Mean Viral Load Research Articles

Wastewater-Based Surveillance Reveals the Effectiveness of the First COVID-19 Vaccination Campaigns in Assisted Living Facilities.

The COVID-19 pandemic has disproportionately affected vulnerable populations, including residents of assisted living facilities (ALFs). This study investigates the impact of non-pharmaceutical interventions (NPIs) and mass vaccination campaigns on SARS-CoV-2 transmission dynamics within four ALFs in Maricopa County, Arizona, United States from January to April 2021. Initial observations reveal a significant SARS-CoV-2 prevalence in Maricopa County, with 7452 new COVID-19 cases reported on 4 January 2021. Wastewater surveillance indicates elevated viral loads within ALFs with peak concentrations reaching 1.35 × 107 genome copies/L at Facility 1 and 4.68 × 105 copies/L at Facility 2. The implementation of NPIs, including isolation protocols, resulted in a rapid decline in viral loads in wastewater. Following mass vaccination campaigns, viral loads reduced across all facilities, except Facility 4. Facility 1 demonstrated a mean viral load decrease from 1.65 × 106 copies/L to 1.04 × 103 copies/L post-vaccination, with a statistically significant U-statistic of 28.0 (p-value = 0.0027). Similar trends are observed in Facilities 2 and 3, albeit with varying degrees of statistical significance. In conclusion, this study provides evidence supporting the role of NPIs and vaccination campaigns in controlling SARS-CoV-2 transmission within ALFs.

Efficacy and safety of two fixed doses of ibalizumab plus optimized background regimen in treatment-experienced HIV-positive individuals.

Sustained viral suppression in patients with multidrug-resistant (MDR) human immunodeficiency virus (HIV) infection remains difficult; accordingly, agents targeting different steps in the HIV life cycle are needed. Ibalizumab, a humanized immunoglobulin G4 monoclonal antibody, is a cluster of differentiation (CD4)-directed post-attachment inhibitor. In this Phase IIb study, 113 individuals with MDR HIV-1 and limited treatment options were assigned an optimized background regimen (OBR) and randomized to either 800 mg ibalizumab every two weeks (q2wk; n=59) or 2,000 mg ibalizumab every four weeks (q4wk; n=54) up to Week 24. Viral loads (VL) below the detection limit were achieved in 44% and 28% of patients in the 800 mg q2wk and 2,000 mg q4wk groups, respectively, at Week 24. Mean (standard deviation) VL (log10 copies/mL) decreased from Baseline (4.6(0.8), 800 mg q2wk; 4.7(0.7), 2,000 mg q4wk) to Week 2, with the reduction maintained through Week 24 (2.9(1.5), 800 mg q2wk; 3.2(1.4), 2,000 mg q4wk). Baseline CD4+ counts were 80.5 and 54.0 cells/μL in the 800 mg q2wk and 2,000 mg q4wk groups, respectively. Mean CD4+ T-cell count was increased at Week 24 in both groups. No serious adverse events were related to ibalizumab. In heavily treatment-experienced patients with HIV (PWH) at a more advanced baseline disease severity, clinically significant response rates at Week 24 were achieved with ibalizumab plus OBR. Ibalizumab's unique mechanism of action and lack of cross-resistance to other antiretroviral agents make it an important component of combination treatment regimens for PWH with limited treatment options.

Clinical-Laboratory Outcome of Cryptococcus sp. Multifocal Choroiditis in Acquired Immunodeficiency Syndrome Patients

ABSTRACT Purpose This study aims to describe the clinical and laboratory outcomes of patients with Acquired Immunodeficiency Syndrome (AIDS) presenting multifocal choroiditis secondary to Cryptococcus sp. infection. Methods We reviewed the clinical records of AIDS patients diagnosed with cryptococcal multifocal choroiditis at the Laboratory of Infectious Disease in Ophthalmology, Oswaldo Cruz Foundation, from January 2022 to March 2024. Patients meeting the criteria of typical fundus appearance associated with disseminated cryptococcosis laboratory diagnosis were included. Data collected included age, gender, symptoms, plasma HIV viral load, serum T-CD4+ cell count (cells/mm3), initial visual acuity (VA), laboratory methods for cryptococcosis diagnosis, treatment, and final VA. All patients underwent multimodal evaluation including spectral domain optical coherence tomography, fluorescein angiography (FA), and indocyanine green angiography (ICG). Results Six patients were evaluated, comprising five males (83%) and one female (17%), with a mean age of 36.5 years (standard deviation, SD: 10,46). Five patients (83%) presented with meningeal syndrome. Mean HIV plasma viral load was 262.959 copies/mm3 (SD: 209.469), and the mean serum T-CD4 count was 13.33 cells/mL (SD: 14,63). All patients tested positive for serum cryptococcal antigen (CRAG). Four patients (67%) had positive cerebrospinal fluid (CSF) culture and CRAG. Treatment included intravenous Amphotericin B and oral Fluconazole. Among twelve eyes (100%), four (33%) achieved visual recovery, while eight (67%) maintained initial VA. Lesions were more numerous and distinct on ICG compared to fundus examination and FA. Conclusion This study underscores the importance of clinical, laboratory, and multimodal evaluations in AIDS patients presenting with cryptococcal multifocal choroiditis.

