Phenotypic Antimicrobial Resistance Profiles of Human Campylobacter Species Isolated in Northwest Italy, 2020-2023.

The spread of antimicrobial resistant Campylobacter strains, linked to antimicrobials use and abuse in humans and food animals, has become a global public health problem. In this study, we determine the prevalence of antimicrobial resistance (AMR) in human Campylobacter isolates (n = 820) collected in Piedmont, Italy, from March 2020 to July 2023. The strains underwent susceptibility testing to determine the minimal inhibitory concentration for erythromycin, ciprofloxacin, gentamicin, streptomycin, and tetracycline: 80.1% of the strains showed resistance to at least one antibiotic. The highest prevalence of AMR was noted for ciprofloxacin and tetracycline (72.1% and 52.9%, respectively) and the lowest for erythromycin and aminoglycosides (streptomycin/gentamicin) (3.2% and 5.4%, respectively). The prevalence of co-resistance against fluoroquinolones and tetracyclines was 41.1%. The prevalence of multidrug resistant strains was 5.7%. Our data support evidence that AMR in human Campylobacter strains is common, particularly against ciprofloxacin and tetracycline, two medically important antimicrobials for humans.

Performance Evaluation of a Commercial Real-Time PCR Method for the Detection of Lupin Traces in Food.

In accordance with U.S. FDA Foods Program Regulatory Science Steering Committee guidelines, with this study, we optimized and validated a commercial real-time PCR method for the detection of low amounts of lupin in four classes of food matrices: chocolate cookies, ragù, Olivier salad, and barley and rice flour. DNA extracted from blank (true negative) samples artificially contaminated with lupin (Lupinus albus) flour at 1000 ppm underwent dilutions with the DNA extracted from the true negative samples up to 0.5 ppm. The limit of detection for real-time PCR was 0.5 ppm in the complex matrices (range, Ct 26-34), making this a specific, robust, and rapid method for lupin allergen detection and labeling. Our validation data support the suitability of this commercially available real-time PCR method for this purpose.

Detection and Characterization of Zoonotic Pathogens in Game Meat Hunted in Northwestern Italy.

Wildlife can represent a reservoir of zoonotic pathogens and a public health problem. In the present study, we investigated the spread of zoonotic pathogens (Salmonella spp., Yersinia enterocolitica, Listeria monocytogenes, Shiga-toxin-producing Escherichia coli (STEC), and hepatitis E virus (HEV)) considering the presence of virulence and antibiotic resistance genes in game meat from animals hunted in northwest Italy. During two hunting seasons (2020 to 2022), samples of liver and/or muscle tissue were collected from chamois (n = 48), roe deer (n = 26), deer (n = 39), and wild boar (n = 35). Conventional microbiology and biomolecular methods were used for the detection, isolation, and characterization of the investigated pathogens. Two L. monocytogenes serotype IIa strains were isolated from wild boar liver; both presented fosfomycin resistance gene and a total of 22 virulence genes were detected and specified in the text. Eight Y. enterocolitica biotype 1A strains were isolated from chamois (2), wild boar (5), and deer (1) liver samples; all showed streptogramin and beta-lactam resistance genes; the virulence genes found were myfA (8/8 strains), ymoA (8/8), invA (8/8), ystB (8/8), and ail (4/8). Our data underscore the potential role of wildlife as a carrier of zoonotic and antibiotic-resistant pathogens in northwest Italy and a food safety risk for game meat consumers.

In Vivo Evaluation of the Potential of Thyme and Lemon Hydrolates as Processing Aids to Reduce Norovirus Concentration during Oyster Depuration.

In this study, we evaluated the use of hydrolates, co-products of essential oil distillation, as processing aids to improve the depuration process of Pacific oysters (Crassostrea gigas) as a post-harvest method aimed at reducing the norovirus (NoV) viral load. Live oysters were kept in water to which hydrolates of Thymus serpyllum and Citrus limon at 1% were added for 24 h. The concentration of NoV was quantified using the ISO 15216-1 quantitative real-time RT-PCR method in the oyster digestive tissue both before and after the treatment. The results showed a significant reduction of 0.2 log in the NoV GII concentration after 24 h of treatment with 1% C. limon hydrolate. Conversely, treatment with T. serpyllum did not appear to reduce the concentration of NoV compared to the control. Additionally, a sensory analysis was conducted through a blind survey comparing untreated and treated oysters. No changes in the sensory and physical characteristics of the oysters were observed, except for a decrease in the marine flavour intensity, which was positively perceived by consumers. These results indicate that the addition of hydrolates of C. limon at 1% during depuration might represent a promising processing aid for enhancing both the safety and acceptability of live oysters.

