Tropical Medicine and Infectious Disease

Background: Dyslipidemia and schistosomiasis are major public health challenges, particularly in endemic regions where their coexistence may influence host metabolism and immune responses. This study aimed to evaluate visceral adipose tissue (AT) remodeling in a murine model of acute Schistosoma mansoni infection combined with diet-induced dyslipidemia. Methodology: Female Swiss Webster mice were fed either a standard or high-fat diet (HFD) for 29 weeks and infected with S. mansoni at week 20. Nine weeks after infection, biochemical, morphometric, histopathological, and immunological analyses were performed. Results: The HFD promoted weight gain and dyslipidemia, while S. mansoni infection alone did not alter lipid profiles but partially mitigated the metabolic effects of the HFD. Morphometric analysis revealed adipocyte hypertrophy and reduced cell number in HFD-fed animals. In HFD-fed infected mice, infection partially reversed hypertrophy, suggesting a modulatory effect on AT remodeling. Histopathological examinations showed that while a HFD induced mild inflammation, infection led to intense leukocyte infiltration, hyperemia, and plasma cell degeneration. Peritoneal lavage confirmed a proinflammatory immune profile. Conclusions: These findings indicate that the interaction between a HFD and S. mansoni infection exacerbates adipose tissue inflammation and metabolic alterations, highlighting the complex interplay between parasitic infection, diet, and immune-metabolic regulation. Full article

Objective: To characterize the profiles of resistance mutations to HIV reverse transcriptase inhibitors in Gabon. Design: Cross-sectional study conducted over 37 months, from October 2019 to October 2022, at the IST/HIV/AIDS Reference Laboratory, a reference center for the biological monitoring of people living with the human immunodeficiency virus (PWHIV) in Gabon. Methods: Plasma from 666 PWHIV receiving antiretroviral treatment was collected, followed by RNA extraction, amplification, and reverse transcriptase gene sequencing. Statistical analyses were performed using Stata^® 14.0 software (USA). Results: Six hundred and sixty-six (666) PWHIV plasma collected from 252 male and 414 female patients were analyzed and 1654 mutations were detected in 388 patients, including 849 (51.3%) associated with nucleoside reverse transcriptase inhibitors (NRTIs) and 805 (48.7%) with non-nucleoside reverse transcriptase inhibitors (NNRTIs). Three of the most prescribed treatment regimens were associated to the appearance of both NRTIs and NNRTIs resistance mutations: TDF + 3TC + EFV (24.02%; 160/666); TDF + FTC + EFV) (17.2%; 114/666) and AZT + 3TC + EFV (14.6%; 97/666). Additionally, stage 3 of CD4 T-lymphocyte deficiency, the higher viral load, and treatment duration are risk factors influencing the appearance of virus mutations. Also, treatment containing TDF-3TC + DTG is more protective against mutations. Conclusions: Drug resistance mutations are common in Gabon and compromise the efficacy of ART. Further study must search for other causes of therapeutic failure in Gabon in PWHIV. Full article

Background: Since the current increase in antimicrobial resistance globally, parasitic infectious diseases have become a greater public health crisis than ever before and an absolute priority. The Egyptian community pharmacist, as a health care provider and advisor, has a potential role to play in deworming. Objective: To evaluate the outcomes of community pharmacist-led deworming-related counseling services. Methods: A prospective randomized controlled study was conducted in Damanhur, Behera, Egypt. The intervention group received community pharmacy counseling, and the control group received the usual care. Both groups were monitored for 12 months for recurrence evaluation. Results: A total of 173 patients were included, of whom 96 (55.5%) received patient counseling. The types of infection included Oxyuris (44.5%), Entamoeba histolytica (28.9%), Ascaris (8.7%), Entamoeba Cyst (8.7%), Giardiasis (4.6%), Schistosomiasis (2.9%), and pinworm (1.7%). A total of 119 participants (68.8%) reported a need for dose repetition. Personal hygiene practices were reported by 71 participants (41%). Recurrence of infection was observed in 101 cases (58.4%). Patient counseling was significantly associated with lower recurrence rates and higher rates of personal hygiene application (p < 0.001). Multivariate logistic regression analysis revealed that patient counseling and personal hygiene measures were the only significant predictors of infection recurrence rate. Conclusions: Community pharmacists’ deworming-related counseling had positive behavioral and clinical outcomes. Full article