The Prevalence of Chronic Hepatitis B Infection and Associated Factors Among Pregnant Women in Southeast of Iran

Background: Hepatitis B virus (HBV) infection is a significant global public health issue. Although infant immunoprophylaxis effectively reduces viral transmission, mother-to-child transmission during pregnancy, often associated with intrauterine infection, remains a major route of HBV infection. Objectives: The aim of this study was to determine the prevalence of maternal HBV infection, identify its associated factors, and measure viral load in Hepatitis B surface antigen (HBsAg)-positive pregnant women in Zahedan, Iran. Methods: This cross-sectional study included data from pregnant women who were referred to the Central Laboratory of Zahedan, Iran, for routine pregnancy tests between August and December 2019. Data were collected during the first prenatal visit along with HBsAg testing. Hepatitis B surface antigen testing was repeated for HBsAg-positive women at 24 - 28 weeks of gestation. If the patient remained HBsAg-positive, serum viral load was determined. Factors associated with HBV infection were identified using multiple logistic regression models. Results: A total of 1,474 pregnant women participated in this study. Among them, 24 (1.6%, 95% CI: 0.9% - 2.2%) were HBsAg-positive, with 17 (71%) of these patients being unaware of their infection. The mean viral load in infected women was (11.1 ± 3.2) × 107 IU/mL. Only 13.3% of pregnant women were vaccinated against HBV. Maternal tobacco use, family history of HBV infection, previous history of HBV infection, and history of nonalcoholic fatty liver disease were significantly associated with HBV infection (P < 0.05). Conclusions: The prevalence of HBV infection remains high among pregnant women, and vertical transmission poses a major challenge in the elimination of HBV infection in southeastern Iran. In addition to active-passive immunoprophylaxis, universal screening of all pregnant women for HBV infection and the use of peripartum antiviral therapy in HBsAg-positive pregnant women with high viral loads are warranted.

Long-term outcomes of congenital cytomegalovirus infection in children early identified by extended hearing-targeted screening

ObjectivesCongenital Cytomegalovirus (cCMV) has been associated with hearing, vision, and neurodevelopmental long-term sequelae. Despite the social burden associated with the disease, a universally accepted consensus on screening, diagnostic, therapeutic and follow-up approaches has not been reached. The present observational retrospective study aims at describing long-term sequelae and radiological abnormalities associated with cCMV in children early identified by extended hearing-targeted screening and evaluated by audiological follow-up in a single III Level Audiological Referral Center for at least 2 years. MethodsAudiological neonatal and follow-up data were available for all subjects. Data collection included clinical neonatal and virological assessment at birth. Ophthalmological, neurodevelopmental and neuroradiological follow-up abnormalities compatible with cCMV sequelae were collected by clinical reports. Spearman's rank correlation coefficient (rho-ρ) was used to evaluate possible correlations among the considered parameters. Results61 newborns were identified by extended hearing-targeted cCMV screening and diagnosed mostly (83.6 %) by PCR viral DNA extraction in urine collected within the 15° day of life. Seventeen babies were born preterm, with a mean gestational age of 33.5 weeks. Sixteen patients (26.2 %) were admitted to an Intensive or sub-Intensive Neonatal Care Unit. At birth, 35 newborns were symptomatic (57.3 %), and 19 of them received antiviral treatment by valganciclovir or ganciclovir. Overall, 20 children (32.7 %) were diagnosed with sensorineural hearing loss (SNHL), among them 17 (85 %) were refer at the newborn hearing screening while 3 (15 %) were Pass. 5/20 children (25 %) presented isolated SNHL, while in 15/20 (75 %) children SNHL was associated to other long-term sequelae. In 5 patients (25 %) a progression of the hearing threshold was observed, with a mean age of progression of 26 months of age. Risk factors for progression were a worse final hearing threshold (Spearman's ρ = 0.434; p = 0.0001) and a worse hearing threshold at birth (Spearman's ρ = 0.298; p = 0.020). Thirteen children were fitted with hearing aids, 8 of whom subsequently underwent cochlear implantation. Concerning long term impairments, 10/61 children (17 %) presented a variety of ophthalmological sequelae, while 16/40 cCMV patients (40 %) were diagnosed with neurodevelopmental abnormalities. Language delays were significantly associated with a worse hearing threshold (ρ = 0.582; p = 0.0001) and with other neurocognitive abnormalities (ρ = 0.677, p = 0.0001). 30 children underwent radiological brain evaluation by Magnetic Resonance Imaging, and 63.3 % of them presented abnormalities compatible with cCMV. Mean viral load at birth did not show significant associations with long-term sequelae. ConclusionsThe study highlights the diverse and significant long-term sequelae of cCMV infection detected through early screening. With a significant proportion of cCMV children developing sensorineural hearing loss, ophthalmological and neurodevelopmental issues, the results emphasize the importance of continuous, multidisciplinary follow-up. Early identification and tailored interventions are crucial for improving the long-term health and quality of life of children affected by cCMV.

