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Serologic studies for swine influenza viruses (SIVs) in humans with occupational exposure to swine have been reported from the Americas but not from Europe. We compared levels of neutralizing antibodies against 3 influenza viruses--pandemic (H1N1) 2009, an avian-like enzootic subtype H1N1 SIV, and a 2007-08 seasonal subtype H1N1--in 211 persons with swine contact and 224 matched controls in Luxembourg. Persons whose profession involved contact with swine had more neutralizing antibodies against SIV and pandemic (H1N1) 2009 virus than did the controls. Controls also had antibodies against these viruses although exposure to them was unlikely. Antibodies against SIV and pandemic (H1N1) 2009 virus correlated with each other but not with seasonal subtype H1N1 virus. Sequential exposure to variants of seasonal influenza (H1N1) viruses may have increased chances for serologic cross-reactivity with antigenically distinct viruses. Further studies are needed to determine the extent to which serologic responses correlate with infection. [ABSTRACT FROM AUTHOR]

We investigated the genetic diversity of measles virus (MV) in Nigeria (2004-2005) and the Democratic Republic of the Congo (DRC) (2002-2006). Genotype B3 strains circulating in Kinshasa, DRC, in 2002-2003 were fully replaced by genotype B2 in 2004 at the end of the second Congo war. In Nigeria (2004-2005), two genetic clusters of genotype B3, both of which were most closely related to 1 variant from 1998, were identified. Longitudinal analysis of MV strain diversity in Nigeria suggested that only a few of the previously described 1997-1998 variants had continued to circulate, but this finding was concomitant with a rapid restoration of genetic diversity, probably caused by low vaccination coverage and high birth rates. In contrast, the relatively low genetic diversity of MV in DRC and the genotype replacement in Kinshasa reflect a notable improvement in local measles control. [ABSTRACT FROM AUTHOR]

In Haiti, >90% of the population descended from African slaves. Of 7,147 Haitian pregnant women sampled, 44% of hepatitis B virus (HBV) infections were caused by genotype A1, which today is found mainly in eastern Africa. Twenty percent belong to a rare subgenotype, A5, which has been found only in the former Bight of Benin, a former primary slave trading post. Haitian A subgenotypes appear to have separated early from the African subgenotypes; the most prevalent genotype and subgenotype in West Africa today (E and A3, respectively) are rare in Haiti. This difference indicates that the dominant subgenotypes in Africa emerged in the general population only after the slave trade and explains the low genetic diversity of genotype E. The high prevalence of HBV genotype E in much of Africa further suggests that HBV hyperendemicity is a recent phenomenon, probably resulting from extensive use of unsafe needles. [ABSTRACT FROM AUTHOR]

We conducted a phylogenetic analysis of 19 hepatitis B virus strains from Laos that belonged to 2 subgenotypes of a new genotype I. This emerging new genotype likely developed outside Southeast Asia and is now found in mixed infections and in recombinations with local strains in a geographically confined region. [ABSTRACT FROM AUTHOR]

Background: The α-Gal syndrome (AGS) is characterized by the presence of specific IgE-antibodies to the carbohydrate galactose-α-1,3-galactose (α-Gal). Sensitization to α-Gal has been associated with tick bites and individuals exposed to ticks have an elevated risk of sensitization. The aim of this study was to analyze IgG and IgE antibody responses to α-Gal in a high-risk cohort of forestry employees (FE) in Luxembourg., Methods: Questionnaires and serum samples of FE from Luxembourg (n = 219) were retrospectively analyzed. α-Gal specific IgE was quantified by ImmunoCAP, α-Gal specific IgG and subclasses IgG 1-4 were determined by ELISA. Additionally, sera from population-based controls (n = 150) and two groups of food-allergic patients, patients with AGS (n = 45) and fish-allergic patients (n = 22) were assessed for IgG antibody responses to α-Gal and cod extract., Results: Twenty-one percent of FE was sensitized to α-Gal (sIgE ≥ 0.1 kU A /L). Both sensitized and non-sensitized FE exhibited high levels of α-Gal specific IgG, IgG1 and IgG3 compared with controls, indicating a stimulation of IgG responses by recurrent tick bites, independent of the sensitization status. AGS patients had the highest levels of IgG1 and IgG2 antibodies, whereas the profile of fish-allergic patients was similar to the profile of the controls for which anti-α-Gal responses were dominated by IgG2 antibodies. α-Gal sIgG4 levels were either very low or undetectable in all groups., Conclusion: Our study provides evidence for a continuous stimulation of α-Gal related immune responses by repeated tick bites, translating into highly elevated levels of IgG1 antibodies directed against α-Gal., (© 2024 The Author(s). Clinical and Translational Allergy published by John Wiley & Sons Ltd on behalf of European Academy of Allergy and Clinical Immunology.)

Intense psychosocial stress during early life has a detrimental effect on health-disease balance in later life. Simultaneously, despite its sensitivity to stress, the developing microbiome contributes to long-term health. Following stress exposure, HPA-axis activation regulates the "fight or flight" response with the release of glucose and cortisol. Here, we investigated the interaction between the oral microbiome and the stress response. We used a cohort of 115 adults, mean age 24, who either experienced institutionalisation and adoption (n = 40) or were non-adopted controls (n = 75). Glucose and cortisol measurements were taken from participants following an extended socially evaluated cold pressor test (seCPT) at multiple time points. The cohort´s oral microbiome was profiled via 16S-V4 sequencing on microbial DNA from saliva and buccal samples. Using mixed-effect linear regressions, we identified 12 genera that exhibited an interaction with host's cortisol-glucose response to stress, strongly influencing intensity and clearance of cortisol and glucose following stress exposure. Particularly, the identified taxa influenced the glucose and cortisol release profiles and kinetics following seCPT exposure. In conclusion, our study provided evidence for the oral microbiome modifying the effect of stress on the HPA-axis and human metabolism, as shown in glucose-cortisol time series data., (© 2024. The Author(s).)

