Common Serotypes Research Articles

Relationship Between Serotypes and Antimicrobial Nonsusceptibility of Streptococcus pneumoniae Clinical Isolates in Tunisia.

Streptococcus pneumoniae remains a significant cause of morbidity and mortality worldwide despite the overall success of the vaccine programs. In Tunisia, pneumococcal conjugate vaccines (PCV)10 was introduced in the national immunization program in April 2019. We sought to determine the relationship between serotypes and antimicrobial nonsusceptibility of S. pneumoniae isolates recovered from clinical samples in the prevaccination period in the south of Tunisia. A total of 504 nonduplicate S. pneumoniae isolates collected between 2012 and 2018 were tested for antimicrobial susceptibility, among them 439 (87.1%) were serotyped. The most common serotypes were 19F (17.8%), 14 (15.3%), 3 (9.1%), 19A (8.2%), and 23F (7.3%). The proportions of isolates with serotypes covered by PCV7, PCV10, and PCV13 were 55.4%, 56.3%, and 77.9%, respectively. Three-quarters (74.4%) of pneumococcal isolates were nonsusceptible to penicillin, and about half (54.8%) were multidrug resistant. Penicillin nonsusceptibility was observed for all 19A and 23F isolates, and was significantly associated with serotypes 19F (odds ratio [OR]: 33.7) and 14 (OR: 8.7). A significant association with multidrug resistance was noted for serotypes 19A (OR: 10), 19F (OR: 9.4), 23F (OR: 8.6), and 6B (OR: 5.2). The alarming rates of pneumococcal antimicrobial nonsusceptibility and the strong association with the most prevalent serotypes compel microbiologists to monitor the impact of the PCV10 introduced recently in our national immunization program.

Is Salmonella enterica shared between wildlife and cattle in cattle farming areas? An 11-year retrospective study in Tokachi district, Hokkaido, Japan.

Background Salmonella enterica in cattle has long been problematic and suspected to be transmitted by wildlife in Tokachi, Hokkaido, a major cattle farming area in Japan. Understanding the role of wildlife in S. enterica transmission would be helpful for developing control strategies of bovine salmonellosis.ObjectivesWe aimed to elucidate the possibility of S. enterica transmission between sympatric wildlife, including raccoons and crows and cattle, in Tokachi from 2008 to 2018 by analysing S. enterica detection records, and the genetic relatedness of serotypes shared between wildlife and cattle.Methods S. enterica detection records were based on the results of a field survey and existing cattle records at relevant organisations, including clinical reports, a monitoring survey and quarantine for introduced calves at growing farms and public calving farms. S. enterica was identified by polymerase chain reaction assay and serotyped by agglutination assay. The detection records were organised chronologically to investigate whether common serotypes in wildlife and cattle were detected in the same year. The isolates corresponding to detection records were assessed for their genetic patterns by pulsed‐field gel electrophoresis.ResultsThe prevalence of S. enterica in raccoons and crows was 10.7% (17/159) and 5.7% (55/967), respectively. The following serotypes were detected from both wildlife and cattle: Braenderup, Dublin, Infantis, Mbandaka, Montevideo, 4,[5],12:i:‐ and Typhimurium. Genetically similar isolates for S. Braenderup, S. Dublin, S. Montevideo and S. 4,[5],12:i:‐ were detected from both species in the same year.ConclusionsOur long‐term retrospective observations supported that S. enterica was shared between wildlife and cattle. Wildlife invasions should be controlled at farms to prevent inter‐species transmission of S. enterica from livestock farms.

Pneumococcal serotypes causing non-invasive pneumonia in adults from a South Indian tertiary care hospital and the impact of the newer conjugate vaccines.

Background Streptococcus pneumoniae is the leading cause of community-acquired pneumonia (CAP) in adults. Ageing, chronic conditions and comorbidities are important risk factors for pneumococcal pneumonia.PurposeThere is lack of data on the pneumococcal serotypes causing non-invasive pneumonia in India. This study aims to determine the prevalent pneumococcal serotypes causing non-invasive pneumonia, the associated comorbidities, and the coverage of both the available pneumococcal vaccines in India and conjugate vaccines that are currently undergoing clinical trials.MethodsA total of 280 subjects (aged >16 years) who had clinical symptoms correlating with radiological findings for non-invasive bacteremic pneumonia and microbiological evidence of S. pneumoniae between 2018 and 2020 were included. The clinical, demographic, radiological and microbiological findings were retrieved from the Hospital Information System (HIS).ResultThe common serotypes in order of prevalence were 19F, 9V, 23F, 6B, 11A, 13, 34, 10A, 19A and 6A. The predominant non-vaccine serotypes were 13, 34, 35B, 31 and 16F. The associated radiological findings were pneumonic consolidation and multi-lobar involvement. Other coinfected bacterial pathogens included H. influenzae, S. aureus, K. pneumoniae and P. aeruginosa.ConclusionThe pneumococcal vaccines: PCV10/GSK, PCV10/SII, PCV13, PCV15, PCV20 and PPSV23 provide an overall serotype coverage of 36, 41, 47, 48, 61 and 69 %, respectively of S. pneumoniae causing non-invasive pneumonia in South India. Increasing catch-up vaccination using PCV10(SII) in pre-school children could have a more significant impact on reducing pneumococcal pneumonia in adults (>50 years) in terms of increased herd immunity at an affordable cost.

