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- Rapid response Task Force: addressing the detection of Aedes albopictus in Lisbon, PortugalPublication . Grau-Pujol, B.; Moreira, A.; Vieira Martins, J; Costa Osório, H; Ribeiro, L.; Dinis, A.; Sousa, C.; Alves, M. João; Leite, P. Pinto; Vasconcelos, P.Issue: The invasive Aedes albopictus mosquitoes are spreading in southern Europe and pose a heightened risk of mosquito-borne diseases, like Dengue and Chikungunya. On September 2023, A. albopictus was first identified in Lisbon through community-based surveillance and a multidisciplinary national Task Force (TF) was convened. Description of the problem: The TF included experts in epidemiology, entomology, environmental health, laboratory diagnosis, prevention and control and communication. The objectives were to i) confirm A.albopictus presence in Lisbon and adjust the risk assessment, ii) raise awareness and prevent mosquitoes’ spread, iii) revise national guidelines on arboviruses surveillance and control. During September-December 2023, the National Network for Vectors Surveillance (REVIVE) and the Hygiene and Tropical Medicine Institute sampled mosquito-prone habitats within 2km-radius of the detection point using QGIS software. They used snowball sampling to identify any new foci. They placed ovitraps for ongoing surveillance and conducted morphological analysis and xenomonitoring for pathogen DNA and RNA detection at the national reference laboratory. Results: By November 2023, A.albopictus was detected in two new foci. No mosquitoes tested positive for arboviruses, and no autochthonous cases of mosquito-borne diseases were identified. The TF developed national guidelines for vector prevention and control and awareness for arboviruses and trained municipality workers. To raise awareness, we communicated with experts and the public through existing channels and media. As of April 2024, no new foci have been detected. Lessons: Community-based surveillance and REVIVE approach were crucial for invasive A.albopictus detection and monitoring in Lisbon. Rapid multidisciplinary TF coordination may have been key for timely vector control.
- World Youth Day 2023 - non-endemic infectious disease surveillance through wastewater analysesPublication . Valério, Elisabete; Palminha, Paula; Neves, Raquel; Cadinha, Luis; Ferreira, João Duarte; Correia, Paulo; Medeiros, InêsThe World Youth Day (WYD) is an event that mobilizes millions of people from all over the world. It is an event promoted by the Catholic Church mainly focused on young people and aims to bring together representatives from all Catholic countries. In 2023, WYD was held in Portugal, hosted in Lisbon between August 1st and 6th. Other satellite events prior to WYD were promoted throughout the country. Portimão Municipality agreed to collaborate with the “Welcome to Paradise” festival between the 26th and 31st of July, before traveling to Lisbon for WYD. THE PROBLEM: 4000 young people were expected to arrive from 27 countries, on 5 continents, that were accommodated in 10 schools in the municipality of Portimão. There was a very diverse epidemiological context, namely the endemic diseases in the origin countries. Taking into account the high proximity of the different groups in provisionary accommodation establishments, we could suppose that the conditions could favor the spread of a non-endemic infectious disease with the potential to pressure available health services beyond their capacity to respond. AIM Monitor possible introduction of non-endemic infectious disease into the community, through analyses of the wastewater. THE APPROACH: The Barlavento Public Health Unit carried out a survey of endemic diseases in each country of origin of the participants. In order to monitor the possible introduction of a non-endemic disease into the community, focused on poliovirus, a protocol was established with the National Institute of Health Doutor Ricardo Jorge to monitor some diseases through the analysis of wastewater from the Portimão WWTP before, during and after the event. Research was carried out on non-endemic Poliovirus, and other endemic agents as control: Non Polio Enterovirus, adenovirus, Hepatitis A virus and Norovirus GI and GII search. RESULTS: Monitoring carried out on wastewater in Portimão before arrival, upon departure and 10 days after departure was always negative for Poliovirus and Hepatitis A virus and always positive for Norovirus I and II, Non Polio Enterovirus and Adenovirus. CONCLUSIONS: This Case Study of the wastewater monitoring performed allowed to discard the worries raised with such a high dimension event. This kind of approach is important, since we have already verified that wastewater analysis might be a relevant early warning system for public health risks.
