Nasal Samples Research Articles

A COVID-19 Case: Isolated and Persistent Headache

COVID-19 effects many systems such as respiratory, nervous, renal and hematologic system. Third of patients have experienced neurological symptoms such as headache, anosmia, ageusia, and dizziness. Headache has been reported as the most common neurological manifestation but cases just headache are extremely rare. This case is a Covid-19 case with no symptoms other than sudden onset headache. The result of the nasal swab PCR sampling was negative however local infiltration areas with ground glass density in the periphery of the middle lobe, lateral segment of the both lungs were detected in the thorax CT.

Whole-Genome Deep Sequencing of the Healthy Adult Nasal Microbiome.

This study aimed to determine shifts in microbial populations regarding richness and diversity from the daily use of a popular over-the-counter nasal spray. In addition, the finding of nasal commensal bacterial species that overlap with the oral microbiome may prove to be potential probiotics for the "gateway microbiomes". Nasal swab samples were obtained before and after using the most popular over-the-counter (OTC) nasal spray in 10 participants aged 18-48. All participants were healthy volunteers with no significant medical histories. The participants were randomly assigned a number by randomizing software and consisted of five men and five women. The sampling consisted of placing a nasal swab atraumatically into the nasal cavity. The samples were preserved and sent to Northwestern University Sequencing Center for whole-genome deep sequencing. After 21 days of OTC nasal spray use twice daily, the participants returned for further nasal microbiome sampling. The microbial analysis included all bacteria, archaea, viruses, molds, and yeasts via deep sequencing for species analysis. The Northwestern University Sequencing Center utilized artificial intelligence analysis to determine shifts in species and strains following nasal spray use that resulted in changes in diversity and richness.

Diagnostic Utility of Multiplex Polymerase Chain Reaction in Patients of Clinically Suspected Pure Neuritic Leprosy by Identifying Mycobacterium leprae in Skin Biopsy Samples and Nasal Swabs.

Pure neuritic leprosy (PNL) often remains underdiagnosed due to the lack of simple, reliable diagnostic tools to detect Mycobacterium leprae. This study aimed to investigate the utility of multiplex polymerase chain reaction (MPCR) in easily accessible and less invasive biopsy sites, including skin biopsy samples and nasal swabs (NSs), to detect M. leprae. A total of 30 (N = 30) clinically suspected and untreated patients with PNL were recruited. Nasal swabs and skin biopsy samples from the innervation territory of an "enlarged nerve" were collected. DNA was extracted and subjected to MPCR (targeting leprae-specific repetitive element [RLEP], 16S rRNA, and SodA genes) and RLEP-PCR (individual gene PCR). The PCR products were analyzed by 3% agarose gel electrophoresis. In 30 patients with clinically suspected PNL, 60% (N = 18) of skin biopsy samples and 53% (N = 16) of NSs were found positive for M. leprae DNA by MPCR, whereas only 23.3% (N = 7) of skin biopsy samples and 10% (N = 3) of NSs were found positive by RLEP-PCR. MPCR demonstrated a greater positivity rate than did RLEP-PCR for detection of M. leprae. Serologic positivity for anti-natural disaccharide-octyl conjugated with bovine serum albumin (ND-O-BSA) antibodies was 80% (16/20), including 35% (7/20) of PNL patients for which the skin MPCR was negative. Both serologic positivity and skin MPCR positivity were observed in 65% of patients (N = 20). Multiplex polymerase chain reaction is a useful tool for detection for M. leprae in skin biopsy samples and NSs in clinically suspected cases of PNL, with the added advantages of being less invasive and technically easier than nerve biopsy.

Serum IgG Immunoglobulin Levels are Associated with Reduced PCR Detection of Mycoplasma bovis in Naturally Infected American Bison (Bison bison).

