Modeling the spatial risk of malaria through probability distribution of Anopheles maculipennis s.l. and imported cases.

AbstractMalaria remains one of the most important infectious diseases globally due to its high incidence and mortality rates. The influx of infected cases from endemic to non-endemic malaria regions like Europe has resulted in a public health concern over sporadic local outbreaks. This is facilitated by the continued presence of competent Anopheles vectors in non-endemic countries.We modeled the potential distribution of the main malaria vector across Spain using the ensemble of eight modeling techniques based on environmental parameters and the Anopheles maculipennis s.l. presence/absence data collected from 2000 to 2020. We then combined this map with the number of imported malaria cases in each municipality to detect the geographic hot spots with a higher risk of local malaria transmission.The malaria vector occurred preferentially in irrigated lands characterized by warm climate conditions and moderate annual precipitation. Some areas surrounding irrigated lands in northern Spain (e.g., Zaragoza, Logroño), mainland areas (e.g., Madrid, Toledo) and in the South (e.g., Huelva), presented a significant likelihood of A. maculipennis s.l. occurrence, with a large overlap with the presence of imported cases of malaria.While the risk of malaria re-emergence in Spain is low, it is not evenly distributed throughout the country. The four recorded local cases of mosquito-borne transmission occurred in areas with a high overlap of imported cases and mosquito presence. Integrating mosquito distribution with human incidence cases provides an effective tool for the quantification of large-scale geographic variation in transmission risk and pinpointing priority areas for targeted surveillance and prevention.

Sources of persistent organic pollutants and their physiological effects on opportunistic urban gulls.

Urbanization is associated with drastic shifts in biodiversity. While some species thrive in urban areas, the impact of inhabiting these human-altered environments on organism physiology remains understudied. We investigated how exposure to polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) affects the physiology of yellow-legged gulls (Larus michahellis) inhabiting a densely populated, industrialized city. We analyzed blood samples from 50 gulls (20 immatures and 30 adults) and assessed 27 physiological parameters and biomarkers related to xenobiotic protection, health, and feeding habits in these same individuals. We also tracked the movements of 25 gulls (15 immatures and 10 adults) to identify potential sources of persistent organic pollutants (POPs). Both adult and immature gulls primarily inhabited urban areas, followed by marine habitats. Immature gulls spent more time in freshwater, landfills, and agricultural areas. Bioaccumulated ΣPCB (median = 92.7 ng g-1 ww, 1.86-592) and ΣPBDE (median = 1.44 ng g-1 ww, 0.022-9.58) showed no significant differences between age and sex groups. Notably, immature males exhibited the highest correlations with POP concentrations, particularly with the activity of carboxylesterases (CEs), suggesting a higher sensitivity than adults. These findings highlight the potential of plasmatic CEs in immature yellow-legged gulls as effective tracers of POPs exposure and effects, offering insights into the anthropogenic impacts on urban biodiversity.

A spatial analysis of urban gulls contribution to the potential spread of zoonotic and antibiotic-resistant bacteria.

Wildlife human interactions within cities are becoming more common with consequences for pathogen transmission and human health. Large gulls are opportunistic feeders, adapted to coexist with humans in urban environments, and are potential vectors for spread and transmission of pathogens, including antimicrobial-resistant bacteria. We investigated the potential role that urban gulls play in the spread and dispersal of these bacteria. We analysed 129 faecal swabs from yellow-legged gulls (Larus michahellis) of different ages (56 adults and 73 immatures) during the breeding period from three years in the highly populated city of Barcelona (northeastern Spain). Thirteen individuals tested positive for the pathogenic bacteria (Escherichia coli, Listeria monocytogenes, Campylobacter jejuni), including antibiotic-resistant strains. We modelled the potential spatial spread of pathogens using the GPS trajectories of 58 yellow-legged gulls (23 adults, 35 immature individuals), which included the thirteen individuals that tested positive for pathogenic bacteria. By overlapping the spatially explicit pathogen dispersal maps with the distribution of urban installations sensitive at risk of possible pathogen spillover (e.g. elder and medical centres, markets, food industries, kindergartens, or public water sources), we identified potential areas at risk of pathogen spillover. Pathogens may be potentially spread to municipalities beyond Barcelona city borders. The results revealed that immature gulls dispersed pathogens over larger areas than adults (maximum dispersal distances of 167 km versus 53.2 km, respectively). Recreational urban water sources were the most sensitive habitats visited by GPS-tagged gulls that tested positive, followed by schools. Combining GPS movement data with pathogen analytics allows spatially explicit maps to be generated using a One Health approach that can help urban and public health management within large cities, such as Barcelona, and identify areas used by humans that are sensitive to pathogen spillover from gulls.