The influenza A virus has been scarcely investigated in pigs in Africa, with rare detection prior to 2009. The spread of A(H1N1)pdm09 changed the epidemiology due to frequent human-to-swine transmission and the emergence of various new reassortants. This study therefore aimed at estimating the level of circulation and characterizing influenza A viruses at the interface between swine workers, who are crucial players in the inter-species transmission of influenza A viruses, and their animals in several farms in Nigeria, a hub for pig production in Africa. This cross-sectional study showed that 24.6% (58/236) of the pig serum samples collected in 2013-2014 had anti-influenza A antibodies in the absence of vaccination programs, but none of the pig swabs ( n = 1193) were positive according to RT-qPCR. Viral RNA was detected in 0.9% (2/229) of swine workers sampled at their place of work, and the strains were characterized as A(H1N1)pdm09 and seasonal A(H3N2). Our results highlight that more awareness of swine workers regarding the consequences of reverse zoonosis for animal and public health is warranted. Annual vaccination and the wearing of masks when experiencing influenza-like symptoms would help decrease influenza inter-species transmission, while surveillance should be adequately supported for early detection.

Introduction: Children requiring multiple blood transfusions are at high risk of transfusion-transmissible infections (TTIs). Lao People's Democratic Republic is a low-resource setting where donor blood screening faces challenges. This study aimed to determine the burden of TTIs in children in Vientiane Capital., Methods: 300 children with transfusion history and 300 controls were recruited. In addition, 49 newly diagnosed transfusion recipients were followed for up to 12 months. Serum was tested for hepatitis B surface antigen and IgG antibodies against parvovirus B19, hepatitis B, C and E viruses., Results: The patients had a similar prevalence of anti-hepatitis B core antibodies (56; 18.7%) and hepatitis B surface antigen (8; 2.7%) as the controls (58; 19.3% and 9; 3.0%, respectively). However, there was a higher prevalence of an antibody profile suggestive of hepatitis B vaccination (anti-hepatitis B surface antibody positive/anti-hepatitis B core antibody negative) in the transfused group (140/299; 46.8%) than in controls (77/300; 25.7%, p<0.01). All other markers were similar in the patients and controls or higher in the controls: anti-hepatitis C virus (2.7% and 3.3%, p=0.6), anti-hepatitis E virus (7.5% and 12.7%, p=0.006) and anti-parvovirus B19 (2.4% and 8.5%, p=0.001). The longitudinal cohort did not show an increase in any marker over time., Conclusion: Our results suggest no significant role of TTIs in Lao children. The higher prevalence of the hepatitis B vaccination profile in transfusion recipients showed that recommendations to vaccinate before commencing transfusions is at least partially implemented, although there is room for improvement., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Introduction: Vaccination has dramatically reduced invasive Haemophilus influenzae type b (Hib) disease worldwide. Hib vaccination was introduced in the Lao PDR in 2009, as part of the pentavalent vaccine. To contribute to the understanding of the epidemiology of Hib in Lao PDR and the protection levels before and after the introduction of the vaccination, we tested serum samples from existing cohorts of vaccine age-eligible children and unvaccinated adolescents for antibodies against Hib., Methods: Serum samples from 296 adolescents born before vaccine introduction and from 1017 children under 5 years (vaccinated and unvaccinated) were tested for anti-Hib antibodies by ELISA. Bivariate analyses were performed to investigate factors associated with long-term protection., Results: The vast majority of all participants showed evidence of short- (42.7%) or long-term (56.1%) protection against Hib. Almost all of the unvaccinated adolescents had antibody titers indicating short-term protection and almost half (45.6%) were long-term protected. Nearly all children (>99.0%) were at least short-term protected, even those that were unvaccinated or whose vaccination status was unknown. Among vaccinated children, participants vaccinated more than 1 or 2 years ago and with a mid-upper arm circumference z-score < -2 were less likely to be long-term protected., Discussion: Nearly all adolescents born before the introduction of Hib vaccination in the Lao PDR had antibody titers corresponding to at least short-term protection, indicating a high burden of Hib disease at that time. After vaccine introduction, all but four children (>99%) showed at least short-term protection. Possible explanations for the proportion of protected, yet apparently unvaccinated children, may be past infections, cross-reacting antibodies or faulty vaccination documentation. Our results highlight the need for robust surveillance and reporting of invasive Hib disease to determine the burden of disease despite vaccination., Competing Interests: The authors have declared that no competing interests exist.

During 2017-2019, a total of 88/753 (11.7%) of patients 5-90 years of age in hospitals in Saravan Province, Laos, were seropositive for hepatitis C virus antibodies. Viral RNA was found in 44 samples. Sequencing showed high diversity within genotype 6. We recommend exposure-risk investigations and targeted testing and treatment.

