionals agreed that it is essential to have the caregiver's support to facilitate the implementation of the CST for the Brazilian population.

  Barriers and facilitators for referrals to non-pharmacological interventions Relevance of the approach Frequency and content of the sessions Adaptation of materials and procedures Validation of outcomes

Table 2 .

 2 Themes and sub-themes identified across focus groups and individual interviews.

	Themes	Sub-themes
	Barriers	More work for the caregivers
		Patients' motivation to go to CST
		Distance from patients' home
		Frequency of the sessions
		Cognitive stimulation uncommon for PwD
	Facilitators	Have the caregivers' support
		Have two sessions per day (once a week)
		Give the caregivers something to do while they wait
		Group activities

Table 3 .

 3 Cultural adaptations made in the CST for Brazil and the rational

	Session themes	Cultural adaptation	Reasons for adaptation
	1. Physical games	Nil			Not applicable
	2. Sounds	Use of familiar Brazilian songs	Adaptation of the material to
					the Brazilian population
	3. Childhood	Suggested sweets replaced with	Unfamiliarity with suggested
		typical Brazilian childhood sweets	sweets
		(e.g. cocada or paçoca)
	4. Food	Suggested foods used as memory	Unfamiliarity with suggested
		triggers replaced with typical	foods
		Brazilian foods (e.g. pudim de pão
		or mariola).
		Names of food items to be
		completed replaced with typical
		Brazilian dishes (e.g. canja de
		galinha or pão de ló)
	5. Current affairs	Removal of royal family-and of	Unfamiliarity	with	those
		charity-related question.		themes.
	6. Faces/scenes	Use of photographs of famous	Adaptation of the material to
		Brazilian faces and places (e.g.	the Brazilian population
		Carmem Miranda or Sugarloaf
		Mountain)
	7. Word associations	Suggested	word	associations	Adaptation of the material to
		replaced by culturally adapted	the Brazilian population
		phrases (e.g. "Açucar … Pão de" or
		"é melhor prevenir … do que
		remediar")
		Use of Brazilian songs, such as
		famous soccer anthems.
	8. Being creative	Removal of the gardening and the Uncommon activities in the

Acknowledgements

We acknowledge the invaluable contributions of all the research participants (health professional, patients and their caregiver) who took part in this study. JL is a Researcher

Acknowledgements

We acknowledge the invaluable contributions of all the research participants (health professional, patients and their caregiver) who took part in this study. JL is a Researcher from Conselho Nacional de Pesquisa (CNPq) and Cientista do Nosso Estado form Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (CNE-FAPERJ). DCM is a Researcher from Conselho Nacional de Pesquisa (CNPq) and Jovem Cientista do Nosso Estado form Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (JCNE-FAPERJ).

from Conselho Nacional de Pesquisa (CNPq) and Cientista do Nosso Estado form Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (CNE-FAPERJ). DCM is a Researcher from Conselho Nacional de Pesquisa (CNPq) and Jovem Cientista do Nosso Estado form Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (JCNE-FAPERJ).

Funding

This work was supported by a Newton Advanced Fellowship provided by the Academy of Medical Sciences, UK, and Royal Society [grant number NAF004\1001].

Disclosure statement

All of the authors have no financial conflicts of interests or other disclosures to report.


Abirami Baleswaran

Christel Couderc

Marine Reyrolle

Mickael Le Bechec

Jean Dayde

Hélène Tormo

Gwénaëlle Jard
email: gwenaelle.jard@purpan.fr

Elaboration and characterisation of a miniature soft lactic goat cheese model to mimic a factory cheese

Keywords: Conceptualization, Methodology, Formal analysis, Investigation, Writing -Original Draft, Review & Editing, Visualization Conceptualization, Writing -Review & Editing, Supervision, Project administration, Funding acquisition Conceptualization, Writing -Review & Editing, Supervision, Project

Although soft lactic goat cheeses are widely produced in France, there is little information available in the literature about them and no miniature model for this kind of cheese has yet been proposed. In this study, a miniature soft lactic goat cheese model was compared with equivalent factory cheese to use it as a reliable model for this cheese family. The composition of the cheeses was compared at four key stages of the production process using microbiological, physicochemical and Sift-MS volatilome analysis. The compositions of the model and factory cheeses were found to be similar until the ripening step, but some differences were observed on ripened cheeses. Moreover, under carefully monitored laboratory conditions, the variability across batches of the model cheese was reduced. This miniature soft lactic goat cheese could therefore be used as a reliable model to study the aspects of the same cheese type.

