Molecular Typing of Adenoviruses Associated with Respiratory Illness Among Humans and Poultry, Pakistan

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Robie, Emily S. Bailey, Nazish Badar, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5811360/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Adenoviruses have caused epidemics among both humans and poultry in Pakistan. There is growing evidence that adenoviruses sometimes spillover to infect other species. In this pilot study, we sought to study the distribution of circulating adenovirus types among humans, poultry and other animals in several areas of Pakistan. Methods From February 2019 to March 2021, patients with influenza-like illness or pneumonia in seven hospitals near Islamabad were invited to participate in this study. Volunteers permitted the collection of an oropharyngeal or a nasopharyngeal swab and epidemiological data. Concomitantly, in Islamabad’s live bird markets and commercial or backyard farms, owners granted permission for oral or nasal swabs of their livestock with signs of respiratory illness to be studied. These specimens were screened with molecular assays for evidence of adenovirus infection. Results Among the 1705 samples collected, 96 (8.8%) of 1084 humans and 15 (4%) of 385 poultry had molecular evidence of adenovirus infection. The odds ratio (OR) of such molecular detections was greatest among participants with wheezing (OR = 10.9, 95% CI 6.0-19.7), coughing (OR = 3.3, 95% CI 1.8–5.8), fever (OR = 3.2, 95% CI1.8-5.7) or sore throat (OR = 3.2, 95% CI 1.8–5.6) compared to nasal congestion. Similarly, odds of positivity were greatest for participants from Sindh (OR = 6.4, 95% CI 2.3–18.0), Baluchistan (OR = 6.4, 95% CI2.3-18.0), Azad Jammu and Kashmir (OR = 4.8, 95% CI, 1.3–16.9), or Federal Capital regions (OR = 3.6, 95% CI, 1.4–9.6) compared to Punjab. Partial hexon gene sequencing identified co-circulation of HAdV-7 and HAdV-3 strains among humans. Further sequencing of adenoviral DNA polymerase identified HAdV-C1, HAdV-5, HAdV-89, HAdV-12, and HAdV-56. Notably, we detected molecular evidence of bovine adenovirus 2 in a sick human’s nasal swab. Additionally, FAdV-11 and FAdV-4 strains were identified among poultry swab specimens. Conclusion Compared to other studies, there was a relatively high prevalence of adenoviruses among sick humans and poultry, with the unusual discovery of molecular evidence of bovine adenovirus in a sick human’s airway. There was considerable diversity among detected adenovirus strains. As adenovirus epidemics have periodically occurred in Pakistan, assessing adenovirus prevalence and genotype distributions is prudent, especially among humans with respiratory illnesses. Adenovirus respiratory infection epidemiology phylogenetic analyses Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Background Adenoviruses are non-enveloped, double-stranded DNA viruses that belong to the Adenoviridae family. The viral capsid comprises an icosahedral hexon, penton, and fibre protein structure. Hypervariable regions of hexon and fibre genes are frequently studied to help classify various adenovirus types [ 1 – 3 ]. Isolated initially from human adenoid glands in 1953 [ 4 , 5 ], the Adenoviridae viral family is subdivided into six recognized genera with numerous species in each genus by the International Committee for the Taxonomy of Viruses. Adenoviruses are known to infect nearly all vertebrate species and are often species-specific. Within the Mastadenovirus genus, human adenoviruses (HAdV) are made up of more than 110 known types separated into seven species (A – G) according to their serological and molecular properties [ 6 ]. HAdV-D has the most members amongst HAdV, while HAdV-C infections are most prevalent among those suffering from respiratory tract infections [ 7 , 8 ]. HAdVs are commonly associated with upper respiratory illness but can manifest in many illnesses, including gastroenteritis, conjunctivitis, and pneumonia [ 9 ]. Many of these will resolve with minimal concern; however, infants, older persons, and immunocompromised populations face a heightened risk of mortality, particularly from pneumonia and diarrhoea [ 10 – 12 ]. HAdVs are responsible for about 5% -10% lower respiratory tract infections in children and newborns [ 13 ]. Transmission primarily occurs through direct contact, aerosols, fomites, and the fecal-oral route, with some infected individuals shown to shed the virus for several weeks. Endemic HAdV infection has historically been a concern for populations living in crowded conditions, like military recruits. However, conjunctival adenovirus and pneumonia outbreaks have been rising in recent years among noncrowded populations [ 14 ], leading to concerns regarding emerging virulent strains. Adenoviruses are thought to have contributed to large outbreaks of human conjunctivitis in 2023 in Pakistan [ 15 ]. Furthermore, adenoviruses infect many other vertebrate hosts, including horses, cattle, pigs, reptiles, and fish. Though many mammals are susceptible to adenoviruses in the Mastadenovirus genus, human adenoviruses are rarely pathogenic to animals. Nevertheless, evidence of occasional cross-species transmission of adenoviruses has been described [ 16 ]. Similarly, antibodies to canine, bovine, and simian adenoviruses have been detected in humans, and HAdV-12 has been identified in simian species [ 17 ]. Some have wondered if adenovirus may be a pandemic threat [ 18 ]. Adenoviruses infect avian species across the Aviadenovirus, Siadenovirus, and Atadenovirus genera [ 19 ]. Acute disease manifestations, such as inclusion body hepatitis and hydropericardium syndrome, can cause rapid mortality, leading to significant economic losses in poultry industries [ 20 ]. For example, an adenovirus outbreak in broiler farm chicks in Pakistan in 1985 resulted in mortality rates of 30–60% [ 21 ]. Pathogenic adenovirus strains have been identified globally, often displaying seasonal variations. These viruses threaten human and animal populations and have shown evidence of recombining into novel forms. Underscoring the importance of surveillance in monitoring circulating adenovirus strains within specific regions [ 22 ]. In Pakistan, reports indicate an increasing [ 23 ] incidence of adenovirus infections across various areas [ 23 , 24 ]. However, comprehensive epidemiological data remain scarce despite this apparent rise, highlighting a gap in understanding the prevalence and distribution of the virus. To address this, we conducted a pilot study investigating adenovirus epidemiology among humans and livestock in and around Islamabad, Pakistan. Methods Sample Collection Between February 2019 and March 2021, a total of 1,705 samples were collected from humans (1,084), poultry (385), and livestock (236) presenting with respiratory symptoms and illness. Human samples were obtained from seven hospitals participating in the influenza surveillance network across regions and cities of Pakistan: Jinnah Hospital, Lahore; Nishter Medical Complex Hospital, Multan; and Allied Hospital, Faisalabad (Eastern Pakistan); Saidu Sharif Hospital, Swat (Northwestern Pakistan); Civil Hospital, Karachi (Southern Pakistan); Bolan Medical Complex Hospital, Quetta (Southwestern Pakistan); and Provincial Headquarters Hospital, Gilgit (Northern Pakistan) (Fig. 1 ). Sampling followed a non-probability convenience approach, targeting patients who met the World Health Organization’s case definition for influenza-like illness (ILI) or severe acute respiratory infection (SARI). [ 25 ]. Outpatients and hospitalized patients of all ages were eligible to participate if they presented with acute respiratory symptoms, a temperature exceeding 38°C, and a cough onset within the past ten days. Participants were asked to complete a brief epidemiological survey and consent to collect an oropharyngeal or nasopharyngeal swab by a licensed clinician. In this study, we exclusively screened samples for adenovirus and did not test for other causes of respiratory illnesses. Poultry samples were collected from live bird markets and commercial or backyard farms within Islamabad using two-stage probability proportional to size (PPS) cluster sampling. Birds with signs of gasping, wheezing sounds, low feed intake, decrease in weight gain and/or sudden mortality at each location were screened for adenoviruses. We targeted 13 sampling areas and obtained written permission from owners prior to sampling poultry. Livestock animals, including buffalo, cattle, goats, sheep, and dogs, were sampled using convenience sampling methods at animal farms and markets in Islamabad Capital Territory and adjacent areas of Punjab province. Nasal or oral swabs were collected from animals exhibiting signs of respiratory distress within 30 days. Owners provided written consent prior to sampling and provided epidemiological data regarding their animals. Human, poultry, and livestock samples were placed in universal transport media and transported in ice coolers to the National Influenza Center at the National Institute of Health in Islamabad, Pakistan. Upon arrival, samples were stored at -80°C until further analysis. Laboratory Methods at Pakistan’s National Influenza Center (PNIC) Human respiratory swabs were first evaluated at the National Institute of Health in Islamabad, Pakistan. According to the manufacturer's instructions, DNA was extracted from 140 µL of the sample using the QIAamp DNA Mini kit (QIAGEN, Hilden, Germany). Conventional polymerase chain reaction (PCR) targeting the conserved hexon HVR 1-6 gene was performed on extracted DNA, following the assay designed by Lu and Erdman [ 26 ]. Briefly, 5 µl of extracted DNA was added to 45 µl of the reaction mixture (10 mM Tris-HCl [pH 8.3], 1.5 mM MgCl2, 50 mM KCl, 200 mM each dNTP, 0.2 mM each of forward and reverse primer, 1 U of Taq DNA polymerase) and amplified in the Veriti Thermal Cycler (Applied Biosystems, Massachusetts, USA). Amplification consisted of five minutes at 94°C followed by 45 cycles of 94°C for 1 minute, 54°C for 45 seconds, and 72°C for 2 minutes and a final step at 72°C for 5 minutes. The resulting product was loaded onto 1% ethidium bromide-stained agarose gel and allowed to run for 1.5 hr at 120V before visualizing with UV trans-illumination. DNA extracted from poultry and animal samples was amplified and analyzed using methods targeting the species-specific hexon gene, as described by Wajid and Balboni [ 27 , 28 ]. Amplicons were subjected to Sanger sequencing, and generated adenoviral sequences were submitted to GenBank with accession numbers PP923957- PP923965 ( Supplemental Table 1 ). On March 25, 2022, aliquots of human, livestock and poultry specimens were shipped on dry ice from the National Institute of Health in Islamabad, Pakistan, to the One Health Laboratory at the University of Texas in Galveston, TX—the shipment aimed to facilitate further characterization of the viruses. Laboratory Methods at the University of Texas Medical Branch (UTMB) At UTMB, a total of 268 samples were received, including human samples (n = 224), poultry specimens (n = 25), and livestock specimens (n = 19). Viral DNA was extracted from human samples using the QIAamp DNA Mini Kit (Qiagen GmbH, Germany) on the QIAcube Connect, following the manufacturer’s protocol. DNA extraction was performed using the Trizol LS method for animal samples, adhering to the recommended guidelines. Adenovirus detection and molecular characterization for both human and animal samples were performed using the pan-pol nested PCR assay described by Wellehan et al. [ 29 ] without the addition of DMSO, as it is already included in the Invitrogen kit. This nested PCR consists of two sequential reactions with different primer sets. The first reaction (primary PCR) uses extracted DNA as the template, while the second reaction (secondary PCR) amplifies the product from the primary PCR.PCR amplification was carried out using Platinum™ Taq DNA Polymerase (Invitrogen™, Thermo Fisher Scientific Corporation, Waltham, MA, USA). For PCR 1 (primary PCR), the reaction mixture contained 0.5 µL of forward primer (25 µM), 0.5 µL of reverse primer (25 µM), 0.5 µL of dNTP (10 mM), 0.75 µL of MgCl2 (50 mM), 15.15 µL of RNase-free water, and 5 µL of extracted DNA, for a total reaction volume of 25 µL. For PCR 2 (secondary PCR), the same reagents were used with the PCR 2 primer set and 2 µL of the PCR 1 product. Amplification was conducted on a PTC Tempo 48/48 Thermal Cycler (Bio-Rad, USA). PCR products were visualized on a 1% agarose gel, and products with the expected molecular weight were sent for Sanger sequencing. Each nested PCR assay included a positive control (standard adenovirus DNA) and a no-template negative control to ensure the validity and specificity of the amplification process. For the phylogenetic analysis, sequences generated at UTMB were aligned with representative sequences of human adenoviruses (types A, B and C), bovine adenovirus and Fowl aviadenovirus (types C and D) were retrieved from NCBI, including those originating from Pakistan. Sequence alignment was performed using MAFFT [ 30 ], and maximum likelihood phylogenetic trees were constructed with IQ-TREE with the best-fit substitution model selected and 1,000 iterations of ultrafast bootstrapping [ 31 ]. The resulting trees were annotated using Figtree v1.4.4 [ 32 ]. Statistical Analysis The adenovirus patients were categorized by age groups and clinical presentation for comparison. Epidemiological and laboratory data were analyzed using Pearson’s chi-squared test to assess differences in epidemiological factors. A p-value of < 0.05 was considered statistically significant. Statistical analysis was performed using SPSS 16 software. Odds ratios were calculated using the StatCalc module of Epi-Info ver. 7.2.5.0.(US Centers for Disease Control and Prevention, Atlanta, GA). Results At PNIC, 1,705 samples from humans (n=1,084), poultry (n=385), and livestock (n=236) presenting signs of respiratory illness were screened for adenoviruses. Among these, 111 samples (6.5%) showed molecular evidence of adenovirus infection, including 96 human samples (8.9%) and 15 poultry samples (3.9%). No adenovirus was detected in the remaining animal samples. To validate the PNIC findings, 224 human samples, 25 poultry specimens, and 19 additional livestock specimens were retested at UTMB using the pan-species assay described by Wellehan et al. [29]. Human Samples Among the 1,084 human study participants, most were enrolled in hospitals in either the Federal Capital (n=390, 33.2%), Punjab (n=271, 23.1%), and Gilgit (n=171, 14.6%) regions ( Table 1 ). The geographical distribution of the 96 adenovirus detections in humans included Sindh 15% (n=15); Khyber Pakhtunkhwa 5.2% (n=5), Punjab 26% (n=25), Baluchistan 7.24% (n=7), Gilgit Baltistan 7.2% (n=7), Azad Jammu and Kashmir 5.2% (n=5) and Federal capital (ICT) 33% (n=32)). The study population was balanced with respect to sex (male: 50.6%) with a median age of 39 years. Molecular evidence of adenovirus infection was significantly more likely among participants reporting wheezing (Odds Ratio [OR] = 10.9, 95% CI: 6.0–19.7), coughing (OR = 3.3, 95% CI: 1.8–5.8), fever (OR = 3.2, 95% CI: 1.8–5.7), or sore throat (OR = 3.2, 95% CI: 1.8–5.6) compared to nasal congestion ( Table 1 ). Regionally, participants from Sindh (OR = 6.4, 95% CI: 2.3–18.0), Baluchistan (OR = 6.4, 95% CI: 2.3–18.0), Azad Jammu and Kashmir (OR = 4.8, 95% CI: 1.3–16.9), and the Federal Capital (OR = 3.6, 95% CI: 1.4–9.6) had significantly higher odds of testing positive compared to those from Punjab ( Table 1 ). Analysis of Sanger sequencing data generated at PNIC revealed the presence of Human adenoviruses A, B, and F in the human samples ( Supplemental Table 1 ). Among these, human adenovirus B strains include HAdV-7 and HAdV-3. One sequence was identified as Human adenovirus A (HAdV-31), while another was Human adenovirus F (HAdV-41). Notably, the HAdV-41 sequence is the only one from Pakistan currently available in NCBI, with 90.8% similarity to a sequence (Accession Number MK296466) isolated in Australia in 2016. At UTMB, 18.3% (41/224) of the human samples tested positive for adenovirus ( Table 2 ) and were subjected to Sanger sequencing. The predominant type was Human mastadenovirus C (HAdV-C), detected in 83% (34/41) of PCR-positive samples. Within this group, 79.41% (27/34) were HAdV-C1, 2.94% (1/34) were HAdV-C5, and 17.65% (6/34) were HAdV-C89 ( Fig. 2 ). Other adenovirus types detected included Human adenovirus B serotype 7 (HAdV-B7) ( Fig. 3 ) in 7.32% (3/41), Human adenovirus A type 12 (HAdV-A12) in 2.44% (1/41), and Human adenovirus D type 56 (HAdV-D56) in 7.32% (3/41). Additionally, bovine adenovirus 2 (BAdV-2) was detected in a nasal swab from a symptomatic human ( Fig. 4 ). The BAdV-2 sequence closely resembled strains isolated from cows in Spain (NCBI Accession Number AY288817.1) and Japan (NCBI Accession Number LC528154.1). Attempts to further characterize this virus using MinION sequencing were unsuccessful, likely due to sample degradation or insufficient sequencing depth. Animal Samples The poultry specimens were collected from various Federal Capital, Islamabad areas. Phulgran (5/15) and Barki Badhal (3/15) had the highest positivity rates. At PNIC, five PCR-positive poultry specimens underwent further characterization using Sanger sequencing. Of these, four (80%) were identified as FAdV-4 species C, while one (20%) was classified as serotype FAdV-11 species D. At UTMB, all five specimens were confirmed as Fowl aviadenovirus C serotype 4 through additional analysis. Phylogenetic analysis grouped all five UTMB-generated sequences, along with four sequences from PNIC, within FAdV-4 species C ( Fig. 5 ). The variation in adenovirus types identified between UTMB and PNIC is likely due to differences in PCR amplification strategies used for sample screening at both sites. Furthermore, at UTMB, we only received and screened poultry samples positive for FAdV-4 species C at PNIC. Sequences generated at UTMB have been deposited in GenBank under accession numbers PV067196–PV067199 for poultry and PV067200–PV067227 and PV171450–PV171463 for human samples (Table 2). Discussion Our study provides valuable insights into the epidemiology and genetic diversity of adenoviruses circulating among humans and poultry in Pakistan. The observed prevalence of adenovirus infections in humans and poultry suggests that these viruses are common causes of respiratory illnesses in these populations [ 33 – 35 ]. Detecting adenoviral strains with zoonotic potential, particularly bovine adenovirus 2 (BoAdV-2) in a human sample, further highlights the need for a One Health approach to understanding adenovirus transmission dynamics. Several studies on the region's human adenoviruses (HAdV) have attempted to subtype circulating strains, identifying a variety of adenoviruses. These include HAdV-1, HAdV-2, and HAdV-5 in China [ 36 ], the emergence of HAdV-54 in India [ 37 ] and more recently, HAdV-B (such as HAdV-B3, HAdV-B66, and HAdV-B68) and HAdV-C (HAdV-C01, HAdV-C05, and HAdV-C104) from a recent study [ 38 ] conducted in Pakistan. The association of adenovirus detection with specific clinical symptoms such as wheezing, coughing, fever, and sore throat reinforces