Introduction: Despite the key role of healthcare workers (HCWs) in tuberculosis (TB) prevention and control, there is a lack of regional data on their knowledge, attitudes, and practices (KAPs) regarding the disease in Atlantic Canada. Objectives: To assess the KAPs of HCWs and identify targets for educational interventions to enhance TB care and control. Methods: A cross-sectional study was conducted among HCWs in Atlantic Canada aged $\ge$19 years from October 2023 to February 2024. Participants were recruited via multiple channels such as social media, collegiate email lists, and snowball sampling. Survey data were collected using an online platform and analyzed using IBM SPSS Statistics v29. KAPs were assessed using Likert-type scales and internal consistency was evaluated using Cronbach’s alpha. Results: A total of 157 HCWs participated in this study (age range: 19 to 69 years); most were women (n = 145, 92%), born in Canada (n = 134, 85.4%), with nearly three-quarters (n = 115, 73.2%) who had never lived outside of Canada. Study participants demonstrated moderately high knowledge (M = 29.32, SD = 3.25) and positive attitudes (M = 3.87, SD = 0.37) towards TB and strong practices (M = 4.24, SD = 0.69) in TB care; however, gaps were identified in HCW abilities to recognize less common TB symptoms (e.g., rash and nausea), as well as inconsistent practices in ventilation and pre-treatment initiation. Internal consistency analysis indicated suboptimal reliability across all three KAP domains, with Cronbach’s alpha values falling below 0.7, thwarting further planned analyses. Conclusions: This study found overall moderate-to-strong TB-related KAPs among HCWs in Atlantic Canada; however, critical gaps in knowledge and practice were noted. This new information can now guide future educational initiatives and targeted training to enhance TB preparedness and ensure equitable care for patients in the region. Full article

Coronavirus disease (COVID-19) is an infectious disease caused by the SARS-CoV-2 virus, whose 2020 outbreak was characterized as a pandemic by the World Health Organization. Restriction measures changed healthcare delivery, with telehealth providing a viable alternative throughout the pandemic. This study analyzed a telemedicine platform database with the goal of developing a diagnostic prediction model for COVID-19 patients. This is a longitudinal study of patients seen on the Conexa Saúde telemedicine platform in 2022. A multiple binary logistic regression model of controls (negative confirmation for COVID-19 or confirmation of other influenza-like illness) versus COVID-19 was developed to obtain an odds ratio (OR) and a 95% confidence interval (CI). In the final binary logistic regression model, six factors were considered significant: presence of rhinorrhea, ocular symptoms, abdominal pain, rhinosinusopathy, and wheezing/asthma and bronchospasm were more frequent in controls, thus indicating a greater chance of flu-like illnesses than COVID-19. The presence of tiredness and fatigue was three times more prevalent in COVID-19 cases (OR = 3.631; CI = 1.138–11.581; p-value = 0.029). Our findings suggest potential predictors associated with influenza-like illness and COVID-19 that may distinguish between these infections. Full article