Introduction

Pilot-scale cheesemaking experiments under controlled conditions are costly and timeconsuming in contrast to experiments at the laboratory scale (Shakeel-Ur-Rehman, [START_REF] Shakeel-Ur-Rehman | Alternatives to pilot plant experiments in cheese-ripening studies[END_REF].

Protocols for the preparation of miniature cheese models, produced in laboratorycontrolled conditions, where key properties of the miniature cheese models resemble those of conventionally manufactured cheeses, have been proposed for fresh cheeses [START_REF] Hynes | Protocol for the manufacture of miniature washed-curd cheeses under controlled microbiological conditions[END_REF], soft cheeses, semi-hard cheeses (Shakeel-Ur-Rehman, [START_REF] Shakeel-Ur-Rehman | Protocol for the manufacture of miniature cheeses[END_REF] and hard cheeses [START_REF] Antonsson | Screening and selection of Lactobacillus strains for use as adjunct cultures in production of semi-hard cheese[END_REF][START_REF] Vélez | Short communication: A new minicurd model system for hard cooked cheeses[END_REF]. However, despite the importance of soft lactic cheeses (simultaneous acidification and rennet coagulation), no comparable procedure has yet been described for them.

The microbiological, physicochemical, and sensorial characteristics of a few factory soft lactic goat cheeses have been described [START_REF] Couderc | Harnessing diversity of Lactococcus lactis from raw goat milk : Design of an indigenous starter for the production of Rocamadour, a French PDO cheese[END_REF][START_REF] Lucas | Respective effects of milk composition and the cheese-making process on cheese compositional variability in components of nutritional interest[END_REF], but not their volatile profiles. Gas chromatography-mass spectrometry (GC-MS) is generally used to identify and quantify volatile compounds, although this approach is costly and time-consuming. Selected ion flow tube-mass spectrometry (Sift-MS), a recently developed alternative approach, allows faster analysis and has already shown promising results for cheese aroma characterisation [START_REF] Langford | Comprehensive odorant analysis for on-line applications using selected ion flow tube mass spectrometry (SIFT-MS)[END_REF][START_REF] Reyrolle | Volatile fingerprint of food products with untargeted SIFT-MS data coupled with mixOmics methods for profile discrimination: Application case on cheese[END_REF].

The purpose of this study was to establish a reliable model for the soft lactic goat cheese family and compare it with its factory equivalent. The compositions of the model and factory cheeses were compared using microbiological, physicochemical and Sift-MS volatilome analysis.

J o u r n a l P r e -p r o o f

Materials and methods

Production of factory and miniature model cheese

Three batches of raw goats' milk were collected from "Les Fermiers du Rocamadour" (Alvignac, France) at different times of the year to make three batches of miniature models (produced in laboratory-controlled conditions) and factory cheeses. Each batch of milk was split into two parts; 180 L were used directly to factory cheese production at "Les Fermiers du

Rocamadour" and 2 L were transferred into 250 mL bottles and frozen at -20 °C for the subsequent production of the miniature models.

Factory cheeses were produced in accordance with Rocamadour PDO cheese specifications (Legifrance, 2014), as followed by [START_REF] Couderc | Harnessing diversity of Lactococcus lactis from raw goat milk : Design of an indigenous starter for the production of Rocamadour, a French PDO cheese[END_REF]. The process is outlined in Fig. 1A.