Prediction of differential Gag versus Env responses to a mosaic HIV-1 vaccine regimen by HLA class I alleles.

HLA class I variation has the strongest effect genome-wide on outcome after HIV infection, and as such, an understanding of the impact of HLA polymorphism on response to HIV vaccination may inform vaccine design. We sought HLA associations with HIV-directed immunogenicity in the phase 1/2a APPROACH vaccine trial, which tested vaccine regimens containing mosaic inserts in Ad26 and MVA vectors, with or without a trimeric gp140 protein. While there were no HLA allelic associations with the overall cellular immune response to the vaccine assessed by ELISpot (Gag, Pol, and Env combined), significant associations with differential response to Gag compared to Env antigens were observed. Notably, HLA class I alleles known to associate with disease susceptibility in HIV natural history cohorts are associated with stronger Env-directed responses, whereas protective alleles are associated with stronger Gag-directed responses. Mean viral loads determined for each HLA allele in untreated individuals correlated negatively with the strength of the Gag response minus the Env response in Black vaccinees based on both ELISpot and CD8+ T cell ICS responses. As the association of T cell responses to conserved Gag epitopes with lower viral load in untreated individuals is well established, our data raise the possibility that the Ad26.Mos.HIV vaccine may induce more effective cellular responses in those with HLA alleles that confer improved virologic control in untreated HIV infection.IMPORTANCENo vaccine tested to date has shown sufficient efficacy against HIV infection. A vaccine that induces robust responses in one individual may fail to do so in another individual due to variation in HLA class I genes, loci central to the immune response. Extensive data have shown the strong effect of HLA variation on outcome after HIV infection, but very little is known about the effect of such variation on HIV vaccine success. Here, we identify a link between the effect of HLA variation on HIV disease outcome and immune responses to an HIV vaccine. HLA variants associated with better HIV control after infection also induce stronger responses against the HIV Gag protein relative to the Env protein after vaccination. Given the virologic control conferred by responses to Gag in natural history of HIV infection, these data suggest that HLA alleles conferring protection after HIV infection may also support a more effective cellular response to HIV vaccination.

Co-relation of hepatitis C RNA load with antiviral therapy and risk factors among hepatitis C seropositive patients

Hepatitis C virus is a bloodborne virus and the major cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma. Viral load is the prognostic marker of the disease progression. The aim of this study is to determine the correlation of hepatitis C RNA load with antiviral therapy and its risk factors among the HCV seropositive patients. This study will be helpful in early assessment of the disease progression and its complications.The blood samples were collected over a period of one year from April 2022-March 2023. The serum was subjected to ELISA for Anti HCV Ab. Viral load quantification was done by MylabPathoDetect HCV Quantitative PCR Kit in HCV seropositive patients.About 18,882 patients were tested for HCV infection over a period of one year. 75 patients were positive for HCV infection. Prevalence of HCV was 0.39. The mean viral load was reduced from 3.08×10 IU/ml to 1.98×10 IU/ml and Sustained Viral Response (SVR12) was achieved in 63(84%) patients after effective treatment with direct acting antiviral agents. High risk sexual behavior was the most common risk factor observed in seropositive patients.The current study determines the reduction in mean viral load and achieving sustained viral response after the effective antiviral therapy at the earliest, which is helpful in prevention of disease progression and its complications.