Mycoplasma bovis (M. bovis) is an important pathogen of American bison (Bison bison), associated with high morbidity and mortality epizootics of respiratory and reproductive disease. Despite the significant negative impact on bison health, little is known about the kinetics of disease and the host immune response to infection. To address these questions, a cohort of bison calves was created and serially sampled 5 times, once every 2-3 mo, over a 12-mo period. At each sampling period nasal swab samples were collected and tested by PCR for the presence of M. bovis. Serum samples were also collected and assessed for M. bovis-specific antibodies using both a commercial and an in-house ELISA. Overall, 19/41 bison (46.3%) had positive PCR tests, and 31/41 (75.6%) were seropositive. Over the course of the study, the frequency of PCR-positive nasal swabs and the ELISA scores decreased, although serum samples remained positive for at least 6 mo following the final positive PCR test. Bison were grouped according to results from the in-house ELISA into high-responder (n=7), low-responder (n=5), and seronegative (n=7) groups. M. bovis-specific IgG antibody levels were significantly elevated in the high-responder group compared to the low-responder and seronegative groups. The differences were statistically significant for 3/5 sampling periods. A trend toward increased IgG2 levels was observed in the high-responder group. High total IgG responses correlated with a decline in positive PCR tests from nasal swabs. These data provide evidence that a strong humoral response is beneficial and is probably involved in the clearance of M. bovis from bison.

Evaluation of Non-Invasive Sampling Techniques for the Molecular Surveillance of Equid Herpesviruses in Yearling Horses.

Equid alphaherpesvirus 1 (EHV-1) is a highly contagious respiratory tract pathogen of horses, and infection may be followed by myeloencephalopathy or abortion. Surveillance and early detection have focused on PCR assays using less tolerated nasal swabs. Here, we assess non-invasive non-contact sampling techniques as surveillance tools in naturally equid gammaherpesvirus 2-shedding horses as surrogates for EHV-1. Horses were individually housed for 10 h periods on 2 consecutive days. Sampling included nasal swabs, nostril wipes, environmental swabs, droplet-catching devices, and air sampling. The latter was completed via two strategies: a combined air sample collected while going from horse to horse and a collective air sample collected at a stationary central point for 6 h. Samples were screened through quantitative PCR and digital PCR. Nine horses on day 1 and 11 horses on day 2 were positive for EHV-1; overall, 90.9% of the nostril wipes, 81.8% of the environmental surfaces, and 90.9% of the droplet-catching devices were found to be positive. Quantitative analysis showed that the mean DNA copies detection per cm2 of nostril wipe sampled concentration (4.3 × 105 per day) was significantly (p < 0.05) comparable to that of nasal swabs (3.6 × 105 per day) followed by environmental swabs (4.3 × 105 per day) and droplet catchers (3.5 × 103 per day), respectively. Overall, 100% of the air samples collected were positive on both qPCR and dPCR. In individual air samples, a mean concentration of 1.0 × 104 copies of DNA were detected in per m3 air sampled per day, while in the collective air samples, the mean concentration was 1.1 × 103. Environmental samples look promising in replacing direct contact sampling. Environmental and air sampling could become efficient surveillance tools at equestrian events; however, it needs threshold calculations for minimum detection levels.

Screening and validation of therapeutic targets for chronic sinusitis with nasal polyps based on proteomics