Model cheese was produced under laboratory conditions based on the factory cheese process (Fig. 1B). Frozen raw goats' milk was thawed for one day at 4 °C, pasteurised (85 °C, 5 min) and cooled to 35 °C. The same concentrations of starter and ripening microorganisms were used as in the factory process. CaCl2 (Durcigel, Coquard, Villefranche-sur-Saône, France) was added at 0.4 mL L -1 and a short maturation period of 45 min at 25 °C was done. Once the pH had decreased by 0.2 units, rennet (100 mL 100 L -1 of 520 mg L -1 chymosin; Coquard) was added.

The milk was then distributed into 50 mL tubes and incubated at 25 °C for 22 h.

As described in the protocols of previously produced model cheeses, centrifugation was used to separate the curd from the whey [START_REF] Hynes | Protocol for the manufacture of miniature washed-curd cheeses under controlled microbiological conditions[END_REF][START_REF] Shakeel-Ur-Rehman | Protocol for the manufacture of miniature cheeses[END_REF][START_REF] Van Tassell | Use of a miniature laboratory fresh cheese model for investigating antimicrobial activities[END_REF]. To be in accordance with

Rocamadour PDO cheese specifications (minimum 31% of dry matter and 0.6-0.8% of salt J o u r n a l P r e -p r o o f content; Legifrance, 2014), three conditions of centrifugation and three conditions of salting had previously been tested to select the following parameters (data not shown). The tube was centrifuged for 3 h 30 at 25 °C at 4430 rpm, the whey then discarded and the curd salted with a 185 mg mL -1 NaCl brine solution. The curd was placed in a custom-built mould (diameter, 38 mm; height, 10 mm, Verres Vagner, Toulouse, France), 3.5 times smaller than those used for the factory cheeses but with the same diameter/height ratio (Fig. 2A).

The resulting cheese was then placed in a miniature (175 cm 3 ) ripening cellar with two air inlets to insure proper ventilation (Fig. 2B). The mini cellar was placed in a temperaturecontrolled refrigerated container. A Petri dish with sterile cotton wool soaked in sterile water was placed at the bottom of the cellar to regulate relative humidity (RH). Temperature and RH were monitored every 2 h during the ripening process using a LogTag recorder (Trix8, LogTag, France) placed in the cellar.

All the equipment used were autoclaved or disinfected and UV-treated. Fig. 1B illustrates the two stages of the ripening process: (i) one day of ripening at 13 ± 0.2 °C and 92.8 ± 0.8 % RH and (ii) 6 days at 10 ± 0.1 °C and 94.5 ± 0.5% RH. The cheeses were turned once a day throughout the ripening process.

Samples of the model and factory cheeses were collected at four key stages of the production process: before salting, after salting, on the first day of ripening and at the end of the ripening stage (indicated with asterisks in Fig. 1).

Cheese composition analysis

Physicochemical analysis

Microbiological analysis

Microbiological analyses were performed on three ripened factory cheeses and three ripened model cheeses stored at 4 °C of three production batches. Aerobic mesophilic bacteria levels were enumerated on plate count agar (PCA; Biokar, France) supplemented with skimmed milk powder and, yeasts and moulds counts were enumerated on oxytetracycline glucose agar (OGA; Biokar). Serial decimal dilutions in buffered peptone (Biomerieux, France) were prepared and 0.1 mL samples of appropriate dilutions were plated on appropriate media. The PCA plates were incubated at 30 °C for 48 h and the OGA plates were incubated at 25 °C for 5 days. The same analyses were performed in Agrolabs (Auch, France) for the factory cheeses and internally for the model cheeses.

J o u r n a l P r e -p r o o f

Sift-MS analysis

Sift-MS analysis was performed on three ripened factory cheeses and three ripened miniature cheeses from a single production batch, that were vacuum-packed and stored at -20 °C.

The sample preparation protocol was inspired by [START_REF] Taylor | Discrimination of swiss cheese from 5 different factories by high impact volatile organic compound profiles determined by odor activity value using selected ion flow tube mass spectrometry and odor threshold[END_REF]. Five grams (± 0.3 g) of ripened cheese was placed in a 1 L bottle and sealed with a Teflon cap. The bottle was then heated at 40 °C for 1 h before Sift-MS analysis to allow the headspace to equilibrate.