Background: Arthropod-borne diseases primarily affect tropical and subtropical regions, exhibiting seasonal patterns that peak during hot and rainy months when conditions favor mosquito vector proliferation. Factors such as high temperatures, elevated humidity, rainfall, urbanization, and the abundance of natural and artificial breeding sites influence Aedes vector dynamics. In this context, arboviruses pose significant public health challenges, likely worsened by global warming. In Brazil, Aedes (Stegomyia) aegypti (Linnaeus, 1762) is the primary vector for yellow fever, dengue, chikungunya, and Zika. Aedes (Stegomyia) albopictus (Skuse, 1894) is an important global arbovirus vector and is considered a potential vector in Brazil. Entomological surveillance of these species often uses oviposition traps targeting immature stages. Evaluating studies that use ovitraps to collect Ae. aegypti and Ae. albopictus egg is essential for improving mosquito surveillance strategies. This study systematically reviewed peer-reviewed articles on ovitrap-based surveillance of Aedes mosquitoes in Brazil, published in Portuguese and English from 2012 to 2022. The findings suggest that ovitraps are an effective method for detecting the presence or absence of Ae. aegypti and Ae. albopictus, serving as a reliable proxy for estimating mosquito abundance in Brazilian contexts. Full article

Neisseria gonorrhoeae, the causative agent of gonorrhea, represents a major public health concern due to its increasing antimicrobial resistance. While often asymptomatic—particularly in extragenital infections—untreated cases can lead to severe complications and further transmission. Despite global efforts to monitor antimicrobial resistance, data on the molecular determinants underlying decreased susceptibility in N. gonorrhoeae remain scarce in Peru. This study aimed to detect mutations in the penA and 23S rRNA genes, which confer decreased susceptibility to cephalosporins and azithromycin resistance. We extracted DNA from 124 N. gonorrhoeae-positive clinical rectal specimens collected in Aptima Combo 2 transport tubes from MSM patients. These DNA samples were then screened using the Mismatch Amplification Mutation Assay-based real-time PCR (MAMA-qPCR) to identify mutations in the 23S rRNA and penA genes. Each sample underwent separate reactions to detect A2059G and C2611T mutations in the 23S rRNA gene, and 86 of these samples were further tested in individual qPCR assays for the penA D345 deletion (D345del) or G545S mutations. Sanger sequencing was performed on all DNA samples positive for 23S rRNA mutations by MAMA-qPCR assay, and on 27 DNA samples that yielded sufficient penA amplicons for additional sequencing. Using the MAMA-qPCR assay for the 23S rRNA gene, 64 of 124 samples amplified in the A2059G reaction: 2 (3.1%) carried the mutation, and 62 were classified as wild type. In the C2611T reaction, 42 of 124 samples amplified, and none of them carried the mutation. Using the MAMA-qPCR assay for the penA gene, we only analyzed 86 samples, as the remaining 38 samples had insufficient DNA yield. A total of 44 of the 86 samples amplified in the D345del reaction: 5 (11.4%) carried the D345del, and 39 were classified as wild type. In the G545S reaction, 4 (6.4%) carried the mutation, and 58 were classified as wild type. Finally, sequencing of the penA gene in the 27 samples revealed mutations related to decreased susceptibility to cephalosporins. This study identified genetic mutations conferring resistance to azithromycin and decreased susceptibility to cephalosporins, providing an overview of the circulating mutations conferring resistance in N. gonorrhoeae strains in Peru. Full article

Weeksella virosa (W. virosa) is a rare, non-saccharolytic Gram-negative bacterium initially described in the 1970s, later proposed as a distinct genus in 1986. The genus Weeksella currently contains two species, namely W. virosa and W. massiliensis. Although primarily considered non-pathogenic, recent evidence has linked W. virosa to a limited number of clinical infections, mostly in immunocompromised patients. This review aims to consolidate the current body of knowledge on W. virosa, encompassing its microbiological and biochemical characteristics, involvement in human and animal infections, antimicrobial susceptibility profiles, and a critical evaluation of existing diagnostic methodologies. This review includes 13 case reports detailing 16 human cases retrieved from multiple databases, highlighting diagnostic inconsistencies and a lack of standardized antimicrobial susceptibility testing. Although W. virosa is generally susceptible to most antibiotics with the exception of aminoglycosides, recent reports seem to suggest a possible emerging resistance trend. The presence of this organism in hospital environments raises concerns about its potential transmission within healthcare settings. While biochemical testing appears to offer reasonably accurate identification of W. virosa, molecular confirmation may be warranted in some cases mainly due to the organism’s rarity. The reliability of MALDI-TOF MS for the identification of W. virosa remains currently uncertain. Further studies, including electron microscopy and genome-wide analysis, are urgently needed to clarify the pathogenic potential of this bacterium and guide clinical management. This review underscores the necessity for awareness among clinicians and microbiologists regarding this underrecognized pathogen. Full article