Objective: To identify potential therapeutic targets of chronic sinusitis with nasal polyps (CRSwNP) through proteomics screening of and verify its effectiveness experimentally. Methods: The nasal tissue samples were collected from patients undergoing surgical treatment in the Department of Otorhinolaryngology, Head and Neck Surgery in Yuhuangding Hospital of Yantai from June 2010 to December 2021, including 69 patients with CRSwNP and 39 patients in the control group. Tissue samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in data-independent acquisition (DIA) mode to find differentially expressed proteins. Bioinformatics tools were employed to analyze the functions of differentially expressed proteins. The expression of hematopoietic cell kinase (HCK) in nasal tissues of patients with CRSwNP was further confirmed by qPCR and western blot. The mouse model of CRSwNP was established and treated with HCK inhibitor. The levels of inflammatory factors IgE, IL-4 and IL-5 in serum of CRSwNP mice, both treated and untreated with HCK inhibitors, were detected by enzyme-linked immunosorbent assay (ELISA) across different experimental groups. The experimental data were analyzed by Graphpad Prism 9 software. Results: DIA analysis identified 1 850 differential proteins, including 760 up-regulated proteins and 1 090 down-regulated proteins. Weighted correlation network analysis (WGCNA) correlation analysis of phenotypic data such as cell count and CT score with the results of genomics indemnified 575 proteins of MEBrown module which intersected with 35 kinases further screened from 1 850 differential proteins, yielding eight protein kinases: HCK, SYK, PDK2, FGR, PRKCB, ROR1, CAMK1 and GRK6. qPCR showed that the expression of HCK in CRSwNP was significantly higher than that in the control group (P<0.05). Further experiments in mice confirmed that the secretion of IgE, IL-4 and IL-5 in the serum of CRSwNP group was significantly higher than the control group (all P<0.05), indicating successful model establishment. The intervention of HCK significantly decreased the secretion of IgE, IL-4 and IL-5 in serum of mice (all P<0.05). Conclusion: The HCK inhibitor can reduce the inflammatory index of mice with CRSwNP, and HCK is a potential therapeutic target of CRSwNP.

Postoperative complications and mortality after thoracic surgery of COVID-19 omicron hospital infections: A retrospective multicenter study

BackgroundPostoperative complications and mortality of Omicron hospital infection, who have undergone thoracic surgery, are scarce. MethodsBased on real-time reverse transcription polymerase chain reaction results in-hospital using nasal or pharyngeal swab samples, the patients who had undergone thoracic surgery were categorized into two groups: G1 (Omicron hospital infection) group and G2 (non- Omicron hospital infection) group. The occurrence of postoperative complications was assessed using the Clavien–Dindo classification. Both groups were compared using propensity score matching (PSM) to evaluate the occurrence of complications and mortality. The trail was enrolled in the clinical trail (NCT06040606). Results119 patients (G1, 53 and G2, 66) were included. The occurrence of postoperative complications was significantly higher in G1 group than G2 group (37.7% vs. 10.6%, respectively) (P < 0.001) and the mortality also increased (7.5% vs. 0). The logistic regression analysis identified Omicron hospital infection as the risk factors for postoperative complications (95% CI, 3.989 to 33.858). The PSM analysis indicated significant increase in postoperative complications and mortality in the G1 vs. G2 group (P < 0.01, NA). But, there was no significant difference in the occurrences of grade ≥3 complications and mortality between lung surgery and upper gastrointestinal operation, as the PSM analysis. ConclusionThe occurrences of complications and mortality significantly increased in thoracic surgery recipients with Omicron hospital infection.

Investigation of the Use of Environmental Samples for the Detection of EHV-1 in the Stalls of Subclinical Shedders.

In populations of healthy show horses, the subclinical transmission and circulation of respiratory pathogens can lead to disease outbreaks. Due to recent outbreaks of equine herpesvirus-1 myeloencephalopathy (EHM) in the USA and Europe, many show organizers have instituted various biosecurity protocols such as individual horse testing, monitoring for early clinical disease and increasing hygiene and cleanliness protocols. The aim of this study was to determine the accuracy of detecting EHV-1 in the various environmental samples collected from the stalls of subclinical shedders. Four healthy adult horses were vaccinated intranasally with a modified-live EHV-1 vaccine in order to mimic subclinical shedding. Three additional horses served as non-vaccinated controls. All the horses were stabled in the same barn in individual stalls. Each vaccinated horse had nose-to-nose contact with at least one other horse. Prior to the vaccine administration, and daily thereafter for 10 days, various samples were collected, including a 6" rayon-tipped nasal swab, an environmental sponge, a cloth strip placed above the automatic waterer and an air sample. The various samples were processed for nucleic acid purification and analyzed for the presence of EHV-1 via quantitative PCR (qPCR). EHV-1 in nasal secretions was only detected in the vaccinated horses for 1-2 days post-vaccine administration. The environmental sponges tested EHV-1 qPCR-positive for 2-5 days (median 3.5 days) in the vaccinated horses and 1 day for a single control horse. EHV-1 was detected by qPCR in stall strips from three out of four vaccinated horses and from two out of three controls for only one day. EHV-1 qPCR-positive air samples were only detected in three out of four vaccinated horses for one single day. For the vaccinated horses, a total of 25% of the nasal swabs, 35% of the environmental stall sponges, 7.5% of the strips and 7.5% of the air samples tested qPCR positive for EHV-1 during the 10 study days. When monitoring the subclinical EHV-1 shedders, the collection and testing of the environmental sponges were able to detect EHV-1 in the environment with greater frequency as compared to nasal swabs, stationary strips and air samples.