the respiratory tropism of these viruses. These findings are consistent with previous reports that linked human adenoviruses (HAdVs), particularly types HAdV-7 and HAdV-3, to respiratory outbreaks worldwide [ 39 ]. Notably, the presence of HAdV-C1, HAdV-5, HAdV-12, HAdV-56, and HAdV-89 in our study underscores the genetic diversity of circulating strains in Pakistan. The geographic variability in adenovirus detection rates, with higher odds observed in Sindh, Baluchistan, Azad Jammu Kashmir, and the Federal Capital regions, may reflect differences in environmental factors, population density, healthcare access, or sampling bias. These findings warrant further investigation to elucidate the drivers of regional disparities in adenovirus prevalence. The most intriguing finding of our study is the molecular evidence of BoAdV-2 in the nasal swab of a 22-year-old male with respiratory illness. His symptoms, which began one week before sample collection, included fever, cough, sore throat, and chest pain. While zoonotic transmission of adenoviruses is rarely reported, this discovery aligns with other adenovirus spillover reports [ 16 , 40 ]. It raises important questions about the potential reservoirs and pathways of adenovirus transmission at the human-animal interface, particularly in settings like live bird markets or mixed-species farms. Detecting FAdV-11 and FAdV-4 in poultry adds to the growing evidence that avian adenoviruses are widespread and potentially pathogenic to commercial and backyard poultry [ 41 ]. These viruses have been associated with outbreaks of inclusion body hepatitis (IBH) and hydropericardium syndrome (HPS), which present significant economic challenges to poultry farmers. In Asian countries, FAdV-4 is the most prevalent serotype, while FAdV-11, recently reported for the first time in Pakistan, has been linked to more cases of IBH than HPS [ 42 ]. We did not detect adenoviruses among other livestock specimens, consistent with a study conducted in Lahore, Pakistan [ 43 ], which identified molecular evidence of canine adenovirus in only two of 56 necropsy specimens collected between April 2009 and June 2010. Unlike humans and poultry, adenovirus infections in other animals are rarely investigated in Pakistan. This study was limited in that the design targeted patients with acute respiratory illness and may have excluded additional adenovirus infections with presentations of conjunctival, gastrointestinal, or genitourinary illnesses. Additionally, true adenovirus prevalence is difficult to quantify through hospital surveillance studies as those experiencing minimal or mild symptoms are less likely to report for medical care. Our study was further limited in that it was not a national study. Despite these limitations, this pilot work contributes valuable epidemiological data on adenovirus detections in humans and poultry with acute respiratory infections in Pakistan. It offers insight into the burden of adenovirus disease in a region where such data has been historically sparse. Conclusions The prevalence of adenovirus identified among ill human and poultry populations in Pakistan was notably higher than other respiratory viruses, excluding COVID-19 and influenza A. This finding aligns with prior reports of human and fowl adenovirus outbreaks, emphasizing the critical need for routine molecular surveillance of adenoviruses in Pakistan to monitor their impact and guide public health interventions. Abbreviations COVID 19-Coronavirus disease 2019 DNA Deoxyribonucleic acid HAdV human adenoviruses ILI influenza-like illness (ILI) or severe acute respiratory infections SARI severe acute respiratory infection PCR polymerase chain reaction Declarations Ethics approval and consent to participate All human study procedures were approved by the National Health Institute of Islamabad Ethical Review Committee and the Duke University Institutional Review Board (Pro00102143). Individual written informed consent for specimen collection, testing, and publication was obtained from the patients or patient's parents or guardians. Written permission was obtained from animal owners prior to sampling any animals. Consent for publication Approved by informed consent. Availability of data and materials The datasets used and/or analyzed in the current study are available from Dr. Jamil Ansari upon reasonable request via email ( [email protected] ). Competing interests The authors declare that they have no conflicts of interest. Authors’ contributions J.A. designed, collected specimens, conducted laboratory analysis and drafted the manuscript. E.R.R revised the manuscript. E.S.B. performed laboratory training, guided laboratory work and revised the manuscript. N.B. performed laboratory analysis in Pakistan. F.T. performed field specimen collections. L.V.M and J.O.U. performed supplemental laboratory work in the USA and revised the manuscript. G.C.G. performed training, guided the work, and revised the manuscript. All authors reviewed the manuscript. Acknowledgements This project was supported by One Health Training Program, Duke Global Health Institute, Duke University, USA, USDA-ARS Agreement 58-3022-4-048, the Agriculture and Food Research Initiative Competitive Grant from the American Rescue Plan Act (award number 2023-70432-39558), and Professor Gray’s startup funding at the University of Texas Medical Branch, USA. The content is solely the authors' responsibility and does not necessarily represent the official views of the National Institutes of Health. The authors thank the contributions of Dr Muhammad Salman, Chief of Public Health Laboratories at the National Institute of Health, Park Road, Chak Shahzad Road, Islamabad, Pakistan for assisting with laboratory analyses. References Liu EB, Ferreyra L, Fischer SL, Pavan JV, Nates SV, Hudson NR, Tirado D, Dyer DW, Chodosh J, Seto D et al : Genetic analysis of a novel human adenovirus with a serologically unique hexon and a recombinant fiber gene . PLoS One 2011, 6 (9):e24491. 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Philomenadin FS, Singh MP, Shastry J, Phukan AC, Nagarajan M, Kaliaperumal S, Ratho RK, Ram J, Sathe MJ, Ingole A et al : Molecular characterization of adenovirus from an ongoingmulti-centric keratoconjunctivitis study in India . J Infect Dev Ctries 2020, 14 (4):404-407. Mahmood K, Ahmed W, Farooq S, Habib G, Sindhu MA, Asif A, Iftner T: Molecular characterization of human adenoviruses associated with pediatric respiratory infections in Karachi, Pakistan . BMC Infect Dis 2024, 24 (1):538. Abdirizak F, Winn AK, Parikh R, Scobie HM, Lu X, Vega E, Almendares O, Kirking HL, Rose EB, Silk BJ: Surveillance of Human Adenovirus Types and the Impact of the COVID-19 Pandemic on Reporting - United States, 2017-2023 . (1545-861X (Electronic)). Lange CE, Niama FR, Cameron K, Olson SH, Aime Nina R, Ondzie A, Bounga G, Smith BR, Pante J, Reed P et al : First evidence of a new simian adenovirus clustering with Human mastadenovirus F viruses . (1743-422X (Electronic)). Sadekuzzaman M, Miah MS, Parvin R, Haque ME, Islam TR, Sigma SH, Hossain MG, Hayat S, Hossain MT, Islam MA: Pathological investigation, molecular characterization and first-time isolation of the predominant serotypes of fowl adenovirus (FAdV-D and E) from commercial poultry in Bangladesh . (1664-302X (Print)). Wajid A, Basharat A, Shahid MA, Muntaham ST, Basit A, Hussain T, Tahir MF, Ahar M, Babar ME, Rehmani SF: Molecular Characterization and Phylogenetic Analysis of Fowl Adenoviruses Isolated from Commercial Poultry Flocks in Pakistan during 2014-15 . Pakistan J Zool 2018, 50 :863-1873. Cheema AH, Ahmed I, Mustafa G, Aslam A: Peracute infectious canine hepatitis . Pak Vet J 2012, 32 (2):277-279. Tables Table 1: Characteristics of humans with respiratory illness, by adenovirus molecular assay results in Pakistan. Human adenovirus testing (n=1084) Characteristics Positive n (%) Negative Sum p-value Odds ratio (95% CI) Overall 96 (8.9%) 988 1084 --- Age groups ≤5 46 (10.3%) 401 447 0.04 1.4 (0.8-2.6) 6-20 years 21 (7.8%) 247 268 1.0 (0.5-2.1) 21-40 years 14 (8.2%) 157 171 1.1 (0.5-2.3) 41 years and above 15 (7.6%) 183 198 ref Sex Male 53 (9.7%) 496 549 0.39 1.2 (0.8-1.9) Female 43 (8.0%) 494 537 ref Clinical Characteristics Wheezing 78 (26.4%) 217 295 0.01 10.9 (6.0-19.7) Cough 92 (9.8%) 849 941 0.023 3.3 (1.8-5.8) Fever 94 (9.6%) 889 983 0.09 3.2 (1.8-5.7) Sore Throat 92 (9.5%) 879 971 0.01 3.2 (1.8-5.6) Shortness of Breath 36 (5.2%) 661 697 0.01 1.6 (0.9-3.1) Nasal Congestion 14 (3.2%) 424 438 0.01 ref Sentinel Site Sindh 15 (15%) 125 140 0.03 6.4 (2.3-18.0) Baluchistan 7 (7.2%) 125 140 6.4 (2.3-18.0) Azad Jammu and Kashmir (AJK) 5 (5.2%) 56 61 4.8 (1.3-16.9) Federal Capital 32 (33%) 365 390 3.6 (1.4-9.6) Gilgit 7 (7.2%) 164 171 2.3 (0.7-7.3) Punjab 25 (26%) 266 271 ref Table 2. Summary of Adenovirus Types Identified in Human and Poultry Samples Analyzed at UTMB Name of specimen Adenovirus species and serotype identified Accession Numbers Hu-2-Human mastadenovirus C/Pakistan/2019 HAdV-C1 PV171450 Hu-4-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067201 Hu-5-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067202 Hu-8-Human mastadenovirus C/Pakistan/2020 HAdV-C89 PV067203 Hu-9-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV171451 Hu-11-Human adenovirus B type 7/Pakistan/2020 HAdV-B7 PV171457 Hu-16-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067204 Hu-17-Human adenovirus A type 12/Pakistan/2020 HAdV-A12 PV171463 Hu-18-Human mastadenovirus C/Pakistan/2020 HAdV-C5 PV067205 Hu-19-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067206 Hu-20-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067207 