1309. Incidence and Epidemiology of Invasive Pneumococcal Disease due to Serotype 3 in South-Central Ontario

BackgroundIn our population, the most common serotype (ST) of S. pneumoniae causing invasive pneumococcal disease (IPD) is now ST 3. We undertook an analysis of population based surveillance for IPD to examine the incidence and epidemiology of ST 3 disease over the last 25 years.MethodsThe Toronto Invasive Bacterial Diseases Network has performed population-based surveillance for IPD in Toronto/Peel region (pop’n 4.5M) since 1995. All sterile site isolates of S. pneumoniae are reported to a central study laboratory, isolates are serotyped, and clinical and vaccination data are collected via patient and physician interview and chart review. Population data are obtained from Statistics Canada.ResultsFrom 1995-2020, 11032 episodes of IPD occurred; 10015 had STs available, and 10484 clinical data. Overall, ST 3 comprised 9.2% of cases (N=931). Compared to other patients with IPD, those with ST 3 IPD were older (median age 65 vs. 58.5, P< .001), more likely to have underlying lung (22.7% v 16.0%, P< .0001) and cardiac (21.7 v 18.4, P=.02) disease and less likely to be immunocompromised (IC) (23.1% v 29.0% P< .0001). ST3 episodes were more likely to be pneumonia (81% v 65%), less likely to be bacteremia without focus (7.6% v 18.9%), and more likely to require ICU admission (42.3% v 25.1%) and to die (27.1% v 16.6%). In multivariable analysis, patients with ST 3 disease remained more likely to die (OR 1.65; 95%CI1.3-2.0). Over time, the proportion of patients with ST 3 IPD who were nursing home (NH) residents (18/171 in 1995-2000 vs. 4/215 in 2016-2020, P=.0002), and who were IC (46/169 in 1995-2000 vs 39/204 in 2016-2020, P=.007) decreased significantly; in IPD due to other STs, the proportion who were NH residents declined, but the proportion IC increased significantly. The case fatality rate (CFR) declined significantly in IPD due to ST3 but not other STs (Figure 1). Changes in incidence are shown in Figure 2.Figure 2: Incidence of serotype 3 IPD over time, Toronto/Peel, 1995-2020 The incidence of ST3 IPD in children and adults under 65 did not change significantly from 1995/96 to 2019/20. In older adults, the annual incidence of disease declined from 4.98 per 100,000 per year in 1995-2000 to 3.53 per 100,000 per year in 2001-2010 (IRR 0.71, 95%CI 0.56-0.90), then to 2.23 per 100,000 per year in 2011-2020 (IRR compared to 2001-2010 0.63, 95%CI 0.50-0.79)FIgure 2: Case fatality rate of IPD due to serotype 3 and other serotypes over time, 1995-2020, Toronto-Peel The case fatality rate of IPD due to ST3 declined from 37.6% (56/149) in 1995-2000 to 50/235 (21.3%) in 2015-2020 (P<.0001). The CFR in other serotypes did not change.ConclusionThe epidemiology of IPD due to ST3 has changed significantly over time and the CFR has declined. The incidence of ST3 disease in children and younger adults has not changed significantly, although the power to detect change is low in children. In older adults the incidence of ST3 disease declined significantly after PPV23 introduction in 1995/6 and again after PCV13 introduction for children.Disclosures All Authors: No reported disclosures

1156. Pneumococcal Colonization in Children with Persistent Asthma and without Asthma

BackgroundThe most common underlying medical condition among children ≥ 5 years of age with invasive pneumococcal disease is asthma. How asthma affects pneumococcal colonization is not fully understood. Our objective was to compare pneumococcal colonization rates in children with persistent asthma vs. without asthma.MethodsThis is a single center retrospective cohort study. We used salvage mid-turbinate samples testing negative for influenza per routine care from 5-18 year-olds with upper respiratory symptoms or febrile illness during 2017-18 and 2018-19 northern hemisphere respiratory seasons (November to April). Analyzed groups were those with persistent asthma or those without asthma. Samples were evaluated for pneumococcal colonization by real-time PCR using CDC lytA primers (positive Ct ≤ 35). Positive samples were further tested with multiplex serotype-specific PCR assays to determine pneumococcal serotype. ResultsOf 363 children (120 with persistent asthma and 243 without asthma), 87.6% were 5-10 years old; and 49.9% were male. Fifty percent of samples were from January-February. Pneumococcal colonization rate was lower in children with persistent asthma (10%) vs. without asthma (18.9%) (p=0.03). The odds of colonization were lower in children with persistent asthma (OR 0.4 [95%CI 0.2-0.9]) after adjusting for age, sex, clinic site, smoking exposure, and number of pneumococcal vaccine doses. Colonized patients without asthma were younger than the other groups (Table 1). Pneumococcal serotype/serogroup was assigned in 45 (77.6%) positive samples; 16 (36%) samples corresponded to PCV13 serotypes and 29 (64%) samples to non-PCV13 serotypes. The most common serotypes were: 19F (n=7), 3 (n=6), 6C/6D (n=5), 23B (n=4), 33F/33A/37 (n=4), 35B (n=3), 22F/22A (n=3), 23A (n=3).Table 1 ConclusionPatients with persistent asthma had lower rates of pneumococcal colonization than patients without asthma during respiratory season. Disclosures Liset Olarte, MD, MSc, GSK (Research Grant or Support)Merck (Research Grant or Support)Pfizer (Research Grant or Support)Sanofi (Research Grant or Support) Douglas S. Swanson, MD, Merck (Research Grant or Support)Pfizer (Research Grant or Support)Sanofi (Research Grant or Support) Brian R. Lee, PhD, MPH , Merck (Grant/Research Support)Pfizer (Grant/Research Support) Christopher J. Harrison, MD, GSK (Grant/Research Support)Merck (Grant/Research Support)Pfizer (Grant/Research Support, Scientific Research Study Investigator, Research Grant or Support)

