Dengue virus seroprevalence study in Bangphae district, Ratchaburi, Thailand: A cohort study in 2012-2015

Background To determine the seroprevalence and transmission dynamics of dengue virus (DENV), age-stratified longitudinal serological surveys were conducted in Bangphae district, Ratchaburi province, Thailand, for 3 years between April 2012 and April 2015. Methodology The surveys enrolled 2012 healthy children and adults between 1 and 55 years-of-age, and a longitudinal serosurvey of six repeated bleeds of the same cohort of individuals was conducted every 8 months for the first 2 years (M0, M8, M16) and every half a year (M24, M30, M36) for the rest of the study period. All samples were tested using in-house indirect sandwich dengue IgG ELISA to determine DENV antibody titer, and 640 paired samples which showed rising of DENV IgG titers in paired serum were further tested using in-house neutralization assay, Plaque Reduction Neutralization Test (PRNT50). Principal findings When compared against the gold standard based on the results of PRNT50, sensitivity and specificity of indirect ELISA were found to be both about 85%. The overall DENV IgG positivity determined by ELISA was 74.3% in 2012 and increased to 79.4% by the final sample collection in 2015. In our study sample, more than 98% of subjects older than 25 years were found to be seropositive. Among 518 IgG negative subjects at enrollment, the seroconversion rates were measured in paired bleeds; the rates (between successive visits, approximately 6 months) ranged between 4.8% (between M16 and M24) and 14.7% (between M0 and M8). The dominant serotype of primary DENV infection cases based on seroconversion was identified from the PRNT results and it was DENV-2. Conclusions Our study documented high levels of seroprevalence and rate of transmission. Given the importance of the serostatus and disease burden in consideration for dengue vaccine introduction, our data could be used in decision-making on implementation of various dengue control and preventive measures.

The serotype distribution of Streptococcus agalactiae (GBS) carriage isolates among pregnant women having risk factors for early-onset GBS disease: a comparative study with GBS causing invasive infections during the same period in Denmark

Background We describe the serotype distribution of Streptococcus agalactiae (GBS) carriage isolates from women in labor and among GBS isolates causing invasive infections during the same period to see if the distribution of carriage serotypes reflects the GBS serotypes causing invasive diseases including early-onset disease (EOGBS). Methods Data on invasive isolates from 2019 including serotype, erythromycin and clindamycin susceptibility was retrieved from the Danish national reference laboratory, Statens Serum Institut. Carriage isolates were collected from women with risk factors for EOGBS enrolled at delivery at the maternity ward at a Danish University Hospital, first half of 2019. Results Among carriage isolates, the dominant serotype was IX (21 %) followed by serotype III (19 %). The resistance to erythromycin and clindamycin was 21 and 26 %, respectively. Among invasive GBS isolates, no case of EOGBS with serotype IX was detected but the distribution of serotypes were otherwise similar to the GBS carrier strains. The corresponding resistance to erythromycin and clindamycin was 23 and 15 %, respectively. Penicillin resistance was not detected among carriage nor invasive isolates. Conclusions The distribution of serotypes among carriage and invasive GBS reflects the assumption that EOGBS occur following transmission of GBS from mother to newborn, with the exception of serotype IX.

Emergence of Dengue 4 as Dominant Serotype During 2017 Outbreak in South India and Associated Cytokine Expression Profile

Dengue virus (DENV) infection is prevalent in tropical and subtropical regions of the world, which is fatal if untreated symptomatically. Emergence of new genotype within serotypes led to enhanced severity. The objective of the study is to identify the molecular characteristics of the DENV circulated during 2017 outbreak in Tamil Nadu, India, and to investigate the role of inflammatory cytokines in different “serotypes” and in “dengue severity”. A total of 135 suspected samples were tested for DENV infection using IgM, IgG, and qPCR assay; where 76 samples were positive for DENV and analyzed for 12 inflammatory cytokines using ELISA. Serotyping shows 14 DENV-1, 22 DENV-2, 7 DENV-3, and 33 DENV-4, where DENV-4 was predominant. Among 76, 42 isolates were successfully sequenced for C-prM region and grouped. A lineage shift was observed in DENV-4 genotype. Irrespective of serotypes, IFNγ was significantly elevated in all serotypes than control as well as in primary infection than secondary, indicating its role in immune response. GM-CSF and IP-10 were significantly elevated in secondary infection and could be used as prognostic biomarkers for secondary infection. Our observation shows differential cytokine expression profile varied with each serotype, indicating serotype/genotype-specific viral proteins might play a major role in dengue severity. DENV-4 as dominant serotype was reported in Tamil Nadu for the first time during an outbreak with a mixed Th1/Th17 cytokine expression profile that correlated with disease severity. We conclude it is essential to identify circulating viral genotype and their fitness by mutational analysis to correlate with disease severity and immune status, as this correlation will be helpful in diagnostics and therapeutics applications.

