Emerging viral outbreaks continue to pose a persistent global health threat, underscoring the urgent need for a shift from reactive to proactive health security strategies. Viral metagenomic next-generation sequencing (mNGS) offers an unbiased, powerful approach to pathogen detection and discovery, yet its utility has been constrained by the computational complexity and slow turnaround time of data analysis during outbreak crises. The integration of artificial intelligence (AI) and mNGS is dismantling these barriers, enabling faster, more scalable outbreak response. This review synthesizes how AI-driven analytics are transforming mNGS applications, from genome assembly to sequence classification, using advanced architectures such as convolutional neural networks, recurrent neural networks, and transformers. Beyond accelerating workflows, AI’s capacity for pattern recognition outperforms traditional homology-based methods, facilitating the discovery of novel viral families and tracing hidden transmission chains through anomaly detection. Nonetheless, critical challenges remain, including limited training data, the interpretability of AI models, and resource-intensive computational demands that risk widening an “AI divide” in global health. We evaluate these obstacles and highlight forward-looking strategies, including federated learning for privacy-preserving data sharing and explainable AI for improving trust and biological insight. Looking ahead, we envision an “AI-first” paradigm for outbreak preparedness, anchored in integrated “Digital Immune Systems” for continuous, global-scale surveillance. By framing the synergy between mNGS and AI as a transformative leap, this review underscores its potential to strengthen resilience against future pandemics.

Introduction: Cholera remains a significant public health challenge in Zambia, particularly in the Copperbelt Province, where antibiotic-resistant Vibrio cholerae strains are increasingly threatening treatment efficacy. This study aimed to determine the prevalence of cholera and the antibiotic resistance patterns of V. cholerae isolates at three tertiary hospitals in the region. Methods: A retrospective cross-sectional study was conducted across three major referral hospitals in the Copperbelt Province (Arthur Davison Children’s Hospital, Kitwe Teaching Hospital, and Ndola Teaching Hospital) during the cholera outbreak from January to April 2024. Clinical samples from suspected cholera cases were analysed, and antimicrobial susceptibility testing was performed following Clinical Laboratory Standards Institute guidelines and the European committee on antimicrobial susceptibility testing methodology for Vibrio cholerae. To isolate Vibrio cholerae, alkaline peptone water and thiosulfate-citrate-bile salt-sucrose agar were utilized. The isolates were identified based on colony morphology, Gram staining, biochemical testing, and serotyping. Antimicrobial susceptibility testing was conducted by using the Kirby-Bauer disk diffusion method. Descriptive statistics were employed to assess the prevalence of Vibrio cholerae, and chi-square tests were applied with p-values of < 0.05 indicating statistical significance. Results: Of the 892 suspected cases, 334 (37.4%) were confirmed as V. cholerae through culture. The highest number of V. cholerae confirmed cases was recorded at Ndola Teaching Hospital 221 (24.8%), followed by Kitwe Teaching Hospital 88 (9.9%), while Arthur Davison Children’s Hospital 25 (2.8%) reported the lowest. High antimicrobial resistance was observed trimethoprim/sulfamethoxazole 69 (74.2%), ampicillin 75 (54.3%), and imipenem 22 (46.8%). In contrast, erythromycin 25 (100%), gentamicin 6 (85.7%) and ciprofloxacin 118 (76.6%) remained highly effective. The overall prevalence of multidrug resistance (MDR) in Vibrio cholerae was 3.7%. Among these, resistance to four or more antibiotics was observed in 3 (1.2%), followed by resistance to the combination of Ciprofloxacin, Ceftazidime, and Tetracycline in 2 (0.8%). All other MDR patterns were detected in a single isolate each (0.4%). Conclusion: The high prevalence of antibiotic-resistant Vibrio cholerae in the Copperbelt Province underscores the urgent need for enhanced antimicrobial stewardship and robust surveillance systems to inform effective cholera control strategies. Sustainable public health impact can be achieved through targeted immunization campaigns in endemic areas combined with strengthened water, sanitation, and hygiene (WASH) interventions, including improved access to clean water, adequate sanitation infrastructure, hygiene promotion, and supportive policies, which are essential for reducing transmission and preventing future outbreaks in vulnerable populations. However, because the number of antibiotics used in antimicrobial susceptibility testing across isolates varies, these results should be interpreted cautiously. Such differences may affect the comparability and overall interpretation of resistance patterns.

Background: Malaria remains a significant global health challenge, particularly in sub-Saharan Africa (SSA), where asymptomatic cases contribute to ongoing transmission and hinder elimination efforts. Asymptomatic individuals act as hidden reservoirs, sustaining onward malaria transmission. This study aimed to determine the prevalence and risk factors of asymptomatic malaria in Mwandi District, Zambia. Methods: A cross-sectional study was conducted between January to May 2024 in Mwandi District. Blood samples were collected for malaria diagnosis and simultaneously tested using rapid diagnostic tests and Giemsa-stained blood smear microscopy techniques to detect Plasmodium infections. Structured questionnaires were administered to gather demographic data and information on potential risk factors. Descriptive statistics were used to summarize the data while categorical variables were compared using the chi-square test or Fisher’s exact test. Logistic regression was used to assess associations between outcomes and independent variables, with statistical significance set at p < 0.05. Results: A total of 370 participants were enrolled in the study, with females comprising the majority (52.4%) and a median age of 24 years (IQR: 9–30). The overall prevalence of asymptomatic malaria was (4.1%), as determined by microscopy. Logistic regression analysis showed that females had significantly lower odds of asymptomatic malaria compared to males (AOR: 0.20, 95% CI 0.05–0.68; p = 0.010). Additionally, participants residing in Matoya and Sikute were more likely to have asymptomatic malaria, with adjusted odds ratios of 4.56 (95% CI 1.10–18.80; p = 0.036) and 4.72 (95% CI 1.03–21.50; p = 0.045), respectively. No significant associations were found with insecticide-treated net use, indoor residual spraying, or socioeconomic status. Conclusion: The findings highlight the need for targeted surveillance and interventions in high-risk groups and locations to curb silent transmission. Despite limitations in diagnostic sensitivity, the study underscores the importance of integrating asymptomatic malaria screening into control programmes to advance elimination efforts in Zambia and similar endemic regions.