Introduction: Leptospirosis is an under-recognized zoonosis that affects both tropical and temperate regions. While it is often associated with exposure to contaminated water or infected animals, its presentation and epidemiology in Mediterranean countries remain incompletely understood. This retrospective cohort study investigates the clinical and epidemiological profile of leptospirosis in Crete, Greece, a region where data are scarce. Methods: All adult patients with laboratory-confirmed leptospirosis admitted to three major public hospitals in Crete, Greece, between January 2019 and December 2023 were included in the analysis. Diagnosis was made through serologic testing along with compatible clinical symptoms. Results: A total of 17 patients were included. Their median age was 48 years, with a predominance of males (70.6%). Notably, more than half of the patients had no documented exposure to classic risk factors such as rodents or standing water. Clinical presentations were varied but commonly included fever, fatigue, acute kidney injury, and jaundice. Of the patients who underwent imaging, most showed hepatomegaly. The median delay from symptom onset to diagnosis was 11 days, underscoring the diagnostic challenge in non-endemic areas. Ceftriaxone was the most frequently administered antibiotic (76.5%), often in combination with tetracyclines or quinolones. Despite treatment, three patients (17.6%) died, all presenting with severe manifestations such as ARDS, liver failure, or shock. A concerning increase in cases was noted in 2023. Conclusions: Leptospirosis can present with severe and potentially fatal outcomes even in previously healthy individuals and in regions not traditionally considered endemic. The relatively high mortality and disease frequency noted emphasize the importance of maintaining a high index of suspicion. Timely diagnosis and appropriate antimicrobial therapy are essential to improving patient outcomes. Additionally, the need for enhanced public health awareness, diagnostic capacity, and possibly environmental surveillance to control this neglected but impactful disease better, should be emphasized. Full article

Background: Scrub typhus (ST) is caused by Orientia tsutsugamushi (OT) infection, which is transmitted to humans through the bites of infected chiggers. The clinical presentations range from mild to life-threatening multi-organ dysfunction. This report describes a cluster of ST cases involving five oil palm estate workers in Pekan district, Pahang, Malaysia. Methods: The clinical history, laboratory, and entomological investigation were conducted on the patients, including the index case and four suspected cases in the cluster. Polymerase chain reaction (PCR) tests for OT and genotyping were performed on the patients’ blood and urine samples. Serological testing by indirect immunoperoxidase (IIP) test against Rickettsial diseases was also conducted. Principal Findings: Patients presented with fever, myalgia, headache, rash, cough, and eschar. The index case developed severe ST complicated by acute kidney injury (AKI) and respiratory distress, requiring intubation and ventilation at the intensive care unit of a tertiary hospital. ST was confirmed through PCR analysis of a urine sample, showcasing a novel diagnostic approach. The other four cases were confirmed by a four-fold rise in immunoglobulin G (IgG) antibody titers. Conclusions: oil palm estate workers are at high risk for chigger exposure in Malaysia. Awareness among clinicians and the public of ST is crucial for effective prevention, accurate diagnosis, and optimal management. Full article