First Report of Food Poisoning Due to Staphylococcal Enterotoxin Type B in Döner Kebab (Italy).

Staphylococcal food poisoning results from the consumption of food contaminated by staphylococcal enterotoxins. In July 2022, the Turin local health board was notified of a suspected foodborne outbreak involving six children who had consumed döner kebab purchased from a takeaway restaurant. The symptoms (vomiting and nausea) were observed 2-3 h later. A microbiological analysis of the food samples revealed high levels (1.5 × 107 CFU/g) of coagulase-positive staphylococci (CPS). The immunoassay detected a contamination with staphylococcal enterotoxins type B (SEB). The whole genome sequencing of isolates from the food matrix confirmed the staphylococcal enterotoxin genes encoding for type B, which was in line with the SEB detected in the food. This toxin is rarely reported in staphylococcal food poisoning, however, because there is no specific commercial method of detection. The involvement of enterotoxin type P (SEP) was not confirmed, though the corresponding gene (sep) was detected in the isolates. Nasal swabs from the restaurant food handlers tested positive for CPS, linking them to the likely source of the food contamination.

Food Safety Monitoring of Salmonella spp. in Northern Italy 2019-2021.

Salmonella is the second most frequent bacterial pathogen involved in human gastrointestinal outbreaks in the European Union; it can enter the food-production chain from animal or environmental sources or from asymptomatic food operators. European food legislation has established microbiological criteria to ensure consumer protection. Salmonella is listed under both process hygiene criteria and food safety criteria. Each EU member state designates an agency to organize or perform controls and other official activities. This paper describes the official control plans performed by competent authorities in Northern Italy in the three-year period 2019-2021. A total of 4413 food samples were delivered to the IZS Food Safety laboratories for Salmonella detection, of which 36 (0.8%) tested positive. Salmonella was most frequently detected in poultry meat samples (25/36 positive samples) followed by other meat products and pork products. The official controls for the protection of consumer health apply the EU's farm-to-fork approach: the samples were collected during production (food production plants), from products on the market, and from collective catering (restaurants, cafeterias, canteens). This manuscript will provide information about the presence of Salmonella in foodstuffs that can help competent authorities to set control plans based on risk assessments.

Soy, Rice and Oat Drinks: Investigating Chemical and Biological Safety in Plant-Based Milk Alternatives.

During the last decades, plant-based milk has become very appreciated by consumers, becoming a staple ingredient, especially for alternative breakfasts. Milk contains lactose, which is a sugar hydrolysed by the lactase enzyme. Lactose intolerance and lactose malabsorption are very common food intolerances among individuals. However, a lot of consumers consider themselves as lactose intolerant on the basis of self-reported intolerance and start to avoid dairy products, ignoring that plant-based milk alternatives are not nutritionally comparable to animal milk, especially in terms of protein intake. The aim of this study is to grow folder knowledge of the security of plant-based drinks, helping competent authorities to issue a risk assessment and to apply national plans about consumer safety. Results show that proper sanitary practices, such as pasteurization, are necessary in plant-based milk alternatives as well as in dairy milk. Chemical analysis has highlighted that there are no pesticide risks for consumers.

First Description of Acaricide Resistance in Populations of Rhipicephalus microplus Tick from the Lower Amazon, Brazil.

There is limited information on the resistance to acaricidal drugs of the cattle tick Rhipicephalus microplus in the Lower Amazon region. Thus, we aimed to determine the efficiency of three widely used acaricide products (Amitraz, Cypermethrin, and Deltamethrin) in the control of this tick species. The adult immersion test was used on engorged female ticks sampled on farms in the Lower Amazon region, Brazil. For the test, homogeneous batches of 10 engorged females were placed in Petri dishes and immersed in the tested acaricidal drugs, using four replicates of each acaricide and three replicates as a negative control, immersed in distilled water. The acaricides were diluted as recommended by the manufacturers, and the ticks were submerged for five minutes. Mortality of engorged females, production of eggs, and percentage of larval hatching were evaluated. Tick mortality was 2.5%, 7.5%, and 0% for Amitraz, Cypermethrin, and Deltamethrin, respectively. The percentage of larval hatching was 53.7% for Amitraz, 88.7% for Cypermethrin, and 80.0% for Deltamethrin. As recommended by the FAO, for the acaricide to be considered effective, it must have a control rate ≥95%. Among the tested acaricides, Amitraz showed an efficacy of 90.5%, Cypermethrin 10.4%, and Deltamethrin 26.6%. Ticks from the lower Amazon showed marked resistance to Cypermethrin and Deltamethrin and mild resistance to Amitraz. This is the first report of acaricidal resistance in the region.