Sensitization to avian and fungal proteins in different work environments.

INTRODUCTION: Hypersensitivity pneumonitis (HP) is usually caused by the inhalation of avian and fungal proteins. The present study assesses a cohort of Urban Pest Surveillance and Control Service (UPSCS) workers with high exposure to avian and fungal antigens, in order to identify their degree of sensitization and the potential risk of developing HP. METHODS: Workers were divided according to their work activity into Nest pruners (Group 1) and Others (Group 2). All individuals underwent a medical interview, pulmonary function tests and the determination of specific IgG antibodies. Antigenic proteins of pigeon sera were analysed using two-dimensional immunoblotting. Proteins of interest were sequenced by liquid-chromatography-mass spectrometry (LC-MS). RESULTS: 101 workers were recruited (76 men, average age: 42 yrs); (Group 1 = 41, Group 2 = 60). Up to 30% of the study population exhibited increased levels of IgGs to pigeon, small parrot and parrot, and up to 60% showed high levels of Aspergillus and Penicillium IgGs. In Group 1, specific parakeet and Mucor IgGs were higher (p = 0.044 and 0.003 respectively) while DLCO/VA% were lower (p = 0.008) than in Group 2. Two-dimensional immunoblotting showed protein bands of 20-30 KDa recognized by HP patients but not by workers. LC-MS analysis identified Ig Lambda chain and Apolipoprotein A-I as candidate proteins for distinguishing HP patients from exposed workers. CONCLUSIONS: Two pigeon proteins were identified that may play a role in the development of pathological differences between HP patients and exposed workers. DLCO/VA may have a predictive value in the development of HP disease.

Decision-support tools to build climate resilience against emerging infectious diseases in Europe and beyond.

Climate change is one of several drivers of recurrent outbreaks and geographical range expansion of infectious diseases in Europe. We propose a framework for the co-production of policy-relevant indicators and decision-support tools that track past, present, and future climate-induced disease risks across hazard, exposure, and vulnerability domains at the animal, human, and environmental interface. This entails the co-development of early warning and response systems and tools to assess the costs and benefits of climate change adaptation and mitigation measures across sectors, to increase health system resilience at regional and local levels and reveal novel policy entry points and opportunities. Our approach involves multi-level engagement, innovative methodologies, and novel data streams. We take advantage of intelligence generated locally and empirically to quantify effects in areas experiencing rapid urban transformation and heterogeneous climate-induced disease threats. Our goal is to reduce the knowledge-to-action gap by developing an integrated One Health-Climate Risk framework.

Changes in the feeding ecology of an opportunistic predator inhabiting urban environments in response to COVID-19 lockdown.

Urban-dwelling species present feeding and behavioural innovation that enable them to adjust to anthropogenic food subsidies available in cities. In 2020, the SARS-CoV-2 virus outbreak resulted in unprecedented reduction in the human activity worldwide associated with the human lockdown. This situation opened an excellent opportunity to investigate the capability of urban wildlife to cope with this anthropopause event. Here, we investigated the effects of the COVID-19 lockdown on the feeding strategies of the urban yellow-legged gull (Larus michahellis) population inhabiting the highly dense city of Barcelona (NE Spain). We compared the diet of chicks (through stomach content and stable isotope analyses) sampled randomly around the city of Barcelona before (2018 and 2019), during (2020) and after (2021) the COVID-19 lockdown. The results revealed that the anthropopause associated with the lockdown had an effect on the diet of this urban-dwelling predator. The diversity of prey consumed during the lockdown was lower, and consumption of urban birds (pigeons and parakeets) and marine prey (fishery discards and natural prey) decreased during the year of lockdown. Although it was not analysed, these diet changes probably were associated with variations in the availability of these resources due to the decrease in human activity during the lockdown. These results demonstrate the trophic flexibility of urban-dwelling species to cope with the changes in the availability of human-related anthropogenic resources in urban marine ecosystems.