- Mark-Release-Recapture of Aedes albopictus in Portugal: the influence of climatic factorsPublication . Soares, Patricia; Carvalho, Danilo O.; Silva, Susana Pereira; Gomez Pacheco, Maylen; Amaro, Fátima; Amaro F.; Osório, HugoAedes albopictus mosquitoes spread diseases like dengue, Zika, yellow fever, and chikungunya. The Sterile Insect Technique (SIT), which is based on production of male sterile mosquitoes through radiation and in the release in target areas, can be used as a prevention and control tool against Ae. albopictus native populations. Mark-Release-Recapture trials (MRR) estimate the number of mosquitoes to release during the SIT, but weather conditions can affect results. Thus, we aimed to understand the influence of climatic factors on mosquito collection. MRRs were conducted in October 2022 in Faro, Southern Portugal. Mosquito sterilisation, marking and transport followed protocols from the International Atomic Energy Agency. Releases occurred weekly for three consecutive weeks at two different sites, and mosquitoes were collected using Human Landing Collections (HLC) one, two, four and six days after release. Climatic data, such as temperature, humidity, wind intensity (moderate 15-35 km/h vs weak <15 km/h) and precipitation, were obtained from the Portuguese Institute of the Sea and the Atmosphere. We used generalised linear models with a negative binomial family and log function to estimate factors associated with the number of captured mosquitoes, estimated prevalence ratios (PRs), and the 95% confidence intervals (CI). We released 84 000 marked sterile male mosquitoes and recaptured 528 marked males (0.7%) by HLC. The prevalence of captured mosquitoes was 23% lower when the wind intensity was moderate compared with a weak wind intensity (PR: 0.77, 95% CI: 0.61−0.98). We did not find a statistically significant association between the number of captured mosquitoes and humidity (PR: 0.98, 95% CI: 0.96−1.00), temperature (PR: 1.03, 95% CI: 0.83−1.28) and precipitation (PR: 1.07, 95% CI: 0.60−2.03). Conclusion: It is crucial to plan MRRs carefully and consider weather conditions during their execution to avoid underestimating the population needed for the SIT and compromising the effectiveness of this control method.
- Comprehensive typing and genetic analysis of L. monocytogenes isolates: implication for food safety and antibiotic resistance surveillancePublication . Silva, Adriana; Silva, Vanessa; Borges, Vítor; Coelho, Anabela; Batista, Rita; Esteves, Alexandra; Igrejas, Gilberto; Saraiva, Cristina; Gomes, João Paulo; Poeta, PatríciaListeria species are commonly found in various environments and contaminated food, with livestock serving as a significant source of foodborne pathogens. Among these species, Listeria monocytogenes (L. monocytogenes) is particularly noteworthy as it can affect both livestock and humans. Antibiotics are frequently used in food animals for disease treatment and prevention on a large scale. This practice can lead to the selection of antibiotic-resistant bacterial strains, which can then spread to humans through the food chain. Consequently, L. monocytogenes, a ubiquitous foodborne pathogen, has been associated with global outbreaks of foodborne illnesses. To address this concern, the aim of the study was to conduct comprehensive typing and genetic analysis of 13 L. monocytogenes isolates obtained from food and food-processing environments.Among the 13 L. monocytogenes isolates, eight sequence types (ST) were identified: two isolates were identified as belonging to ST9; one as ST155; four as ST3, two as ST121, one as ST8; one as ST87; one as ST1; and one new ST belonging to CC121. Core-genome clustering analysis of L. monocytogenes was made to assess the genetic relatedness among the isolates. The core genome Multilocus Sequence Typing (cgMLST) analysis revealed three genetic clusters of high closely related isolates (≤7 allelic differences (ADs)): cluster 1. Regarding L. monocytogenes typing, ST3 was the most prevalent among the isolates, found in 4 isolates, followed by ST9 and ST121. Some of these isolates, like ST1, ST9 and ST87, were previously associated with human clinical cases. We used Whole Genome Sequencing (WGS) alongside epidemiological data to link strains to human illnesses and potential food sources. Through cgMLST analysis, we identified genetic clusters of closely related isolates, all linked to the same producers. This approach helped us pinpoint common sources of contamination and gain insights into the transmission dynamics of L. monocytogenes in the context of food safety and public health. The escalating antibiotic resistance in Listeria species, particularly in L. monocytogenes, emphasizes the need for heightened surveillance and improved hygiene practices in the food industry to curb the spread of antibiotic resistance and ensure food safety.