Background: While efforts have been made to control meningococcal disease or carriage during mass gatherings (MGs), it is still a significant problem. This meta-analysis aims to assess the prevalence and predictors of meningitis carriage during MGs and travel. Methodology: PubMed, Scopus, Embase, and Cochrane were searched from their conception to January 2025. Cohort and cross-sectional studies assessing the prevalence of meningitis carriage and its serotype related to MGs and/or travel, and risk factors associated with its spread, were considered. The Newcastle–Ottawa scale was used for the quality assessment of studies. Results: Out of 1301 studies, 25 were considered for this meta-analysis. The largest geographic area involved was Saudi Arabia. A meta-analysis of 24 studies identified a pooled prevalence rate of meningococcal disease or carriage of 15.9% (95%CI: 4.45–27.4%) and the most frequent infecting organisms to be Serotype C (13.9%; 95%CI: −14.7 to 42.5; 4 studies) and A (11.5%; 95%CI: −2.13 to 25.2; 9 studies) among those at MGs or traveling. Age, gender, smoking history, and the vaccination status did not affect the infection risk. Conclusions: There is an increased prevalence of meningococcal disease and carriage, especially Serogroups A and C, associated with MGs and travel. New interventions and methodologies should be undertaken to control and prevent meningococcal disease or carriage transmission during such events. Full article

Extra-pulmonary tuberculosis (EPTB) can affect any organ of the body, producing a wide variety of clinical manifestations that make the diagnosis and treatment of EPTB challenging. The optimum treatment varies depending on the site of EPTB, its severity, and response to treatment. There is often uncertainty about the best management practices, with a significant departure from national guidelines. This study aims to identify gaps and barriers in adhering to the national guidelines for the diagnosis and treatment of EPTB. We included 433 patients having EPTB and followed up at predefined intervals of 2 months, 6 months, 9 months, and 12 months. Questionnaire-based interviews of the treating physician and the patients in different departments were conducted. For confirmatory diagnosis, heavy dependence on clinical-radiological diagnosis without microbiological support was observed, which is a deviation from National Tuberculosis Elimination Programme (NTEP) guidelines and raises concerns about the potential for misdiagnosis and overtreatment. Apart from patient delays, long health system delays in EPTB were observed. The median patient delay, health system delay, and total treatment delay times were 4.2, 4, and 10.1 weeks, respectively. To enhance EPTB diagnosis and management, there is a pressing need for improved access to microbiological testing, enhanced physician training on adherence to NTEP guidelines, and greater utilisation of imaging and histopathological techniques. Full article

Background: Rickettsioses are diseases caused by obligate intracellular non-motile coccobacilli transmitted via arthropods. The most common rickettsioses are scrub typhus (ST), typhus group rickettsioses (TGR), and spotted fever group rickettsioses (SFGR). This study aims to provide information and insight into rickettsioses seropositivity among suspected patients in East and Peninsular Malaysia over a six-year period from 2016 to 2021. Methodology/Principal Findings: Data obtained from four state hospitals and one national research institute providing rickettsial serological testing were analyzed using the IBM SPSS (Statistical Package for the Social Sciences) software program. The six-year analysis revealed that ST had the highest number of seropositivity cases, followed by TGR, and SFGR, for both immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies. Of the 3228 samples, 21.6%, 16.1%, and 13.9% of suspected patients were IgM seropositive for ST, TGR, and SFGR, respectively. IgG seropositivity for ST was 21.9%, followed by TGR at 21.4%, and SFGR at 17.2% among suspected rickettsioses cases. All regions in Malaysia were significantly associated with IgM seropositivity for ST, TGR, and SFGR. IgM seropositivity for SFGR was significantly higher in females. Age group 41–65 years was highly associated with IgG seropositivity for ST, TGR, and SFGR. Conclusions/Significance: Analysis of six-year data on ST, TGR, and SFGR seropositivity in Malaysia revealed variations across regions, age groups, and genders. This seropositivity study underscores ST, TGR, and SFGR as possible causes of acute febrile illness among patients suspected of rickettsial disease in Malaysia. The findings contributed to the awareness of reemerging rickettsioses and warrant public health interventions that may reduce the incidence of rickettsioses in Malaysia. Abstract summary: Scrub typhus (ST), typhus group rickettsioses (TGR), and spotted fever group rickettsioses (SFGR) are significant global public health concerns. Our results showed that the highest number of IgM and IgG seropositivity cases was observed for ST, followed by TGR and SFGR. All regions in Malaysia were significantly associated with IgM seropositivity for ST, TGR, and SFGR. East Malaysia exhibited significantly higher seropositivity for ST, TGR, and SFGR than other regions in Malaysia. IgM seropositivity for SFGR was significantly higher in females. The age group 41–65 years was highly associated with IgG seropositivity for ST, TGR, and SFGR. This study highlights the value of serological data in uncovering the hidden burden of disease in Malaysia. In addition, the findings contributed to bridging knowledge gaps on the limited data from Malaysia spanning extended periods, despite being one of the countries in the endemic Tsutsugamushi Triangle. The findings from this study may direct future research on rickettsioses and warrant public health interventions in Malaysia. Full article