Molecular surveillance of Dengue Virus (DENV) and its co-infection with Chikungunya Virus (CHIKV) among febrile patients: A comparative study from South Delhi, India

Dengue and Chikungunya are two major arboviral infections transmitted worldwide by the mosquitoes, Aedes aegypti and Ae. albopictus. India suffers enormously with both Dengue and Chikungunya as they pose a great public health challenge. The present study aims to evaluate the prevalence of Dengue Virus (DENV), Chikungunya Virus (CHIKV) and DENV/CHIKV co-infection (by Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR)/Enzyme Linked Immunosorbent Assay (ELISA), their clinical features, DENV serotypes and CHIKV specific Immunoglobulin G (IgG) within a 7 years gap in the Delhi population. The study sample included clinically suspected febrile patients (?7 days) sera collected during 2017-2018 (n=87) and during 2008-2010 (n=623) from Delhi. Captured ELISA was performed for CHIKV IgG screening and nested PCR was done for DENV serotyping. The percentage prevalence for DENV was significantly higher than CHIKV with 41.38% (n=87) and 16.1% (n=87), respectively; interestingly, DENV/CHIKV co-infection was detected in 10.34% (n=9/87) cases during 2017-2018. Similarly, a high DENV prevalence was observed during 2008-2010 with the prevalence rate of 38.3% (69/180), 34.65% (35/101) and 47.07% (161/342), respectively. DENV 1 and DENV 3 were dominant serotype during 2008-2010 and 2017-2018 respectively. We have noticed a high prevalence (36.67%, 22/60) of the CHIKV IgG antibody in the 2017-2018 samples. Joint pain was more preferential to CHIKV mono-infection and DENV/CHIKV co-infection compared to DENV mono-infection. The present study highlights the need for active surveillance simultaneously for both DENV and CHIKV and to evaluate the role of CHIKV/DENV co-infections in disease severity in the endemic regions.

Sero-prevalence and serotypes of infectious bronchitis virus in free-range chicken in Plateau state, Nigeria

Globally, infectious bronchitis (IB) is an important respiratory viral disease responsible for enormous economic losses to poultry farmers. In Nigeria, limited reports on the prevalence and serotypes of the IB virus are available. Here, we investigated the prevalence and serotypes of infectious bronchitis virus (IBV) in chicken in Plateau State. A descriptive cross-sectional study was carried out involving 440 apparently healthy free-range local chickens sampled from eleven villages in four Local Government Areas (LGA) of Plateau State. Sera collected from the birds were screened for the presence of four IBV serotypes namely; Massachusetts (Mass), Arkansas (Ark), Connecticut (Con) and Delaware (De-072) using haemagglutinationinhibition (HI) test. In all, a prevalence of 82.95% (n = 365) was recorded. At LGAlevel, prevalence of 79.50%, 47.37%, 95.45% and 100% were recorded in Kanam, Mangu, Qua’an pan and Bassa LGAs, respectively. Based on serotype prevalence, Mass had 89.30% (n = 326); Ark 79.70% (n = 291); Con 88.20% (n = 322) while De-072 was 42.70% (n = 156). There were statistically significant associations between dominant serotype and the LGAs (p≤0.001). This study shows high prevalence of IBwith at least four strains of IBV present in free-range chicken flocks in Plateau State requiring attention for control measures. Keywords: Free-range chicken; Infectious bronchitis virus; Plateau state, Serosurvey, Serotype

Construction and Immunogenicity Comparison of Three Virus-Like Particles Carrying Different Combinations of Structural Proteins of Avian Coronavirus Infectious Bronchitis Virus

