Enteric protozoan parasites, such as Blastocystis sp., Balantioides coli, Cryptosporidium spp., and Giardia duodenalis, may have implications for both animal and human health.Transmitted through the fecal–oral route, these parasites cause symptoms such as diarrhea, abdominal pain, and weight loss. This study investigated the presence of these enteric protozoan parasites and genetically characterized them in hedgehogs from Portugal. A total of 110 hedgehog stool samples were collected. Molecular detection methods showed an overall occurrence of protozoa in 1.82% (2/110 95% CI: 0.22–6.41) of hedgehogs, with Blastocystis being found in one hedgehog and Cryptosporidium being found in another. No evidence for the presence of B. coli or G. duodenalis was found. This study suggests that there is a need to stay aware of hedgehogs as potential hosts of enteric protozoa. Ongoing research and surveillance efforts are recommended to explore practical prevention and control strategies. The results contribute to the limited knowledge of these parasites in Portuguese hedgehog populations and underscore their potential relevance to both veterinary and public health.

Enterocytozoon bieneusi is a microsporidia commonly found in the gastrointestinal tract of humans and a wide range of other animals, constituting a major cause of microsporidiosis in humans. Although E. bieneusi has been detected in humans, domestic, and wild animals in Portugal, and its presence in bats has been linked to zoonotic characteristics, its occurrence in bats within the country has not been reported. In this study, we investigated the presence of E. bieneusi in 380 bat fecal samples collected in mainland Portugal through a nested PCR assay targeting the internal transcribed spacer region and the flanking small and large subunit of the ribosomal RNA. Enterocytozoon bieneusi was detected in one bat sample (i.e., 0.26%; Pipistrellus pipistrellus). Additionally, another sample tested positive for Enterocytozoon sp. Phylogenetic analysis of the obtained ITS sequence of E. bieneusi revealed clustering within the potentially zoonotic Group 1. This study represents the first report of E. bieneusi in a bat from Europe. Findings presented here contribute to an enhanced understanding of E. bieneusi epidemiology. Enterocytozoon bieneusi is the most frequent cause of microsporidiosis in humans. In this study, E. bieneusi, belonging to a potentially zoonotic Group, was detected in 0.26% bat samples from Portugal, highlighting bats' potential role in transmitting this microsporidia to humans and other animals.

Porcine circovirus type 2 (PCV2) is a major viral pathogen affecting swine worldwide, causing significant economic losses through its association with porcine circovirus-associated diseases (PCVD), which can lead to reproductive failure, growth retardation, and increased mortality in affected herds. Although vaccination against the PCV2a and PCV2b genotypes has reduced clinical disease, the PCV2d genotype is increasingly prevalent in many regions. Molecular detection was performed on fecal samples from collected from a slaughterhouse using PCR targeting the ORF2 gene to detect PCV2 genotypes. Of 400 samples tested, 8.5% (17/200; 95% CI: 5.03–13.26) of samples from Portugal were positive for PCV2d, while none from Spain tested positive. This study reports the first confirmed detection of PCV2d in domestic pigs in Portugal in indexed journals, indicating circulation within the Portuguese swine population. The absence of PCV2 in Spanish samples may reflect differences in regional epidemiology or biosecurity practices. These findings highlight the importance of ongoing molecular surveillance and farm-level monitoring to inform vaccination strategies and control measures in response to evolving PCV2 genotypes. Integrated approaches should also consider wildlife reservoirs and environmental factors to improve management of PCV2 spread in domestic swine.

Enterocytozoon bieneusi, an intracellular eukaryote closely related to fungi, is recognized as a significant pathogen affecting humans, particularly those with compromised immune systems. While its transmission routes are still not fully elucidated, fecal–oral transmission remains the primary one. With a wide host range, the zoonotic potential of E. bieneusi is a concern, albeit direct evidence of animal-to-human transmission remains scarce. Genotyping based on the internal transcribed spacer (ITS) region facilitates the delineation of genetic diversity, with potentially zoonotic genotypes predominantly associated with Groups 1 and 2. Despite the broad spectrum of susceptible animal hosts, research into microsporidian infection among zoo animals remains limited. This study aimed to evaluate the occurrence of E. bieneusi infection across diverse captive animals, focusing on zoo settings in Portugal. Fecal samples were collected from a variety of animals, and molecular detection of E. bieneusi was conducted using nested PCR targeting the ITS region. Of 127 fecal samples, 1.57% (95% CI: 0.19–5.57) tested positive for E. bieneusi, with non-human primates (NHP’s) exhibiting an 18.18% (95% CI: 2.28–51.78) occurrence. Phylogenetic analysis revealed clustering within Group 2 genotypes, indicating potential zoonotic implications. This study highlights the need for further research to understand the epidemiology of E. bieneusi in zoo environments and its potential transmission pathways to humans.

Enterocytozoon bieneusi is a widespread endoparasite, commonly found in the gastrointestinal tract of humans and various vertebrate and invertebrate animals. It is the most common cause of microsporidiosis in humans, particularly among immunosuppressed individuals. Given the limited epidemiological data in Portugal, this study aimed to estimate the occurrence of E. bieneusi, perform genetic characterization, and explore its circulation dynamics in small mammal populations of Trás-os-Montes region. A total of 294 faecal samples from rodents and shrews non-invasively collected were analysed using a nested polymerase chain reaction (PCR) assay targeting the internal transcribed spacer region and flanking ribosomal RNA coding sequences of E. bieneusi. The parasite DNA was detected in two rodent samples: wood mouse (Apodemus sylvaticus) and Lusitanian pine vole (Microtus lusitanicus), corresponding to an overall occurrence of 0.68%. Our findings represent the first report of E. bieneusi in Lusitanian pine vole and the first detection of this parasite in small mammals from Portugal. Both samples belonged to the zoonotic Group 1, with one identified as genotype C. These findings provide baseline data on the distribution and host range of E. bieneusi in Portuguese wildlife and may guide future surveillance efforts within a One Health framework.

Enteric protozoa such as Balantioides coli , Giardia duodenalis , and Cryptosporidium scrofarum are of veterinary importance and may pose occupational exposure risks, yet data on their occurrence in pigs ( Sus scrofa domesticus ) raised for human consumption in the Iberian Peninsula are limited. A total of 400 stool samples were collected at a slaughterhouse in northern Portugal, originating from farms in Portugal ( n = 200) and Spain ( n = 200), and screened by PCR and qPCR. B. coli was highly prevalent in both countries (Portugal: 93%, Spain: 89.5%), with variant B predominating and variant A detected only in Portugal. C. scrofarum was detected exclusively in Portuguese pigs (4.5%), whereas G. duodenalis occurred more frequently in Portugal (8.5%) than in Spain (0.5%). These findings indicate that enteric protozoa of veterinary and potential occupational relevance are widespread in Iberian pig production, highlighting the importance of biosecurity measures for workers in contact with pigs and slaughterhouse environments.