Hu-22-Human mastadenovirus C/Pakistan/2020 HAdV-C89 PV067208 Hu-23-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067209 Hu-24-Human mastadenovirus C/Pakistan/2020 HAdV-C89 PV067210 Hu-25-Human mastadenovirus C/Pakistan/2020 HAdV-C89 PV067211 Hu-26-Human mastadenovirus C/Pakistan/2020 HAdV-C89 PV067212 Hu-28-Human adenovirus B type 7/Pakistan/2020 HAdV-B7 PV171458 Hu-29-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067213 Hu-30-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067214 Hu-31-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067215 Hu-32-Human mastadenovirus C/Pakistan/2020 HAdV-C89 PV171452 Hu-33-Human adenovirus B type 7/Pakistan/2020 HAdV-B7 PV171459 Hu-34-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV171453 Hu-35-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067216 Hu-36-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067217 Hu-37-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067218 Hu-38-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067219 Hu-39-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067220 Hu-40-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067221 Hu-41-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV171454 Hu-42-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV171455 Hu-43-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067222 Hu-44-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067223 Hu-47-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067224 Hu-48-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067225 Hu-49-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067226 Hu-50-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV171456 Hu-51-Human mastadenovirus C/Pakistan/2020 HAdV-C1 PV067227 Hu-76-Human adenovirus D/Pakistan/2019 HAdV-D56 PV171460 Hu-80-Human adenovirus D/Pakistan/2019 HAdV-D56 PV171461 Hu-90-Human adenovirus D/Pakistan/2019 HAdV-D56 PV171462 P-54 Fowl aviadenovirus C type 4/Pakistan/2020 FAdV-4 PV067196 P-55 Fowl aviadenovirus C type 4/Pakistan/2020 FAdV-4 PV067197 P-56 Fowl aviadenovirus C type 4/Pakistan/2020 FAdV-4 PV067198 P-57 Fowl aviadenovirus C type 4/Pakistan/2020 FAdV-4 PV067199 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5811360","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":439381758,"identity":"195d0d95-4d36-4d2b-af5a-012d2d0e7042","order_by":0,"name":"Jamil Ansari","email":"","orcid":"","institution":"Field Epidemiology and Disease Surveillance Division, National Institute of Health, Park Road, Chak Shahzad Road, Islamabad, Pakistan","correspondingAuthor":false,"prefix":"","firstName":"Jamil","middleName":"","lastName":"Ansari","suffix":""},{"id":439381759,"identity":"80047808-33d7-413e-abc2-18113f758838","order_by":1,"name":"Emily R. Robie","email":"","orcid":"","institution":"Duke University","correspondingAuthor":false,"prefix":"","firstName":"Emily","middleName":"R.","lastName":"Robie","suffix":""},{"id":439381760,"identity":"fac3cc0a-c343-41de-a1e6-0e538abb6c52","order_by":2,"name":"Emily S. Bailey","email":"","orcid":"","institution":"Campbell University","correspondingAuthor":false,"prefix":"","firstName":"Emily","middleName":"S.","lastName":"Bailey","suffix":""},{"id":439381761,"identity":"3ac417f0-45a9-4e0f-b044-e24e77d6975e","order_by":3,"name":"Nazish Badar","email":"","orcid":"","institution":"Public Health Laboratories, National Institute of Health, Islamabad, Pakistan","correspondingAuthor":false,"prefix":"","firstName":"Nazish","middleName":"","lastName":"Badar","suffix":""},{"id":439381764,"identity":"0b8feca3-ef43-456d-a623-727a4d68ea87","order_by":4,"name":"Farooq Tahir","email":"","orcid":"","institution":"Poultry Research Institute, Rawalpindi Pakistan","correspondingAuthor":false,"prefix":"","firstName":"Farooq","middleName":"","lastName":"Tahir","suffix":""},{"id":439381765,"identity":"28623f90-cbd6-43c9-8953-bd1c9b212bbc","order_by":5,"name":"Lyudmyla V. Marushchak","email":"","orcid":"","institution":"The University of Texas Medical Branch at Galveston","correspondingAuthor":false,"prefix":"","firstName":"Lyudmyla","middleName":"V.","lastName":"Marushchak","suffix":""},{"id":439381766,"identity":"71180815-4d56-435f-b0d6-f1a0efab0d5f","order_by":6,"name":"Judith U. Oguzie","email":"","orcid":"","institution":"The University of Texas Medical Branch at Galveston","correspondingAuthor":false,"prefix":"","firstName":"Judith","middleName":"U.","lastName":"Oguzie","suffix":""},{"id":439381767,"identity":"c1556074-e66d-47ea-a3ea-0743c635b345","order_by":7,"name":"Gregory C. Gray","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAs0lEQVRIiWNgGAWjYPACGyDmAWI24rWkATWQqOUwCVrk3Y9f+/Cz7XzifrGzBxg+lB0mrMXwTE7xzN6224k90nkJjDPOEaNlBk8yAy9YS44BM28bkVoY/7adg2j5S4wWeQn2w0DDD0C0MBKjxYAnh5lZ5lyycc/tvISDPefSibCl/fhjxjdldrLts3MPPvhRZk2ELQd4DBgYodFxgLB6kC0N7A8YGP4QpXYUjIJRMApGKgAAcrE654shpZoAAAAASUVORK5CYII=","orcid":"","institution":"The University of Texas Medical Branch at Galveston","correspondingAuthor":true,"prefix":"","firstName":"Gregory","middleName":"C.","lastName":"Gray","suffix":""}],"badges":[],"createdAt":"2025-01-11 23:53:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5811360/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5811360/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":80284193,"identity":"a69f20f8-974c-4073-a27b-445751032b7c","added_by":"auto","created_at":"2025-04-10 06:28:46","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":221823,"visible":true,"origin":"","legend":"\u003cp\u003eGeographic Distribution of Influenza Sentinel Sites in Pakistan\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/21dfc240194844e4d123638d.png"},{"id":80284832,"identity":"56b01d03-18bd-43fb-a606-b233668c3485","added_by":"auto","created_at":"2025-04-10 06:36:47","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":386988,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum likelihood phylogenetic tree of Human Mastadenovirus C illustrating the relationships between sequences from this study (highlighted in red) and related sequences obtained from NCBI. Abbreviation: NCBI, National Center for Biotechnology Information.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/32ac693e908b7de22941a706.png"},{"id":80283272,"identity":"576162fb-1af5-4f7b-9006-4523f700714a","added_by":"auto","created_at":"2025-04-10 06:20:46","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":133880,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of Human Mastadenovirus B. This maximum likelihood phylogenetic tree illustrates the relationships between sequences from this study (highlighted in red) and related sequences obtained from NCBI.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/a058132cfd0e92b2ad71a815.png"},{"id":80283285,"identity":"cdda598d-6517-49dc-badb-a5e1ed6a581c","added_by":"auto","created_at":"2025-04-10 06:20:47","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":166014,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of Bovine Adenovirus Type 2B. This maximum likelihood phylogenetic tree illustrates the relationships between sequences from this study (highlighted in red) and related sequences from NCBI.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/791e33817b56f9b4a441235d.png"},{"id":80284833,"identity":"ee83c3f6-154b-4c5c-8f0e-fcca859db78e","added_by":"auto","created_at":"2025-04-10 06:36:47","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":198712,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic Tree of Fowl Aviadenovirus C and D. This maximum likelihood phylogenetic tree illustrates the relationships between sequences from this study, highlighted in red (Fowl Aviadenovirus C) and blue (Fowl Aviadenovirus D), along with related sequences from NCBI.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/4a4810df4898cbe46ae5b31b.png"},{"id":90845280,"identity":"1c1bc6c0-bc6b-4101-af68-505034f1c865","added_by":"auto","created_at":"2025-09-08 23:16:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3511065,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/58ba24a7-48d4-48f3-99ae-667db6102bbd.pdf"},{"id":80283268,"identity":"4e4ccadd-6e59-472c-a5b0-10b1c4fd087b","added_by":"auto","created_at":"2025-04-10 06:20:46","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":15765,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementalTable1.docx","url":"https://assets-eu.researchsquare.com/files/rs-5811360/v1/67c5b9d839939bd3d8deabc1.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eMolecular Typing of Adenoviruses Associated with Respiratory Illness Among Humans and Poultry, Pakistan\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003eAdenoviruses are non-enveloped, double-stranded DNA viruses that belong to the \u003cem\u003eAdenoviridae\u003c/em\u003e family. The viral capsid comprises an icosahedral hexon, penton, and fibre protein structure. Hypervariable regions of hexon and fibre genes are frequently studied to help classify various adenovirus types [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Isolated initially from human adenoid glands in 1953 [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], the \u003cem\u003eAdenoviridae\u003c/em\u003e viral family is subdivided into six recognized genera with numerous species in each genus by the International Committee for the Taxonomy of Viruses. Adenoviruses are known to infect nearly all vertebrate species and are often species-specific.