(1) Background: Murine typhus, caused by Rickettsia typhi, is a neglected rickettsial disease and an underdiagnosed cause of acute febrile illness (AFI), particularly in endemic regions such as Indonesia. (2) Case description: We report a case series of four patients presenting with AFI of less than seven days in duration. Three patients were admitted with moderate disease, while one presented with septic shock with the macrophage activation-like syndrome (MALS) phenotype. Common clinical features included myalgia and headache; additional symptoms included cough, sore throat, and abdominal pain. Laboratory findings revealed bicytopenia, elevated transaminases, and raised inflammatory and bacterial infection markers. Common tropical infections—dengue, typhoid fever, and leptospirosis—and other potential sources of infection were excluded early during hospitalization. Diagnosis was confirmed by nucleic acid amplification testing (NAAT), which detected R. typhi in all patients. Doxycycline was initiated following confirmation, leading to defervescence within 36–48 h. (3) Conclusions: Murine typhus remains an underrecognized cause of febrile illness in Indonesia. In the near future, the inclusion of rickettsial testing in the diagnostic protocol of AFI will be crucial, as it enables timely administration of effective, low-cost treatment. Full article

Localised cutaneous leishmaniasis (LCL) is a common neglected tropical disease in Ethiopia, which is mainly treated with intralesional (IL) pentavalent antimonial such as sodium stibogluconate (SSG) and/or cryotherapy. Both treatments are painful, and studies are lacking on the pain associated with these or affected individuals’ experiences of them. A cross-sectional, observational study was conducted at ALERT Comprehensive Specialized Hospital, Addis Ababa/Ethiopia. The socio-demographic and clinical data of individuals affected by LCL receiving IL SSG and/or cryotherapy was gathered, and their treatment was observed. Participants quantified their treatment-associated pain using the Wong–Baker Pain Scale. Health-related quality of life was measured using the (Children’s) Dermatology Life Quality Index. Adverse effects, participant experiences with local therapies, and dermatologists’ experiences and opinions of local LCL treatment were assessed using structured questionnaires. Of the thirty-six individuals with LCL included (64% male, 14% children), 52% reported a treatment-associated pain score ≥ 8. Cryotherapy administered with a cotton bud was associated with lower pain scores ≤ 6 (odds ratio: 0.15, 95% confidence interval: 0.03–0.89) compared to a cryotherapy spray device. There was wide variation in treatment administration. Local LCL treatment is painful, and most individuals experience significant pain. This study highlights the need for less painful but effective treatments, structured training, and clear standard operating procedures. Full article