The epidemiology of typhoid fever in Lao People`s Democratic Republic is poorly defined. Estimating the burden of typhoid fever in endemic countries is complex due to the cost and limitations of population-based surveillance; serological approaches may be a more cost-effective alternative. ELISAs were performed on 937 serum samples (317 children and 620 adults) from across Lao PDR to measure IgG antibody titers against Vi polysaccharide and the experimental protein antigens, CdtB and HlyE. We measured the significance of the differences between antibody titers in adults and children and fitted models to assess the relationship between age and antibody titers. The median IgG titres of both anti-HylE and CdtB were significantly higher in children compared to adults (anti-HylE; 351.7 ELISA Units (EU) vs 198.1 EU, respectively; p<0.0001 and anti-CdtB; 52.6 vs 12.9 EU; p<0.0001). Conversely, the median anti-Vi IgG titer was significantly higher in adults than children (11.3 vs 3.0 U/ml; p<0.0001). A non-linear trend line fitted to the anti-CdtB and anti-HlyE IgG data identified a peak in antibody concentration in children <5 years of age. We identified elevated titers of anti-HlyE and anti-CdtB IgG in the serum of children residing in Lao PDR in comparison to adults. These antigens are associated with seroconversion after typhoid fever and may be a superior measure of disease burden than anti-Vi IgG. This approach is scalable and may be developed to assess the burden of typhoid fever in countries where the disease may be endemic, and evidence is required for the introduction of typhoid vaccines., Competing Interests: The authors have declared that no competing interests exist.

Introduction: Hepatitis B is endemic in Lao PDR and about 9% of the adult population is chronically infected. In this study, we investigated regional, occupational, age and sex-related differences in hepatitis B epidemiology in Lao blood donors., Methods: 5017 voluntary blood donors from 8 different provinces were tested for hepatitis B markers by ELISA. Predictors for the prevalence of hepatitis B surface antigen (HBsAg) and antibodies against the core antigen (anti-HBc) were assessed by bivariate and multivariable analyses., Results: In total, 41% of the participants were positive for anti-HBc; the HBsAg prevalence was estimated at 6.9% among all participants (9.2% among first-time donors and 3.9% among repeat donors). Among first-time donors, HBsAg positivity was associated independently with being male (p<0.001), being from the North (p<0.001) and being soldier (p<0.001). Participants were more likely to be anti-HBc positive when they were male (p<0.001), from the Northern provinces (p<0.001) and older than 20 years (p<0.01)., Conclusion: In conclusion, our study confirmed an overall high HBsAg and anti-HBc prevalence in Lao PDR, albeit with considerable regional variation. The identification of a sizeable number of HBsAg positives among repeat donors warrants a thorough investigation of current blood screening, record keeping, donor identification and counselling practises., Competing Interests: The authors have declared that no competing interests exist.

The early-life microbiome (ELM) interacts with the psychosocial environment, in particular during early-life adversity (ELA), defining life-long health trajectories. The ELM also plays a significant role in the maturation of the immune system. We hypothesised that, in this context, the resilience of the oral microbiomes, despite being composed of diverse and distinct communities, allows them to retain an imprint of the early environment. Using 16S amplicon sequencing on the EpiPath cohort, we demonstrate that ELA leaves an imprint on both the salivary and buccal oral microbiome 24 years after exposure to adversity. Furthermore, the changes in both communities were associated with increased activation, maturation, and senescence of both innate and adaptive immune cells, although the interaction was partly dependent on prior herpesviridae exposure and current smoking. Our data suggest the presence of multiple links between ELA, Immunosenescence, and cytotoxicity that occur through long-term changes in the microbiome.

Introduction: Measles is an endemic but largely neglected disease in Lao People's Democratic Republic. New-borns are protected by maternal antibodies, but antibody waning before measles vaccination at 9 months of age leaves infants susceptible to infection. In this study, the susceptibility window of infants was determined to generate scientific evidence to assess the national measles immunization strategy., Methods: Between 2015 and 2016, demographic data, medical history, and blood samples were collected from 508 mother-child pairs at the provincial hospital in Vientiane. The samples were screened with a commercial kit detecting anti-measles IgG antibodies., Results: The large majority (95.7%) of the mothers were seropositive for anti-measles IgG and antibody titers of the mothers and infants were highly correlated ( p < 0.01). While at birth 97.7% of the infants were seropositive, seropositivity rates decreased to 74.2% two months later to reach only 28.2% four months after birth ( p < 0.01). Just before the first dose of the measles-rubella vaccine, scheduled at 9 months of age, was actually given, less than 14% of the infants were seropositive., Conclusion: This alarmingly wide susceptibility gap due to rapid maternal antibody decay leaves infants at risk of measles infection and serious disease complications. A high herd immunity is crucial to protect young infants and can be achieved through improved routine vaccination coverage and (expanded age group) supplementary immunization activities.

Respiratory infections are one of the most frequent reasons for medical consultations in children. In low resource settings such as in Lao People's Democratic Republic, knowledge gaps and the dearth of laboratory capacity to support differential diagnosis may contribute to antibiotic overuse. We studied the etiology, temporal trends, and genetic diversity of viral respiratory infections in children to provide evidence for prevention and treatment guidelines. From September 2014 to October 2015, throat swabs and nasopharyngeal aspirates from 445 children under 10 years old with symptoms of acute respiratory infection were collected at the Children Hospital in Vientiane. Rapid antigen tests were performed for influenza A and B and respiratory syncytial virus. Real-time reverse-transcription polymerase chain reactions (RT-PCRs) were performed to detect 16 viruses. Influenza infections were detected with a higher sensitivity using PCR than with the rapid antigen test. By RT-PCR screening, at least one pathogen could be identified for 71.7% of cases. Human rhinoviruses were most frequently detected (29.9%), followed by influenza A and B viruses combined (15.9%). We identify and discuss the seasonality of some of the infections. Altogether these data provide a detailed characterization of respiratory pathogens in Lao children and we provide recommendations for vaccination and further studies., (© 2021 The Authors. Journal of Medical Virology Published by Wiley Periodicals LLC.)