\u003c/p\u003e \u003cp\u003eWithin the \u003cem\u003eMastadenovirus\u003c/em\u003e genus, human adenoviruses (HAdV) are made up of more than 110 known types separated into seven species (A \u0026ndash; G) according to their serological and molecular properties [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. HAdV-D has the most members amongst HAdV, while HAdV-C infections are most prevalent among those suffering from respiratory tract infections [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. HAdVs are commonly associated with upper respiratory illness but can manifest in many illnesses, including gastroenteritis, conjunctivitis, and pneumonia [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Many of these will resolve with minimal concern; however, infants, older persons, and immunocompromised populations face a heightened risk of mortality, particularly from pneumonia and diarrhoea [\u003cspan additionalcitationids=\"CR11\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. HAdVs are responsible for about 5% -10% lower respiratory tract infections in children and newborns [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Transmission primarily occurs through direct contact, aerosols, fomites, and the fecal-oral route, with some infected individuals shown to shed the virus for several weeks. Endemic HAdV infection has historically been a concern for populations living in crowded conditions, like military recruits. However, conjunctival adenovirus and pneumonia outbreaks have been rising in recent years among noncrowded populations [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], leading to concerns regarding emerging virulent strains. Adenoviruses are thought to have contributed to large outbreaks of human conjunctivitis in 2023 in Pakistan [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFurthermore, adenoviruses infect many other vertebrate hosts, including horses, cattle, pigs, reptiles, and fish. Though many mammals are susceptible to adenoviruses in the Mastadenovirus \u003cem\u003egenus, human\u003c/em\u003e adenoviruses are rarely pathogenic to animals. Nevertheless, evidence of occasional cross-species transmission of adenoviruses has been described [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Similarly, antibodies to canine, bovine, and simian adenoviruses have been detected in humans, and HAdV-12 has been identified in simian species [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Some have wondered if adenovirus may be a pandemic threat [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAdenoviruses infect avian species across the \u003cem\u003eAviadenovirus, Siadenovirus, and Atadenovirus\u003c/em\u003e genera [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Acute disease manifestations, such as inclusion body hepatitis and hydropericardium syndrome, can cause rapid mortality, leading to significant economic losses in poultry industries [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. For example, an adenovirus outbreak in broiler farm chicks in Pakistan in 1985 resulted in mortality rates of 30\u0026ndash;60% [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePathogenic adenovirus strains have been identified globally, often displaying seasonal variations. These viruses threaten human and animal populations and have shown evidence of recombining into novel forms. Underscoring the importance of surveillance in monitoring circulating adenovirus strains within specific regions [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. In Pakistan, reports indicate an increasing [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] incidence of adenovirus infections across various areas [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. However, comprehensive epidemiological data remain scarce despite this apparent rise, highlighting a gap in understanding the prevalence and distribution of the virus. To address this, we conducted a pilot study investigating adenovirus epidemiology among humans and livestock in and around Islamabad, Pakistan.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eSample Collection\u003c/h2\u003e \u003cp\u003eBetween February 2019 and March 2021, a total of 1,705 samples were collected from humans (1,084), poultry (385), and livestock (236) presenting with respiratory symptoms and illness. Human samples were obtained from seven hospitals participating in the influenza surveillance network across regions and cities of Pakistan: Jinnah Hospital, Lahore; Nishter Medical Complex Hospital, Multan; and Allied Hospital, Faisalabad (Eastern Pakistan); Saidu Sharif Hospital, Swat (Northwestern Pakistan); Civil Hospital, Karachi (Southern Pakistan); Bolan Medical Complex Hospital, Quetta (Southwestern Pakistan); and Provincial Headquarters Hospital, Gilgit (Northern Pakistan) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eSampling followed a non-probability convenience approach, targeting patients who met the World Health Organization\u0026rsquo;s case definition for influenza-like illness (ILI) or severe acute respiratory infection (SARI). [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Outpatients and hospitalized patients of all ages were eligible to participate if they presented with acute respiratory symptoms, a temperature exceeding 38\u0026deg;C, and a cough onset within the past ten days. Participants were asked to complete a brief epidemiological survey and consent to collect an oropharyngeal or nasopharyngeal swab by a licensed clinician. In this study, we exclusively screened samples for adenovirus and did not test for other causes of respiratory illnesses.\u003c/p\u003e \u003cp\u003ePoultry samples were collected from live bird markets and commercial or backyard farms within Islamabad using two-stage probability proportional to size (PPS) cluster sampling. Birds with signs of gasping, wheezing sounds, low feed intake, decrease in weight gain and/or sudden mortality at each location were screened for adenoviruses. We targeted 13 sampling areas and obtained written permission from owners prior to sampling poultry.\u003c/p\u003e \u003cp\u003eLivestock animals, including buffalo, cattle, goats, sheep, and dogs, were sampled using convenience sampling methods at animal farms and markets in Islamabad Capital Territory and adjacent areas of Punjab province. Nasal or oral swabs were collected from animals exhibiting signs of respiratory distress within 30 days. Owners provided written consent prior to sampling and provided epidemiological data regarding their animals.\u003c/p\u003e \u003cp\u003eHuman, poultry, and livestock samples were placed in universal transport media and transported in ice coolers to the National Influenza Center at the National Institute of Health in Islamabad, Pakistan. Upon arrival, samples were stored at -80\u0026deg;C until further analysis.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eLaboratory Methods at Pakistan’s National Influenza Center (PNIC)\u003c/h3\u003e\n\u003cp\u003eHuman respiratory swabs were first evaluated at the National Institute of Health in Islamabad, Pakistan. According to the manufacturer's instructions, DNA was extracted from 140 \u0026micro;L of the sample using the QIAamp DNA Mini kit (QIAGEN, Hilden, Germany). Conventional polymerase chain reaction (PCR) targeting the conserved hexon HVR\u003csub\u003e1-6\u003c/sub\u003e gene was performed on extracted DNA, following the assay designed by Lu and Erdman [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Briefly, 5 \u0026micro;l of extracted DNA was added to 45 \u0026micro;l of the reaction mixture (10 mM Tris-HCl [pH 8.3], 1.5 mM MgCl2, 50 mM KCl, 200 mM each dNTP, 0.2 mM each of forward and reverse primer, 1 U of Taq DNA polymerase) and amplified in the Veriti Thermal Cycler (Applied Biosystems, Massachusetts, USA). Amplification consisted of five minutes at 94\u0026deg;C followed by 45 cycles of 94\u0026deg;C for 1 minute, 54\u0026deg;C for 45 seconds, and 72\u0026deg;C for 2 minutes and a final step at 72\u0026deg;C for 5 minutes. The resulting product was loaded onto 1% ethidium bromide-stained agarose gel and allowed to run for 1.5 hr at 120V before visualizing with UV trans-illumination.\u003c/p\u003e \u003cp\u003eDNA extracted from poultry and animal samples was amplified and analyzed using methods targeting the species-specific hexon gene, as described by Wajid and Balboni [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Amplicons were subjected to Sanger sequencing, and generated adenoviral sequences were submitted to GenBank with accession numbers PP923957- PP923965 (\u003cb\u003eSupplemental Table\u0026nbsp;1\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eOn March 25, 2022, aliquots of human, livestock and poultry specimens were shipped on dry ice from the National Institute of Health in Islamabad, Pakistan, to the One Health Laboratory at the University of Texas in Galveston, TX\u0026mdash;the shipment aimed to facilitate further characterization of the viruses.\u003c/p\u003e\n\u003ch3\u003eLaboratory Methods at the University of Texas Medical Branch (UTMB)\u003c/h3\u003e\n\u003cp\u003eAt UTMB, a total of 268 samples were received, including human samples (n\u0026thinsp;=\u0026thinsp;224), poultry specimens (n\u0026thinsp;=\u0026thinsp;25), and livestock specimens (n\u0026thinsp;=\u0026thinsp;19). Viral DNA was extracted from human samples using the QIAamp DNA Mini Kit (Qiagen GmbH, Germany) on the QIAcube Connect, following the manufacturer\u0026rsquo;s protocol. DNA extraction was performed using the Trizol LS method for animal samples, adhering to the recommended guidelines.