The VOCs released by the cheese samples were analysed using a Sift-MS Voice 200 Ultra device (Syft Technologies Ltd., New Zealand) using positive soft ionising precursor ions (H3O + , NO + and O2 + ). After each analysis, a blank test was performed using an empty bottle.

Immediately after incubation, the headspace of the sample was introduced via a calibrated capillary at a sampling flowrate of 20 nmL min -1 into the reaction chamber (a flow tube at 120 °C and 0.07 kPa) with nitrogen (180 nmL min -1 ) as the carrier gas. The measurements were performed in full scan mode from 15 to 250 m/z with count rates (signal intensity in counts per second) calculated for each m/z unit.

The Sift-MS data were acquired and analysed using LabSyft v.1.6.2. (Syft Technologies, New Zealand). Four full mass scans were recorded for each sample but only the last three were analysed.

The data were co-added and pre-processed as described by [START_REF] Reyrolle | Volatile fingerprint of food products with untargeted SIFT-MS data coupled with mixOmics methods for profile discrimination: Application case on cheese[END_REF]. The VOC profiles of the model and factory cheese samples were compared based on the reagent ionproduct ion combinations of 36 VOCs previously identified in goat cheeses (Supplementary material Table S1) [START_REF] Bontinis | Study of proteolysis, lipolysis and volatile profile of a traditional Greek goat cheese (Xinotyri) during ripening[END_REF][START_REF] Delgado | Formation of the aroma of a raw goat milk cheese during maturation analysed by SPME-GC-MS[END_REF][START_REF] Hayaloglu | Characterizing volatile compounds and proteolysis in Gokceada artisanal goat cheese[END_REF].

J o u r n a l P r e -p r o o f

The volatile fingerprints of the model and factory cheeses were compared using the mixOmics R package [START_REF] Rohart | MixOmics : An R package for J o u r n a l P r e -p r o o f omics feature selection and multiple data integration[END_REF].

Statistical analysis

Gross cheese compositions were expressed as means and standard deviations over the three production runs and four sampling stages and compared between the factory and model cheeses using Kruskal-Wallis tests.

VOC product ion intensities were compared between the factory and the model cheeses with Wilcoxon signed-rank tests. Homogeneity of variance was verified using Levene's test. The threshold for statistical significance were set at p <0.05.

Results and discussion

Regarding physicochemical parameters (Table 1), fat and salt contents of the model cheeses were similar to those of the factory cheeses at all stages. For the other parameters, they were similar until the first day of ripening. Indeed, during ripening, few differences were recorded between the two types of cheeses concerning the dry matter contents, the moisture on a fat-free basis and the pH values. The lower values of the moisture on a fat-free basis (higher dry matter content) of the model cheeses could be explained by lower relative humidity during ripening. At the end of ripening, the pH value of the model cheeses was also significantly higher than those of the factory cheeses, showing more intense deacidifying microbial activity in the model cheeses. In fact, increases in pH value during ripening can be due to the production of ammonia and various amino acids by proteolysis, and by the consumption of the lactic acid by J o u r n a l P r e -p r o o f moulds and yeasts (which are the main deacidifying microorganisms in cheeses) [START_REF] Bonaiti | Deacidification by Debaryomyces hansenii of smear soft cheeses ripened under controlled conditions : relative humidity and temperature influences[END_REF][START_REF] Fox | Proteolysis during cheese manufacture and ripening[END_REF]. This was confirmed by the fact that after ripening, the factory cheeses had substantially lower levels of yeasts, such as Debaryomyces hansenii, than the model cheeses (6.60 ± 0.47 versus 2 log10 cfu g -1 , respectively, Table 2). Nonetheless, the model cheeses' levels of aerobic mesophilic bacteria and Geotrichum candidum were comparable with those of the factory cheeses. The different microbiological compositions of cheeses could explain the rise of the pH value that was more important in the model cheeses during ripening (+0.7 versus +0.3 units of pH for the model and factory cheeses, respectively). Moreover, this observation was not surprising since in the case of model cheeses, the microbial contamination of milk and environment are almost negligible (pasteurised milk, sterile mini cellar for the ripening) compared with factory production conditions. These microorganisms, and especially G.

