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INTRODUCTION: Since the mpox outbreak in 2022, it was unclear if and how often infections with mpox virus (MPXV) were clinically inapparent, i.e. not presenting to clinical care with mpox symptoms. Moreover, it was hypothesized that MPXV circulated in the affected communities before the outbreak was officially detected. METHODS: We retrospectively tested rectal and urethral swabs, and pooled samples for presence of MPXV. Samples were obtained from routine STI testing of three anonymous Community Based Voluntary Counselling and Testing (CBVCT) centres in Berlin, in 2022 and 2023. Testing results were linked to anonymously provided behavioural data. RESULTS: Overall, 9,053 samples from 6,600 client visits were included. Clinically inapparent MPXV infections were detectable in 1.1% of the samples. We did not find MPXV infections in the month before the first cases appeared in Berlin or between October 2022 and January 2023 when case numbers were low in Germany. However, during the outbreak period in 2022, we found clinically inapparent MPXV infections among 2.2% of the clients and during summer/autumn 2023 among 0.3%. The number of condomless anal/vaginal intercourse partners within the previous 6 months and PrEP use were identified as predictors of clinically inapparent MPXV infection. CONCLUSION: Clinically inapparent MPXV infections occurred during the mpox outbreak in Berlin in 2022 and post-outbreak in summer/autumn 2023. Unrecognized MPXV circulation in Berlin before the recognition of the outbreak in May 2022 appears unlikely. However, low-level sustained circulation of clinically inapparent MPXV infections need to be acknowledged in mpox prevention strategies.

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Tilapia lake virus ('TiLV-MH-2022') was recently recovered from the naturally infected farmed tilapia. Reverse transcription-polymerase chain reaction (RT-PCR) using segment 1 specific primers, followed by Sanger sequencing, confirmed the infection. The pairwise sequence homology of segment 1 showed its close relationship with the previous isolates. The virus was successfully detected from the mucus, which emphasised the possibility of non-invasive screening of tilapia on a large scale. The virus inoculum prepared from the infected tissues was tested for in vivo and in vitro pathogenicity. Around 100-140 nm-sized electron-dense virus particles were observed in the infected OnlL cells. Based on the onset of symptoms and lesions, all RT-PCR-positive fish were categorised into two groups, 'clinical' and 'subclinical'. A lesion-scoring technique was developed for assessing the pathogenicity of the virus isolate. The external and internal gross lesions and histopathological alterations in the critical organs of the fish, such as the brain, kidney, gills, and liver, were assessed on a scale of 0 (no gross lesion) to 5 (most severe lesions). Overall lesion score was significantly high in the clinical and subclinical groups for gross and histopathology, respectively. This study is the first such attempt to standardise a semi-quantitative lesion scoring technique for TiLV infection, which establishes a clinical relevance and prognostic ability to distinguish between the apparent and inapparent infection.

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Tick-borne encephalitis virus (TBEV) and Powassan virus (POWV) are neurotropic tick-borne orthoflaviviruses. They cause mostly asymptomatic infections in hosts, but severe forms with CNS involvement can occur. Studying the early stages of viral infections in humans is challenging, and appropriate animal models are essential for understanding the factors determining the disease severity and for developing emergency prophylaxis and treatment options. In this work, we assessed the model of the early stages of TBEV and POWV mono- and co-infections in Macaca fascicularis. Serological, biochemical, and virological parameters were investigated to describe the infection, including its impact on animal behavior. Viremia, neutralizing antibody dynamics, and viral load in organs were chosen as the main parameters distinguishing early-stage orthoflavivirus infection. Levels of IFNα, monocyte count, and cognitive test scores were proposed as additional informative indicators. An assessment of a tick-borne encephalitis vaccine using this model showed that it provided partial protection against POWV infection in Macaca fascicularis without signs of antibody-dependent enhancement of infection.