Differential viral load of surgical masks worn by patients infected with respiratory viruses

Background: This study investigated viral load in surgical masks worn by adult patients infected with respiratory viruses. Method: Surgical masks were dissected into eight pieces at pre-selected sites: inner, middle, and outer layers at the nose (N1-N3) and mouth (M1-M3), as well as full-thickness on the right (RS) and left (LS) sides. Viral load was detected, correlated with nasopharyngeal specimens and patients' demographics. Result: Among 230 patients infected with influenza A virus (n=91), respiratory syncytial virus (RSV) (n=61), and SARS-CoV-2 (n=78) from April 1 to August 31, 2023, 90.9% (209/230) were from the medical specialty. Of the 230 surgical masks collected, viral RNA was detected in 79.6% at one or more sites, with 75.7% positive at N1 or M1, 55.2% positive at N3 or M3, and 22.6% exhibiting viral RNA at all sites. Pearson correlation showed viral load correlation between nasopharyngeal specimens and N1 (0.244, p=0.002) and M1 (0.174, p=0.031). The mean viral load at N1 (4.14 ± 1.46 log10 copies/ml) was significantly higher than M1 (3.74 ± 1.32 log10 copies/ml, p=0.014) and N3 (3.58 ± 1.27 log10 copies/ml, p=0.003). Significant differences in viral load were observed across N1-N3 and M1-M3 in RSV patients, but not in influenza A or SARS-CoV-2 patients. SARS-CoV-2 patients exhibited significantly lower viral load at RS and LS sites compared to influenza A or RSV patients. Conclusion: Viral RNA was detected in N3 or M3 sites in our masks, highlighting the potential risk associated in these areas. Differential viral load across various sites in surgical masks worn by patients infected with different respiratory viruses warrants further investigation.

Differential Gene Expression in the Upper Respiratory Tract following Acute COVID-19 Infection in Ambulatory Patients That Develop Long COVID.

Background: Post-acute sequelae of COVID-19, or long COVID, is a condition characterized by persistent COVID-19 symptoms. As long COVID is defined by clinical criteria after an elapsed period, an opportunity for early intervention may aid in future prophylactic approaches; however, at present, the pathobiological mechanisms are multifactorial. By analyzing early virally infected upper respiratory tract tissue prior to eventual clinical diagnosis, it may be possible to identify biomarkers of altered immune response to facilitate future studies and interventions. Methods: This is a sub-group analysis of samples collected from those with confirmed COVID-19. RNA extraction from nasopharyngeal/mid-turbinate samples, sequencing, and bioinformatic analysis were performed to analyze long COVID and non-long COVID cohorts at day 14 post infection. Differences in mean viral load at various timepoints were analyzed as well as serological data. Results: We identified 26 upregulated genes in patients experiencing long COVID. Dysregulated pathways including complement and fibrinolysis pathways and IL-7 upregulation. Additionally, genes involved in neurotransmission were dysregulated, and the long COVID group had a significantly higher viral load and slower viral clearance. Conclusions: Uncovering early gene pathway abnormalities associated with eventual long COVID diagnosis may aid in early identification. We show that, post acute infection, in situ pathogenic deviations in viral response are associated with patients destined to meet consensus long COVID diagnosis that is entirely dependent on clinical factors. These results identify an important biological temporal window in the natural history of COVID-19 infection and long COVID pathogenesis amenable to testing from standard-of-care upper respiratory tract specimens.

Liver function tests, CD4+ counts, and viral load among people living with HIV on dolutegravir compared to efavirenz-based cART; a comparative cross-sectional study

BackgroundRecently, dolutegravir-based therapy has become the first-line treatment when compared to others. However, dolutegravir-associated side effects in the liver and levels of efficacy haven't been addressed yet in underdeveloped countries such as Ethiopia. ObjectiveThe purpose of this study was to compare liver function tests, CD4+ counts, and viral load among people living with HIV on dolutegravir and efavirenz-based antiretroviral regimens at Debre Markos Comprehensive Specialized Hospital in Northwest Ethiopia. MethodsAn institutional-based comparative cross-sectional study was carried out from May 20 to July 10, 2020. An equal number of dolutegravir and efavirenz-prescribed patients (n = 53 each) for 6 months and above were included, and a judgmental sampling technique was used. A comparison of categorical and continuous parameters was analyzed with chi-square and an independent t-test, respectively, using SPSS version 26. A multivariable logistic regression was conducted and considered statistically significant at a p-value of <0.05. ResultsThe magnitude of liver enzyme (AST/ALT) abnormalities was 22.4 % (12/53) and 30.2 % (16/53) among dolutegravir- and efavirenz-prescribed patients, respectively. The dolutegravir group had significantly higher mean CD4+ counts than the efavirenz group (589.40 ± 244.38 vs. 450.64 ± 203.54 cell/mm3; p = 0.002). The efavirenz group had a significantly higher mean viral load than the dolutegravir group (783.83 ± 476.82 vs. 997.98 ± 439.11 cp/ml; p = 0.032). There was a statistically insignificant difference in AST (p = 0.709) or ALT (p = 0.687) between dolutegravir and efavirenz-based regimens. The multivariable logistic regression analysis revealed that BMI ≥25 kg/m2 was associated with liver enzyme abnormalities (AOR = 6.60, 95 % CI: 1.17, 42.82). ConclusionA dolutegravir-based regimen was more likely to result in patients achieving higher efficacy for viral suppression and a CD4+ count increase. Although the differences were statistically insignificant, the mean AST and ALT levels were marginally higher in efavirenz-treated groups than in dolutegravir-treated groups.