Identification of bla OXA-23 gene in resistant Pseudomonas aeruginosa strains isolated from cows and humans in Basra province, Iraq.

Pseudomonas aeruginosa is an infectious agent of great importance for animals and humans. It causes serious infections that show high resistance to antibiotics. This study investigated the molecular detection of blaOXA-23 gene in antibiotic-resistant P. aeruginosa strains isolated from cows and humans. In total, 120 samples, comprised 60 from cows (30 milk and 30 nasal discharge) and 60 from their owners (30 urine and 30 sputum), were individually collected, cultured, and tested for P. aeruginosa through molecular analysis targeting the blaOXA-23 gene. P. aeruginosa antibiotic-resistant isolates were identified by performing antibiotic susceptibility testing and detecting biofilm formation. In total, 74.17% positive P. aeruginosa isolates, including 66.67% and 81.67% for cows and humans, respectively. Subsequently, positive cow isolates were detected in 60% of milk samples and 73.33% of nasal discharge samples; while positive human isolates were detected in 76.67% of urine samples and 86.66% of sputum samples. Targeting blaOXA-23 gene, 58.43% of cultured isolates were positive for P. aeruginosa by polymerase chain reaction. Respectively, positive isolates were detected in 66.67% and 45.46% of cow milk and nasal discharges as well as in 60.87% and 61.54% of human urine and sputum. The antibiotic susceptibility test revealed that all isolates were resistant to all applied antibiotics, particularly imipenem. Results of biofilm formation revealed 67.31% total positives, including 51.43% strong, 34.285% moderate, and 14.285% weak reactions. In addition, although values of the total positive cows and humans differed insignificantly, total positives showed insignificant variation between values of milk and nasal discharges of cows as well as between urine and sputum of humans; however, significant differences were identified in the distribution of strong, moderate, and weak positivity of these samples. Antibiotic overuse contributes extensively to increasing the prevalence of resistant P. aeruginosa isolates carrying the blaOXA-23 gene in both cows and humans. Furthermore, studies in other Iraqi areas are necessary to support our findings. The main limitations include that the number of tested samples is relatively low, and there is a need to use a large number of samples from different sources. Also, the current methods for detection of resistant isolates are still culture-based approaches.

Molecular Identification of Mycobacterium leprae in the Leprosy Patients.

Several discoveries about leprosy indicate that Mycobacterium leprae transmission mainly occurs by inhalation, and the nose is a major port of entry and exit. Molecular probes have shown certain potential for the detection and identification of M. leprae in patients. The aim of this study was to identify M. leprae in nasal swab specimens using polymerase chain reaction (PCR)-based assays followed by gene sequencing methods. This observational study examines 64 anterior nasal swab samples taken from pretreatment leprosy patients, on-treatment and completed leprosy treatment in Bulukumba, South Sulawesi, Indonesia. samples were analyzed by molecular detection methods according to the standard methods at the Clinical Microbiology Laboratory of Hasanuddin University. Descriptive statistics were utilized to summarize patient demographics and outcomes. This study uses PCR to detect the M. leprae deoxyribonucleic acid (DNA) from nasal swab specimens. Data were collected from 64 patients with a percentage of male patients 51.54%. Based on the age category, the group 45-46 years was the most frequent (39.05%). PCR detection proline-rich antigen gene of a 531 bp DNA fragment from M. leprae, was positive in eight patients, and they were multibacillary. Furthermore, PCR was positive in 5 (31.25%) of 16 new leprosy patients, 2 (8.69%) of 23 on-treatment patients, and 1 (4%) of 25 treatment completed patients. Based on the results of the phylogenetic tree and analysis of 8 positive results detected by M. leprae from leprosy patients, almost all samples have a level of similarity, except for sample Ua7. M. leprae cannot grow in vitro, so molecular diagnostic tools were used to confirm the disease. This study predominantly of males with the age above 45 years of age being the most common. Eight M. leprae were positive from nasal swab leprosy patients. The sequencing findings provide insight into the genetic diversity of the genus M. leprae, so it is necessary to consider the detection of whole-genome sequence.