\u003c/p\u003e \u003cp\u003eAdenovirus detection and molecular characterization for both human and animal samples were performed using the pan-pol nested PCR assay described by Wellehan et al. [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] without the addition of DMSO, as it is already included in the Invitrogen kit. This nested PCR consists of two sequential reactions with different primer sets. The first reaction (primary PCR) uses extracted DNA as the template, while the second reaction (secondary PCR) amplifies the product from the primary PCR.PCR amplification was carried out using Platinum\u0026trade; Taq DNA Polymerase (Invitrogen\u0026trade;, Thermo Fisher Scientific Corporation, Waltham, MA, USA). For PCR 1 (primary PCR), the reaction mixture contained 0.5 \u0026micro;L of forward primer (25 \u0026micro;M), 0.5 \u0026micro;L of reverse primer (25 \u0026micro;M), 0.5 \u0026micro;L of dNTP (10 mM), 0.75 \u0026micro;L of MgCl2 (50 mM), 15.15 \u0026micro;L of RNase-free water, and 5 \u0026micro;L of extracted DNA, for a total reaction volume of 25 \u0026micro;L. For PCR 2 (secondary PCR), the same reagents were used with the PCR 2 primer set and 2 \u0026micro;L of the PCR 1 product. Amplification was conducted on a PTC Tempo 48/48 Thermal Cycler (Bio-Rad, USA). PCR products were visualized on a 1% agarose gel, and products with the expected molecular weight were sent for Sanger sequencing. Each nested PCR assay included a positive control (standard adenovirus DNA) and a no-template negative control to ensure the validity and specificity of the amplification process.\u003c/p\u003e \u003cp\u003eFor the phylogenetic analysis, sequences generated at UTMB were aligned with representative sequences of human adenoviruses (types A, B and C), bovine adenovirus and Fowl aviadenovirus (types C and D) were retrieved from NCBI, including those originating from Pakistan. Sequence alignment was performed using MAFFT [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], and maximum likelihood phylogenetic trees were constructed with IQ-TREE with the best-fit substitution model selected and 1,000 iterations of ultrafast bootstrapping [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The resulting trees were annotated using Figtree v1.4.4 [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eThe adenovirus patients were categorized by age groups and clinical presentation for comparison. Epidemiological and laboratory data were analyzed using Pearson\u0026rsquo;s chi-squared test to assess differences in epidemiological factors. A p-value of \u0026lt;\u0026thinsp;0.05 was considered statistically significant. Statistical analysis was performed using SPSS 16 software. Odds ratios were calculated using the StatCalc module of Epi-Info ver. 7.2.5.0.(US Centers for Disease Control and Prevention, Atlanta, GA).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eAt PNIC, 1,705 samples from humans (n=1,084), poultry (n=385), and livestock (n=236) presenting signs of respiratory illness were screened for adenoviruses. Among these, 111 samples (6.5%) showed molecular evidence of adenovirus infection, including 96 human samples (8.9%) and 15 poultry samples (3.9%). No adenovirus was detected in the remaining animal samples. To validate the PNIC findings, 224 human samples, 25 poultry specimens, and 19 additional livestock specimens were retested at UTMB using the pan-species assay described by Wellehan et al. [29].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eHuman Samples\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;Among the 1,084 human study participants, most were enrolled in hospitals in either the Federal Capital (n=390, 33.2%), Punjab (n=271, 23.1%), and Gilgit (n=171, 14.6%) regions (\u003cstrong\u003eTable 1\u003c/strong\u003e). The geographical distribution of the 96 adenovirus detections in humans included Sindh 15% (n=15); Khyber Pakhtunkhwa 5.2% (n=5), Punjab 26% (n=25), Baluchistan 7.24% (n=7), Gilgit Baltistan 7.2% (n=7), Azad Jammu and Kashmir 5.2% (n=5) and Federal capital (ICT) 33% (n=32)). The study population was balanced with respect to sex (male: 50.6%) with a median age of 39 years.\u003c/p\u003e\n\u003cp\u003eMolecular evidence of adenovirus infection was significantly more likely among participants reporting wheezing (Odds Ratio [OR] = 10.9, 95% CI: 6.0–19.7), coughing (OR = 3.3, 95% CI: 1.8–5.8), fever (OR = 3.2, 95% CI: 1.8–5.7), or sore throat (OR = 3.2, 95% CI: 1.8–5.6) compared to nasal congestion (\u003cstrong\u003eTable 1\u003c/strong\u003e). Regionally, participants from Sindh (OR = 6.4, 95% CI: 2.3–18.0), Baluchistan (OR = 6.4, 95% CI: 2.3–18.0), Azad Jammu and Kashmir (OR = 4.8, 95% CI: 1.3–16.9), and the Federal Capital (OR = 3.6, 95% CI: 1.4–9.6) had significantly higher odds of testing positive compared to those from Punjab (\u003cstrong\u003eTable 1\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003eAnalysis of Sanger sequencing data generated at PNIC revealed the presence of Human adenoviruses A, B, and F in the human samples\u0026nbsp;(\u003cstrong\u003eSupplemental Table 1\u003c/strong\u003e). Among these, human adenovirus B strains include HAdV-7 and HAdV-3. One sequence was identified as Human adenovirus A (HAdV-31), while another was Human adenovirus F (HAdV-41). Notably, the HAdV-41 sequence is the only one from Pakistan currently available in NCBI, with 90.8% similarity to a sequence (Accession Number MK296466) isolated in Australia in 2016.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; At UTMB,\u0026nbsp;18.3% (41/224) of the human samples tested positive for adenovirus (\u003cstrong\u003eTable 2\u003c/strong\u003e) and were subjected to Sanger sequencing. The predominant type was Human mastadenovirus C (HAdV-C), detected in 83% (34/41) of PCR-positive samples. Within this group, 79.41% (27/34) were HAdV-C1, 2.94% (1/34) were HAdV-C5, and 17.65% (6/34) were HAdV-C89 (\u003cstrong\u003eFig. 2\u003c/strong\u003e). Other adenovirus types detected included Human adenovirus B serotype 7 (HAdV-B7) (\u003cstrong\u003eFig. 3\u003c/strong\u003e) in 7.32% (3/41), Human adenovirus A type 12 (HAdV-A12) in 2.44% (1/41), and Human adenovirus D type 56 (HAdV-D56) \u0026nbsp; in 7.32% (3/41). Additionally, bovine adenovirus 2 (BAdV-2) was detected in a nasal swab from a symptomatic human (\u003cstrong\u003eFig. 4\u003c/strong\u003e). The BAdV-2 sequence closely resembled strains isolated from cows in Spain (NCBI Accession Number AY288817.1) and Japan (NCBI Accession Number LC528154.1). Attempts to further characterize this virus using MinION sequencing were unsuccessful, likely due to sample degradation or insufficient sequencing depth.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAnimal Samples\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe poultry specimens were collected from various Federal Capital, Islamabad areas. Phulgran (5/15) and Barki Badhal (3/15)\u0026nbsp;had the highest positivity rates. At PNIC, five PCR-positive poultry specimens underwent further characterization using Sanger sequencing. Of these, four (80%) were identified as FAdV-4 species\u0026nbsp;C, while one (20%) was classified as serotype FAdV-11 species D. At UTMB, all five specimens were confirmed as Fowl aviadenovirus C serotype 4 through additional analysis. Phylogenetic analysis grouped all five UTMB-generated sequences, along with four sequences from PNIC, within FAdV-4 species C (\u003cstrong\u003eFig. 5\u003c/strong\u003e). The variation in adenovirus types identified between UTMB and PNIC is likely due to differences in PCR amplification strategies used for sample screening at both sites. Furthermore, at UTMB, we only received and screened poultry samples positive for FAdV-4 species C at PNIC. Sequences generated at UTMB have been deposited in GenBank under accession numbers PV067196–PV067199 for poultry and PV067200–PV067227 and PV171450–PV171463 for human samples (Table 2).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eOur study provides valuable insights into the epidemiology and genetic diversity of adenoviruses circulating among humans and poultry in Pakistan. The observed prevalence of adenovirus infections in humans and poultry suggests that these viruses are common causes of respiratory illnesses in these populations [\u003cspan additionalcitationids=\"CR34\" citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Detecting adenoviral strains with zoonotic potential, particularly bovine adenovirus 2 (BoAdV-2) in a human sample, further highlights the need for a One Health approach to understanding adenovirus transmission dynamics.\u003c/p\u003e \u003cp\u003eSeveral studies on the region's human adenoviruses (HAdV) have attempted to subtype circulating strains, identifying a variety of adenoviruses. These include HAdV-1, HAdV-2, and HAdV-5 in China [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e], the emergence of HAdV-54 in India [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] and more recently, HAdV-B (such as HAdV-B3, HAdV-B66, and HAdV-B68) and HAdV-C (HAdV-C01, HAdV-C05, and HAdV-C104) from a recent study [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e] conducted in Pakistan.\u003c/p\u003e \u003cp\u003eThe association of adenovirus detection with specific clinical symptoms such as wheezing, coughing, fever, and sore throat reinforces the respiratory tropism of these viruses. These findings are consistent with previous reports that linked human adenoviruses (HAdVs), particularly types HAdV-7 and HAdV-3, to respiratory outbreaks worldwide [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Notably, the presence of HAdV-C1, HAdV-5, HAdV-12, HAdV-56, and HAdV-89 in our study underscores the genetic diversity of circulating strains in Pakistan.