SARS-CoV-2 Viral Load in the Nasopharynx at Time of First Infection Among Unvaccinated Individuals

SARS-CoV-2 viral load (VL) in the nasopharynx is difficult to quantify and standardize across settings, but it may inform transmission potential and disease severity. To characterize VL at COVID-19 diagnosis among previously uninfected and unvaccinated individuals by evaluating the association of demographic and clinical characteristics, viral variant, and trial with VL, as well as the ability of VL to predict severe disease. This secondary cross-protocol analysis used individual-level data from placebo recipients from 4 harmonized, phase 3 COVID-19 vaccine efficacy trials sponsored by Moderna, AstraZeneca, Janssen, and Novavax. Participants were SARS-CoV-2 negative at baseline and acquired COVID-19 during the blinded phase of the trials. The setting included the US, Brazil, South Africa, Colombia, Argentina, Peru, Chile, and Mexico; start dates were July 27, 2020, to December 27, 2020; data cutoff dates were March 26, 2021, to July 30, 2021. Statistical analysis was performed from November 2022 to June 2023. Linear regression was used to assess the association of demographic and clinical characteristics, viral variant, and trial with polymerase chain reaction-measured log10 VL in nasal and/or nasopharyngeal swabs taken at the time of COVID-19 diagnosis. Among 1667 participants studied (886 [53.1%] male; 995 [59.7%] enrolled in the US; mean [SD] age, 46.7 [14.7] years; 204 [12.2%] aged 65 years or older; 196 [11.8%] American Indian or Alaska Native, 150 [9%] Black or African American, 1112 [66.7%] White; 762 [45.7%] Hispanic or Latino), median (IQR) log10 VL at diagnosis was 6.18 (4.66-7.12) log10 copies/mL. Participant characteristics and viral variant explained only 5.9% of the variability in VL. The independent factor with the highest observed differences was trial: Janssen participants had 0.54 log10 copies/mL lower mean VL vs Moderna participants (95% CI, 0.20 to 0.87 log10 copies/mL lower). In the Janssen study, which captured the largest number of COVID-19 events and variants and used the most intensive post-COVID surveillance, neither VL at diagnosis nor averaged over days 1 to 28 post diagnosis was associated with COVID-19 severity. In this study of placebo recipients from 4 randomized phase 3 trials, high variability was observed in SARS-CoV-2 VL at the time of COVID-19 diagnosis, and only a fraction was explained by individual participant characteristics or viral variant. These results suggest challenges for future studies of interventions seeking to influence VL and elevates the importance of standardized methods for specimen collection and viral load quantitation.

Risk assessment of wild fish as environmental sources of red sea bream iridovirus (RSIV) outbreaks in aquaculture.

Red sea bream iridovirus (RSIV) causes substantial economic damage to aquaculture. In the present study, RSIV in wild fish near aquaculture installations was surveyed to evaluate the risk of wild fish being an infection source for RSIV outbreaks in cultured fish. In total, 1102 wild fish, consisting of 44 species, were captured from 2 aquaculture areas in western Japan using fishing, gill nets, and fishing baskets between 2019 and 2022. Eleven fish from 7 species were confirmed to harbor the RSIV genome using a probe-based real-time PCR assay. The mean viral load of the RSIV-positive wild fish was 101.1 ± 0.4 copies mg-1 DNA, which was significantly lower than that of seemingly healthy red sea bream Pagrus major in a net pen during an RSIV outbreak (103.3 ± 1.5 copies mg-1 DNA) that occurred in 2021. Sequencing analysis of a partial region of the major capsid protein gene demonstrated that the RSIV genome detected in the wild fish was identical to that of the diseased fish in a fish farm located in the same area in which the wild fish were captured. Based on the diagnostic records of RSIV in the sampled area, the RSIV-infected wild fish appeared during or after the RSIV outbreak in cultured fish, suggesting that RSIV detected in wild fish was derived from the RSIV outbreak in cultured fish. Therefore, wild fish populations near aquaculture installations may not be a significant risk factor for RSIV outbreaks in cultured fish.