Influence of Season and Food Type on Bacterial and Entero-Toxigenic Prevalence of Staphylococcus aureus.

Staphylococcus (S.) aureus is a coagulase-positive pathogen of interest for human health and food safety in particular. It can survive in a wide environmental temperature range (7-48 °C, optimum 37 °C). Its enterotoxins are thermostable, which increases the risk of potential contamination in a variety of food products. Here we investigated the influence of seasonality and food type on bacterial count and presence of S. aureus enterotoxins. To do this, we analyzed 3604 food samples collected over a 5-year period (2016-2020). Ordinal logistic regression showed an influence of both seasonality and food type on the bacterial count. Regarding bacterial counts, winter was found to be the season with the highest risk, while with regards to enterotoxin production, the highest risk was found in autumn, specifically in October. The risk of contamination with S. aureus was greatest for dairy products. Our findings may inform food epidemiologists about foodborne illness prevention and risk to human health.

Biofilm formation and genomic features of Listeria monocytogenes strains isolated from meat and dairy industries located in Piedmont (Italy).

Listeria monocytogenes is considered a major challenge for the food industry as it can persist for long periods in food processing plants by forming biofilms. The aims of this study were: i) to assess the biofilm producing ability of 57 Listeria monocytogenes isolates previously subjected to whole-genome sequencing (WGS); ii) to compare the levels of biofilm formation with the presence or absence of biofilm associated genes. To determine the presence or absence of a known set of biofilm associated genes, a comparative genomic analysis was performed on each strain. Among Listeria monocytogenes isolates, 58 %, 38.5 % and 3.5 % exhibited weak, moderate or strong biofilm production, respectively. No difference in biofilm production was observed between food and environmental isolates. The percentage of Listeria monocytogenes strains isolated from meat products (57 %) classified as moderate or strong biofilm producers was higher than the percentage obtained for strains isolated from dairy products (28 %). The presence of the Stress Survival Islet 1, the arsD stress gene and the truncated inlA protein was significantly associated with increased levels of biofilm. Combining biofilm phenotype with molecular and genotyping data may provide the opportunity to better understand the relationship between genes linked to biofilm formation in Listeria monocytogenes.

Detection of Peanut Traces in Food by an Official Food Safety Laboratory.

Food safety laboratories rely on validated methods that detect hidden allergens in food to ensure the safety and health of allergic consumers. Here we present test results for the validation and accreditation of a real-time PCR assay for the detection of peanut traces in food products. The method was tested on five classes of food matrices: bakery and pastry products, meats, ready-to-eat and dairy products, and grains and milling products. Blank samples were spiked starting with the peanut samples (Arachis hypogaea) at a concentration of 1000 ppm. Serial dilutions were then prepared with the DNA extracted from the blank samples to a final concentration of 0.5 ppm. The limit of detection in grains and milling products, ready-to-eat, meats, bakery and pastry products was 0.5 ppm (range, Ct 27-34) and 2.5 ppm in dairy products (range, Ct 25-34). In order to determine the exclusivity parameter of the method, the ragù matrix was contaminated with Prunus dulcis (almonds), Glycine max (soy), Sinapis alba (mustard), Apium graveolens (celery), Allium cepa (onion), Pisum sativum (peas), Daucus carota (carrots), and Theobroma cacao (cocoa); no cross-reactions were observed. The method was rated satisfactory for sensitivity (98%), specificity (100%), robustness, and repeatability and it was fully validated and accredited.

Membrane electrical properties of mouse hippocampal CA1 pyramidal neurons during strong inputs.

In this work, we highlight an electrophysiological feature often observed in recordings from mouse CA1 pyramidal cells that has so far been ignored by experimentalists and modelers. It consists of a large and dynamic increase in the depolarization baseline (i.e., the minimum value of the membrane potential between successive action potentials during a sustained input) in response to strong somatic current injections. Such an increase can directly affect neurotransmitter release properties and, more generally, the efficacy of synaptic transmission. However, it cannot be explained by any currently available conductance-based computational model. Here we present a model addressing this issue, demonstrating that experimental recordings can be reproduced by assuming that an input current modifies, in a time-dependent manner, the electrical and permeability properties of the neuron membrane by shifting the ionic reversal potentials and channel kinetics. For this reason, we propose that any detailed model of ion channel kinetics for neurons exhibiting this characteristic should be adapted to correctly represent the response and the synaptic integration process during strong and sustained inputs.