candidum that could be present in the factory ripening room's environment, may play an important role in the inoculation of the fresh cheese. G. candidum's high and early presence in factory cheeses may make it harder for other yeasts to develop during ripening. Indeed, it is known that G. candidum could inhibit D. hansenii during ripening [START_REF] Mounier | Microbial interactions within a cheese microbial community[END_REF].

The volatile fingerprints of the factory and model cheeses, given by the average intensities of 119 product ions (associated with the 36 selected VOCs, Supplementary material Table S1), are compared in Fig. 3. Only 21/119 product ions intensities differed significantly between the factory and model cheeses (p <0.05, Wilcoxon tests, indicated by asterisks in Fig. 3 and Supplementary material Table S1), showing a relatively similar cheese VOCs composition.

Moreover, a significant heterogeneity of variance between samples was observed for 3/119 model cheeses product ions and 9/119 factory cheeses product ions (p <0.05, Levene test, Supplementary material Table S2). Therefore, the level of repeatability is higher on model J o u r n a l P r e -p r o o f cheeses than on factory cheeses, certainly due to pasteurised milk used to make the model cheeses.

The gross composition of the ripened model cheese presented in our study is similar to that of soft lactic cheeses previously characterised [START_REF] Gaborit | Impact of ripening strains on the typical flavour of goat cheeses[END_REF]Raynalljutovac, Pape, Gaborit, & Barrucand, 2011;[START_REF] Sablé | Microbiological changes in a soft raw goat's milk cheese during ripening[END_REF] and could therefore be used as a reliable model for this cheese type, for which a model has never been described in literature.

Conclusion

In this study, a miniature soft lactic goat cheese model was elaborated and compared with a factory cheese, based on microbiological, physicochemical and Sift-MS volatilome analysis.

The model cheese has a similar composition to the factory cheese until ripening and differed on non-Geotrichum candidum yeasts, moisture on a fat-free basis levels and pH values for ripened cheese. During ripening, less variability was recorded in the model cheese between production batches compared with the factory cheese. This model should therefore facilitate future studies on the impact of various factors on quality of soft lactic cheeses. J o u r n a l P r e -p r o o f

  J o u r n a l P r e -p r o o f Physicochemical analyses were performed on three factory cheeses and three model cheeses stored at 4 °C at all four stages of all three production batches. Dry matter (DM), fat (FC) and salt (S) contents were analysed according to NF EN ISO 5534, NF V04-287 and NF EN ISO 5943 standards, respectively. The salt-in-moisture (S/M) and moisture on a fat-free basis (MFFB) contents were calculated as follows: S/M = (%S) / (100 -%DM) MFFB = [(100 -%DM) / (100 -%FC)] × 100 All analyses for the factory cheeses and the fat and salt content measurements for the model cheeses were performed by Agrolabs (Auch, France). The DM and pH measurements for the model cheeses were performed internally. The pH values were measured using a HQ11d pH meter (Hach, France).

Fig. 1 .

 1 Fig. 1. Production flow charts for (A) factory and (B) miniature model soft lactic goat

Fig. 2 .Fig. 3 .

 23 Fig. 2. Photographs of (A) a miniature model cheese and (B) the miniature cheese ripening

Acknowledgments

The authors thank Sebastien Fau and Benoit Bonizzoni from "Les Fermiers du Rocamadour" for their technical support during the cheesemaking experiments, Leticia Vitola Pasetto for her help with Sift-MS analysis, and to Occitanie Region for funding the project.

J o u r n a l P r e -p r o o f

Declaration of interests

☒ The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: