Henrique Borges da Silva Grisard

Bivalve mollusks are recognized as bioaccumulators of enteric viruses and, therefore, represent important environmental sentinels for microbiological contamination. The contamination of these organisms is primarily associated with the exposure of coastal waters to human and agricultural effluents, which introduce pathogens into the aquatic environment. Due to their filter-feeding activity, bivalves can bioaccumulate these pathogens, and because they are often consumed raw or undercooked, contaminated bivalves may pose a public health risk. Hepatitis A virus (HAV), hepatitis E virus (HEV), and norovirus genogroups I and II (NoV GI and NoV GII) are recognized by the European Food Safety Authority and the U.S. Food and Drug Administration as major etiological agents responsible for numerous foodborne outbreaks worldwide. The absence of established regulatory limits for viral contamination in food has driven the development of molecular approaches to generate scientific evidence and support risk assessment and control strategies. In this context, the present study aimed to investigate the occurrence of HAV, HEV, NoV GI, and NoV GII in commercially available mussels (Mytilus galloprovincialis). A total of 78 pooled samples, each comprising digestive tissues from four to five mussels, were collected from retail markets between February and December 2025 and during January and February 2026 and were analyzed by reverse transcription quantitative PCR (RT-qPCR). NoV GII RNA was detected in 5 pooled samples, corresponding to an overall pool-level positivity of 6.4% (5/78; exact 95% confidence interval [CI]: 2.1–14.3%), while HAV, HEV, and NoV GI were not detected (0/78; exact 95% CI: 0.0–4.6%). RNA concentrations in positive pooled samples were low, with a mean of 143.3 ± 132.4 copies/g, a median of 120.0 copies/g, and cycle threshold (Ct) values consistent with low-level RNA detection. Overall, the findings indicate a low but detectable pool-level positivity of NoV GII in commercially available depurated mussels. The detection of viral RNA in retail mussels indicates previous exposure to viral contamination and supports the importance of continued virus-specific surveillance of commercially available bivalves.

The expansion of e-commerce and last-mile food delivery has created an increasingly relevant but understudied context for food safety surveillance. However, the occurrence of enteric viruses in ready-to-eat (RTE) foods within this distribution context remains largely unexplored. This study aimed to investigate the presence of enteric viruses in multi-component RTE salad bowls obtained through last-mile delivery. The study was designed to characterize the virological status of products received through this distribution pathway, without attributing any detected contamination specifically to the delivery process itself. A total of 30 salad samples were collected in two independent batches over a two-month interval via a third-party e-commerce platform and delivered directly to the laboratory. Virus concentration was performed using a method based on ISO 15216-1:2017, followed by RNA extraction and reverse transcription quantitative polymerase chain reaction (RT-qPCR) detection of norovirus (NoV) genogroups I (GI) and II (GII), hepatitis A virus (HAV), hepatitis E virus (HEV), human astrovirus (HAstV), and rotavirus (RV). HEV-target RT-qPCR signals were observed in 5 of 30 samples (16.7%); however, all positive signals corresponded to levels below the estimated limit of quantification (LOQ). No RT-qPCR amplification was observed for the other targeted viruses. This study provides initial occurrence data for enteric viruses in multi-component RTE meals obtained through last-mile delivery and supports the inclusion of this food category and emerging distribution pathways in foodborne virus surveillance.

Meat products represent important vehicles for foodborne viral transmission, yet standardized detection methodologies for these complex matrices remain limited. This study systematically compared three viral recovery methods across nine meat products representing chicken, pork, and processed deli meat matrices: polyethylene glycol 6000 (PEG) precipitation, glycine buffer-chloroform (GBC) phase separation, and β-mercaptoethanol (B-M) chaotropic lysis. Using Murine Norovirus 1 (MNV-1) and Murine Hepatitis Virus 3 (MHV-3) as process controls, recovery efficiency was assessed by RT-qPCR. The B-M method demonstrated the most consistent performance, successfully detecting MNV-1 across all tested matrices and significantly outperforming PEG and GBC for MNV-1 in chicken and pork. Applying the B-M method to 25 commercial retail products revealed the genetic material of Hepatitis E virus (HEV) in four samples and Porcine Circovirus type 2 (PCV-2) in one sample, exclusively within pork products. These findings provide critical comparative performance data for method selection in viral surveillance of meat and document the occurrence of HEV and PCV-2 genetic material in commercial pork products from Santa Catarina, Brazil, highlighting a significant food and agricultural biosafety concern in this major meat-producing and exporting state.

Mycoplasma genitalium is an emerging sexually transmitted pathogen with increasing antimicrobial resistance worldwide. In Brazil, molecular testing for M. genitalium and resistance-associated mutations is not routinely available, and national data remain scarce. We analyzed 118 M. genitalium-positive urethral and vaginal swab samples collected between 2018 and 2023 from sites across all five Brazilian geographic regions. PCR amplification and sequencing of 23S rRNA, parC, and gyrA identified macrolide and fluoroquinolone resistance-associated mutations. Among 79 samples successfully sequenced for 23S rRNA, 40.5% harbored macrolide resistance-associated mutations (A2058G, A2059G, or A2058T) detected in samples from all five regions at different frequencies. A total of 6.1% of sequenced samples presented parC mutations, including two with combined 23S rRNA and parC mutations. No resistance-associated mutations were detected in gyrA. While resistance-associated mutations occurred in both symptomatic men and pregnant women, these findings are not representative estimates for the Brazilian population since the study used convenience samples from two distinct surveillance populations. Nonetheless, they provide multicenter molecular data that may inform future surveillance, diagnostic strategies, antimicrobial stewardship, and resistance-guided management of M. genitalium in Brazil.

Hepatites virais entéricas causadas pelos vírus da Hepatite A (HAV) e E (HEV) se apresentam como um grande problema para a saúde pública mundial, afetando países subdesenvolvidos, em desenvolvimento e desenvolvidos. Ambas as infecções apresentam sintomatologia diversa, indo desde quadros assintomáticos até falha hepática fulminante; no entanto, seu quadro clínico mais frequente é uma hepatite aguda autolimitante. A Hepatite E pode se tornar crônica, além de apresentar um maior risco em pacientes imunossuprimidos e gestantes. Atualmente, o diagnóstico de ambas as doenças é feito por imunoensaios, mas a ascensão de testes moleculares é promissora visando a melhora na detecção dessas infecções. Enquanto as vacinas para a Hepatite A são eficazes e amplamente utilizadas, a vacina para a Hepatite E está licenciada apenas na China. Para a Hepatite E, não existe um medicamento com uso autorizado para o seu tratamento, o que, casado com a ausência de uma vacina disponível, torna-se uma ameaça alarmante, especialmente para os seus grupos de risco. Dessa maneira, o aprimoramento de testes diagnósticos, o aumento na vigilância de ambas as doenças e o desenvolvimento de tratamentos eficazes demonstram-se como importantes objetivos no enfrentamento de hepatites entéricas.

Since December 2019, the COVID-19 pandemic caused by SARS-CoV-2 has reached approximately 769 million people, leading to more than 7 million deaths worldwide. Faced with the possibility of other respiratory pathogens co-infecting patients and modifying their clinical response to SARS-CoV-2, some researchers have explored this line of investigation. The relationship between these co-infections remains unclear, underscoring the need to deepen our understanding of interactions among pathogens and between pathogens and the host. Thus, the present study employed RT-qPCR to assess the presence of Human Adenovirus (HAdV), Influenza A (Flu A), Influenza B (Flu B), Human Metapneumovirus (HMPV), Respiratory Syncytial Virus (RSV), Human Rhinovirus (HRV), and Parainfluenza Virus (PIV). Nasopharyngeal samples (187) from adult patients exhibiting respiratory symptoms were collected between February 2021 and November 2022 at the University Hospital Polydoro Ernani de São Thiago in Florianópolis, SC, Brazil. The present findings revealed that 25.16% of samples tested positive for non-SARS-CoV-2 respiratory viruses (29.8%—HRV; 5.3%—PIV; 4.3%—RSV; and 1.1%—HMPV). In the 74.84% of SARS-CoV-2-positive patients, co-infection was observed in 9.7% of patients, with 7.5% being HRV, 1.1% HAdV, and 1.1% Influenza A. Since co-infections can potentially alter patient prognoses and impact local epidemiological dynamics, this study highlights the significance of ongoing monitoring and epidemiological assessment through genomic surveillance of other clinically relevant respiratory pathogens.