Gulls as potential sentinels for urban litter: combining nest and GPS-tracking information.

The production of urban waste has increased in the past decades leading to its mishandling. The effects on public health, economy, and wildlife that waste mismanagement can have are forcing governments to increase their efforts in detecting and mitigating the presence of waste. Identifying and monitoring sentinel species to assess the presence of urban litter could be a cost-effective option. Thus, analyzing the nest composition of yellow-legged gulls from an urban population inhabiting a very high populated city (Barcelona, Spain), and combining this information with accurate GPS tracking data, provides a potential tool to monitor the presence of marine and terrestrial litter over time. The results revealed the highest presence of debris in the nests of a seabird ever recorded. All the nests examined contained anthropogenic waste, with plastic items present in all of them. Crossing the nest composition with GPS tracking movements confirmed that the waste to build the nests was collected in the urban area and not in other environments surrounding the city. Then, the nest waste composition may be a good indicator of waste mismanagement and advise the municipalities to improve waste management and recycling strategies for the different types of litter. Using gulls breeding in cities as sentinel species and, in particular, the study of their nest composition, may provide essential data to decision-making stakeholders to adopt a One Health approach and help improve not only the environment's health but also the health of those who live in it.

Multiple invasions, Wolbachia and human-aided transport drive the genetic variability of Aedes albopictus in the Iberian Peninsula.

The Asian tiger mosquito, Aedes albopictus, is one of the most invasive species in the world. Native to the tropical forests of Southeast Asia, over the past 30 years it has rapidly spread throughout tropical and temperate regions of the world. Its dramatic expansion has resulted in public health concerns as a consequence of its vector competence for at least 16 viruses. Previous studies showed that Ae. albopictus spread has been facilitated by human-mediated transportation, but much remains unknown about how this has affected its genetic attributes. Here we examined the factors that contributed to shaping the current genetic constitution of Ae. albopictus in the Iberian Peninsula, where the species was first found in 2004, by combining population genetics and Bayesian modelling. We found that both mitochondrial and nuclear DNA markers showed a lack of genetic structure and the presence of worldwide dominant haplotypes, suggesting regular introductions from abroad. Mitochondrial DNA showed little genetic diversity compared to nuclear DNA, likely explained by infection with maternally transmitted bacteria of the genus Wolbachia. Multilevel models revealed that greater mosquito fluxes (estimated from commuting patterns and tiger mosquito population distribution) and spatial proximity between sampling sites were associated with lower nuclear genetic distance, suggesting that rapid short- and medium-distance dispersal is facilitated by humans through vehicular traffic. This study highlights the significant role of human transportation in shaping the genetic attributes of Ae. albopictus and promoting regional gene flow, and underscores the need for a territorially integrated surveillance across scales of this disease-carrying mosquito.

Climate Change and Health in Urban Areas with a Mediterranean Climate: A Conceptual Framework with a Social and Climate Justice Approach.

The consequences of climate change are becoming increasingly evident and highlight the important interdependence between the well-being of people and ecosystems. Although climate change is a global phenomenon, its causes and consequences vary dramatically across territories and population groups. Among settings particularly susceptible to health impacts from climate change are cities with a Mediterranean climate. Here, impacts will put additional pressure on already-stressed ecosystems and vulnerable economies and societies, increasing health inequalities. Therefore, this article presents and discusses a conceptual framework for understanding the complex relationship between climate change and health in the context of cities with Mediterranean climate from a social and climate justice approach. The different elements that integrate the conceptual framework are: (1) the determinants of climate change; (2) its environmental and social consequences; (3) its direct and indirect impacts on health; and (4) the role of mitigation and adaptation policies. The model places special emphasis on the associated social and health inequalities through (1) the recognition of the role of systems of privilege and oppression; (2) the distinction between structural and intermediate determinants of climate change at the root of health inequalities; (3) the role of individual and collective vulnerability in mediating the effects of climate change on health; and (4) the need to act from a climate justice perspective to reverse health inequities.

Widespread resistance to anticoagulant rodenticides in Mus musculus domesticus in the city of Barcelona.

