Introduction: Balamuthia mandrillaris is an opportunistic free-living amoeba that causes granulomatous amoebic encephalitis, a rare but often fatal disease with delayed diagnosis. Laboratory identification is challenging because of low organism burden, slow growth, and morphological similarity to other free-living amoebae. Consequently, several laboratory techniques have been developed for detection, isolation, or cultivation, though their performance and feasibility vary across studies. This systematic review evaluates and compares these methods. Methods: Following PRISMA 2020 guidelines, a systematic search (24/09/2025) was conducted in Medline, Web of Science, ScienceDirect, and ProQuest using “Balamuthia mandrillaris” combined with “Detection”, “Isolation”, “Culture Techniques”, and “In Vitro Techniques”. Experimental and observational laboratory studies assessing detection, isolation, or cultivation methods in clinical, veterinary, or environmental samples were included. Outcomes comprised methodological performance, feasibility, and cultivation success. Results: Of 350 articles screened, 10 studies met the inclusion criteria. Molecular methods, particularly PCR, demonstrated the highest analytical performance, with sensitivities up to 84% in formalin-fixed tissues and limits of detection as low as ~1 amoeba per sample. In environmental studies, PCR showed higher detection rates than culture (e.g., 47.4% vs 18.4% for free-living amoebae). Culture-based methods enabled isolation but required prolonged incubation (up to 60 days) and showed low recovery (e.g., 4.5% in water samples). Multi-step protocols combining environmental culture, mammalian cell co-culture, and axenization improved isolation but remained technically demanding. In vitro cultivation advances demonstrated sustained growth in human fibroblast and endothelial cell lines and axenic media, achieving yields up to 1.3 × 10⁶ amoebae/mL with long-term viability. Immunological assays were useful for exposure assessment but insufficient as standalone diagnostics. Conclusions: Molecular methods provide the most sensitive detection, while culture remains essential for isolation despite low efficiency. Integrated workflows and standardized protocols are needed to improve diagnostic reliability and support future research.

Introduction

Leptospirosis is a neglected zoonotic disease with increasing global incidence, particularly in tropical urban environments. Rapid urbanization, climate change, inadequate sanitation, and expanding rodent populations have driven the re-emergence of leptospirosis, leading to recurrent outbreaks and substantial morbidity and mortality. We conducted a systematic review to synthesize current evidence on the climatic, environmental, and socioeconomic determinants of urban leptospirosis.

Methods

A systematic search of PubMed, Embase, Web of Science, and Scopus was conducted from January 2000 to December 2025 following PRISMA guidelines. Eligible studies included observational, ecological, and modeling studies evaluating environmental, climatic, and social drivers of leptospirosis transmission in urban settings. Primary outcomes were incidence, outbreak occurrence, and mortality. Secondary outcomes included environmental risk factors and socioeconomic vulnerability indicators. Qualitative synthesis and random-effects meta-analysis were performed when feasible.

Results

Forty-eight studies from 32 countries were included. Heavy rainfall, flooding events, and elevated temperatures were consistently associated with increased leptospirosis incidence, with pooled relative risks ranging from 1.6 to 3.4 following extreme precipitation events. Informal housing, poor sanitation infrastructure, high rodent density, and occupational exposure were major socioeconomic determinants. Urban slums accounted for a disproportionate burden of severe disease and mortality. Climate anomalies, including El Niño-related rainfall surges, were strongly linked to large-scale outbreaks. Integrative One Health surveillance models demonstrated improved outbreak prediction and early warning capacity.

Conclusions

Leptospirosis is rapidly re-emerging in urban tropical environments due to the synergistic effects of climate change, environmental degradation, and social vulnerability. Integrated One Health approaches combining environmental monitoring, rodent control, infrastructure investment, and climate-adaptive public health policies are urgently needed to reduce disease burden and prevent future epidemics.

Introduction

Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira, prevalent in tropical regions. Transmission is facilitated by heavy rainfall, inundation, and poor sanitation. In 2024, Rio Grande do Sul experienced widespread flooding. Therefore, this study aimed to evaluate socio-demographic distribution and temporal trend of leptospirosis in southern Brazil, with particular attention to the 2024 event.