The Hepatitis E virus (HEV) is responsible for causing Hepatitis E, a zoonotic disease that has emerged as a significant global health concern, accounting for about 20 million infections and 70,000 deaths annually. Although it is often recognized as a disease that is acute in low-income countries, HEV has also been recognized as a zoonotic disease in high-income countries. The zoonotic transmission requires flexible approaches to effectively monitor the virus, vectors, and reservoirs. However, the environmental monitoring of HEV presents additional challenges due to limitations in current detection methods, making it difficult to accurately assess the global prevalence of the virus. These challenges hinder efforts to fully understand the scope of the disease and to implement effective control measures. This review will explore these and other critical concerns, addressing gaps in HEV research and highlighting the need for improved strategies in the monitoring, prevention, and management of Hepatitis E using a One Health approach.

Com a crescente resistência à terapias convencionais para o tratamento de infecções ocasionadas por agentes microbianos, o uso de bacteriófagos para esse fim vem cada vez se tornando mais atrativo. O uso dessa modalidade terapêutica traz como benefício a sua alta especificidade com as espécies alvo, além de poder ser utilizada na forma de coquetéis com diversos bacteriófagos que já vêm demonstrando uma alta eficiência contra biofilmes bacterianos. Dentre o ramo da medicina, ensaios clínicos de fase 1 e 2 evidenciaram potencial terapêutico de bacteriófagos no tratamento de patologias bacterianas; enquanto na odontologia estudos in vivo indicam que para o tratamento de cáries dentárias, doenças periodontais e peri-implantares o uso de bacteriófagos também vem trazendo resultados favoráveis, apesar de estudos clínicos ainda não terem sido autorizados. No entanto para que esses bacteriófagos possam ser cogitados para o uso clínico ainda são necessários mais estudos envolvendo a segurança e padronização dos protocolos de uso dessa terapia, como a avaliação da sua farmacocinética e farmacodinâmica, explorando assim a sua absorção e seus mecanismos de ação em o nosso organismo.

Over the past four years, angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) have been extensively studied, given their important role in SARS-CoV-2 replication; however, most studies have failed to compare their behavior in the face of different SARS-CoV-2 genomic variants. Therefore, this study evaluated the influence of different variants in ACE2/TMPRSS2 expressional and genomic profiles. To achieve this, 160 nasopharyngeal samples, previously detected with SARS-CoV-2 via RT-qPCR (June 2020–July 2022), were quantified for ACE2/TMPRSS2 expression levels, also using RT-qPCR; SARS-CoV-2 genomic variants, along with polymorphisms in the ACE2/TMPRSS2 coding genes, were identified using nanopore sequencing. In order of appearance, the B.1.1.28, Zeta, Gamma, and Omicron variants were identified in this study. The ACE2 levels were higher when B.1.1.28 was present, possibly due to the ACE2/spike binding affinity; the TMPRSS2 levels were also higher in the presence of B.1.1.28, probably attributable to inefficient usage of the TMPRSS2 pathway by the other variants, as well as to the decrease in protease transcription factors when in the presence of Omicron. The rs2285666 (ACE2) polymorphism was less frequent when B.1.1.28 was present, which is befitting, since rs2285666 increases ACE2/spike binding affinity. In conclusion, SARS-CoV-2 genomic variants appear to exhibit varying impacts in regards to ACE2/TMPRSS2 genomic and expressional behavior.