Control of rodent populations is a big challenge because of the rapid evolution of resistance to commonly used rodenticides and the collateral negative impacts that these products may have on biodiversity. Second-generation anticoagulants are very efficient but different single nucleotide polymorphisms (SNPs) in the Vkorc1 gene may confer resistance in rodents. We sequenced exons 1, 2 and 3 of the Vkorc1 gene from 111 mice (Mus musculus domesticus) captured across the city of Barcelona and found SNPs associated with resistance to first- and second-generation anticoagulants in all of them. Although most of the SNPs were associated with resistance to bromadiolone, we also found SNPs associated with resistance to brodifacoum. Out of all the individuals analyzed, 94.59 % carried mutations associated to introgression events with Mus spretus, a sympatric rodent species. Currently most of the chemical products for rodent control commercialized in the area are based on bromadiolone, although recent public control campaigns have already shifted to other products. Thus, the widespread occurrence of resistant mice to bromadiolone represents a challenge for rodent control in Barcelona and may increase the risk of secondary poisoning of animals preying on this species. Public health managers, pest control companies and citizens should be aware that the use of bromadiolone based products is ineffective and represents a risk for the environment, including human and animal health.

Molecular detection of Leishmania infantum in rats and sand flies in the urban sewers of Barcelona, Spain.

BACKGROUND: Classically, dogs have been considered to be the only reservoir of leishmaniasis in urban areas. However, in a previous study, we found a 33.3% prevalence of Leishmania infantum in the spleens of Norway rats (Rattus norvegicus) sampled in the underground sewer system of the city of Barcelona (Spain). The aim of the present study was to verify, using molecular methods, the potential reservoir role of these rats in the same sewer system. METHODS: A sensitive real-time PCR (qPCR) assay, DNA sequencing and phylogenetic analysis were carried out to identify and quantify the presence of L. infantum DNA in sand fly individuals captured in the same underground sewer system of Barcelona as in our previous study and in the spleens and ears of rats captured in the same sewer system. RESULTS: Leishmania infantum DNA was found in 14 of the 27 (51.9%) sand flies identified as Phlebotomus perniciosus, and 10 of the 24 (41.7%) rats studied were infected. Leishmania infantum was found in the spleens (70%) and in the ears (40%) of the infected rats. Quantitative results revealed the presence of high loads of L. infantum in the rats studied (> 3 × 106 parasites/g ear tissue) and among the sand flies (> 34 × 106 parasites in 1 individual). CONCLUSIONS: The molecular methods used in this study demonstrated a high prevalence of L. infantum in the underground sewer populations of both R. norvegicus and P. perniciosus. These results suggest that sewer rats, in addition to dogs, are likely to act as reservoirs of leishmaniasis in cities, where sewer systems seem to offer the ideal scenario for the transmission of leishmaniasis. Therefore, to achieve the WHO 2030 target on the elimination of leishmaniasis as a public health problem successfully, an efficient control strategy against leishmaniasis in rats and sand flies should be implemented, particularly in the sewer systems of urban areas of endemic countries.

Un sistema de vigilancia de salud pública para el cambio climático en las ciudades / A public health surveillance system applied to climate change for cities

Se presenta una primera propuesta de sistema de vigilancia de salud pública aplicado al cambio climático en el contexto urbano, y el proceso que llevó a su definición. Tras varios años de seguimiento de los diversos aspectos del cambio climático y de su impacto, desde los servicios de salud pública de Barcelona se formuló una propuesta preliminar y se reunió un grupo de trabajo de personas expertas que discutieron y valoraron su contenido. Se partió de cuatro categorías de componentes: datos de clima, impactos en salud del cambio climático y sus determinantes, contribuciones de la ciudad a la mitigación (especialmente con cobeneficios para la salud) y acciones para reducir la vulnerabilidad ante eventos extremos. Se desglosaron en 12 componentes, con indicadores para cada uno. El esquema se refinó con aportaciones posteriores y se ha incorporado al trabajo de los servicios de salud pública de la ciudad implicados en este campo. (AU)

This paper makes a first proposal for a public health surveillance system for climate change in cities, and describes the process that led to its definition. After several years of monitoring different aspects related to climate change and its impact, the public health services of Barcelona made a preliminary proposal and gathered a working group of experts to discuss and review it. Four categories of components were defined: climate data, health impacts of climate change and its determinants, contributions of the city to mitigation (especially those with health co-benefits), and actions to reduce vulnerability to extreme events. They were broken in twelve components, with indicators for each. The proposal was further refined with subsequent reviews, and is being used by the city public health services involved in this field. (AU)

Effectiveness of the Modification of Sewers to Reduce the Reproduction of Culex pipiens and Aedes albopictus in Barcelona, Spain.