\u003c/p\u003e \u003cp\u003eThe geographic variability in adenovirus detection rates, with higher odds observed in Sindh, Baluchistan, Azad Jammu Kashmir, and the Federal Capital regions, may reflect differences in environmental factors, population density, healthcare access, or sampling bias. These findings warrant further investigation to elucidate the drivers of regional disparities in adenovirus prevalence.\u003c/p\u003e \u003cp\u003eThe most intriguing finding of our study is the molecular evidence of BoAdV-2 in the nasal swab of a 22-year-old male with respiratory illness. His symptoms, which began one week before sample collection, included fever, cough, sore throat, and chest pain. While zoonotic transmission of adenoviruses is rarely reported, this discovery aligns with other adenovirus spillover reports [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. It raises important questions about the potential reservoirs and pathways of adenovirus transmission at the human-animal interface, particularly in settings like live bird markets or mixed-species farms.\u003c/p\u003e \u003cp\u003eDetecting FAdV-11 and FAdV-4 in poultry adds to the growing evidence that avian adenoviruses are widespread and potentially pathogenic to commercial and backyard poultry [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. These viruses have been associated with outbreaks of inclusion body hepatitis (IBH) and hydropericardium syndrome (HPS), which present significant economic challenges to poultry farmers. In Asian countries, FAdV-4 is the most prevalent serotype, while FAdV-11, recently reported for the first time in Pakistan, has been linked to more cases of IBH than HPS [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWe did not detect adenoviruses among other livestock specimens, consistent with a study conducted in Lahore, Pakistan [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], which identified molecular evidence of canine adenovirus in only two of 56 necropsy specimens collected between April 2009 and June 2010. Unlike humans and poultry, adenovirus infections in other animals are rarely investigated in Pakistan.\u003c/p\u003e \u003cp\u003eThis study was limited in that the design targeted patients with acute respiratory illness and may have excluded additional adenovirus infections with presentations of conjunctival, gastrointestinal, or genitourinary illnesses. Additionally, true adenovirus prevalence is difficult to quantify through hospital surveillance studies as those experiencing minimal or mild symptoms are less likely to report for medical care. Our study was further limited in that it was not a national study. Despite these limitations, this pilot work contributes valuable epidemiological data on adenovirus detections in humans and poultry with acute respiratory infections in Pakistan. It offers insight into the burden of adenovirus disease in a region where such data has been historically sparse.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe prevalence of adenovirus identified among ill human and poultry populations in Pakistan was notably higher than other respiratory viruses, excluding COVID-19 and influenza A. This finding aligns with prior reports of human and fowl adenovirus outbreaks, emphasizing the critical need for routine molecular surveillance of adenoviruses in Pakistan to monitor their impact and guide public health interventions.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCOVID\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003e19-Coronavirus disease 2019\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eDNA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eDeoxyribonucleic acid\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eHAdV\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ehuman adenoviruses\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eILI\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003einfluenza-like illness (ILI) or severe acute respiratory infections\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eSARI\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003esevere acute respiratory infection\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003ePCR\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003epolymerase chain reaction\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll human study procedures were approved by the National Health Institute of Islamabad Ethical Review Committee and the Duke University Institutional Review Board (Pro00102143).\u0026nbsp;Individual written informed consent for specimen collection, testing, and publication was obtained from the patients or patient's parents or guardians. Written permission was obtained from animal owners prior to sampling any animals.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eApproved by informed consent.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed in the current study are available from Dr. Jamil Ansari upon reasonable request via email ([email protected]).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors’ contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;J.A. designed, collected specimens, conducted laboratory analysis and drafted the manuscript. E.R.R revised the manuscript. E.S.B. performed laboratory training, guided laboratory work and revised the manuscript. N.B. performed laboratory analysis in Pakistan. F.T. performed field specimen collections. L.V.M and J.O.U. performed supplemental laboratory work in the USA and revised the manuscript. G.C.G. performed training, guided the work, and revised the manuscript. All authors reviewed the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis project was supported by One Health Training Program, Duke Global Health Institute, Duke University, USA, USDA-ARS Agreement 58-3022-4-048, the Agriculture and Food Research Initiative Competitive Grant from the American Rescue Plan Act (award number 2023-70432-39558), and Professor Gray’s startup funding at the University of Texas Medical Branch, USA. The content is solely the authors' responsibility and does not necessarily represent the official views of the National Institutes of Health. The authors thank the contributions of Dr Muhammad Salman, Chief of Public Health Laboratories at the National Institute of Health, Park Road, Chak Shahzad Road, Islamabad, Pakistan for assisting with laboratory analyses. \u0026nbsp;\u003cstrong\u003e\u003cbr\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eLiu EB, Ferreyra L, Fischer SL, Pavan JV, Nates SV, Hudson NR, Tirado D, Dyer DW, Chodosh J, Seto D\u003cem\u003e\u0026nbsp;et al\u003c/em\u003e: \u003cstrong\u003eGenetic analysis of a novel human adenovirus with a serologically unique hexon and a recombinant fiber gene\u003c/strong\u003e. \u003cem\u003ePLoS One\u0026nbsp;\u003c/em\u003e2011, \u003cstrong\u003e6\u003c/strong\u003e(9):e24491.\u003c/li\u003e\n \u003cli\u003eMatsushima Y, Shimizu H, Kano A, Nakajima E, Ishimaru Y, Dey SK, Watanabe Y, Adachi F, Suzuki K, Mitani K\u003cem\u003e\u0026nbsp;et al\u003c/em\u003e: \u003cstrong\u003eNovel human adenovirus strain, Bangladesh\u003c/strong\u003e. \u003cem\u003eEmerg Infect Dis\u0026nbsp;\u003c/em\u003e2012, \u003cstrong\u003e18\u003c/strong\u003e(5):846-848.\u003c/li\u003e\n \u003cli\u003eRobinson CM, Singh G, Lee JY, Dehghan S, Rajaiya J, Liu EB, Yousuf MA, Betensky RA, Jones MS, Dyer DW\u003cem\u003e\u0026nbsp;et al\u003c/em\u003e: \u003cstrong\u003eMolecular evolution of human adenoviruses\u003c/strong\u003e. \u003cem\u003eSci Rep\u0026nbsp;\u003c/em\u003e2013, \u003cstrong\u003e3\u003c/strong\u003e:1812.\u003c/li\u003e\n \u003cli\u003eHilleman MR, Werner JH: \u003cstrong\u003eRecovery of new agent from patients with acute respiratory illness\u003c/strong\u003e. \u003cem\u003eProc Soc Exp Biol Med\u0026nbsp;\u003c/em\u003e1954, \u003cstrong\u003e85\u003c/strong\u003e(1):183-188.\u003c/li\u003e\n \u003cli\u003eRowe WP, Huebner RJ, Gilmore LK, Parrott RH, Ward TG: \u003cstrong\u003eIsolation of a cytopathogenic agent from human adenoids undergoing spontaneous degeneration in tissue culture\u003c/strong\u003e. \u003cem\u003eProc Soc Exp Biol Med\u0026nbsp;\u003c/em\u003e1953, \u003cstrong\u003e84\u003c/strong\u003e(3):570-573.\u003c/li\u003e\n \u003cli\u003eGonzalez G, Hayes M, Balansay M, Underwood R, Balagot C, Pan R, Pineda K, Iniguez-Stevens E, Kriner P, Estrada I\u003cem\u003e\u0026nbsp;et al\u003c/em\u003e: \u003cstrong\u003eTwo novel recombinant human mastadenovirus D genotypes associated with acute respiratory illness\u003c/strong\u003e. \u003cem\u003eJ Med Virol\u0026nbsp;\u003c/em\u003e2023, \u003cstrong\u003e95\u003c/strong\u003e(3):e28653.\u003c/li\u003e\n \u003cli\u003eDhingra A, Hage E, Ganzenmueller T, Bottcher S, Hofmann J, Hamprecht K, Obermeier P, Rath B, Hausmann F, Dobner T\u003cem\u003e\u0026nbsp;et al\u003c/em\u003e: \u003cstrong\u003eMolecular Evolution of Human Adenovirus (HAdV) Species C\u003c/strong\u003e. \u003cem\u003eSci Rep\u0026nbsp;\u003c/em\u003e2019, \u003cstrong\u003e9\u003c/strong\u003e(1):1039.\u003c/li\u003e\n \u003cli\u003eLynch JP, 3rd, Kajon AE: \u003cstrong\u003eAdenovirus: Epidemiology, Global Spread of Novel Types, and Approach to Treatment\u003c/strong\u003e. 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(1545-861X (Electronic)).\u003c/li\u003e\n \u003cli\u003eLange CE, Niama FR, Cameron K, Olson SH, Aime Nina R, Ondzie A, Bounga G, Smith BR, Pante J, Reed P\u003cem\u003e\u0026nbsp;et al\u003c/em\u003e: \u003cstrong\u003eFirst evidence of a new simian adenovirus clustering with Human mastadenovirus F viruses\u003c/strong\u003e. (1743-422X (Electronic)).\u003c/li\u003e\n \u003cli\u003eSadekuzzaman M, Miah MS, Parvin R, Haque ME, Islam TR, Sigma SH, Hossain MG, Hayat S, Hossain MT, Islam MA: \u003cstrong\u003ePathological investigation, molecular characterization and first-time isolation of the predominant serotypes of fowl adenovirus (FAdV-D and E) from commercial poultry in Bangladesh\u003c/strong\u003e. (1664-302X (Print)).