183. Decrease in Invasive Pneumococcal Disease in 7 United States Children’s Hospitals during the COVID-19 Pandemic

BackgroundDuring the 2020 SARS-CoV-2 pandemic, physical distancing and mask use guidelines were implemented resulting in a decline in the number of infections caused by influenza, respiratory syncytial virus and otitis media. A surveillance analysis from England and Taiwan showed a decline in invasive pneumococcal disease (IPD) (Clin Infect Dis. 2021;72: e65-75 and J Infect. 2021;82:296-297). We hypothesized that COVID mitigation efforts resulted in a decrease in incidence of pediatric IPD within the U.S. during 2020 compared to previous years.MethodsWe reviewed all cases of IPD among 7 children’s hospitals from the U.S. Pediatric Multicenter Pneumococcal Surveillance Group from 2017-2020. IPD was defined by the isolation of Streptococcus pneumoniae from normally sterile sites (eg. blood, cerebrospinal, pleural, synovial or peritoneal fluid). Pneumococcal pneumonia was defined as an abnormal chest radiograph in the presence of a positive blood, pleural fluid or lung culture. Mastoiditis was identified by positive middle ear, subperiosteal abscess or mastoid bone culture. Serotypes were determined by the capsular swelling method. Hospital admission numbers were obtained for incidence calculations. Statistical analyses were performed using STATA11. A p< 0.05 was considered significant.ResultsA total of 410 IPD cases were identified. The cumulative incidence of IPD (0-22 years of age) decreased from 99.2/100,000 admissions in 2017-2019 to 53.8/100,000 admissions in 2020 (risk ratio 0.54, CI: 0.40-0.72, p< 0.00001). Pneumococcal bacteremia and pneumonia decreased significantly in 2020 (p< 0.05), and although not statistically significant, there were fewer cases of meningitis and mastoiditis when compared to previous years (p=0.08) (Figure 1). Sex, race, age or presence of comorbidities were not significantly different between groups. Most common serotypes in 2020 were 35B, 3 and 15B/C (Figure 2). ConclusionThe observed decline in IPD cases during the first year of the SARS-CoV-2 pandemic is likely associated with mask use and physical distancing limiting transmission of S. pneumoniae via droplets and viral infections frequently preceding IPD. These precautions might be useful in the future to decrease IPD, especially in high-risk patients.Disclosures Sheldon L. Kaplan, MD, Pfizer (Research Grant or Support) Tina Q. Tan, MD, GSK (Individual(s) Involved: Self): Advisor or Review Panel member, Grant/Research Support; ILiAD (Individual(s) Involved: Self): Advisor or Review Panel member; Merck (Individual(s) Involved: Self): Advisor or Review Panel member, Grant/Research Support; Moderna (Individual(s) Involved: Self): Advisor or Review Panel member; Pfizer (Individual(s) Involved: Self): Advisor or Review Panel member Pia S. Pannaraj, MD, MPH, Pfizer (Grant/Research Support)Sanofi-Pasteur (Advisor or Review Panel member)Seqirus (Advisor or Review Panel member) Larry Givner, MD, AstraZeneca (Advisor or Review Panel member) Kristina G. Hulten, PhD, Pfizer (Research Grant or Support)

1181. Serotype Distribution by Age of Remaining Invasive Pneumococcal Disease After Long-Term PCV10/13 Use: The PSERENADE Project