Mosquitoes breeding in urban sewage infrastructure are both a source of nuisance to the local population and a public health risk, given that biting mosquitoes can transmit pathogenic organisms to humans. The increasing presence of the invasive mosquito species Aedes albopictus in European cities has further exacerbated the problems already caused by native Culex pipiens. We tested the effectiveness of modifications to sewage structures as an alternative to the use of biocides to prevent mosquito breeding. The placing of a layer of concrete at the bottom of sand sewers to prevent water accumulation completely eliminated mosquito reproduction, and so eliminates the need for biocides in modified structures. Sewer modification is thus a valid low-cost alternative for mosquito control.

Gulls living in cities as overlooked seed dispersers within and outside urban environments.

The yellow-legged gull is an opportunistic and generalist bird that has colonised urban areas, where it has found very favourable trophic resources but also causes disturbance to humans and damage to infrastructure. Here, we investigated the potential role that gulls play in the dispersal of plants in Barcelona, a highly populated city of north-eastern Spain. We analysed the stomach contents of 145 chicks collected in urban nests and reported the presence of seeds of 27 plant taxa. We then developed a plant dispersal model based on the movements of 20 GPS-tracked yellow-legged gulls breeding in the city of Barcelona. We estimated seed dispersal distances, seed shadows and percentage of seeds reaching habitats suitable for seeds regurgitated in pellets and those excreted in faeces. Seven of the 27 plant taxa found in the stomachs were alien taxa to Spain. Average dispersal distances of plant seeds by gulls were around 700 m, but maximum dispersal distances reached up to 35 km. Dispersal distances and seed spatial patterns did not differ between faeces and pellet models, as most strongly depended on gull movements. About 95% of the seeds were dispersed within urban environments and between 20 and 30% reached suitable habitats for seed deposition (urban woodlands, green urban parks and urban grasslands). Urban gulls frequently dispersed seeds (including alien species) within urban habitats, both via direct consumption or via secondary dispersal after consuming granivorous birds that had ingested the seeds, such as pigeons or parakeets. Urban planning for Barcelona is based on native plant species, and thus, special attention should be paid to alien plants dispersed by birds, which could pose a risk to native biodiversity in urban ecosystems.

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases.

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named "AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108". AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species.

[A public health surveillance system applied to climate change for cities]. / Un sistema de vigilancia de salud pública para el cambio climático en las ciudades.

This paper makes a first proposal for a public health surveillance system for climate change in cities, and describes the process that led to its definition. After several years of monitoring different aspects related to climate change and its impact, the public health services of Barcelona made a preliminary proposal and gathered a working group of experts to discuss and review it. Four categories of components were defined: climate data, health impacts of climate change and its determinants, contributions of the city to mitigation (especially those with health co-benefits), and actions to reduce vulnerability to extreme events. They were broken in twelve components, with indicators for each. The proposal was further refined with subsequent reviews, and is being used by the city public health services involved in this field.

A rapid test for the environmental detection of pigeon antigen.

INTRODUCTION: Avoidance of inhaled bird antigens is essential to prevent hypersensitivity pneumonitis disease progression. The aim of the present study was to develop a sandwich enzyme link immunoassay (ELISA) and an immunochromatographic test (ICT) and compare their ability to detect pigeon antigens in environmental samples. METHODS: An amplified sandwich ELISA using pigeon serum as a calibration standard and a ICT using gold-labeled anti-pigeon serum antibodies for the rapid detection of pigeon antigens in environmental samples were developed. Twenty-two different airborne samples were collected and analysed using both methods. Strip density values obtained with ICT were calculated and compared with the concentrations determined by the ELISA method for pigeon antigens. Strips results were also visually analysed by five independent evaluators. RESULTS: The ELISA method to quantify pigeon antigen had a broader range (58.4 and 10,112.2 ng/ml), compared to the ICT assay (420 to 3360 ng/ml). A kappa index of 0.736 (p < 0.0001) was obtained between the observers evaluating the ICT strips. The results of the ELISA and the relative density of the ICT showed a highly significant correlation (rs:0.935; p < 0.0001). Bland-Altman plot also confirmed excellent agreement between the two methods (mean difference: -1.626; p < 0.0001). CONCLUSIONS: Since there was a good correlation between both assays, we can conclude that the rapid and simple ICT assay is a good and valid alternative, which does not require expensive equipment, for the validated ELISA technique.