Laniakea@ReCaS: exploring the potential of customisable Galaxy on-demand instances as a cloud-based service.

BACKGROUND: Improving the availability and usability of data and analytical tools is a critical precondition for further advancing modern biological and biomedical research. For instance, one of the many ramifications of the COVID-19 global pandemic has been to make even more evident the importance of having bioinformatics tools and data readily actionable by researchers through convenient access points and supported by adequate IT infrastructures. One of the most successful efforts in improving the availability and usability of bioinformatics tools and data is represented by the Galaxy workflow manager and its thriving community. In 2020 we introduced Laniakea, a software platform conceived to streamline the configuration and deployment of "on-demand" Galaxy instances over the cloud. By facilitating the set-up and configuration of Galaxy web servers, Laniakea provides researchers with a powerful and highly customisable platform for executing complex bioinformatics analyses. The system can be accessed through a dedicated and user-friendly web interface that allows the Galaxy web server's initial configuration and deployment. RESULTS: "Laniakea@ReCaS", the first instance of a Laniakea-based service, is managed by ELIXIR-IT and was officially launched in February 2020, after about one year of development and testing that involved several users. Researchers can request access to Laniakea@ReCaS through an open-ended call for use-cases. Ten project proposals have been accepted since then, totalling 18 Galaxy on-demand virtual servers that employ ~ 100 CPUs, ~ 250 GB of RAM and ~ 5 TB of storage and serve several different communities and purposes. Herein, we present eight use cases demonstrating the versatility of the platform. CONCLUSIONS: During this first year of activity, the Laniakea-based service emerged as a flexible platform that facilitated the rapid development of bioinformatics tools, the efficient delivery of training activities, and the provision of public bioinformatics services in different settings, including food safety and clinical research. Laniakea@ReCaS provides a proof of concept of how enabling access to appropriate, reliable IT resources and ready-to-use bioinformatics tools can considerably streamline researchers' work.

Comparison of Clostridioides difficile strains from animals and humans: First results after introduction of C. difficile molecular typing and characterization at the Istituto Zooprofilattico Sperimentale of Piemonte, Liguria e Valle d'Aosta, Italy.

PCR ribotypes (RTs027 and 078) are known causes of Clostridioides difficile infection (CDI) in humans. Molecular typing and characterization of 39 C. difficile strains isolated from samples from humas and animals in 2016-2018 indicated an overlap of RTs between community-acquired patients (CA-CDI) and domestic animals from the same geographical area; 14 RTs were identified: 12 RTs were positive for toxins A/B; RT078, RT080 and RT126 were also positive for binary toxin (CDT). Most of the RTs from the animals (RTs020, 078, 106, 126) were also detected in the samples from humans. Strains grouped into three clusters: cluster I included prevalently human strains, mainly RT 018; clusters II and III included strains from humans and animals, mainly RT078 and RT020. The CA-CDI strains suggested animals as a reservoir of C. difficile isolated together with other microorganisms from animals, highlighting the association of enteric pathogens as a cause of infection and death.

Abordagem das reações de hipersensibilidade perioperatória: Orientações da Sociedade Brasileira de Anestesiologia e da Associação Brasileira de Alergia e Imunologia - Parte II: etiologia e diagnóstico / Approach to perioperative hypersensitivity reactions: Guidelines from the Brazilian Society of Anesthesiology and the Brazilian Association of Allergy and Immunology - Part II: etiology and diagnosis

A anafilaxia perioperatória é manifestação importante no contexto de eventos adversos relacionados à cirurgia. Embora frequentemente relacionada à indução anestésica, pode ocorrer por outros agentes administrados por outras vias. A anafilaxia pode se apresentar como colapso cardiovascular, obstrução da via aérea e/ou insuficiência respiratória com ou sem manifestação cutânea, com consequências fatais em muito casos. Apesar de considerada inevitável em alguns casos, a sua incidência poderia (e deveria) ser reduzida através da busca por fármacos mais seguros. A avaliação abrangente de um episódio é um dos elementos primordiais para tornar a exposição subsequente mais segura, com orientações derivadas dessa investigação. Entretanto, representa um desafio estatístico por ser reação rara, randômica e muitas vezes independente de exposições sucessivas dos pacientes a procedimentos de baixo risco. Neste documento são revisados os mecanismos fisiopatológicos, agentes desencadeantes (adultos e crianças), assim como a abordagem diagnóstica durante a crise e após o episódio. Uma avaliação abrangente, a identificação das medicações, antissépticos e outras substâncias usadas em cada região, registros detalhados e nomenclatura padronizada são pontos fundamentais para a obtenção de dados epidemiológicos mais fidedignos sobre a anafilaxia perioperatória.