Infectious bronchitis virus (IBV) poses massive economic losses in the global poultry industry. Here, we firstly report the construction and immunogenicity comparison of virus-like particles (VLPs) carrying the S, M and E proteins (SME-VLPs); VLPs carrying the S and M proteins (SM-VLPs); and VLPs carrying the M and E proteins (ME-VLPs) from the dominant serotype representative strain GX-YL5 in China. The neutralizing antibody response induced by the SME-VLPs was similar to that induced by the inactivated oil vaccine (OEV) of GX-YL5, and higher than those induced by the SM-VLPs, ME-VLPs and commercial live vaccine H120. More importantly, the SME-VLPs elicited higher percentages of CD4+ and CD8+ T lymphocytes than the SM-VLPs, ME-VLPs and OEV of GX-YL5. Compared with the OEV of GX-YL5, higher levels of IL-4 and IFN-γ were also induced by the SME-VLPs. Moreover, the mucosal immune response (sIgA) induced by the SME-VLPs in the tear and oral swabs was comparable to that induced by the H120 vaccine and higher than that induced by the OEV of GX-YL5. In the challenge experiment, the SME-VLPs resulted in significantly lower viral RNA levels in the trachea and higher protection scores than the OEV of GX-YL5 and H120 vaccines, and induced comparable viral RNA levels in the kidneys, and tear and oral swabs to the OEV of GX-YL5. In summary, among the three VLPs, the SME-VLPs carrying the S, M and E proteins of IBV could stimulate the strongest humoral, cellular and mucosal immune responses and provide effective protection, indicating that it would be an attractive vaccine candidate for IB.

Assessing the impact of the 13 valent pneumococcal vaccine on childhood empyema in Australia

BackgroundEmpyema is a serious complication of pneumonia frequently caused by Streptococcus pneumoniae (SP). We assessed the impact of the 13-valent pneumococcal conjugate vaccine (13vPCV) on childhood pneumonia and empyema after inclusion in the Australian National Immunisation Program.MethodsFor bacterial pneumonia and empyema hospitalisations, we ascertained incidence rates (IRs) using the National Hospital Morbidity Database International Statistical Classification of Disease discharge codes and relevant population denominators, and calculated incidence rate ratios (IRR) comparing the 13vPCV period (June 2012–May 2017) with the 7vPCV period (June 2007–May 2011). Blood and pleural fluid (PF) cultures and PF PCR of 401 children with empyema from 11 Australian hospitals during the 13vPCV period were compared with our previous study in the 7vPCV period.FindingsAcross 7vPCV and 13vPCV periods, IRs per million children (95% CIs) were 1605 (1588 to 1621) and 1272 (1259 to 1285) for bacterial pneumonia, and 14.23 (12.67 to 15.79) and 17.89 (16.37 to 19.42) for empyema hospitalisations. IRRs were 0.79 (0.78 to 0.80) for bacterial pneumonia and 1.25 (1.09 to 1.44) for empyema. Of 161 empyema cases with SP serotypes, 147 (91.3%) were vaccine types. ST3 accounted for 76.4% of identified serotypes in the 13vPCV period, more than double than the 7vPCV period (p<0.001); ST19A decreased from 36.4% to 12.4%. No cases of ST1 empyema were identified in the 13vPCV period versus 14.5% in the 7vPCV period.Interpretation13vPCV resulted in a significant reduction in all-cause hospitalisations for bacterial pneumonia but empyema hospitalisations significantly increased, with emergence of pneumococcal ST3 as the dominant serotype in empyema.Trial registration numberAustralian and New Zealand Clinical Trial Registry ACTRN 12614000354684.

Group B Streptococcus serotypes associated with different clinical syndromes: Asymptomatic carriage in pregnant women, intrauterine fetal death, and early onset disease in the newborn

Objectives To study Group B Streptococcus (GBS) isolates associated with different clinical syndromes: asymptomatic carriage in pregnant women, intrauterine fetal death (IUFD), and early onset disease (EOD) in the newborn. Methods GBS isolates were collected from asymptomatic pregnant women admitted for labor, IUFD cases, and neonates with EOD. Serotypes and antibiotic susceptibilities were determined. Multilocus sequence typing (MLST) was performed to assess genetic epidemiology. Results GBS carriage rate was 26.1% (280/1074). The dominant serotype among asymptomatic pregnant women was VI [98/240 women (40.8%)], followed by serotypes III, V and IV in 42/240 (17.5%), 30/240 (12.5%) and 28/240 (11.7%) women, respectively. The dominant serotype in IUFD cases was serotype VI [10/13 (76.9%)]. In contrast the prevalent serotype among EOD cases was III [16/19 (84.2%)]. ST-1 was associated with IUFD [7/13 (53.8%)], ST-17 was associated with serotype III and EOD in the newborn 14/19 (73.7%)]. Erythromycin and clindamycin resistance reached 36.8%, 7.7% and 20.0%among EOD, vaginal carriage and IUFD, respectively. Conclusions Serotypes VI and ST-1 were dominant among asymptomatic pregnant women and in IUFD cases while EOD was associated with serotype III and ST-17. Invasive mechanisms thus may differ between IUFD and EOD in the newborn and virulence may be related to capsule serotype. Resistance rates to erythromycin and clindamycin were high in EOD cases.