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Inapparent infection plays an important role in the disease spread, which is an infection by a pathogen that causes few or no signs or symptoms of infection in the host. Many pathogens, including HIV, typhoid fever, and coronaviruses such as COVID-19 spread in their host populations through inapparent infection. In this paper, we formulated a degenerated reaction-diffusion host-pathogen model with multiple infection period. We split the infectious individuals into two distinct classes: apparent infectious individuals and inapparent infectious individuals, coming from exposed individuals with a ratio of (1-p) and p, respectively. Some preliminary results and threshold-type results are achieved by detailed mathematical analysis. We also investigate the asymptotic profiles of the positive steady state (PSS) when the diffusion rate of susceptible individuals approaches zero or infinity. When all parameters are all constants, the global attractivity of the constant endemic equilibrium is established. It is verified by numerical simulations that spatial heterogeneity of the transmission rates can enhance the intensity of an epidemic. Especially, the transmission rate of inapparent infectious individuals significantly increases the risk of disease transmission, compared to that of apparent infectious individuals and pathogens in the environment, and we should pay special attentions to how to regulate the inapparent infectious individuals for disease control and prevention, which is consistent with the result on the sensitive analysis to the transmission rates through the normalized forward sensitivity index. We also find that disinfection of the infected environment is an important way to prevent and eliminate the risk of environmental transmission.

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BACKGROUND: The epidemiological importance of asymptomatic individuals who would never develop illness, compared to those who eventually develop symptoms, has yet to be fully clarified. METHODS: The very first cluster data in Tokyo and Kanagawa (n = 36) were analyzed. Movement of all close contact was restricted for 14 days and they underwent laboratory testing with polymerase chain reaction. The reproduction numbers of symptomatic and asymptomatic cases were estimated. RESULTS: The reproduction number for symptomatic cases was estimated to be 1.2 (95% confidence interval (CI): 0.5-2.9). The relative infectiousness of asymptomatically infected cases was estimated to be 0.27 (95% CI: 0.03-0.81) of symptomatic cases. CONCLUSION: The relative transmissibility of asymptomatic cases is limited. Observing clusters starting with symptomatic transmission might be sufficient for the control.

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BACKGROUND: Tick-borne encephalitis virus (TBEV) is an important human pathogen that can cause the serious illness tick-borne encephalitis (TBE). Patients with clinical symptoms can suffer from severe meningoencephalitis with sequelae that include cognitive disorders and paralysis. While less than 30% of patients with clinical symptoms develop meningoencephalitis, the number of seropositive individuals in some regions indicates a much higher prevalence of TBEV infections, either with no or subclinical symptoms. The functional relevance of these subclinical TBEV infections and their influence on brain functions, such as learning and memory, has not been investigated so far. METHODS: To compare the effect of low and high viral replication in the brain, wildtype and Irf-7-/- mice were infected with Langat virus (LGTV), which belongs to the TBEV-serogroup. The viral burden was analyzed in the olfactory bulb and the hippocampus. Open field, elevated plus maze, and Morris water maze experiments were performed to determine the impact on anxiety-like behavior, learning, and memory formation. Spine density of hippocampal neurons and activation of microglia and astrocytes were analyzed. RESULTS: In contrast to susceptible Irf-7-/- mice, wildtype mice showed no disease signs upon LGTV infection. Detection of viral RNA in the olfactory bulb revealed CNS infections in wildtype and Irf-7-/- mice. Very low levels of viral replication were detectable in the hippocampus of wildtype mice. Although wildtype mice develop no disease signs, they showed reduced anxiety-like behavior and impaired memory formation, whereas Irf-7-/- mice were not affected. This impairment was associated with a significant decrease in spine density of neurons in the hippocampal CA1 region of wildtype mice. Microglia activation and astrogliosis were detected in the hippocampus. CONCLUSION: In this study, we demonstrate that subclinical infections by viruses from the TBEV-serogroup affected anxiety-like behavior. Virus replication in the olfactory bulb induced far-reaching effects on hippocampal neuron morphology and impaired hippocampus-dependent learning and memory formation.

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This study was designed to detect mutations that occur within the "a" determinant in the S gene of the hepatitis B virus (HBV) in patients with occult hepatitis B (OHB), and to analyze the influence of these mutations on expression and reactivity of the hepatitis B surface antigen (HBsAg). Twenty-three certified OHB samples were compared to 32 HBsAg positive samples from patients with chronic hepatitis B. The median HBV DNA levels in the OHB group were significantly lower than those in the control group (P < 0.0001). Mutations within the "a" determinant were analyzed by gene amplification and sequencing. This revealed mixed infections in which clones within a sample displayed either different mutations or mutations in association with clones that exhibited wild type amino acid patterns. Sequencing analysis also showed a significant difference between the proportions of amino acid mutations observed in the OHB and control groups. Seven recombinant S (rS) proteins with corresponding OHB mutations and three wild type alleles were expressed and purified in the Pichia pastoris expression system to preserve conformational attributes, and their reactivity analyzed using six commercial HBsAg assays. The OHB sera were HBsAg nonreactive while the rS proteins with corresponding OHB mutations were universally reactive. Thus, we postulate that the reduced binding affinity between mutated HBsAg and its antibody may not be as important in defining OHB as is the effect of specific mutations in the preS/S region of the genome that affect the synthesis and secretion of the S protein and/or the virion.