Perioperative anaphylaxis is an important manifestation in the context of surgery-related adverse events. Although often related to anesthetic induction, it may be caused by other agents administered by other routes. Anaphylaxis may manifest as cardiovascular collapse, airway obstruction and/or respiratory failure with or without skin manifestation, resulting often in death. Although this reaction is considered inevitable in some cases, its incidence could (and should) be reduced by the search for safer drugs. Comprehensive assessment of an allergic reaction is a key element to make subsequent exposure safer, with guidance derived from this investigation. However, surveillance of perioperative anaphylaxis represents a statistical challenge because this is a rare, random reaction and often independent of successive patient exposures to low-risk procedures. This paper reviews pathophysiological mechanisms, triggering agents (adults and children), as well as therapeutic and diagnostic approach during and after an allergic reaction. Comprehensive assessment, identification of medications/antiseptics used in each region and detailed records with standardized terminology are key points for obtaining more reliable epidemiological data on perioperative anaphylaxis.

Genetic Characterization of a Listeria monocytogenes Serotype IVb Variant 1 Strain Isolated from Vegetal Matrix in Italy.

We report the chromosome and plasmid sequences of a strain of Listeria monocytogenes IVb variant 1, a recently emerging serotype, isolated in Italy from ready-to-eat vegetables.

An Italian survey of undeclared allergens in food over the years 2014-2018.

Large population studies estimated that the frequency of food allergies is increasing worldwide. In the last two decades, a 'second wave' of the allergy epidemic has emerged as a growing public health problem. EU regulation strengthened information to consumers about allergens in pre-packed food, since December 2014 it has been extended to non-prepacked foods by the Regulation (EU) No 1169/2011 of the European Commission. The present work aims to present a five-year survey on the presence of nine types of allergen in several foods, including food of animal origin and baby food. The concentration found for each irregular allergen is generally low, but some samples with no gluten indication contained a concentration above 10 mg kg-1 with the highest value of 138.5 mg kg-1 in a dietetic food, during the screening in 2017. These data underline the importance and the necessity of a complete food labelling to protect the majority of allergic individuals.

Atualização sobre reações de hipersensibilidade perioperatória: Documento conjunto da Sociedade Brasileira de Anestesiologia e Associação Brasileira de Alergia e Imunologia ­ Parte II: etiologia e diagnóstico / Update on perioperative hypersensitivity reactions: Joint document from the Brazilian Society of Anesthesiology and Brazilian Association of Allergy and Immunology ­ Part II: Etiology and diagnosis

A anafilaxia perioperatória é manifestação importante no contexto de eventos adversos relacionados à cirurgia. Embora frequentemente relacionada à indução anestésica, pode ocorrer por outros agentes administrados por outras vias. A anafilaxia pode se apresentar como colapso cardiovascular, obstrução da via aérea e/ou insuficiência respiratória com ou sem manifestação cutânea, com consequências fatais em muito casos. Apesar de considerada inevitável em alguns casos, a sua incidência poderia (e deveria) ser reduzida através da busca por fármacos mais seguros. A avaliação abrangente de um episódio é um dos elementos primordiais para tornar a exposição subsequente mais segura, com orientações derivadas dessa investigação. Entretanto, representa um desafio estatístico por ser reação rara, randômica e muitas vezes independente de exposições sucessivas dos pacientes a procedimentos de baixo risco. Neste documento são revisados os mecanismos fisiopatológicos, agentes desencadeantes (adultos e crianças), assim como a abordagem diagnóstica durante a crise e após o episódio. Uma avaliação abrangente, a identificação das medicações, antissépticos e outras substâncias usadas em cada região, registros detalhados e nomenclatura padronizada são pontos fundamentais para a obtenção de dados epidemiológicos mais fidedignos sobre a anafilaxia perioperatória.