BackgroundPneumococcal conjugate vaccines (PCV) have reduced invasive pneumococcal disease (IPD) (see other PSERENADE abstract), of which > 70% was vaccine-type pre-PCV. We described the serotype (ST) distribution of remaining IPD in countries with mature infant PCV10/13 programs.MethodsIPD ST distribution data were obtained directly from surveillance sites, supplemented with published literature. Mature programs were defined as exclusive use of PCV10 or PCV13 for at least 5-7 years (depended on if prior PCV7 use and/or PCV10/13 catch-up) with primary series uptake > 70%. The distribution was estimated using a multinomial Dirichlet regression, stratified by PCV product and age (< 5 years, ≥ 50 years).ResultsSerotyped IPD cases from 42 PCV13- (n=78,912) and 12 PCV10-using sites (n=8,429) in 41 countries were analyzed. Most sites were from high-income countries (67%) and used a booster dose schedule (81%). For low- and middle-income countries, only 5 and 7 sites had more than 20 eligible cases for children and adults, respectively. In PCV10 sites, 10.0% (95% CI: 6.3-12.9%) and 15.5% (95% CI: 13.4-19.3%) of the remaining IPD during the mature period was PCV10-type among children and adults, respectively (Figure 1). For PCV13 sites, PCV13-type was 26.4% (95% CI: 21.3-30.0%) among children and 29.5% (95% CI: 27.5-33.0%) among adults. PCV20-, PCV24-, and PPV23-type cases ranged from 62-72% across all age and PCV-use groups. ST 19A was the leading ST at PCV10 sites, though more so for children (30.6%, 95% CI: 18.2-43.1%) than adults (14.8%, 95% CI: 11.9-17.8%; Figure 2). ST 3 was a top ST in both PCV10 and PCV13 sites, causing about 9% of cases in children and 14% in adults. ST 6C was the third most common ST in PCV10 sites, causing 6% of cases in both age groups. Some top non-PCV13 STs are included in higher-valent investigational PCVs (15BC, 12F, 22F, 8, 9N) but others are not (24F, 23B, 23A, 15A). Figure 1. Percentage of IPD cases in the mature PCV10/13 period due to serotypes included in current and upcoming products. Serotype (ST) 3 is illustrated separately in lighter purple in the bars corresponding to products that include ST3 due to the uncertain effectiveness against ST3 in current products. ST6C is illustrated in grey above the bars where ST6A is included. Although ST6C is not included in PCV10 or PCV13, PCV13 offers cross-protection through ST6A. ST6A also benefits from cross-protection with ST6B, included in both PCV10 and PCV13. Therefore, ST6A causes a very small fraction of disease in both settings and age groups, and it is not shown. Confidence intervals do not include ST6C, as this serotype is not included in PCV10/13. PCV13 is Pfizer’s Prevnar13/Prevenar13; PCV10 is GSK’s Synflorix.Figure 2. Serotype-specific distribution of IPD in the mature PCV10/13 period. Serotypes are colored by the lowest valency PCV product they are included in. The “x” in the PCV legend represents the extra serotypes included in that product relative to the next lower product (i.e., PCV13x includes serotypes 3, 6A, and 19A not in PCV10). Serotype (ST) 6C is colored separately because, although it is not included in any product, it is covered through cross-protection with PCV13-type serotype 6A. PCV13 is Pfizer’s Prevnar13/Prevenar13; PCV10 is GSK’s Synflorix.ConclusionIPD due to vaccine STs was low for both children and adults in countries with mature PCV programs. ST distribution of remaining IPD differed between PCV10 and PCV13 sites and between age groups. Higher-valency PCVs under evaluation target over half of remaining IPD cases, but some prevalent STs are not included in known investigational products.Disclosures Maria Deloria Knoll, PhD, Merck (Research Grant or Support)Pfizer (Research Grant or Support)

Serotype distribution and incidence of invasive early onset and late onset group B streptococcal disease amongst infants in Singapore

BackgroundThe current group B streptococcal (GBS) preventive measures had reduced invasive GBS early onset disease (EOD) incidences worldwide, but the late onset disease (LOD) incidences had remained unchanged. Administration of a safe and effective GBS vaccine in addition to the current strategies were thought to be the next steps in reducing the incidences of invasive GBS infection especially LOD. In this study, we aimed to examine the causative GBS serotypes in invasive GBS disease, determine the incidences of EOD and LOD, and compare the risk factors between EOD and LOD.MethodsA retrospective study of infants ≤ 90-day-old over an 8-year period (2010–2017). The incidences of EOD and LOD were obtained by using patients with EOD and LOD who were born in our institution as the numerator and the live births in our institution per year of the study period as the denominator. Available GBS isolates were serotyped by the National Public Health Laboratory using capsular serotyping methods. The risk factors of EOD and LOD were compared.ResultsA total of 71 infants were identified; 16 (22.5%) and 55 (77.5%) of them had EOD and LOD, respectively. Serotype III (n = 42, 71.2%) was the most common serotype amongst the 59 isolates available for serotyping. Serotypes Ia, Ib, II, III, and V accounted for 98.3% (n = 58) of the invasive GBS diseases. The overall incidence was 0.42 per 1000 live births. The mean incidences of EOD and LOD were 0.13 per 1000 live births and 0.29 per 1000 live births, respectively. On multivariate analysis, risk factors for LOD as compared to EOD were: Chinese ethnicity (OR 27.1, 95% CI 3.0–243.1, p = 0.003) and negative/unknown maternal GBS status (OR 20.0, 95% CI 2.0–250.0, p = 0.012). Prematurity and intrapartum risk factors (peripartum maternal pyrexia, prolonged rupture of membrane) of EOD were not associated with LOD.ConclusionsThe LOD incidence had remained higher than EOD incidence in our cohort. A GBS vaccine that covers the major causative serotypes found in our cohort can potentially reduce the overall GBS disease burden in the country.

Serotyping and Antibiotic Susceptibility of Invasive Streptococcus agalactiae in Egyptian Patients with or without Diabetes Mellitus.