Early Life Adversity (ELA) is closely associated with the risk for developing diseases later in life, such as autoimmune diseases, type-2 diabetes and cardiovascular diseases. In humans, early parental separation, physical and sexual abuse or low social-economic status during childhood are known to have great impact on brain development, in the hormonal system and immune responses. Maternal deprivation (MD) is the closest animal model available to the human situation. This paradigm induces long lasting behavioral effects, causes changes in the HPA axis and affects the immune system. However, the mechanisms underlying changes in the immune response after ELA are still not fully understood. In this study we investigated how ELA changes the immune system, through an unbiased analysis, viSNE, and addressed specially the NK immune cell population and its functionality. We have demonstrated that maternal separation, in both humans and rats, significantly affects the sensitivity of the immune system in adulthood. Particularly, NK cells' profile and response to target cell lines are significantly changed after ELA. These immune cells in rats are not only less cytotoxic towards YAC-1 cells, but also show a clear increase in the expression of maturation markers after 3h of maternal separation. Similarly, individuals who suffered from ELA display significant changes in the cytotoxic profile of NK cells together with decreased degranulation capacity. These results suggest that one of the key mechanisms by which the immune system becomes impaired after ELA might be due to a shift on the senescent state of the cells, specifically NK cells. Elucidation of such a mechanism highlights the importance of ELA prevention and how NK targeted immunotherapy might help attenuating ELA consequences., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Fernandes, Patil, Meriaux, Theresine, Muller, Leenen, Elwenspoek, Zimmer and Turner.)

Background: Even though measles vaccination was introduced in the Lao PDR in 1984, coverage rates remain consistently low and outbreaks continue to occur frequently. This study was performed to investigate the seroprevalence of measles and rubella antibodies in vaccinated and unvaccinated children from Central Lao PDR., Methods: Antibody titres of 1090 children aged 8-29 months who were vaccinated at different levels of the health care system were assessed by ELISA. Bivariate and multivariable analyses were performed to identify factors affecting seropositivity against measles and rubella., Results: Among the vaccinated children, 67.5% in Vientiane Province and 76.4% in Bolikhamxay Province were double positive/borderline for measles and rubella IgG. A high proportion of unvaccinated children at both study sites (24.4% and 38.4%) were positive/borderline for measles and/or rubella. Time since vaccination <180 days, more than two siblings, and a mother who is a farmer/labourer were negatively associated with seropositivity., Conclusions: A high prevalence of measles and rubella antibodies was found in unvaccinated children, indicating widespread circulation of both viruses and underreporting of cases. The high proportion of vaccinated children still susceptible to measles suggests problems with vaccine immunogenicity, emphasizing the need for regular evaluations of vaccine efficacy and management., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Foot and mouth disease (FMD) virus remains enzootic in Lao People's Democratic Republic (Lao PDR) due to insufficient control measures, including low vaccination coverage. We assessed virus epidemiology and evaluated knowledge, attitude and practice of Lao farmers and animal health stakeholders towards FMD to support politics in devising evidence-based control measures. Sera were collected from 972 domestic ruminants in three provinces of Lao PDR: Vientiane Capital, Vientiane and Xiengkhouang provinces. Seroprevalence of antibodies directed against non-structural proteins of FMD virus was assessed using a commercial ELISA. Positive sera were further characterized by detecting antibodies directed against the structural proteins of FMD serotypes O, A and Asia 1. Information about farm management, biosecurity practices, livestock trade, and past FMD outbreaks was obtained. Overall 35% (340/972) of the ruminants had antibodies against FMD virus with a similar seroprevalence in each province. Seroprevalence depended significantly on the ruminant species (p < .001): 61% of buffaloes, but only 41% of cattle and 15% of goats were seropositive. While antibodies against FMD serotype Asia 1 were absent, 87% (297/340) of the seropositive animals had antibodies against FMD serotype O and 32% (110/340) against FMD serotype A. Many seropositive animals (31%) had antibodies against both serotypes O and A. The majority of the farmers could name the symptoms of FMD and the susceptible animal species. Although many had likely observed FMD outbreaks in their herd and were aware of FMD vaccines, only few vaccinated their animals. This study confirms that FMD remains enzootic in at least three provinces of Lao PDR where vaccination coverage is low. It also shows the relevance of nationwide active and passive disease surveillance, as well as of vaccination campaigns using bivalent FMD vaccines and targeting all susceptible animal species., (© 2020 Wiley-VCH GmbH.)

Competing Interests: The author declares no competing interests.