Association between suction tip culture and surgical site infection in elective clean orthopedic lower limb surgeries

Background: Surgical site infections (SSIs) can be caused by various factors, including contamination of the surgical field. Perioperative cultures can be employed in clean orthopedic procedures to detect bacterial contamination that may occur during surgery. Aims and Objectives: The study aimed to evaluate the correlation between suction tip cultures and surgical wounds in clean orthopedic surgeries. Materials and Methods: A cross-sectional study was conducted at Yenepoya Medical College Hospital, Mangalore, Karnataka, with 250 patients in whom drain tips were sent for culture and followed up for 3 months to assess for SSI. Skin commensals from the nasal and groin samples were analyzed for the presence of bacteria. Results: A total of 250 patients meeting the inclusion and exclusion criteria were included. Thirty patients were positive for drain tip culture, and 12 (4.8%) patients had SSI during the 3rd month follow-up period, showing a statistically significant relationship between drain tip culture and SSIs (P=0.001). Skin commensals constituted 67% (nasal) and 100% in the groin of perioperative contaminants, accounting for 4.8% of SSIs. Conclusion: Suction drain tip culture and skin commensal analysis may be good predictors of SSIs. Intraoperative surgical site contaminants can be identified using perioperative cultures. Timely administration of suitable antibiotics and local wound care for perioperative contamination can help minimize the incidence of SSI.

A Novel Multiplex LAMP Assay for the Detection of Respiratory Human Adenoviruses.

Human adenoviruses (HAdVs) are common pathogens that are associated with a variety of diseases, including respiratory tract infections (RTIs). Without reliable, fast, and cost-effective detection methods for HAdVs, patients may be misdiagnosed and inappropriately treated. To address this problem, we have developed a multiplex loop-mediated isothermal amplification (LAMP) assay for the detection of the species Human adenovirus B (HAdV-B), Human adenovirus C (HAdV-C) and Human adenovirus E (HAdV-E) that cause RTIs. This multiplexing approach is based on the melting curve analysis of the amplicons with a specific melting temperature for each HAdV species. Without the need for typing of HAdVs, the LAMP results can be visually detected using colorimetric analysis. The assay reliably detects at least 375 copies of HAdV-B and -C and 750 copies of HAdV-E DNA per reaction in less than 35 min at 60 °C. The designed primers have no in silico cross-reactivity with other human respiratory pathogens. Validation on 331 nasal swab samples taken from patients with RTIs showed a 90-94% agreement rate with our in-house multiplex quantitative polymerase chain reaction (qPCR) method. Concordance between the quantitative and visual LAMP was 99%. The novel multiplexed LAMP could be an alternative to PCR for diagnostic purposes, saving personnel and equipment time, or could be used for point-of-care testing.

Upper Airway Alarmin Cytokine Expression in Asthma of Different Severities.

Background: The secretion of alarmin cytokines by epithelial cells, including thymic stromal lymphopoietin (TSLP), interleukin (IL)-25, and IL-33, initiates inflammatory cascades in asthma. However, alarmin cytokine expression in the upper airways in asthma remains largely unknown. Methods: We recruited 40 participants with asthma into four groups as per the Global Initiative for Asthma (GINA) steps (10 in each group of GINA 1/2, 3, 4, and 5). Cells were derived from nasal, buccal, and throat brushings. Intracellular cytokine expression (TSLP, IL-25, and IL-33) was assessed by flow cytometry in cytokeratin 8+ (Ck8+) epithelial cells immediately following collection. Results: TSLP was significantly increased (p < 0.001) in GINA 5 patients across nasal, buccal, and throat Ck8+ epithelial cells, while IL-25 was elevated in nasal and throat samples (p < 0.003), and IL-33 levels were variable, compared with GINA 1-4 patients. Individual GINA subgroup comparison showed that TSLP levels in nasal samples from GINA 5 patients were significantly (p = 0.03) elevated but did not differ between patients with and without nasal comorbidities. IL-25 and IL-33 (obtained from nasal, buccal, and throat samples) were not significantly different in individual groups. Conclusions: Our study demonstrates for the first time that Ck8+ nasal epithelial cells from GINA 5 asthma patients express elevated levels of TSLP.

Pathogenic and Antibiotic-Resistance Genes of Pasteurella multocida Isolated from Goats in the Mekong Delta, Vietnam

Pasteurella multocida (P. multocida) is one of the predominant pathogens that mostly cause respiratory diseases in domestic animals, such as goats. To determine P. multocida serotypes and the prevalence of pathogenic and antibiotic-resistance genes the PCR method was used. A total of 143 isolated P. multocida strains were collected from 289 healthy hybrid Boer-Saanen goats’ nasal samples in the Mekong Delta, Vietnam, from March to June 2023. A total of 143 P. multocida strains, serotype B accounted for the highest proportion (51.05%), followed by serotype A (14.69%), and the lowest was serotype E (0.70%) while (39.86%) of strains could not be determined serotypes. Among the six virulence genes surveyed, the sodA gene (56.64%) had the highest presence, while the ompH gene (4.20%) had the lowest presence. Pathogenic genes were present mainly in serotypes A and B; tbpA was frequently detected in serotype A (66.67%), and sodA was commonly detected in serotype B (56.16%). There were 14 virulence gene combinations in 59/109 (54.13%) serotyped P. multocida strains, and the pattern of sodA + toxA + tbpA was prevalent at the highest rate (12.84%). Moreover, among the eight investigated antibiotic resistance genes, the sulII gene had the highest presence rate (74.13%), compared to the tetA gene with the lowest presence rate (13.29%). Gene sulII was mainly detected on strains belonging to serotypes A (80.95%), B (83.56%), and F (77.78%). A total of (77.98%) of serotyped P. multocida strains indicated multi-harbor from two to six antibiotic-resistance genes, and the most common pattern was aadB + sulII (10.09%). The prevalence of five pathogenic P. multocida serotypes harboring diverse antibiotic-resistance genes isolated from nasal samples could be a critical issue in treating and preventing the respiratory diseases caused by P. multocida in goats in the Mekong Delta.

Does the Addition of Oral Sampling Improve Waveform Capnography When Compared to Traditional Nasal Sampling in Open-Airway Patients?

BackgroundEnd-tidal carbon dioxide (EtCO2) is an invaluable anesthesia measure due to minimal delay in monitoring ventilation. Oral and Maxillofacial Surgery (OMS) presents special challenges because oral exhalations are not sampled effectively via nasal cannula normally used to avoid interfering with procedures. PurposeThe purpose of the study was to compare EtCO2 waveforms obtained in subjects using nasal monitoring and combined nasal and oral monitoring under simulated ventilatory conditions. Study design, setting, sampleA single-blinded, randomized crossover controlled study was conducted at the University of Illinois Chicago with healthy volunteers, who were blinded to the monitoring system used. Inclusion criteria required subjects be >18 years of age, be consentable and English-speaking. Exclusion criteria required no airway abnormalities, no edentulism, no conditions that preclude undergoing an OMS procedure under sedation. Predictor variableThe primary predictor variable was the EtCO2 monitoring system: nasal with a standard nasal cannula, and combined with a nasal cannula and oral device. The secondary predictor variable evaluated three ventilatory states (nasal breathing, oral breathing, apnea) in each arm, standardizing by maintaining consistent breath counts across observation periods. Main outcome variableThe main outcome variable was the number of waveforms recorded over 30 seconds for nasal and combined monitoring. Each recording was standardized by dividing it by the baseline waveforms and multiplying by 20. Higher values, closer to baseline, were considered more accurate depictions of ventilation. CovariatesCovariates were body mass index (BMI), age, sex, Mallampati score, and breathing method. AnalysesANOVA, ANCOVA, and MANOVA were performed. P-value<0.05 was considered statistically significant. ResultsThe sample included 25 subjects (18 male, 7 female) with mean age 29.9+7.98 years and BMI 23.9+3.51 kg/m2. Combined sampling detected more waveforms during nasal and oral breathing (p<0.001). Higher BMI correlated with reduced waveform capture during nasal sampling in oral breathing (p=0.013). Combined sampling detected more waveforms during nasal breathing (p=0.005) in subjects with BMI < 23.5 kg/m2. Mallampati score correlated with increased waveforms during nasal breathing. Conclusions and RelevanceOral EtCO2 sampling may improve accuracy of capnography waveform capture. Further clinical studies in sedated subjects undergoing OMS procedures are needed.

Pneumococcal Colonization in Children With Persistent Asthma: A Retrospective Cohort.

Asthma is the most common chronic medical condition among children ≥5 years of age with invasive pneumococcal disease. How asthma or its management affects pneumococcal colonization is not fully understood. Our objective was to compare pneumococcal colonization rates between children with persistent asthma and children without asthma, and to characterize the pneumococcal serotype distribution. We used nasal mid-turbinate samples obtained per routine care from 5- to 18-year-old children with upper respiratory symptoms from November to April (respiratory seasons) of 2017 to 2018 and 2018 to 2019 in Kansas City, United States. Pneumococcal immunization status, prior antibiotic use and other clinical data were collected. Samples were tested for pneumococcal colonization by real-time polymerase chain reaction targeting lytA gene. Positive samples underwent multiplex serotype-specific polymerase chain reaction assays to determine the serotype. Of 363 children (120 with persistent asthma and 243 without asthma), 87.6% were 5 to 10 years old, 50.1% were female and 74.1% received ≥3 doses of a pneumococcal conjugate vaccine. The pneumococcal colonization rate was lower in children with persistent asthma than in children without asthma (10% versus 18.9%, P = 0.03). The odds of colonization were lower in children with persistent asthma [OR 0.4 (95% confidence interval: 0.2-0.9)] after adjusting for demographic and clinical data. Pneumococcal serotype was confirmed in 77.6% of positive samples; 35.6% of those samples corresponded to PCV13 serotypes and 64.4% to non-PCV13 serotypes. The most common serotypes were 19F (15%), 3 (13%) and 6C/6D (11%). Children with persistent asthma had lower rates of pneumococcal colonization than children without asthma when seeking care for respiratory symptoms.

Immunological characteristics of CD103+CD161+ T lymphocytes on chronic rhinosinusitis with nasal polyps

Chronic rhinosinusitis with nasal polyps (CRSwNPs) is a heterogeneous disease characterized by local inflammation of the upper airway and sinus mucosa. T cell-mediated immune responses play irreplaceable roles in the pathogenesis of nasal polyps. CD161+ T cells have been implicated in the pathology of several diseases through cytokine production and cytotoxic activity. However, the immunological characteristics of CD161+ T cells in nasal mucosa are still not well understood, particularly in CRSwNPs. Our research revealed a notable enrichment of CD161+ T cells in nasal tissues compared to peripheral blood, with a significantly more infiltration of CD161+ T cells in CRSwNPs compared to control nasal samples. Phenotypical analysis found that CD161+ T cells predominantly co-expressed tissue-resident memory surface markers CD103, CD69, and CD45RO. CD161+CD103+ T cells demonstrated complicated effector functions, marked by elevated levels of PD-1, CTLA-4, IL-17, and IFN-γ and diminished expression of FoxP3 and CD25. Interestingly, despite CD161+ T cells was more abundant in polyp tissues compared to normal control tissues, and then further categorizing polyp samples into distinct groups based on clinical characteristics, only the recurrent CRSwNP group showed a significant reduction in CD161+CD8+ T cells compared to the primary CRSwNP group. This finding suggested the necessity for further research to comprehensively understand the underlying mechanisms and the broader significance of CD161+ T cells in the advancement and relapse of CRSwNPs.