Molecular Analysis and Ex Vivo Infectivity of Seronegative Occult Hepatitis C Virus: A Study in Single Haemodialysis Centre.

In occult hepatitis C virus infection (OCI), hepatitis C virus ribonucleic acid (HCV RNA) is detectable in peripheral blood mononuclear cells (PBMCs) but is not evident in serum or plasma. Understanding of OCI in patients with seronegative anti-HCV antibodies is limited. In this study, six HCV isolates from haemodialysis (HD) patients with seronegative OCI were identified by molecular assays and phylogenetic analysis. The virus infectivity was assessed ex vivo using a primary naïve PBMC culture system. HCV isolates obtained from the PBMCs of 10 patients with chronic HCV infection (CCI) were characterised concurrently and used as positive controls in the cell culture. Sequence analysis of the 5' untranslated region (UTR) and non-structural 5B (NS5B) region revealed that HCV genotype 3 was the most prevalent virus type in both the OCI and CCI groups. One of the occult HCV isolates was identified as a mixed type. The mean viral load (log10 RNA copies/106 cells) in the PBMC samples of the OCI group (M = 3.4, SD = 0.7) was lower than that of the CCI group (M = 4.6, SD = 1.7). Upon culture, de novo OCI-HCV replicates were detected in five out of six naïve PBMC cultures. Analysis of the replicates showed a single guanine addition in the domain III of 5'-UTR but the overall molecular structure was retained. Seronegative OCI is an active form of infection that replicates at a low level in PBMCs. Seronegative OCI may share the same route of transmission as CCI. The retained viral competency may have an implication for its persistence.

HIV risk behaviour, viraemia, and transmission across HIV cascade stages including low-level viremia: Analysis of 14 cross-sectional population-based HIV Impact Assessment surveys in sub-Saharan Africa.

As antiretroviral treatment (ART) coverage for people living with HIV (PLHIV) increases, HIV programmes require up-to-date information about evolving HIV risk behaviour and transmission risk, including those with low-level viremia (LLV; >50 to ≤1000 copies/mL), to guide prevention priorities. We aimed to assess differences in sexual risk behaviours, distribution of viral load (VL) and proportion of transmission across PLHIV subgroups. We analysed data from Population-based HIV Impact Assessment surveys in 14 sub-Saharan African countries during 2015-2019. We estimated adjusted prevalence ratios (aPR) of self-reported HIV high-risk behaviour (multiple partners and condomless sex) across cascade stages via generalised estimation equations. We modelled the proportions of transmission from each subgroup using relative self-reported sexual risk, a Hill function for transmission rate by VL, and proportions within cascade stages from surveys and UNAIDS country estimates for 2010-2020. Compared to PLHIV with undetectable VL (≤50 copies/mL), undiagnosed PLHIV (aPR women: 1.28 [95% CI: 1.08-1.52]; men: 1.61 [1.33-1.95]) and men diagnosed but untreated (2.06 [1.52-2.78]) were more likely to self-report high-risk sex. High-risk behaviour was not significantly associated with LLV. Mean VL was similar among undiagnosed, diagnosed but untreated, and on ART but non-suppressed sub-groups. Across surveys, undiagnosed and diagnosed but untreated contributed most to transmission (40-91% and 1-41%, respectively), with less than 1% from those with LLV. Between 2010 and 2020, the proportion of transmission from individuals on ART but non-suppressed increased. In settings with high ART coverage, effective HIV testing, ART linkage, and retention remain priorities to reduce HIV transmission. Persons with LLV are an increasing share of PLHIV but their contribution to HIV transmission was small. Improving suppression among PLHIV on ART with VL ≥1000 copies/mL will become increasingly important.

Human papillomavirus (HPV) load is higher in HPVDNA/p16 positive than in HPVDNA positive/p16 negative oropharyngeal squamous cell carcinoma but does not differ significantly between various subsites or correlate to survival

ObjectivePatients with human papillomavirus DNA positive (HPVDNA+) and p16ink4a overexpressing (p16+) oropharyngeal squamous cell carcinoma (OPSCC), especially those with cancer in the tonsillar and base of tongue subsites as compared to other OPSCC subsites have a better outcome than those with only HPVDNA+ or only p16+ cancer. Likewise having a high viral load has been suggested to be a positive prognostic factor. We therefore hypothesized, that HPV viral load could vary depending on OPSCC subsite, as well as with regard to whether the cancer was HPVDNA+ and p16+, or only HPVDNA+, or only p16+ and that this affected outcome. Material and methodsTo address these issues HPV viral load was determined by HPV digital droplet (dd) PCR in tumor biopsies with previously known HPVDNA/p16 status from 270 OPSCC patients diagnosed 2000–2016 in Stockholm, Sweden. More specifically, of these patients 235 had HPVDNA+/p16+, 10 had HPVDNA+/p16-, 13 had HPVDNA-/p16+ and 12 had HPVDNA-/p16- cancer. ResultsWe found that HPVDNA+/p16+ OPSCC had a significantly higher viral load than HPVDNA+/p16- OPSCC. Moreover, there was a tendency for a higher viral load in the tonsillar and base of tongue OPSCC subsites compared to the other subsites and for a low viral load to correlate to a better clinical outcome but none of these tendencies reached statistical significance. ConclusionTo conclude, the mean viral load in HPVDNA+/p16+ OPSCC was higher than in HPVDNA+/p16- OPSCC, but there was no statistically significant difference in viral load depending on OPSCC subsite or on clinical outcome.