Background: Lao People's Democratic Republic has frequent outbreaks of vaccine-preventable diseases (VPD). This study aimed to determine susceptibility and exposure to VPD in Saravan, a rural province with high ethnic diversity and some of the poorest health indicators nationwide., Methods: Patients from three district hospitals and one provincial hospital were enrolled. Serum was tested by ELISA for IgG against hepatitis B virus (HBV), tetanus, diphtheria, measles, and rubella., Results: The study enrolled 2463 participants aged 5-90 years. Exposure to HBV was 33.2% and increased with age up to 62.4% of those aged >50 years. HBV surface antigen prevalence was 5.7% in males and 2.4% in females; 9.9% had serology compatible with vaccination. Seroprevalence of protective anti-tetanus antibodies was 46.3% overall. Protective anti-diphtheria seroprevalence was 40.5%. Anti-measles seroprevalence increased from 16.7% in those aged 5-10 years to 97.7% in those aged >50 years. Anti-rubella seroprevalence was 93.0% overall. There were differences in seroprevalences between sex, districts and ethnicity., Conclusions: Routine infant vaccination needs strengthening in order to close the immunity gaps. High burden of HBV infection should be addressed by increasing birth dose vaccine coverage. Regional and ethnic differences need to be investigated to enable a targeted approach to vaccination., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background and Aims: Respiratory syncytial virus (RSV) is one of the main viral pathogens causing acute respiratory infections in children under 5 years of age but has seldom been studied in Central African Republic (CAF). Taking advantage of the national influenza surveillance network in CAF, this study aimed at providing the first insights into RSV prevalence and seasonality over 4 years of surveillance and the clinical manifestations of RSV in this population in CAF., Methods: A total of 3903 children under 5 years matching the influenza-like illness (ILI, 68.5%) or severe acute respiratory infection (SARI, 31.5%) case definitions were recruited from January 2015 to December 2018. The presence of RSV viral RNA in nasopharyngeal samples was assessed by RT-PCR, followed by RSV-A and RSV-B typing and Sanger sequencing on a subset of samples. Phylogenetic analyses were carried on partial G protein sequences. Associations between RSV and demographic or clinical manifestations were investigated by statistical analyses., Results: RSV prevalence was significantly higher in infants <6 months (13.4%), in hospitalized children (13.3% vs 5.5%) and in male patients (9.5% vs 6.4%). An overall prevalence of RSV of 8.0% in the period of 2015 to 2018 was shown, with significant annual (6.4%-10.6%) and seasonal (12.7% in rainy season vs 3.0% in dry season) fluctuations. While RSV seasons in 2015, 2016, and 2018 were relatively similar, 2017 showed deviations from the overall patterns with significantly higher RSV circulation and an outbreak peak 3 to 5 months earlier. Concomitant circulation of RSV-A and RSV-B with an alternating predominance of RSV-A and RSV-B strains and temporal RSV-A genotype replacement from NA1 to ON1 was observed., Conclusion: This study represents the first in-depth epidemiological analysis of RSV in CAF and provides first insights into RSV genetic diversity and seasonality in the country., Competing Interests: The authors declare that they have no conflict of interests., (© 2021 The Authors. Health Science Reports published by Wiley Periodicals LLC.)

Competing Interests: The author declares no competing interests.

Introduction and Objectives: Hepatitis B virus (HBV) is endemic in Lao PDR, with up to 10% chronic infections in adults. Dentists have high risk of exposure and transmission to their patients. The aim was to investigate the serological profiles of vaccination, exposure and susceptibility to HBV. In addition, we determined the knowledge, attitude and practice (KAP) of Lao dentists toward HBV., Materials and Methods: Dentists and dental assistants were recruited from 186 private dental clinics in Vientiane Capital. They completed a KAP questionnaire and serum was tested by ELISA for HBV markers (anti-HB surface antigen, anti-core antigen, HBV surface antigen)., Results: 206 dentists and 111 dental workers aged between 18-63 years were included. Serology data showed that 37.8% had previous exposure (49.0% of males and 32.4% of females) and 5.0% were chronically infected (7.7% of males and 3.8% of females). Less than 15% had serological evidence of vaccination and 66.8% were unprotected against infection. Knowledge of HBV was mostly low, for example 61.8% were aware of the childhood HBV vaccine. Just over half said that they should give dental care to HBV carriers. About one quarter reported accidental blood exposure during the last 6 months but knowledge about procedures after exposure was low., Conclusions: Lao dentists are at high risk of occupational exposure to HBV but have low awareness and serological protection. There is a need to introduce mandatory vaccination, to strengthen occupational health in health care providers and to further develop the academic curricula of dental students., (Copyright © 2020 Fundación Clínica Médica Sur, A.C. Published by Elsevier España, S.L.U. All rights reserved.)

The potent neurotoxicity of benzo[a]pyrene (B[a]P) has been suggested to be a susceptibility factor accelerating the onset of brain tumours and the emergence of neurobehavioural disturbances. B[a]P has been shown to be neurotoxic, acting directly on both the central and peripheral nervous systems, as well as indirectly via peripheral organs like liver and gut. By using a realistic B[a]P exposure scenario (0.02-200 mg/kg/day, 10 days) in mice, we elucidated brain-specific B[a]P metabolism and at identified hydroxylated B[a]P metabolites in serum which could be used as markers of cognitive impairment. Repeated oral administration of B[a]P led to, at the doses of 20 and 200 mg/kg/day, significant overexpression of Cyp1a1/Cyp1b1 in 2 out of the 3 brain regions considered, thereby suggesting the ability of the brain to metabolize B[a]P itself. At the same doses, mice exhibited a reduction in anxiety in both the elevated plus maze and the hole board apparatus. Concomitantly, B[a]P triggered dose-dependent changes in Nmda subunit expression (Nr1 and Nr2a/Nr2b) in areas involved in cognition. We detected 9-OH-B[a]P and 7,8-diol-B[a]P in serum at the level for which cognitive impairment was observed. We suggest that these metabolites may, in the future be exploited as potent biomarkers of B[a]P-induced cognitive impairments.