HIV-induced apoptosis is a contradictory complicated phenomenon that occurs at the intersection of viral persistence and host defense. HIV primarily affects CD4 T cells during an infection, causing widespread immune cell death through both direct infection and indirect (bystander) mechanisms. This immunopathologic process is caused by viral proteins such as Tat, Nef, Env, and Vpr, which modify host signaling cascades such as the PI3K/Akt, p53, NF-κB, and mitochondrial pathways. Dysregulation of pro- and anti-apoptotic mediators, particularly Bax, Bcl-2, and caspase activation, which results in mitochondrial depolarization, oxidative stress, and cytochrome c release, exacerbates immune depletion. Although apoptosis serves as a host antiviral mechanism to limit viral replication and spread, HIV exploits it to evade immune surveillance and establish chronic infection. HIV pathogenesis, which includes lymphoid tissue destruction, microbial translocation, and persistent inflammation, is significantly influenced by apoptosis of both infected and bystander cells. Furthermore, alterations in death receptor signaling (Fas/FasL and TNF pathways) and mitochondrial dysfunction highlight the delicate balance between immune defense and viral manipulation. Despite considerable progress in antiretroviral therapy, immune restoration is still incomplete due to ongoing apoptotic loss and immune exhaustion. This review examines the biological mechanisms underlying HIV-induced apoptosis, evaluates the dual role of cell death in host defense versus viral persistence, and highlights novel therapeutic targets intended to restore immune homeostasis and reduce HIV-associated immunopathology.

Introduction: Monkeypox (Mpox), caused by the Mpox virus, is an emerging zoonotic disease of global public health significance. In Zambia, limited data exist on laboratory professionals’ (LPs) preparedness to manage and prevent Mpox outbreaks. This study aimed to assess knowledge, attitudes, and practices (KAP) regarding Mpox among LPs in Zambia. Methods: A cross-sectional online survey was conducted among 293 LPs across Zambia from April to August 2025. A structured, validated questionnaire assessed socio-demographic data and KAP toward Mpox. Descriptive statistics, chi-square tests, and logistic regression analyses were performed using Stata version 15. Results: Of the 293 respondents, 52.2% demonstrated good knowledge, 47.4% had a positive attitude, and 7.5% exhibited good Mpox-related practices. Significant knowledge gaps were observed, including limited awareness of Mpox transmission (62.5%), complications (76.1%). Additionally, 72% of LPs expressed willingness or support for mandatory Mpox vaccination. LPs working in health centres/clinics (AOR = 0.07; 95% CI: 0.009–0.54; p = 0.11), private hospitals (AOR = 0.05; 95% CI: 0.003–0.74; p = 0.030), public hospitals (AOR = 0.06; 95% CI: 0.011–0.39; p = 0.003), and research or academic institutions (AOR = 0.10; 95% CI: 0.01–0.77; p = 0.027) were significantly less likely to report good Mpox-related practices compared to those working in district or provincial health offices. Conclusion: The study highlights a low knowledge, a relatively low positive attitude, and poor practices toward Mpox among Zambian LPs. Identified gaps underscore the important need for targeted education and capacity-building initiatives to enhance Mpox preparedness and response.

Background: Anaemia remains a significant comorbidity among people living with HIV (PLHIV) on antiretroviral therapy (ART) in Sub-Saharan Africa (SSA), exacerbated by biological, sociocultural, and clinical factors. Despite known sex disparities, limited data exist on sex-specific determinants of anaemia in Zambia. This study evaluates anaemia prevalence, sex differences, and associated factors among PLHIV in Southern Province, Zambia. Methods: A retrospective cohort study analyzed 2,840 PLHIV aged ≥15 years from 12 districts in Southern Zambia. Data were abstracted from medical records (November–December 2024). Anaemia was defined as haemoglobin <13 g/dL (men) and <12 g/dL (non-pregnant women). Multivariable logistic regression identified factors associated with anaemia. Results: Anaemia prevalence was 15.7% (446/2,840), with marked sex disparity: females exhibited 21.8% prevalence (380/1,741) versus 6.0% in males (66/1,099). Adjusted odds of anaemia were 3.24-fold higher in females (95% CI: 1.98–5.31, p < 0.0001). Widowed individuals had the highest prevalence (20.2%). Cotrimoxazole (CTX) prophylaxis reduced anaemia likelihood (14.3% vs. 18.3%, p = 0.005). Participants on INSTI-based ART had lower anaemia prevalence (11.9%) compared to protease inhibitors (26.5%) or other regimens (30.0%). Viral load was independently associated with anaemia (AOR: 1.00, p = 0.021). Advanced WHO stages (Stage 3: 50% anaemia) and lower creatinine levels correlated with increased risk. Conclusions: Significant sex-based disparities in anaemia among Zambian PLHIV highlight the need for gender-responsive interventions, including nutritional support and INSTI-based ART. CTX prophylaxis demonstrates protective effects, advocating for broader integration into HIV care. Retrospective design and unmeasured confounders limit causal inference. Future research should prioritize longitudinal studies to refine anaemia management in high-burden settings.