241. Molecular patterns of Streptococcus agalactiae (GBS) Strains Associated with Different Clinical Syndromes: Early-Onset Disease in Neonates, Intrauterine Infection, and Vaginal Colonization, an Orthodox Jewish Community (OJC) Residing in Bney Brak

Background Rectovaginal colonization during pregnancy with Group B Streptococcus (GBS) is a risk factor for early neonatal sepsis, and may also cause chorioamnionitis and fetal death. In Israel, the reported colonization rate in pregnant women is low, and therefore routine screening of pregnant women for GBS colonization is not recommended. We noticed higher rates of early-onset disease (EOD) due to GBS in newborns of women hospitalized in Maayaney Hayeshua Medical Center, which serves an Orthodox Jewish Community (OJC) in Israel. Therefore, our aim was to investigate molecular patterns of GBS strains from mothers and neonates hospitalized in Maayaney Hayeshua. Methods During 2017, GBS isolates were collected from asymptomatic pregnant women (280/1,074), neonates with EOD (n = 7), and intrauterine fetal death remains (IUFD) (n = 7). We serotyped isolates from vaginal carriage (n = 203), EOD (n = 7), IUFD (n = 7) and EOD isolates obtained from the Ministry of Health (n = 11). Multilocus sequence typing (MLST) was performed on isolates from asymptomatic pregnant women (n = 14), EOD (n = 7) and IUFD (n = 7). Antibiotic susceptibilities were determined. Results GBS carriage rate was 26.1%. In asymptomatic pregnant women the dominant serotype was VI [84 women (41.3%)], followed by III, IV and V in 32 (15.7%), 23 (11.3%) and 21 (10.3%) women, respectively. The dominant serotype in EOD was III [15/18 (83.3%)] and in IUFD VI [5 (71.4%)]. ST-17 was expressed mainly by serotype III, and was associated with EOD. ST-1, expressed mainly by serotype VI, was associated with IUFD. See Tables 1 and 2 and Figure 1. Resistance to erythromycin and clindamycin was 19.3% and 18.2% while resistance in invasive isolates was 45.5% to both antibiotics. Conclusion GBS vaginal colonization rate in an OJC was significantly higher than the reported carriage rate of 21.6% reported in Israeli pregnant women. Serotypes VI was dominant in carriage and in cases of IUFD while EOD was exclusively associated with serotype III. Resistance rates to erythromycin clindamycin were high, particularly in invasive disease. These results advocate routine GBS screening in this population and caution against empirical treatment with macrolides or clindamycin in penicillin-allergic women. Disclosures All authors: No reported disclosures.

Impact of 13-Valent Pneumococcal Conjugate Vaccine on Colonization and Invasive Disease in Cambodian Children

Background Cambodia introduced the 13-valent pneumococcal conjugate vaccine (PCV13) in January 2015 using a 3 + 0 dosing schedule and no catch-up campaign. We investigated the effects of this introduction on pneumococcal colonization and invasive disease in children aged &lt;5 years. Methods There were 6 colonization surveys done between January 2014 and January 2018 in children attending the outpatient department of a nongovernmental pediatric hospital in Siem Reap. Nasopharyngeal swabs were analyzed by phenotypic and genotypic methods to detect pneumococcal serotypes and antimicrobial resistance. Invasive pneumococcal disease (IPD) data for January 2012–December 2018 were retrieved from hospital databases. Pre-PCV IPD data and pre-/post-PCV colonization data were modelled to estimate vaccine effectiveness (VE). Results Comparing 2014 with 2016–2018, and using adjusted prevalence ratios, VE estimates for colonization were 16.6% (95% confidence interval [CI] 10.6–21.8) for all pneumococci and 39.2% (95% CI 26.7–46.1) for vaccine serotype (VT) pneumococci. There was a 26.0% (95% CI 17.7–33.0) decrease in multidrug-resistant pneumococcal colonization. The IPD incidence was estimated to have declined by 26.4% (95% CI 14.4–35.8) by 2018, with a decrease of 36.3% (95% CI 23.8–46.9) for VT IPD and an increase of 101.4% (95% CI 62.0–145.4) for non-VT IPD. Conclusions Following PCV13 introduction into the Cambodian immunization schedule, there have been declines in VT pneumococcal colonization and disease in children aged &lt;5 years. Modelling of dominant serotype colonization data produced plausible VE estimates.