Background: The timely administration of vaccines is considered to be important for both individual and herd immunity. In this study, we investigated the timeliness of the diphtheria-tetanus-whole cell pertussis-hepatitis B-Haemophilus influenzae type b (pentavalent) vaccine, scheduled at 6, 10 and 14 weeks of age in the Lao People's Democratic Republic. We also investigated factors associated with delayed immunization., Methods: 1162 children aged 8-28 months who had received the full course of the pentavalent vaccine at different levels of the health care system were enrolled. Vaccination dates documented in hospital records and/or immunisation cards were recorded. Age at vaccination and time intervals between doses were calculated. Predictors for timely completion with the pentavalent vaccine at 24 weeks were assessed by bivariate and multivariable analyses., Results: Several discrepancies in dates between vaccination documents were observed. In general, vaccination with the pentavalent vaccine was found to be delayed, especially in health care settings below the provincial hospital level. Compared to the central hospital level, less participants who were vaccinated at the district/health center level received the third dose by 16 (48% at the central hospital level vs. 7.1% at the district and 12.4% at the health center level) and 24 weeks of age (94.4% at the central hospital level vs 64.6% at the district-outreach and 57.4% at the health center level) respectively. In logistic regression analyses, lower education level of the mother as well as vaccination by outreach service, were independently associated with delayed completion of vaccination., Conclusion: We observed a general delay of vaccination, especially at lower ranked facilities, which correlated with indicators of poor access to health services. This highlights the need for further improving health equity in rural areas. Age-appropriate vaccination should become a quality indicator for the national immunization programme. In addition, we recommend further training of the health care staff regarding the importance of reliable documentation of dates., Competing Interests: Dr. Nouanthong has been a member of the independent National Immunization Technical Advisory Group in Lao PDR since August 2017. The other authors declared no conflict of interest. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Early life adversity (ELA) has been associated with inflammation and immunosenescence, as well as hyporeactivity of the HPA axis. Because the immune system and the HPA axis are tightly intertwined around the glucocorticoid receptor (GR), we examined peripheral GR functionality in the EpiPath cohort among participants who either had been exposed to ELA (separation from parents and/or institutionalization followed by adoption; n = 40) or had been reared by their biological parents (n = 72).Expression of the strict GR target genes FKBP5 and GILZ as well as total and 1F and 1H GR transcripts were similar between groups. Furthermore, there were no differences in GR sensitivity, examined by the effects of dexamethasone on IL6 production in LPS-stimulated whole blood. Although we did not find differences in methylation at the GR 1F exon or promoter region, we identified a region of the GR 1H promoter (CpG 1-9) that showed lower methylation levels in ELA.Our results suggest that peripheral GR signaling was unperturbed in our cohort and the observed immune phenotype does not appear to be secondary to an altered GR response to the perturbed HPA axis and glucocorticoid (GC) profile, although we are limited in our measures of GR activity and time points.

Next-generation sequencing (NGS) has revolutionized the scale and depth of biomedical sciences. Because of its unique ability for the detection of sub-clonal variants within genetically diverse populations, NGS has been successfully applied to analyze and quantify the exceptionally-high diversity within viral quasispecies, and many low-frequency drug- or vaccine-resistant mutations of therapeutic importance have been discovered. Although many works have intensively discussed the latest NGS approaches and applications in general, none of them has focused on applying NGS in viral quasispecies studies, mostly due to the limited ability of current NGS technologies to accurately detect and quantify rare viral variants. Here, we summarize several error-correction strategies that have been developed to enhance the detection accuracy of minority variants. We also discuss critical considerations for preparing a sequencing library from viral RNAs and for analyzing NGS data to unravel the mutational landscape., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Despite several recent reports of outbreaks of hepatitis A, little is known about the disease burden in Lao People's Democratic Republic (PDR). We conducted a cross-sectional age-stratified seroprevalence study of anti-hepatitis A virus (HAV) IgG and risk factors in a rural province (Xiengkhouang) and Vientiane capital in Lao PDR. Overall, 62% of participants were anti-HAV positive in Xiengkhouang Province compared with 45.5% in Vientiane capital. In Xiengkhouang, 23.7% of 5- to 10-year-olds were already seropositive compared with 5% in Vientiane. A dramatic increase in seroprevalence occurred between 15- to 20-year and 21- to 30-year age-groups (35.7-62.4%, Xiengkhouang, and 11.5-69.7%, Vientiane) until essentially all older adults were positive in both locations. The main risk factors for HAV antibodies were age, non-Lao-Tai ethnicity, and food-related risk factors. In conclusion, Lao children seem to be exposed very early to HAV, particularly in rural settings, and exposure continues throughout their lives, mostly without being reported. In the older birth cohorts, the high seroprevalence may largely reflect poor sanitation and exposure during childhood. In Vientiane capital, the lower seroprevalence at young ages may reflect better water sanitation since the late 1990s. A comparison with neighboring Thailand indicates that the impact of improved sanitation in Lao PDR began only two decades later. Further improvements in water sanitation, in particular in the rural districts, and better food hygiene are warranted. Our study also suggests that HAV infections are underreported. Improved reporting would provide guidance for targeted interventions to further reduce HAV infections.