Methods

A retrospective ecological study was conducted including all confirmed leptospirosis cases in Rio Grande do Sul from January 2014 to December 2024, obtained from TABNET/DATASUS. Analyses were performed in R (v.4.4.2) and RStudio. To compare 2024 with the 2014-2023 period, quasi-Poisson regression with population offset was applied due to overdispersion. Cases were further stratified by educational attainment as a proxy for socioeconomic position. As an ecological study, causal inferences at the individual level cannot be established.

Results

A total of 5,569 cases were analyzed. Annual incidence rates remained stable between 2 and 6 cases/100,000 inhabitants from 2014 to 2023, but increased in 2024, exceeding 10 cases per 100,000. In regression analysis adjusted for population size, incidence in 2024 was higher compared to 2014–2023 (IRR = 4.88; 95% CI: 1.90–12.54; p = 0.009). The increase was temporally associated with extreme rainfall and flooding in May 2024. Regarding educational attainment, individuals with primary education consistently represented the majority of cases throughout the decade. Still, the 2024 outbreak was characterized by a substantial increase across all educational levels compared to preceding years.

Conclusions

The increase in leptospirosis incidence in 2024 highlights its sensitivity to extreme flooding events. Although individuals with lower educational attainment accounted for a large proportion of cases, this likely reflects differences in exposure rather than intrinsic risk. Further analytical studies are needed to clarify social and environmental determinants.

The Chlamydiaceae family includes zoonotic pathogens like Chlamydia psittaci, with wild animals acting as reservoirs. As chlamydial diversity expands, surveillance in rehabilitation centers is vital to assess pathogen prevalence and mitigate potential occupational risks under a One Health approach. We analyzed 158 swabs from 105 animals—88 birds (139 swabs) and 17 reptiles (19 swabs)—admitted to CEMPAS (Botucatu, Brazil). Samples were screened using a 23S rRNA Chlamydiaceae real-time PCR. Positive samples underwent species-specific qPCRs for C. psittaci, C. avium, C. gallinacea, C. abortus, Ca. C. testudinis, C. pecorum, and Ca. C. emydidae. Chlamydiaceae DNA was detected in nine animals, resulting in an overall prevalence of 8.5% (9/105). When analyzed by group, reptiles showed a higher positivity rate of 35.3% (6/17) compared to 3.4% (3/88) in birds. C. psittaci was identified in a Zenaida auriculata and C. gallinacea in a Ramphastos toco. All PCR-positive reptile samples tested negative for the targeted species. While this may suggest uncharacterized chlamydial agents—common in chelonians—it could also reflect low DNA load or primer incompatibility; thus, sequencing is required for confirmation. Our findings for birds (3.4%) are below the 16.5% average for South American birds, while the reptile prevalence (35.3%) exceeds the global estimate of 23.5% (Inchuai et al., 2021). The identification of C. psittaci and C. gallinacea aligns with the emerging diversity reported in wild fauna (Zhang et al., 2021). These results indicate a potential occupational risk for veterinarians and staff. Although no human exposure data was collected, the presence of these pathogens highlights the need for strict biosafety protocols and personal protective equipment (PPE) to prevent spillover at the human–wildlife interface.

Coxiella burnetii is a Gram-negative bacterium capable of infecting a wide range of hosts. Even though domestic ruminants are the main reservoirs, wild animals can also play this role, being the source of some documented human outbreaks. In farm animals, it is associated with reproductive disorders, while in humans, it causes Q Fever, a febrile illness that can progress to endocarditis, vascular complications, and persistent infections. Although most cases are asymptomatic, it is essential to track infected animals, as even in the absence of clinical signs, they can shed the pathogen, facilitating transmission and posing a significant risk to human and livestock health. This study screened 182 birds from 11 distinct orders at a wildlife rehabilitation center. Oral and cloacal swabs were collected for DNA extraction and subsequent analysis by real-time PCR (qPCR) using specific primers and a probe targeting the IS1111 gene of C. burnetii. Results identified C. burnetii DNA in 1.6% (3/182) of the subjects. Positivity was confirmed in oral swabs from two Passeriforms (Pitangus sulphuratus and Gnorimopsar chopi) and a cloacal swab of an unidentified Cathatiform. In conclusion, these findings underscore that birds can participate actively in the epidemiology of C. burnetii, and the presence of bacterial DNA in swabs suggests that wild birds may contribute to environmental dissemination and, due to their high mobility and migratory patterns, may contribute to the long-distance environmental dissemination of C. burnetii, potentially infecting wild and domestic animals as well as human populations.