- Implementation of a Nationwide Surveillance Network of Respiratory Syncytial Virus in Children < 2 years old in PortugalPublication . Melo, Aryse; Torres, Ana Rita; Lança, Miguel; Gaio, Vânia; Rodrigues, Ana Paula; Guiomar, Raquel; Bandeira, Teresa; Azevedo, Inês; VigiRSV networkHuman respiratory syncytial virus (RSV) is associated with substantial morbidity and mortality in infants, young children and elderly. Monoclonal antibodies (MAb) therapy is the available method to prevent and combat severe disease in infants, nevertheless there is a global effort in the development of vaccines and new generation of MAb. In this sense, RSV surveillance is essential to estimate the burden of RSV infection, evaluate the impact of preventive measures and to support public health decisions. Following European recommendations, a nationwide hospital-based RSV sentinel network denominated VIGIRSV was set up in Portugal. The aim of this work is to describe the implementation of VIGIRSV and report preliminary results obtained in this surveillance. VIGIRSV was implemented in 2021 with the initiative of the National Institute of Health Dr.outor Ricardo Jorge (INSA) and the Portuguese Paediatrics Society (PPS), and in 2023, 20 Hospitals collaborate in the surveillance. The surveillance is based on the recruitment of children <2 years-old hospitalized for, at least, 24 hours, that fulfill the case definition fordue to an RSVAcute Respiratory Infection (ARI). At recruitment, the paediatrician fills an epidemiological clinical questionnaire, and biological samples are collected for laboratorial diagnosis. Positive samples for RSV are forwarded to INSA for complementary virological analyses such as genetic characterization of the virus. Preliminary results from the first 2 years of surveillance are presented in table 1 and show an early RSV activity with high intensity in the 2022/23 season.. The data obtained from VIGIRSV´s results integrate the weekly published “Epidemiological surveillance bulletin of Influenza and other respiratory viruses” which can be foundavailable at https://www.insa.min-saude.pt/category/informacao-e-cultura-cientifica/publicacoes/atividade-gripal/, as well as the surveillance at European level. In addition, epidemiological and virological results are disclosed in scientific publications. The maintenance of such important surveillance is possible due to effort of distinct organizations and professionals, and has impact on the public health service.
- Epidemiology of invasive meningococcal disease in Portugal from 2012 to 2022Publication . Bettencourt, Célia; Nunes, Alexandra; VigLab-DM – Network for the Laboratory Surveillance of Meningococcal Disease; Simões, Maria JoãoBackroud and aim: Surveillance of invasive meningococcal disease (IMD) is essential to monitor changes in the epidemiology of the disease and for more effective infection control. In Portugal, a surveillance system based on mandatory clinical and laboratory notifications was implemented in late 2002, with the laboratory component under the responsibility of the National Reference Laboratory (NRL) for Neisseria meningitidis. In addition, since 2017, whole genome sequencing (WGS) has been routinely implemented at the NRL as a reference typing method for IMD surveillance. This work aims to analyze both the epidemiology of IMD and the genetic diversity of Neisseria meningitidis strains during the last 11 years (2012-2022) in Portugal. Conclusion: Although the incidence of meningococcal disease has decreased over the past 11 years in Portugal, MenB meningococci are still an important cause of meningitis and septicaemia. While serogroup C IMD was rare and restricted to adults, serogroup Y IMD affected all age groups. On the other hand, serogroup W IMD has been increasing since 2017, initially in adults and later (2019) in children under 4 years of age. WGS analysis revealed a high genetic diversity among N. meningitidis isolates, especially in the population of MenB isolates. The increased circulation of MenB cc213 strains observed over these nine years is particularly important and deserves close monitoring, given the predicted low coverage of this strain by the 4CMenB vaccine [6]. In addition, the slight increase in IMD cases due to the MenW cc11 strains, previously identified as clustered into two sublineages the “Original UK” and “UK 2013” strains [7], should also be monitored, as these strains are associated with an unusual clinical presentation and a higher lethality rate compared to other IMD serogroups [8, 9]. Our results underline the need for continuous surveillance of N. meningitidis infections susceptible to changes in their pattern, in order to promptly adapt IMD control strategies in Portugal.
- Are Food Producing animals a source of multidrug-resistant E. coli and Salmonella spp.?Publication . Pista, Angela; Silveira, Leonor; Gonçalves, Carlota; Costa, Sara; Carolino, Catarina; Santo, Bianca; Rosa, Rute; Penteado, Margarida; Alves, Margarida; Belas, Adriana; Santos, Isabel; Lima, Ana; Mota, Joana; Pedroso, Laurentina; Gomes, João Paulo; Nunes, Alexandra; Ramos, SóniaObjectives: To assess the role of food-producing animals as potential transmission vehicles of Salmonella spp. and E. coli to Humans, in order to understand the epidemiology and population structure of these zoonotic agents in Portugal.