Streptococcus agalactiae serotype distribution and its antibiotic susceptibility affect disease prevention strategies, but the serotype distribution varies among patient groups. The objectives of this study were to establish the group B Streptococcus (GBS) serotype distribution in patients from Egypt and to assess antibiotic sensitivity of invasive GBS isolates. A total of 490 patients participated in this multicenter study; 160 had urinary tract infection, 115 complained of diabetic foot ulcers, 125 men had genital tract infections, and 30 women females had genital tract infections. Others had bronchopneumonia, otitis media, synovitis, or meningitis. Serotyping of the isolated GBS was performed at the CDC in the United States. Antibiotic sensitivity patterns were determined using the disk diffusion method. In men, the most common serotypes were II, III, and V, whereas types Ia, II, III, and V were isolated from women. Macrolides (erythromycin) resistance occurred in 4.1% of the isolates; 10.2% were resistant to both clindamycin and inducible resistance of macrolides, lincomycin, and streptogramin; 17.3% were resistant to quinolones; and 95.9% were resistant to tetracyclines. GBS primarily infected the urinary tract, skin, soft tissue, and genital tract in both genders. Isolates were sensitive to beta-lactam drugs, vancomycin, and linezolid; 14.0% were resistant to macrolides with or without clindamycin. Only 6.0% of the strains were sensitive to tetracyclines. Although GBS causes invasive infections in Egyptian adults, it rarely causes neonatal meningitis or sepsis. Future studies should determine whether GBS isolates are transmitted sexually, by performing a follow-up study of the partner of the infected patient.

Phenotypic and genotypic characteristics of non-invasive S. pneumoniae isolates recovered from PCV10-vaccinated children in Bulgaria

PurposeThe non-invasive pneumococcal disease (NIPD) is a common infection during childhood. We aimed to define the clonal spread of pediatric non-invasive isolates recovered during the PCV10-period in Bulgaria concerning the serotype and antimicrobial susceptibility. Materials and methodsSerogrouping/serotyping were performed using latex agglutination and capsular swelling reaction. Serogroup 6 strains were subjected to serotype-specific PCR's. The antibiotic susceptibilities were assessed by broth microdilution. MLST was performed to define the clonal composition. ResultsWe analyzed 154 pediatrics non-invasive S. pneumoniae isolates. The PCV10-vaccinated children were 94.1%. We disclosed 88% non-vaccine serotypes (NVTs) and 12% PCV10 - serotypes. All common serotypes among PCV10-vaccinated children (n ​= ​145) were non-vaccine types (NVTs): 19A (13.8%), 6C (11.7%), 3 (9.6%), 15A (8.3%) and 23A (5.5%). Antimicrobial non-susceptibility showed highest levels in erythromycin (50.0%), oral penicillin (49.4%), clindamycin (45.4%), trimethoprim-sulfamethoxazole (43.5%), tetracycline (42.2%), and ceftriaxone (14.3%). The multidrug-resistant strains (MDR) were 51.3%. MDR-serotypes were 6C (20.2%), 19A (17.7%), 15A (11.4%), 19F (10.1%), and 23A (8.9%). MLST presented 17 clonal complexes (CCs) with prevalence of CC320, CC386, CC505, CC8029 and CC2613 clustered 83% MDR isolates. ConclusionsAll emergent pediatric non-invasive serotypes in our geographic area during the studied PCV10-period were NVTs (19A, 6C, 3, 15A, and 23A). The fifth widespread CCs: CC320, CC386, CC505, CC8029 and CC2613 clustered 83% MDR isolates. Future surveillance of vaccine-induced changes in the clonality and the antimicrobial resistance of the pneumococcal population is needed.

Regional Variation in Dengue Virus Serotypes in Sri Lanka and Its Clinical and Epidemiological Relevance.

Dengue is a significant health concern in Sri Lanka, but diagnosis of the infecting dengue virus (DENV) serotype has hitherto been largely restricted to the Colombo district in the western province. Salinity tolerant Aedes vectors are present in the island’s northern Jaffna peninsula, which is undergoing rapid groundwater salinization. Virus serotypes were determined by RT-qPCR in 107 and 112 patients diagnosed by NS1 antigen positivity from the Jaffna district in 2018 and 2019, respectively, and related to clinical characteristics. DENV1 and DENV2 were the most common serotypes in both years. Infections with multiple serotypes were not detected. DENV1 was significantly more prevalent in 2019 than 2018, while DENV3 was significantly more prevalent in 2018 than 2019 among the Jaffna patients. Limited genomic sequencing identified DENV1 genotype-I and DENV3 genotype-I in Jaffna patients in 2018. Dengue was more prevalent in working age persons and males among the serotyped Jaffna patients. DENV1 and DENV2 were the predominant serotypes in 2019 in the Colombo district. However, DENV1 and DENV3 were significantly more prevalent in Colombo compared with Jaffna in 2019. The differences in the prevalence of DENV1 and DENV3 between the Jaffna and Colombo districts in 2019 have implications for dengue epidemiology and vaccination. Salinity-tolerant Aedes vector strains, widespread in the Jaffna peninsula, may have contributed to differences in serotype prevalence compared with the Colombo district in 2019. Significant associations were not identified between virus serotypes and clinical characteristics among Jaffna patients.

Development and Evaluation of Alternative Methods to Identify the Three Most Common Serotypes of Salmonella enterica Causing Clinical Infections in Kazakhstan.