Objectives: Pertussis is a debilitating vaccine-preventable infection. The aim of this study was to determine susceptibility and exposure to pertussis in Lao PDR in different age groups and subpopulations., Methods: A total 3072 serum samples were obtained from different cohorts: children with documented vaccination, pre-schoolers, schoolchildren, blood donors, healthcare workers (HCWs), and pregnant women and paired cord blood. Samples were tested for anti-pertussis toxin IgG antibodies. A history of Bordetella pertussis exposure was defined according to antibody titres. Four hundred and seventy-five throat swabs and nasopharyngeal aspirates were analysed by PCR for the presence of B. pertussis in symptomatic children at the Children's Hospital in Vientiane., Results: Overall pertussis seroprevalence was 57.5%. The prevalence of titres indicating acute infection or recent vaccination or infection/vaccination within the last 12 months ranged from 7.4% (100/1356) in adults to 21.4% (25/117) in pre-schoolers (age 1-5 years). B. pertussis was detected in 1.05% (5/475) of children with respiratory symptoms in Vientiane Capital., Conclusions: It is suggested that routine childhood vaccination, in particular outreach, as well as vaccination of HCWs should be strengthened. A childhood booster and vaccination of pregnant mothers should be considered. There is also a need to improve reporting and to introduce pertussis testing in at least one central facility., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objectives: Hepatitis B is endemic in Lao PDR with 8-10% of the adult population being chronically infected. We investigated the impact of hepatitis B vaccination on infection in adolescents born shortly before and after the introduction of the vaccine in 2001., Methods: 779 students from Vientiane Capital and Bolikhamxay province were tested for HBV markers by ELISA. Socio-demographic information was collected with a standardized questionnaire. Predictors/risk factors for seroprotection or exposure to hepatitis B infection were assessed by bivariate and multivariable analyses., Results: The prevalence of a serological vaccination profile increased significantly after the introduction of the vaccine (13.2%-21.9%, p < 0.05, in Vientiane; 3.0%-19.7%, p < 0.001, in Bolikhamxay), which translated into at least a 2-times lower prevalence of past infection. In logistic regression, older students in Bolikhamxay were less likely to be vaccinated and more likely to have been infected by HBV in the past., Conclusion: Even though this study documented a sizable and lasting reduction in past hepatitis B infections in adolescents born after the introduction of infant hepatitis B vaccination, the overall levels of protective anti-HBs were low and warrant at least the introduction of a booster for adolescents. Furthermore, we suggest improving the coverage of the hepatitis B birth dose., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Maternal neonatal tetanus (MNT) was eliminated from Lao People's Democratic Republic (PDR) in 2014. WHO recommends 80% coverage of 2 or more tetanus vaccinations (TT2+) in pregnancy to maintain MNT control. Vaccination coverage in Lao PDR varies among regions although the reasons are not clear., Methods: 185 pregnant women giving birth in three district hospitals in Savannakhet province, Lao PDR were recruited. A questionnaire was administered to determine factors associated with seroprotection and blood was taken from mother and cord blood to be tested for anti-tetanus antibodies by ELISA., Results: 77% of mothers and 79% of newborns had sufficiently protective antibody titres (>0.5 IU/ml) against tetanus. Only 70% of the mothers received one dose of TT vaccination during antenatal care (ANC) consultation and 45% received the recommended two injections. Although most of the vaccination took place during ANC 1 and 2, many were missed at these time-points. Anti-tetanus seroprotection in the mothers was associated with maternal age, number of ANC visits, number of TT vaccinations during and before pregnancy and gestational age., Conclusion: Seroprevalence of anti-tetanus antibodies in mothers and newborns was intermediate but TT2+ coverage was low in healthcare settings in Lao PDR. TT2+ coverage during ANC is likely to be significantly lower in settings with less robust ANC practices. Missed opportunities to vaccinate in ANC 1 and 2 suggest a need to promote vaccine awareness and vaccination at first ANC visit. A booster dose of TT containing vaccine should be considered for children aged between 4 and 7 years old., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In backyard farms of Lao People's Democratic Republic, mixed-species rearing of poultry is a breeding-ground for cross-species transmission. Here, the epidemiology of viruses circulating among backyard poultry in Vientiane Province was assessed to guide future control strategies. Oral/tracheal and cloacal swabs, collected from 605 poultry (308 ducks, 297 chickens) between 2011 and 2015, were screened by PCR for Newcastle disease virus (NDV), coronavirus (CoV) and chicken anaemia virus (CAV). Chicken sera were screened for anti-NDV antibodies by ELISA. Statistical and phylogenetic analyses revealed transmission patterns and relationships. Closely related strains co-circulated in chickens and ducks. While CoV RNA was detected in oral/tracheal swabs of 9.3% of the chickens and 2.4% of the ducks, rates were higher in faecal swabs of both species (27.3% and 48.2%). RNA of infectious bronchitis virus (IBV) and duck CoV was found in faecal swabs of chickens (19.7% and 7.1%) and ducks (4.1% and 44.1%). Moreover, DNA of the generally chicken-specific CAV was detected in oral/tracheal swabs of chickens (18.1%) and, sporadically, of ducks (2.4%). Despite serological evidence of NDV circulation or vaccination (86.9%), NDV RNA was not detected. We found a high prevalence and indication for cross-species transmission of different CoV strains in backyard poultry. Interestingly, ducks served as biological, or at least mechanical, carriers of viral strains closely related not only to IBV, but also to CAV. Bird containment and poultry species separation could be first steps to avoid cross-species transmission and emergence of novel strains with broad host range and enhanced pathogenicity. RESEARCH HIGHLIGHTS High rates of avian viruses were detected by PCR in backyard poultry from Lao PDR. Diverse coronavirus and chicken anemia virus strains co-circulated. Phylogenetic analyses suggested virus transmission between chickens and ducks. Serological evidence of Newcastle disease was found, but viral RNA was not detected.