Urogenital schistosomiasis, caused by Schistosoma haematobium and transmitted by Bulinus snails, affects approximately 190 million individuals globally and remains a major public health concern. Effective surveillance of snail vectors is critical for disease control, but traditional identification methods are time-intensive and require specialized expertise. Environmental DNA (eDNA) detection using qPCR has emerged as a promising alternative for large-scale vector surveillance. To prevent eDNA degradation, benzalkonium chloride (BAC) has been proposed as a preservative, though its efficacy with schistosomiasis snail vectors has not been evaluated. This study tested the impact of BAC (0.01%) on the stability of Bulinus truncatus eDNA under simulated field conditions. Water samples from aquaria with varying snail densities (0.5–30 snails/L) were stored up to 42 days with BAC. eDNA detection via qPCR and multivariable linear mixed regression analysis revealed that BAC enhanced eDNA stability. eDNA was detectable up to 42 days in samples with ≥1 snail/L and up to 35 days at 0.5 snails/L. Additionally, a positive correlation between snail density and eDNA concentration was observed. These findings support the development of robust eDNA sampling protocols for field surveillance, enabling effective monitoring in remote areas and potentially distinguishing between low- and high-risk schistosomiasis transmission zones. Full article

Anticytokine autoantibodies (AAbs), particularly anti-interferon-gamma (anti-IFN-γ) AAbs, disrupt cytokine functions, leading to infections, autoimmune-like diseases, and conditions resembling interleukin-12 (IL-12)/IFN-γ pathway defects. Advances in genetic testing have clarified overlaps between autoinflammatory, autoimmune disorders, and primary immunodeficiencies but reveal complex phenotypes and pathways. While these insights deepen our understanding of immune mechanisms, they also complicate diagnosis and treatment, with limited options for IFN-γ deficiencies caused by genetic mutations. The adult-onset immunodeficiency with disseminated lymphadenitis due to nontuberculous mycobacteria (NTM) and other opportunistic infections has been linked to high levels of anti-IFN-γ AAbs. This syndrome, initially identified in HIV-negative Asian patients, frequently affects individuals of Asian descent and may be associated with specific human leukocyte antigen (HLA) alleles. The presence of neutralizing anti-IFN-γ AAbs impairs the IFN-γ-dependent immune response, likely contributing to the persistent NTM infection. This study underscores the potential for late-onset anti-IFN-γ AAb syndrome to manifest with disseminated NTM (dNTM) infections, highlights the importance of timely diagnosis and considers rituximab as a potential therapeutic option. Full article

Persistent high-risk human papillomavirus (HR-HPV) infection is closely associated with the development of cervical intraepithelial neoplasia (CIN) and cervical cancer. In recent years, the potential impact of viral co-infections on this process has also been investigated. This study investigated the presence of HR-HPV, HSV-1/2, and HHV-8 DNA in formalin-fixed paraffin-embedded (FFPE) cervical biopsy samples, as well as their association with lesion severity. A total of 276 FFPE cervical tissue samples were evaluated. Viral DNA was detected by real-time PCR. The samples were histopathologically classified as normal/non-dysplastic, low-grade (LSIL), and high-grade (HSIL) lesions. HR-HPV DNA was detected in 112 samples (40.6%), with the highest prevalence observed in the 30–39 age group (51.2%). Among the HPV-positive cases, 46.5% (52/112) had single-type infections, 32.1% (36/112) had multiple-type infections, and 21.4% (24/112) were untypable. Together, these categories accounted for all HPV-positive samples. The most common genotype was HPV-16 (16.7%). HHV-8 and HSV-2 DNA were not detected. HSV-1 DNA was detected in only three non-dysplastic, HPV-negative cervical samples. In conclusion, HR-HPV DNA was detected in 40.6% of cervical biopsy samples and showed a significant association with increasing histological severity, highlighting its critical role in the progression of cervical lesions. Although the absence of HHV-8 and HSV-2 suggests a limited contribution of these viruses to cervical disease, the use of a single real-time PCR assay limits the ability to draw generalized conclusions regarding their clinical relevance. Further large-scale, multicenter studies employing both tissue-based and serological approaches are needed to validate these findings and to better understand the dynamics of viral co-infections in cervical disease. Full article