High prevalence and concomitant infection of Ranavirus and Eustrongylides sp. in the invasive American Bullfrog in Brazil

American Bullfrogs, Aquarana catesbeiana, are invasive anuran species distributed worldwide. One of the adverse impacts that this species causes in native communities is as a reservoir host for pathogens and parasites. Here, we report the coinfection of two pathogenic organisms in A. catesbeiana: Ranavirus and the nematode Eustrongylides. Bullfrogs were collected in the wild in a pond close to the urban area of São Paulo, Brazil. The prevalence of both pathogens was high: 77% were infected with ranavirus with a mean viral load of 1010.3 viral copies, and 100% of the bullfrogs were infected by Eustrongylides sp. with a mean intensity of infection of 13.4 specimens per host. Four host specimens (31%) presented pathological signs that seemed to be related to the Eustrongylides sp. infection, such as internal organs adhered to each other due to high intensity and large size of the nematodes, ulcers, and raw flesh wounds caused by the nematode. The pathogenic and concomitant infections have potential zoonotic implications and raise concerns about human infection risks for Eustrongylides infection. Moreover, such infections may represent an additional level of threat to native communities through the potential shifts in patterns of parasite and pathogen transmission. Future research involving the native anuran community is essential to ascertain whether invasive bullfrogs are attenuating or exacerbating diseases such as ranavirosis and eustrongylidiosis.

Assessment of Gastroenteric Viruses in Marketed Bivalve Mollusks in the Tourist Cities of Rio de Janeiro, Brazil, 2022.

This study investigated the prevalence and genetic diversity of gastroenteric viruses in mussels and oysters in Rio de Janeiro, Brazil. One hundred and thirty-four marketed bivalve samples were obtained between January and December 2022. The viral analysis was performed according to ISO/TS 15216, and the screening revealed the detection of norovirus GII/GI (40.3%), sapovirus (SaV; 12.7%), human mastadenovirus (7.5%), and rotavirus A (RVA; 5.9%). In total, 44.8% (60) of shellfish samples tested positive for one or more viruses, 46.7% (28/60) of the positive samples tested positive for a single viral agent, 26.7% (16) tested positive for two viral agents, 8.3% (5) for three viral agents, and 13.3% (8) for four viral agents. Additionally, three mussel samples were contaminated with the five investigated viruses (5%, 3/60). Norovirus GII showed the highest mean viral load (3.4 × 105 GC/g), followed by SaV (1.4 × 104 GC/g), RVA (1.1 × 104 GC/g), human mastadenovirus (3.9 × 103 GC/g), and norovirus GI (6.7 × 102 GC/g). Molecular characterization revealed that the recovered norovirus strains belonged to genotypes GII.2, GII.6, GII.9, GII.17, and GII.27; SaV belonged to genotypes GI.1 and GIV.1; RVA to genotypes G6, G8, P[8]-III, and human mastadenovirus to types F40 and F41. The GII.27 norovirus characterized in this study is the only strain of this genotype reported in Brazil. This study highlights the dissemination and diversity of gastroenteric viruses present in commercialized bivalves in a touristic area, indicating the potential risk to human health and the contribution of bivalves in the propagation of emerging pathogens.

Age and Epstein-Barr viral load at diagnosis of post-transplant lymphoproliferative disease are associated with patient survival in kidney transplant recipients.