Perioperative anaphylaxis is an important manifestation in the context of surgery-related adverse events. Although often related to anesthetic induction, it may be caused by other agents administered by other routes. Anaphylaxis may manifest as cardiovascular collapse, airway obstruction and/or respiratory failure with or without skin manifestation, resulting often in death. Although this reaction is considered inevitable in some cases, its incidence could (and should) be reduced by the search for safer drugs. Comprehensive assessment of an allergic reaction is a key element to make subsequent exposure safer, with guidance derived from this investigation. However, surveillance of perioperative anaphylaxis represents a statistical challenge because this is a rare, random reaction and often independent of successive patient exposures to low-risk procedures. This paper reviews pathophysiological mechanisms, triggering agents (adults and children), as well as therapeutic and diagnostic approach during and after an allergic reaction. Comprehensive assessment, identification of medications/antiseptics used in each region and detailed records with standardized terminology are key points for obtaining more reliable epidemiological data on perioperative anaphylaxis.

Possibilidades da Clínica Gestáltica no Atendimento de Crianças Enlutadas / Possibilities of the Gestalt Clinic in the Care of Bereaved Children / Posibilidades de la Clínica Gestaltáltica en el Cuidado de Duelo en la Infancia

O luto é compreendido como o processo psíquico de elaboração de uma perda que rompe com toda a constituição de mundo interno e segurança da criança, exigindo que novas formas de ajustamento sejam criadas. A presença de um outro-suporte é fundamental para que a criança possa percorrer seus ciclos de contato e alcançar a satisfação de suas necessidades no enfrentamento do luto. A Gestalt-terapia é uma abordagem fenomenológico-existencial, focada na construção dialógica, na qual a relação terapeuta-cliente ganha um eixo central. Em termos de prática clínica, o terapeuta se coloca em abertura à espera do outro-humano que o busca, na expectativa de que se revele, acolhendo os conteúdos, reconhecendo sua manifestação, em um processo de aceitação, confirmação e inclusão. Neste sentido, a intervenção terapêutica acompanha a expressão do brincar como linguagem para a criança comunicar, compreender e assimilar o vivido. O caminho escolhido para realização deste estudo foi a revisão bibliográfica de teóricos da Gestalt-terapia e estudos em luto na contemporaneidade, com enfoque nas bases relacionais destas teorias associadas a vinhetas de casos clínicos para ampliar a compreensão e possibilidades de intervenção com crianças enlutadas na clínica gestáltica. (AU)

Grief is understood as the psychic process of elaboration of a loss that breaks with the entire constitution of the inner world and the child's safety, requiring that new forms of adjustment be created. The presence of another support is fundamental for the child to be able to go through their cycles of contact and achieve the satisfaction of their needs in facing grief. Gestalt therapy is a phenomenological-existential approach, focused on dialogic construction, where the therapist-client relationship takes on a central axis. In terms of clinical practice, the therapist stands open waiting for the other human who seeks him, expecting to reveal himself, welcoming the contents, recognizing their manifestation, in a process of acceptance, confirmation and inclusion. In this sense, therapeutic intervention accompanies the expression of play as a language for the child to communicate, understand and assimilate the lived experience. The path chosen for this study was the bibliographic review of Gestalt therapy theorists and studies in mourning in contemporaneity, focusing on the relational bases of these theories associated with clinical case vignettes to broaden the understanding and possibilities of intervention with bereaved children in the clinic gestalt. (AU)

El duelo se entiende como el proceso psíquico de elaboración de una pérdida que rompe con toda la constitución del mundo interior y la seguridad del niño, requiriendo la creación de nuevas formas de ajuste. La presencia de otro apoyo es fundamental para que el niño pueda pasar por sus ciclos de contacto y alcanzar la satisfacción de sus necesidades al enfrentar el dolor. La terapia gestáltica es un enfoque fenomenológico-existencial, centrado en la construcción dialógica, donde la relación terapeuta-cliente adquiere un eje central. En términos de práctica clínica, el terapeuta permanece abierto esperando al otro ser humano que lo busca, esperando revelarse, acogiendo los contenidos, reconociendo su manifestación, en un proceso de aceptación, confirmación e inclusión. En este sentido, la intervención terapéutica acompaña la expresión del juego como lenguaje para que el niño se comunique, comprenda y asimile la experiencia vivida. El camino elegido para este estudio fue la revisión bibliográfica de los teóricos de la Terapia Gestalt y los estudios contemporáneos de duelo, centrándose en las bases relacionales de estas teorías asociadas con viñetas de casos clínicos para ampliar la comprensión y las posibilidades de intervención con niños en duelo en la clínica gestáltica. (AU)