- Characterization of Haemophilus influenzae non-invasive disease in children, in Portugal: 2015-2022Publication . Duarte, Sara; Bajanca Lavado, Maria Paula; Portuguese Group for the Study of Haemophilus influenzae Invasive InfectionBackground: Haemophilus influenzae is a Gram-negative bacterium that colonizes the human upper respiratory tract, where it can remain asymptomatic. It can also progress from colonizer to pathogen and cause acute mucosal infections, such as otitis and conjunctivitis, particularly in children. These infections are frequently associated to NTHi H. influenzae. Empirical treatment with antibiotics is of concern due to possible emergence and dissemina-tion of resistant strains. Aims / Methods: We aim to characterize H. influenzae isolates from two epidemiologically relevant non-invasive diseases, otitis media and conjunctivitis, in Portugal, from 2015 to 2022 and compare this data with results from invasive disease. From January 2015 to December 2022, 134 H. influenzae isolates (78-ear-swab; 56-eye-swab) were collected in the National Reference Laboratory for Haemophilus influenzae, based at the NIH, in Lisbon. Most isolates were from children (99.5%; 132/134). Capsular status was characterized by conventional PCR. Beta-lactamase producers were identified with nitrocefin. Antimicrobial susceptibility was determined by microdilution, according to EUCAST guidelines, for several antibiotics of interest. Genetic diversity was studied by MLST and ST assigned in PubMLST (https://pubmlst.org/organisms/haemophilus-influenzae/). Results: Among 134 H. influenzae isolates, 99.3% were NTHi (133/134), whereas only one encapsulated isolate was found, and characterized as Hia (0.8%, 1/134). Beta- lactamase producers accounted for 6.7% (9/134). Antibiotic susceptibility results (n=113) showed that most isolates were susceptible to the antibiotics tested, with the exception of 34.5% resistance to trimethoprim-sulfamethoxazole (39/113). In the course of this study, we highlight the characterization of a beta-lactamase negative, NTHi isolate, resistant to ampicillin, cefotaxime, cefuroxime, amoxicillin-clavulanic acid, cefepime, and trimethoprim/sulfamethoxazole. This is the first time that we characterized resistance to cefotaxime in H. influenzae, in our country. The isolate, from a 67 years old man with multi-microorganisms corneal ulcer, was characterized as BLNAR group III-like, ST3 (confirmed by WGS). High genetic diversity was observed among NTHi, as expected, with 22 different STs assigned for 31 isolates (71% 22/31), although ST12 and ST34 included three isolates each. When comparing the MLSTs results of isolates from both invasive and non-invasive diseases, we observed that 41% (9/22) of the STs were shared among both diseases: ST-12, ST-34, ST-142, ST-160, ST-262, ST-367, ST-396, ST-1034, and ST-1411.
- Multidrug-resistant Escherichia coli and Salmonella spp. in pigs slaughtered for human consumption, a potential source for Humans?Publication . Pista, Angela; Silveira, Leonor; Costa, Sara; Rosa, Rute; Gonçalves, Carlota; Alves, Margarida; Belas, Adriana; Santos, Isabel; Lima, Ana; Pedroso, Laurentina; Gomes, João Paulo; Ramos, Sónia; Nunes, AlexandraObjective: To assess the role of pig reservoir as potential transmission vehicle of Salmonella spp. and E. coli to humans, in order to understand the epidemiology and population structure of these zoonotic agents in Portugal.
- Raw milk cheeses from Beira Baixa, Portugal: A contributive study for the microbiological quality assessmentPublication . Mendonça, Rita; Furtado, Rosália; Coelho, Anabela; Belo Correia, Cristina; Suyarco, Elena; Pista, Ângela; Batista, RitaIn Portugal, traditional cheeses are part of the gastronomic culture, with the accessibility to products of high microbiological quality being valued to guarantee the safety of the consumer's population. These artisanal cheeses are normally made from raw milk, having a complex microbiota that will influence the characteristics, quality and shelf life of the final manufactured products. In this study we evaluated the presence of Escherichia coli, Coagulase Positive Staphylococci (CPS), Listeria monocytogenes and Salmonella spp. in 98 cured raw milk cheeses produced in the Beira Baixa region. Conventional methods (based on ISO standards) and alternative methods (TEMPO®, VIDAS® and VITEK®2) for the quantification, detection or identification of each bacterium were used. Subsequent phenotypic characterization (serotyping and antimicrobial resistance – AMR), as well as genotyping by Whole-Genome Sequencing (WGS) were also performed. The results obtained indicate the presence of E. coli, CPS, L. monocytogenes and Salmonella spp. in 64.3%, 51.0%, 4.1% and 1.0% of the samples, respectively. AMR was present in 41.5% of the E. coli isolates, from which 5.1% were multidrug resistant. In the 2 samples with CPS>4.9x104 cfu/g, staphylococcal enterotoxins (A, B, C, D, E) were not detected in 25 g. Salmonella (S. enterica enterica ser. Duisburg) was detected in one sample. Furthermore, WGS and bioinformatics analysis of L. monocytogenes isolates unveiled clusters of high closely related isolates and correlated links to cheese processing facilities. The findings of this study may indicate, among other factors, a lack of adherence to good hygiene and manufacturing practices along one or more phases of the food chain, e.g. in the milk collection/ storage, in the cheesemaking process and in the distribution, or during shelf-life whilst placed in the market. The results also highlight for the importance of the information provided by the WGS data integration, helping in public health surveillance while revealing possible infection contamination routes and epidemiological links, which contributes to the investigation of potential foodborne outbreaks.