Background

Lymphatic Filariasis (LF) is a neglected tropical disease that affects millions of people worldwide and impacts mental, physical, and socioeconomic health globally. India carries a significant portion of this burden, with cases endemic in 21 states and union territories. The Government of India had a nationwide Mass Drug Administration/IDA (triple drug therapy) across various districts, which includes Varanasi, as part of the Global Programme to Eliminate Lymphatic Filariasis.

Objective

This study aims to assess administration, transmission, and intervention outcomes in the Varanasi district from the years 2019 to 2025. This comprises MDA/IDA coverage for the duration from 2019 to 2021 and was evaluated for its impact on LF transmission through night blood surveys and Transmission Assessment Surveys conducted in 2022. The coverage of MDA/IDA was recorded, and administration and evaluation were repeated in regions with more than 1% Mf rate.

Results

Mf rates were observed to have decreased in sentinel sites from 0.94% to 0.15% and in spot check sites from 0.46% to 0.10% from 2019 to 2021. The TAS conducted in 2022 with 19,852 eligible participants revealed an overall Mf rate of 0.1%. After the successful completion and evaluation of TAS, areas like Jaitpura (urban) and Cholapur (rural) exhibited Mf rates greater than 1% and thus continued MDA/IDA for two more years. By 2025, the prevalence of Mf in both regions was less than 1%, signifying a prolonged cessation of transmission.

Conclusion

This study shows that the MDA/IDA successfully lowered LF transmission in areas with high infection rates in the Varanasi district, but ongoing monitoring and strategic interventions are still needed to reach and maintain LF elimination goals through 2027.

Q fever is a globally distributed zoonosis caused by the obligate intracellular Gram-negative bacterium Coxiella burnetii, a highly infectious pathogen with remarkable environmental persistence. Domestic ruminants are the primary source of human infection through inhalation of contaminated aerosols. Clinical manifestations range from acute febrile illness, hepatitis, and pneumonia to severe persistent focalized complications such as endocarditis, osteomyelitis, and vascular infections. Given the limited data available in Brazil, further investigation into the circulation and shedding of C. burnetii is needed. Here, we describe a prospective cohort study evaluating the association between pre-parturition serological status and periparturient shedding of C. burnetii. For serology, an indirect ELISA (ID Screen® Q fever Indirect Multi-species) was used to detect antibodies in serum samples. Vaginal swabs were collected for DNA extraction and subsequent analysis by real-time PCR (qPCR) using specific primers and a probe targeting the IS1111 gene of C. burnetii. Sixty-five percent (67/103) of the herd was seropositive during the prepartum period, indicating prior exposure to C. burnetii. Among the 42 parturient animals, 29 were seropositive, 10 seronegative, and 1 was not serologically evaluated due to logistical constraints. Of the animals that gave birth within 2 months, 28.6% (12/42) were qPCR-positive, with shedding rates of 24.1% (7/29) and 40% (4/10) for seropositive and seronegative animals, respectively. Notably, some animals exhibited Ct values below 25, indicating high bacterial loads and suggesting increased potential for environmental contamination. Shedding in seronegative animals indicates that serology alone underestimates the risk of C. burnetii dissemination. Active circulation in a dairy herd near Americana’s urban area suggests endemic infection and ongoing zoonotic risk via windborne aerosols. Although limited by single-point sampling and total serology, these findings underscore the need for One Health surveillance to protect workers and neighboring populations at the peri-urban interface.

Rabies is a lethal viral zoonosis that requires intensive surveillance and prophylaxis efforts in several countries. Our objective was to describe the epidemiological profile of human anti-rabies consultations recorded in the database of a public regional health division of the most populous state in Brazil between 2014 and 2023, to evaluate the prophylactic measures instituted, and to estimate the direct costs associated with different post-exposure treatment regimens. This is a retrospective, descriptive study based on 54,977 notifications of human anti-rabies consultations from a region comprising approximately 68 municipalities in the State of São Paulo, analyzed using descriptive statistics. The measures were classified as adequate, insufficient, or excessive, according to the guidelines of the Brazilian Ministry of Health. Costs were estimated based on the number of treatments and the average values of the immunobiologicals administered. More than half of the prophylaxis procedures were carried out in situations considered to be low-risk. Children between 1 and 19 years of age comprise the age group with the highest number of consultations. Approximately 42.4% of the measures instituted were classified as inadequate, with a predominance of excessive measures in low-risk situations, such as indirect contact and aggressions by healthy animals. Dogs (80.1%) and cats (14.1%) cited as aggressors accounted for nearly all recorded consultations. The measures in severe exposures and those involving wild animals showed greater adherence to the guidelines of the official protocol. Treatment was discontinued in 20.6% of cases, mainly due to abandonment. The total estimated cost was approximately BRL 15.5 million, of which about 20% corresponded to inadequate measures, representing potentially avoidable expenditures. The adoption of measures more aligned with the guidelines may improve clinical effectiveness and the efficiency of public resource use.