Evaluation of bovine coronavirus in Korean native calves challenged through different inoculation routes

Bovine coronavirus (BCoV) is a pneumoenteric virus that can infect the digestive and respiratory tracts of cattle, resulting in economic losses. Despite its significance, information regarding BCoV pathogenesis is limited. Hence, we investigated clinical signs, patterns of viral shedding, changes in antibody abundance, and cytokine/chemokine production in calves inoculated with BCoV via intranasal and oral. Six clinically healthy Korean native calves (< 30 days old), initially negative for BCoV, were divided into intranasal and oral groups and monitored for 15 days post-infection (dpi). BCoV-infected calves exhibited clinical signs such as nasal discharge and diarrhea, starting at 3 dpi and recovering by 12 dpi, with nasal discharge being the most common symptoms. Viral RNA was detected in nasal and fecal samples from all infected calves. Nasal shedding occurred before fecal shedding regardless of the inoculation route; however, fecal shedding persisted longer. Although the number of partitions was very few, viral RNA was identified in the blood of two calves in the oral group at 7 dpi and 9 dpi using digital RT-PCR analysis. The effectiveness of maternal antibodies in preventing viral replication and shedding appeared limited. Our results showed interleukin (IL)-8 as the most common and highly induced chemokine. During BCoV infection, the levels of IL-8, monocyte chemoattractant protein-1, and macrophage inflammatory protein-1β were significantly affected, suggesting that these emerge as potential and reliable biomarkers for predicting BCoV infection. This study underscores the importance of BCoV as a major pathogen causing diarrhea and respiratory disease.

Absence of SARS-CoV-2 in wildlife of northeastern Minnesota and Isle Royale National Park.

We investigated the presence of SARS-CoV-2 in free-ranging wildlife populations in Northeastern Minnesota on the Grand Portage Indian Reservation and Isle Royale National Park. One hundred twenty nasal samples were collected from white-tailed deer, moose, grey wolves and black bears monitored for conservation efforts during 2022-2023. Samples were tested for viral RNA by RT-qPCR using the CDC N1/N2 primer set. Our data indicate that no wildlife samples were positive for SARS-CoV-2 RNA. Continued surveillance is therefore crucial to better understand the changing landscape of zoonotic SARS-CoV-2 in the Upper Midwest.

Prevalence of coronaviruses in European bison (Bison bonasus) in Poland

Coronaviruses have been confirmed to infect a variety of species, but only one case of associated winter dysentery of European bison has been described. The study aimed to analyze the prevalence, and define the impact on the species conservation, the source of coronavirus infection, and the role of the European bison in the transmission of the pathogen in Poland. Molecular and serological screening was performed on 409 European bison from 6 free-ranging and 14 captive herds over the period of 6 years (2017–2023). Presence of coronavirus was confirmed in one nasal swab by pancoronavirus RT-PCR and in 3 nasal swab samples by bovine coronavirus (BCoV) specific real time RT-PCR. The detected virus showed high (> 98%) homology in both RdRp and Spike genes to BCoV strains characterised recently in Polish cattle and strains isolated from wild cervids in Italy. Antibodies specific to BCoV were found in 6.4% of tested samples, all originating from free-ranging animals. Seroprevalence was higher in adult animals over 5 years of age (p = 0.0015) and in females (p = 0.09). Our results suggest that European bison play only a limited role as reservoirs of bovine-like coronaviruses. Although the most probable source of infections in the European bison population in Poland is cattle, other wild ruminants could also be involved. In addition, the zoonotic potential of bovine coronaviruses is quite low.