Trends in the Epidemiology of Leishmaniasis in the City of Barcelona (1996-2019).

Background: Leishmaniasis is a neglected zoonosis produced by 20 different flagellated parasites of the Leishmania genus, a protozoan transmitted to humans and other vertebrates by the bite of dipteran insects of the Phlebotominae subfamily. It is endemic in Mediterranean countries and the number of cases is expected to increase due to climate change and migration. Prioritizing public health interventions for prevention and control is essential. The objective was to characterize the epidemiology and temporal trends in the incidence of human leishmaniasis in the city of Barcelona, between the years 1996 and 2019. Methods: A population-based, analytical observational study among residents in the city of Barcelona was conducted of all the cases of leishmaniasis reported between 1996 and 2019 to the Public Health Agency. The epidemiological survey contains clinical, diagnostic, and epidemiological data, including contact with suspicious mammals or insects. Annual incidence-rates were calculated by sex, age, and country of origin. Chi-square tests were used to assess association between studied risk factors, periods of time and type of leishmaniasis. Results: During the study period a total of 177 cases of leishmaniasis were reported in Barcelona, being 74.6% (n = 132) of the total cases in Spanish born, although within the foreign-born population the incidence was higher. Median age was 34 years (IQR = 10-48) and 121 (66.8%) were male. The main type was cutaneous (46%) followed by visceral (35.1%). The cumulative incidence was 0.47 per 100,000 inhabitants, with the highest incidence found in 2017 (1.60 per 100,000 inhabitants). A higher incidence was observed in the 0-4-year-old group (1.73 per 100,000 inhabitants), but increased during the study period for all age groups. There was an increase of foreign origin cases, and a decrease in the number of cases associated to any immunosuppression. Conclusion: In Barcelona, leishmaniasis incidence continues to be higher in people under 5 years of age, and 25-64 years old males, but it has also increased in population from foreign country of birth. There is an increase of the cases since 2016, probably due to the changes in the notification system, increasing the diagnosis of cutaneous leishmaniasis. Improvements in the current surveillance system are needed. Notification of the disease, vector, and reservoir control activities are also essential for the control of the disease.

One Health Approach to Zoonotic Parasites: Molecular Detection of Intestinal Protozoans in an Urban Population of Norway Rats, Rattus norvegicus, in Barcelona, Spain.

Rattus norvegicus, the brown or Norway rat, is the most abundant mammal after humans in urban areas, where they live in close proximity to people. Among rodent-borne diseases, the reservoir role of Norway rats of zoonotic parasites in cities has practically been ignored. Considering the parasitic diseases in the One Health approach, we intended to identify and quantify the zoonotic intestinal protozoans (ZIP) in an urban population of R. norvegicus in the city of Barcelona, Spain. We studied the presence of ZIP in 100 rats trapped in parks (n = 15) as well as in the city's sewage system (n = 85) in the winter of 2016/17. The protozoans were molecularly identified by means of a multiplex PCR (AllplexTM Gastrointestinal Panel-Parasite Assay). We also investigated the presence of co-infections among the species found. Four ZIP were identified, presenting significant prevalences in sewers, specifically Blastocystis (83.5%), Giardia duodenalis (37.7%), Cryptosporidium spp. (34.1%), and Dientamoeba fragilis (14.1%). Several co-infections among the detected ZIP were also detected. The reservoir role of ZIP that Norway rats play in cities as well as the role rats may play as sentinels of zoonotic parasites affecting humans in urban areas are strongly backed up by our findings. The increasing worldwide urbanization, climate change, and the COVID-19 pandemic are factors that are producing an increase in human-rat interactions. Our results should be considered a warning to the authorities to intensify rat control and surveillance in public health interventions.