This study investigated variables associated with mortality in kidney transplant recipients (KTRs) diagnosed with post-transplant lymphoproliferative disease (PTLD) and a simultaneous Epstein-Barr virus (EBV) viremia. This was a retrospective cohort study enrolling KTRs diagnosed with PTLD between 2018 and 2020. Outcome: death within two years after diagnosis. Among 1,625 KTRs who collected EBV viremia (by PCR, 2018-2020) for any reason, 238 (14.6%) had a positive viral load and 41 (17.2%) simultaneous PTLD. These 41 patients were 40.1 years old at diagnosis and 8.6 years after transplantation; 26.8% were induced with rATG and 92.7% were maintained on tacrolimus and azathioprine (TAC/AZA) as immunosuppressive regimen. Lymph nodes (75.6%) was the most common site of PTLD, followed by the gastrointestinal tract (48.8%), with 61.0% at Lugano stage IV and 80.5% monomorphic PTLD. The mean EBV viral load was 12,198 IU/mL. One- and two-year patient survival post-diagnosis was 60.4% and 46.8%, respectively. In the Cox regression analysis, age at PTLD diagnosis (HR for each year = 1.039; p < 0.001) and EBV viral load (HR for each log = 1.695; p = 0.026) were associated with risk of death. This study suggests that in patients predominantly on TAC/AZA, PTLD with simultaneous EBV positive viral load is a late event, and worse survival is associated with older age and EBV viral load at diagnosis.

Detection of hepatitis B virus genotypes in a group of hepatitis B virus-infected patients in central and northern Sri Lanka.

Introduction. Hepatitis B infection causes a spectrum of clinical diseases varying from asymptomatic infection to severe or fulminant acute hepatitis, chronic liver disease, cirrhosis and hepatocellular carcinoma. Hepatitis B virus (HBV) genotypes appear to influence transmission dynamics, clinical outcomes and responses to antiviral therapy. However, hepatitis B genotyping has been poorly investigated in Sri Lanka. This study intended to determine hepatitis B genotypes in a group of HBV-infected people in central and northern Sri Lanka. Methodology. The study was a laboratory-based descriptive cross-sectional study. Initial detection of HBV DNA in 100 EDTA blood samples was done by using a commercially validated quantitative real-time PCR kit. Hepatitis B genotyping was performed by in-house conventional semi-nested multiplex PCR using genotype-specific primers (for genotypes A-F). The serological profile was determined using a commercially validated ELISA/chemiluminescence immunoassay. The results were evaluated for genotype prevalence, viral load association and hepatitis B e antigen (HBeAg) expression in the study population. Results and conclusion. The study detected that genotype C (n=38) is most prevalent and infections with multiple genotypes (n=52, 52%) were commoner than mono-genotype (n=23, 23%) infections. In total, 25% of patients had no detectable genotype among genotypes A-F. The mean viral load in asymptomatic patients with a single genotype was 3.28 log10 copies ml-1 and in multiple genotypes was 4.18 log10 copies ml-1 before treatment. Statistical significance was not detected in mean viral loads and HBeAg expression in these two groups. In the future, chronic HBV infection may be effectively treated and managed according to the infected genotype.

Relationship between Stress and Neuroimmunological Responses and Health Literacy in Newly Diagnosed HIV‐Infected Patients: An Exploratory Study

Objectives. We aimed to study self‐rated health and psycho‐neuroimmunological responses during the initial 6 months after testing positive for human immunodeficiency virus (HIV) and its relationship to health literacy, that is, the ability to take in and understand information about one’s illness. Health literacy plays a critical role in patients’ ability to adhere to antiretroviral treatment (ART). However, there is a lack of studies on the possible impact of HIV‐induced proinflammatory processes on health literacy. Methods. Twelve patients with newly diagnosed HIV attending an urban Ryan White‐funded HIV clinic responded to a questionnaire and had blood samples drawn at baseline (first visit) and after 1, 3, and 6 months, respectively. The questionnaire measured stress, depression, and health literacy. Blood was analyzed for HIV RNA plasma viral load, CD4 cell count, pro‐ and antistress, and inflammatory markers. Results. Complete data for the entire 4 collection periods were available for nine patients. Over the 6‐month period, mean viral load decreased from 353,714.83 (standard deviation 870,334.61) to 35.89 (14.04) copies/mL (p &lt; 0.001). CD4 cell count increased from 321.08 (167.96) to 592.44 (300.06) cells/mm3 (p &lt; 0.001). Self‐rated stress decreased from a baseline mean of 7.33 (2.29) to 3.56 (3.21), on a 0–10 visual analogue scale, at the 6‐month follow‐up (p &lt; 0.01). C‐reactive protein (CRP) decreased from 5757.05 (3146.86) to 2360.84 (2277.33) ng/mL (p &lt; 0.05). Mean health literacy score at baseline was 17.67 (3.50; scale range 0–20) and did not change during the follow‐up period. However, increased stress and decreased CRP (p = 0.05) during the 6‐month follow‐up predicted higher health literacy scores at 6 months. Conclusion. Both stress and proinflammatory processes in newly diagnosed HIV‐infected patients might adversely impact patients’ health literacy and thus their capacity to align with treatment guidance.