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Some international guidelines recommend evaluating the need to confirm positive anti-hepatitis C virus (HCV) antibody screening results by means of a more specific antibody or molecular biology test on the basis of a screening threshold value (such as the sample signal/cut-off ratio) that can predict the positivity of additional antibody testing in at least 95% of cases. The aim of this study was to determine the threshold value of the DiaSorin LIAISON XL chemiluminescence test. Two hundred and twenty-eight routine laboratory samples that were chemiluminescence positive for anti-HCV antibodies but had different signal/cut-off ratios were assayed using immunoblotting, which indicated that 155 (68.0%) were positive, 40 (17.5%) were negative, and 33 (14.5%) were indeterminate. When the samples were divided on the basis of their signal/cut-off ratios, 95.5% of the samples with a ratio of ≥3.5 were positive as against 74.1% of the positive or indeterminate samples with a ratio of <3.5. Statistical analysis using Youden's index and a receiver operating characteristic curve showed that the optimum cut-off value was 3.65. These findings indicate that, when using the LIAISON XL system for anti-HCV antibody screening, a signal/cut-off ratio of ≥3.65 makes further confirmatory tests unnecessary.

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Highly pathogenic avian influenza subtype H5N1 represents a threat to the poultry industry and human health worldwide. Inapparently infected birds are suspected to play an essential role in the spread of avian influenza virus. In the current study, a total of 25,646 samples (16,185 chicken, 4696 ducks, 1633 geese and 3132 turkeys) from apparently healthy birds were screened for the presence of positive samples for H5N1 during 2009-2014. The samples were examined by reverse transcriptase real-time polymerase chain reaction (rRT-PCR) for M, H5 and N1 genes of avian influenza viruses. The results revealed that the HPAI H5N1 existed in an inapparent manner in ducks (4.68 %), geese (4.10 %), chickens (2.48 %) and turkeys (2.29 %). The current finding highlights the serious impact of such type on birds in the epidemiology of H5N1 in birds, animals and humans. It also highlights the existence of another reason other than vaccination that contributes to the widespread of inapparent infection of H5N1 in Egypt.

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Inapparent infections by Trypanosoma cruzi were detected, for the first time, in symptomless seropositive healthy individuals from a Yukpa ethnic community in western Venezuela where Chagas disease had not been previously reported. Seropositivity was detected in 24 out of 173 (13.9%) sera samples taken from asymptomatic people by using three serological methods (DAT, IFAT, ELISA). Complementary analyses by IFAT revealed a low level of anti-T. cruzi specific IgM and IgG in all seropositive people when compared with detected levels in acute and chronic chagasic patients included as positive controls. In addition, 100% of the sampled dogs (8) were seropositive showing a level of anti-T. cruzi IgG similar to that detected in humans. Genotyping of a T. cruzi isolate obtained from an infected wild specimen of Rhodnius pictipes, revealed the circulation of lineage1 (DTU1) in the study area. The importance of the detection of asymptomatic T.cruzi-infections in this indigenous community and its potential epidemiological implications are discussed.

Infecciones inaparentes por Trypanosoma cruzi fueron detectadas, por vez primera, en individuos seropositivos asintomáticos muestreados en una comunidad Yukpa del occidente de Venezuela donde no se ha registrado hasta el presente enfermedad de Chagas. De un total de 173 muestras de suero examinadas 24 (13,9%) fueron registradas como seropositivas utilizando tres pruebas serológicas. Análisis complementarios por IFI revelaron bajos niveles de IgM e IgG cuando las muestras de individuos seropositivos asintomáticos fueron comparadas con casos agudos y crónicos incluidos como controles positivos. La totalidad de perros examinados (8) reveló seropositividad, registrándose niveles de IgG similares a los detectados en el grupo humano. La caracterización molecular de un aislado de T. cruzi obtenido de un espécimen silvestre de Rhodnius pictipes reveló la circulación del linaje 1 (DTU1) en el área de estudio. Se discute la importancia de infecciones asintomáticas por T. cruzi y sus implicaciones epidemiológicas en la comunidad indígena estudiada.

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