In this study, we aimed to compare the performance of conventional PCR and real-time PCR assays as screening methods for identification of three frequent, clinically significant Salmonella serovars in Kazakhstan. We determined the diagnostic efficacy of three molecular methods for detection of S. enterica subsp. enterica and typing S. Typhimurium, S. Enteritidis, and S. Virchow. A total of 137 clinical samples and 883 food samples were obtained in Almaty in 2018–2019. All tests showed high analytical specificity for detecting S. enterica and its corresponding serovariants (100%). The sensitivity of real-time PCR for each of the tested targets was 1–10 microbial cells and in conventional PCR 10–100 microbial cells. The trials with conventional PCR and real-time PCR had a diagnostic efficacy (DE) of 100% and 99.71%, respectively. The DE of real-time PCR and conventional PCR for detecting S. Enteritidis and S. Typhimurium was 99.90%, while the DE of conventional PCR and real-time PCR for detecting S. Virchow was 99.31% and 99.80%, respectively. The RAPD-PCR analysis of the genomic DNA of Salmonella enterica showed the genetic kinship of S. Enteritidis isolates, and the genetic heterogeneity of S. Typhimurium and S. Virchow isolates. Thus, the developed methods can be considered as alternatives to classical serotyping using antisera.

Immunogenicity of Conjugated and Polysaccharide Pneumococcal Vaccines Administered During Pregnancy or Postpartum to Women With HIV.

Pneumococcal vaccination is recommended in people with HIV, prioritizing PCV. We compared the immunogenicity of PCV-10 and PPV-23 administered antepartum or postpartum. This double-blind study randomized 346 pregnant women with HIV on antiretrovirals to PCV-10, PPV-23, or placebo at 14-34 weeks gestational age. Women who received placebo antepartum were randomized at 24 weeks postpartum to PCV-10 or PPV-23. Antibodies against 7 serotypes common to both vaccines and 1 serotype only in PPV-23 were measured by ELISA/chemiluminescence; B- and T-cell responses to serotype 1 by FLUOROSPOT; and plasma cytokines/chemokines by chemiluminescence. Antibody responses were higher after postpartum versus antepartum vaccination. PCV-10 generated lower antibody levels than PPV-23 against 4 and higher against 1 of 7 common serotypes. Additional factors associated with high postvaccination antibody concentrations were high prevaccination antibody concentrations and CD4+ cells; low CD8+ cells and plasma HIV RNA; and several plasma cytokines/chemokines. Serotype 1 B- and T-cell memory did not increase after vaccination. Antepartum immunization generated suboptimal antibody responses, suggesting that postpartum booster doses may be beneficial and warrant further studies. Considering that PCV-10 and PPV-23 had similar immunogenicity, but PPV-23 covered more serotypes, use of PPV-23 may be prioritized in women with HIV on antiretroviral therapy. NCT02717494.

Age-Dependent Serotype-Associated Case-Fatality Rate in Invasive Pneumococcal Disease in the Autonomous Community of Madrid between 2007 and 2020.

The aim of this study was to investigate the serotype-associated fatality rate in cases of invasive pneumococcal disease (IPD) in the Spanish region of Madrid between 2007 and 2020. Serotyping was performed by Pneumotest Latex and the Quellung reaction using commercial antisera. Case-fatality rate was estimated as the ratio between the number of deaths at hospital discharge and the number of cases attributable to each serotype. To evaluate the association measures, the odds ratios with a 95% confidence interval were calculated. Twenty five pneumococcal serotypes were associated to mortality and comprised 87.8% of the total number of isolates characterized. Serotypes 8, 3, 19A, 1, 7F, 22F, 12F, and 11A were the most prevalent (≥3% each). Serotypes 31, 11A, and 19F were significantly associated to high case-fatality rates (>20% each). The lower significantly associated case-fatality rate (<10% each) was found in serotypes 5, 1, 12B, 7F, 12F, 8, 33, and 10A. The serotypes with higher mortality levels (≥0.04 per 100,000 population) were 11A (fatality 24.0%), 3 (fatality 18.7%), 19A (fatality 12.5%), and 8 (fatality 7.2%). Serotype 3 was worrisome because it is associated with important fatality levels combined with very high incidence and mortality rates. Serotype 11A also showed a high fatality with marked incidence and mortality levels. Some few frequent serotypes as 31, 19F, and 15A despite its high fatality had low levels of mortality. By contrast other serotypes as 8 showing low fatality had high mortality ranges because it shows a wide extended distribution. Finally, common serotypes, such as 1 and 5, presented small mortality length, due to their low case-fatality rates.

Concurrent circulation of dengue serotype 1, 2 and 3 among acute febrile patients in Cameroon.

Acute febrile patients presenting at hospitals in Douala, Cameroon between July and December 2020, were screened for dengue infections using real time RT-PCR on fragments of the 5' and 3' UTR genomic regions. In total, 12.8% (41/320) of cases examined were positive for dengue. Dengue virus 3 (DENV-3) was the most common serotype found (68.3%), followed by DENV-2 (19.5%) and DENV-1 (4.9%). Co-infections of DENV-3 and DENV-2 were found in 3 cases. Jaundice and headache were the most frequent clinical signs associated with infection and 56% (23/41) of the cases were co-infections with malaria. Phylogenetic analysis of the envelope gene identified DENV-1 as belonging to genotype V, DENV-2 to genotype II and DENV-3 to genotype III. The simultaneous occurrence of three serotypes in Douala reveals dengue as a serious public health threat for Cameroon and highlights the need for further epidemiological studies in the major cities of this region.