Background: The Lao People's Democratic Republic continues to sustain a considerable burden of vaccine-preventable diseases because of incomplete vaccine coverage and weak vaccine responses. We have assessed seroconversion after routine vaccination with the pentavalent vaccine to capture weaknesses of vaccine management at the different levels of the healthcare system., Methods: A total of 1151 children (aged 8-28 months) with 3 documented doses of the pentavalent vaccine delivered at central hospitals in Vientiane and the provincial hospital, 3 district hospitals, and 10 health centers in Bolikhamxay province were enrolled. Sociodemographic information was collected with a standardized questionnaire. Serum samples were analyzed for antibodies against vaccine components, and bivariate and multivariable analyses were performed to identify risk factors for low vaccine responses., Results: Seroprotection rates at the provincial, district, and health center level were as high as in central hospitals, but seroprotection rates in areas covered by remote health centers were significantly lower. Protective levels also rapidly decreased with age at sampling. Seroprotection rates in Bolikhamxay against the different components reached 70%-77% and were up to 20% higher than in previous studies in the same region; 18.8% more children received the hepatitis B vaccine birth dose and the hepatitis B virus infection rate was 4 times lower., Conclusions: Vaccine immunogenicity has dramatically improved in a central province, likely due to training and investment in the cold chain. Nevertheless, there remains a need to focus on the "last mile" in remote areas were most children are vaccinated through outreach activities., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America.)

In Southeast Asia, the large majority of the population remains affected by parasitic worms despite longstanding mass treatment and health education campaigns. Soil-transmitted helminths and also the fish-borne liver fluke negatively affect development during early childhood. Here, the prevalence of helminth infections in stool samples of 610 mother-child pairs from Khammouane, Bolikhamxay and Vientiane provinces in Lao People's Democratic Republic was determined by formalin-ethyl acetate concentration technique. Overall, 15.1% of the children and 46.9% of the mothers were positive for at least one helminth species. Helminth detection rates varied significantly by province with the highest prevelance in Khammouane and the lowest in Bolikhamxay province. Mothers that were positive for soil-transmitted helminths were significantly more likely to have children positive for the same helminth species (p < 0.01) but this was not the case for the liver fluke Opisthorchis viverrini. A protective effect of breastfeeding against soil-transmitted helminths was revealed. Our data reconfirm the generally high helminth burden among mother-child pairs who likely share a number of risky lifestyle behaviors also with other family members. To reduce maternal burden of helminths, we propose that anti-helmintic treatment of women of childbearing age and of mothers during postnatal care should be included in the national strategy., (© 2019 The Author(s).)

The Republic of Serbia is a country with ongoing endemic transmission of measles. The aim of this study is to summarize the main characteristics of the measles resurgence that occurred in Serbia in 2014-2015. The national surveillance data on measles was analysed in relation to the clinical, epidemiological and laboratory data. Between November 2014 and December 2015 a measles resurgence with 420 cases was observed in Serbia. Measles virus was initially introduced by and spread among citizens of Bosnia and Herzegovina with temporary residence in Serbia, before spreading to the resident population. Of the 223 patients with available medical records, 173 (77.6%) were unvaccinated. The overall measles incidence during the outbreak was 5.8/100.000. The highest age-specific incidence rate was recorded in children aged ≤4 years (25.9/100.000), but most cases (67.9%) were ≥20 years old. Hospitalization rate was high (32.9%) and included two cases of encephalitis associated with measles. In total, 42 health-care workers and 22 related cases including hospitalized patients (n = 13) contracted measles. The overall percentage of laboratory confirmed cases was 81.7% (n = 343/420). All measles virus sequences except one (D9) belonged to genotype D8, suggesting interruption of transmission after the previous outbreak in 2010-2011 caused by genotype D4 viruses. The growing number of adult patients as compared to previous epidemics, suggests an urgent need for supplementary immunization activities targeting susceptible health care workers, unvaccinated or incompletely vaccinated adults as well as people without vaccination records. The comprehensive investigation of the 2014/2015 measles resurgence will contribute to decisions about appropriate countermeasures to stop the future measles resurgences in Serbia., Competing Interests: The authors have declared that no competing interests exist.

Several Avian paramyxoviruses 1 (synonymous with Newcastle disease virus or NDV, used hereafter) classification systems have been proposed for strain identification and differentiation. These systems pioneered classification efforts; however, they were based on different approaches and lacked objective criteria for the differentiation of isolates. These differences have created discrepancies among systems, rendering discussions and comparisons across studies difficult. Although a system that used objective classification criteria was proposed by Diel and co-workers in 2012, the ample worldwide circulation and constant evolution of NDV, and utilization of only some of the criteria, led to identical naming and/or incorrect assigning of new sub/genotypes. To address these issues, an international consortium of experts was convened to undertake in-depth analyses of NDV genetic diversity. This consortium generated curated, up-to-date, complete fusion gene class I and class II datasets of all known NDV for public use, performed comprehensive phylogenetic neighbor-Joining, maximum-likelihood, Bayesian and nucleotide distance analyses, and compared these inference methods. An updated NDV classification and nomenclature system that incorporates phylogenetic topology, genetic distances, branch support, and epidemiological independence was developed. This new consensus system maintains two NDV classes and existing genotypes, identifies three new class II genotypes, and reduces the number of sub-genotypes. In order to track the ancestry of viruses, a dichotomous naming system for designating sub-genotypes was introduced. In addition, a pilot dataset and sub-trees rooting guidelines for rapid preliminary genotype identification of new isolates are provided. Guidelines for sequence dataset curation and phylogenetic inference, and a detailed comparison between the updated and previous systems are included. To increase the speed of phylogenetic inference and ensure consistency between laboratories, detailed guidelines for the use of a supercomputer are also provided. The proposed unified classification system will facilitate future studies of NDV evolution and epidemiology, and comparison of results obtained across the world., (Crown Copyright © 2019. Published by Elsevier B.V. All rights reserved.)