ABSTRACT

The effort to eliminate malaria is hampered not only by the lack of effective drugs, but also by the lack of sensitive diagnostic tools to detect infections with low parasite density. As a result, more sensitive and specific high-throughput molecular diagnostic approaches are needed for accurate malaria diagnosis in order to destroy potential Plasmodium reservoirs.

In the present study, the performance of the EXP1 based RT-PCR assay was evaluated for the detection of Plasmodium falciparum infection. After obtaining ethical clearance N° 2018/09/1104/CE/CNERSH/SP from the Research Ethics Committee for Human health and Authorization of Academic Research N°810/ARA/JO5-02/SP from the Divisional officer of Esse subdivision, 134 Samples were taken in three government schools at Esse from children aged between five and fifteen suspected of having malaria for the population screening, and a parasite culture at the Malaria Research Unit of Centre Pasteur of Cameroon for analytical testing.

The detection limit of the test RT-PCR in terms of Concentration and in terms of parasitaemia is 1415×10⁻⁷ ng and 0.0012 parasites/µl respectively. For the PCR multiplex, the limit of detection in terms of concentration is 1815×10⁻⁶ ng and in terms of parasitaemia 1.2 parasites/µl. Plasmodium falciparum was detected in 125 individuals (93.28%) by RT-PCR and in 122 individuals (91.04%) by PCR multiplex. In comparison to the PCR multiplex test, the RT-PCR test has a sensitivity of 90.35%, a specificity of 5.00%, a Positive Predictive Value of 84.43% and a Negative Predictive Value of 8.33%.

These results demonstrate the importance of using the EXP1 based RT-PCR assay as a tool for the rapid diagnosis and early detection of malaria. The test uses primers specific to Plasmodium falciparum. It is reliable, simple, rapid and highly sensitive. It can be used on the field in malaria-endemic areas with limited resources.

Background: Rotavirus is a double-stranded RNA virus of the Reoviridae family and the most common cause of diarrhea in children. This study determined the prevalence and genotypes of Rotavirus in humans and animals in Calabar, Nigeria.

Methods: A total of 1000 faecal samples were collected (400 from children ≤ 5 years and 600 from animals—dogs, pigs and cattle). Samples were tested for Rotavirus using ELISA and genotyping was performed using Multiplex RT-PCR. Questionnaires were used to obtain demographic data from human participants. Data was analysed using SPSS version 25 and significant variations were determined using the Chi Square test.

Results: Of the 1,000 samples analysed, 64 tested positive, yielding an overall prevalence of 6.4%. The distribution of PCR-confirmed Rotavirus infection across species included the following: pigs (20/200, 10.0%); dogs (4/200, 2.0%); cattle (0/200, 0.0%) and humans (40/400, 10.0%). The prevalence of Rotavirus varied markedly among host species with higher rates in humans and pigs (χ² = 33.12, df = 3, p < 0.0001). Eight different Rotavirus A gene (G and P) combinations were detected in humans while only two were detected in animals. Of the 64 genotypes, 24 (37.5%) were obtained from animals (pigs and dogs) and 40 (62.5%) from humans. The predominant genotype in humans was (G10,P8) (9, 22.5%), and in animals, it was (G9,P8) (13, 65%). Meanwhile, two genotypes (G9,P8) and (P8) were common in both humans and animals. Monthly analysis showed that the highest prevalence of Rotavirus was observed in January (26.3%), while the lowest was in June (0%). There was no significant difference in the distribution of rotavirus by month (χ²=0.450; P=0.994).

Conclusion: Rotavirus A is common in both humans and animals in the study area. The presence of similar genotypes highlights the potential for zoonotic transmission to humans. There is a need to improve rotavirus vaccine coverage for children in the study area.