Report on the presence of Salmonella and Campylobacter spp. in fresh meat marketed in Italy

Abstract Introduction Salmonella (S.) and Campylobacter (C.) are the most common agents of foodborne infections and a risk for immunocompromised patients. The present study reports the prevalence and characterization of S. spp. in imported poultry, beef and pork meat marketed in Italy over six years. Furthermore, we present data of prevalence and species identification for C. spp. isolated from poultry samples collected in the years 2016 and 2018. Methods Poultry (chicken and turkey), beef and pork meat samples were collected by Health Local Authorities all over Italy. The presence of C. was tested according to the ISO 10272 and the species identification was assessed using Multiplex PCR (C. coli e C. jejuni). S. was tested according ISO 6579 and serovars were identified through serum agglutination using S. antisera. Results The prevalence of S. in chicken and turkey was respectively 1.31% (4/306) and 0% in 2016, 7.89% (24/304) and 2.33% (7/300) in 2018, 14.4% (37/257) and 3.59% (9/251) in 2020; S. was never detected in beef meat but it was found in 1.05% (3/287) of pork samples in 2015, 1.98% (5/253) in 2017, 1.74% (5/287) in 2019. Among all the available samples, the most common serotypes identified were S. Infantis and S. Bredeney. The presence of C. was detected in 4.25% (13/306) of chicken samples and in 0.74% (2/272) of turkey samples in 2016, increasing respectively to 17% (17/100) and 1.06% (1/94) in 2018. Similar frequencies were found for coli and jejuni species. Conclusions The results show that poultry meat is the most relevant source of C. and S. Uncooked or not well-cooked meat can still be a risk for the consumers, despite the adoption of biosecurity measures suited to prevent S. to enter in farms. Source attribution and prevalence studies like this can be a useful tool to set up monitoring plans, identify priorities for food safety interventions and risk communication. Key messages The crucial role of source attribution studies to help Authorities to plan further controls. Public agencies and stakeholders should give clear communication about the risk of not well-cooked meat.

Burden of Pneumococcal Meningitis and Bacteremia, Serotype Distribution and Antibiotic Resistance in Healthy Children After Conjugated Pneumococcal Vaccine Implementation: Single Center Experience

In Turkey, the seven-valent pneumococcal conjugated vaccine (PCV7) was included in the childhood national immunization programme in April 2008 and was replaced by the 13-valent pneumococcal conjugated vaccine (PCV13) in April 2011. In this retrospective, single-center study, it was aimed to determine the serotype distribution and antimicrobial resistance in Streptococcus pneumoniae isolates of pediatric patients with invasive pneumococcal disease (IPD) after the introduction of PVC7 and PVC13. Fifty pediatric patients diagnosed with meningitis and sepsis/bacteremia between October 2009 and October 2019 were included in the study. The pediatric patient group consisted of previously healthy patients diagnosed with meningitis and sepsis/bacteremia with S.pneumoniae isolated in their blood or cerebrospinal fluids. Patients with pneumonia-associated bacteremia and empyema were not included in the study. Serotyping of the isolates was performed by Quellung reaction using specific antisera (Statens Serum Institute, Denmark) and antibiotic (penicillin and ceftriaxone) susceptibility was determined by antibiotic gradient method based on Clinical Laboratory Standards Institute (CLSI) criteria. Of the children, 29 (58%) were boys and 21 (42%) were girls. The median age of the patients was 19 months (1 month-18 year). When the children under the age of five were evaluated, it was found that 30 (79%) patients were diagnosed with occult bacteremia/sepsis and 8 (21%) with meningitis. The overall annual incidence rate of IPD among the healthy children aged <5 years decreased significantly from 9.35/100000 to 0.83/100000 (p< 0.001). Serotype identification was determined for 44 of 50 pneumococcal isolates . However, since six patients with underlying disease were not included in the evaluation, the remaining 38 isolates were found to be one of the serotypes included in PCV7 and PCV13 at a rate of 28.9% (n= 11) and 44.7% (n= 17), respectively. While the rate of PCV13 serotypes seen in the PCV7 period was 81.8%, this rate decreased to 29.6% within eight years after PCV13 administration. The rate of non-vaccine serotypes was determined as 54.5% in PCV7 period and 70.3% in PCV13 period. The rate of non-vaccine serotypes in patients under 5 years was 60% in the period of PCV7 and 75% in the period of PCV13. The proportion of non-vaccine serotypes has increased over time. However, this difference was not statistically significant (p> 0.05). The most common serotypes detected in isolates were 19F, 23F, 7F, 31 and 24B. According to the minimum inhibitory concentration values of the isolates recovered from patients with meningitis, penicillin and ceftriaxone resistance rates were found as 43.9% and 9.8%, respectively. In conclusion, our study showed that there was a 91.1% decrease in the incidence of IPD in healthy children aged under five years after the implementation of PCV7 and PCV13. It was determined that while the rate of serotypes in vaccine content decreased, there was an increase in non-vaccine serotypes. In addition no significant change was observed in antibiotic resistance rates over the years.

Molecular and epidemiological characterization of human adenoviruses infection among children with acute diarrhea in Shandong Province, China

BackgroundHuman adenovirus (HAdV) had been recognized as one of the most common enteric viruses associated with acute diarrhea in children. The present study was carried out to demonstrate the molecular and epidemiological characterization of HAdV Infections among children in Shandong province in China between July 2017 and June 2018.MethodsFecal specimens were collected from children under 5 years old with acute diarrhea. DNA was extracted from the stool specimens and adenovirus DNA was detected by PCR amplification with specific primers. The amplification products were subjected to electrophoresis and visualized on a UV transilluminator. All positive RT-PCR amplification products were sequenced and the obtained sequences analyzed by MEGA (version 7.0). Demographic information and clinical manifestation data were also analyzed.ResultsIn total, 656 fecal specimens were collected and the overall positive rate of HAdV was 7.47%. HAdV infections were detected in all age groups, in which children aged 13–24 months presented the highest positive rate. Seasonal pattern could be observed with a peak in December, January and February. Diarrhea, vomiting, dehydration and fever were the main clinical manifestations, in which vomiting was the most common accompanied symptom. By phylogenetic analysis, four species (A, B, C, and F) were detected and seven different serotypes were identified. HAdV-41 (48.98%, 24/49) was the most common serotype followed by HAdV-3 (18.37%, 9/49), HAdV-31 (14.29%, 7/49), HAdV-7 (8.16%, 4/49), HAdV-40 (4.08%, 2/49), HAdV-1 (4.08%, 2/49) and HAdV-2 (2.04%, 1/49).ConclusionThis study indicated that HAdV infection was an important cause of acute diarrhea among children under 5 years old in Shandong province. The results will contribute to (a) increase understanding of the role of HAdV in diarrheal children and enhance identification of the predominant diarrhea pathogen for diagnosis; (b) avoid abuse of antibiotics; (c) monitor the change of prevalent HAdV serotypes and promote vaccine development and vaccination.

Isolation and Molecular Characterization of Fowl Adenoviruses Associated with Inclusion Body Hepatitis-hydropericardium Syndrome in Broiler Chickens in Egypt

Avian adenoviruses are an extremely diversified group of pathogens that recently triggering a variety of problems for poultry production. In particular, Inclusion Body Hepatitis-Hydropericardium Syndrome (IBH-HPS), which has been observed in broiler birds from 3 to 6 weeks of age and is associated with anemia, hemorrhagic disorders, hydropericardium, and high mortality. The disease has been reported worldwide, and recently it was reported in many Egyptian farms, causing severe economic losses. Therefore, the current study aimed to isolate, and genetically type the most common Adenovirus serotypes associated with this syndrome in Egyptian farms. A total of 50 broiler chicken farms (3-6 weeks old) located in different Egyptian governorates were examined. Macroscopically, the diseased flock revealed hydropericardium, enlarged friable livers with ecchymotic hemorrhages, and varying mortality rates (1 to 7.5%). Histopathologically, severe diffuse necrotizing enteritis, hepatitis, pericarditis, and diffuse lymphoid depletion of the spleen were the most prominent lesions. Liver tissues and cloacal swabs were collected from all examined flocks for FAdVs detection by conventional polymerase chain reaction (PCR) targeting the L1 loop in the hexon gene. The PCR products were sequenced for typing of the detected viruses. It was found that 10 out of 50 flocks examined were PCR positive for FAdVs (20%). Phylogenetic analysis of the sequenced genes revealed that the obtained viruses clustered with reference strains belonging to FAdV type D and E serotype 2, 11, and 8a respectively. The isolation of both FAdV type D and FAdV type E were carried out on a primary cell culture chicken embryo liver cell (CEL) and the presence of these viruses was confirmed by PCR after the appearance of cytopathic effect (CPE). From this study, it could be concluded that both FAdVs types D and E are the most common adenoviruses circulating in poultry farms suffering from hydropericardium and inclusion body hepatitis.

Dengue and Chikungunya virus co-infection in major metropolitan cities of provinces of Punjab and Khyber Pakhtunkhwa: A multi-center study.

Dengue has become endemic in Pakistan with annual recurrence. A sudden increase in the dengue cases was reported from Rawalpindi in 2016, while an outbreak occurred for the first time in Peshawar in 2017. Therefore, a multi-center study was carried out to determine the circulating dengue virus (DENV) serotypes and Chikungunya virus (CHIKV) co-infection in Lahore, Rawalpindi, and Peshawar cities in 2016–18. A hospital-based cross-sectional study was carried out in Lahore and Rawalpindi in 2016–18, while a community-based study was carried out in Peshawar in 2017. The study participants were tested for dengue NS1 antigen using an immunochromatographic device while anti-dengue IgM/IgG antibodies were detected by indirect ELISA. All NS1 positive samples were used for DENV serotyping using multiplex real-time PCR assay. Additionally, dengue samples were tested for CHIKV co-infection using IgM/IgG ELISA. A total of 6291 samples were collected among which 8.11% were NS1 positive while 2.5% were PCR positive. DENV-2 was the most common serotype (75.5%) detected, followed by DENV-1 in 16.1%, DENV-3 in 3.9% and DENV-4 in 0.7% while DENV-1 and DENV-4 concurrent infections were detected in 3.9% samples. DENV-1 was the predominant serotype (62.5%) detected from Lahore and Rawalpindi, while DENV-2 was the only serotype detected from Peshawar. Comorbidities resulted in a significant increase (p-value<0.001) in the duration of hospital stay of the patients. Type 2 diabetes mellitus substantially (p-value = 0.004) contributed to the severity of the disease. Among a total of 590 dengue positive samples, 11.8% were also positive for CHIKV co-infection. Co-circulation of multiple DENV serotypes and CHIKV infection in Pakistan is a worrisome situation demanding the urgent attention of the public health experts to strengthen vector surveillance.