\u003c/li\u003e\n \u003cli\u003eWajid A, Basharat A, Shahid MA, Muntaham ST, Basit A, Hussain T, Tahir MF, Ahar M, Babar ME, Rehmani SF: \u003cstrong\u003eMolecular Characterization and Phylogenetic Analysis of Fowl Adenoviruses Isolated from Commercial Poultry Flocks in Pakistan during 2014-15\u003c/strong\u003e. \u003cem\u003ePakistan J Zool\u0026nbsp;\u003c/em\u003e2018, \u003cstrong\u003e50\u003c/strong\u003e:863-1873.\u003c/li\u003e\n \u003cli\u003eCheema AH, Ahmed I, Mustafa G, Aslam A: \u003cstrong\u003ePeracute infectious canine hepatitis\u003c/strong\u003e. \u003cem\u003ePak Vet J\u0026nbsp;\u003c/em\u003e2012, \u003cstrong\u003e32\u003c/strong\u003e(2):277-279.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cdiv align=\"left\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"742\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 742px;\"\u003e\n \u003cp\u003eTable 1: \u0026nbsp;Characteristics of humans with respiratory illness, by adenovirus molecular assay results in Pakistan.\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"bottom\" style=\"width: 219px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHuman adenovirus testing\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(n=1084) \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 331px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 192px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;Characteristics \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003en (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 106px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSum\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 124px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOdds ratio\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(95% CI)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 742px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eOverall\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 120px;\"\u003e\n \u003cp\u003e96 (8.9%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 99px;\"\u003e\n \u003cp\u003e988\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 106px;\"\u003e\n \u003cp\u003e1084\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e---\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 742px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003e\u0026le;5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e46 (10.3%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e401\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e447\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e1.4 (0.8-2.6)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003e6-20 years\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e21 (7.8%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e247\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e268\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e1.0 (0.5-2.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003e21-40 years\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e14 (8.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e157\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e171\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e1.1 (0.5-2.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003e41 years and above\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e15 (7.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e198\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003eref\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 742px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSex\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eMale\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e53 (9.7%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e496\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e549\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e1.2 (0.8-1.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eFemale\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e43 (8.0%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e494\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e537\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003eref\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 742px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eClinical Characteristics\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eWheezing\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 120px;\"\u003e\n \u003cp\u003e78 (26.4%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e217\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e295\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 100px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 124px;\"\u003e\n \u003cp\u003e10.9 (6.0-19.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eCough\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e92 (9.8%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e849\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e941\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e3.3 (1.8-5.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eFever\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e94 (9.6%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e889\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e983\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e3.2 (1.8-5.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eSore Throat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e92 (9.5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e971\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e3.2 (1.8-5.6)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eShortness of Breath\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e36 (5.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e661\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e697\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e1.6 (0.9-3.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eNasal Congestion\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e14 (3.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e424\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e438\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003eref\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 742px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSentinel Site\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eSindh\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e15 (15%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e140\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\" style=\"width: 100px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e6.4 (2.3-18.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eBaluchistan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e7 (7.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e140\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e6.4 (2.3-18.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192px;\"\u003e\n \u003cp\u003eAzad Jammu and Kashmir (AJK)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 120px;\"\u003e\n \u003cp\u003e5 (5.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 99px;\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 124px;\"\u003e\n \u003cp\u003e4.8 (1.3-16.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eFederal Capital\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e32 (33%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e365\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e390\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e3.6 (1.4-9.6)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003eGilgit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e7 (7.2%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e164\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e171\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003e2.3 (0.7-7.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 192px;\"\u003e\n \u003cp\u003ePunjab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e25 (26%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e271\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 124px;\"\u003e\n \u003cp\u003eref\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 2. Summary of Adenovirus Types Identified in Human and Poultry Samples Analyzed at UTMB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eName of specimen\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdenovirus species and serotype identified\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAccession Numbers\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-2-Human mastadenovirus C/Pakistan/2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171450\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-4-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067201\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-5-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067202\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-8-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067203\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-9-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171451\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-11-Human adenovirus B type 7/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-B7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171457\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-16-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067204\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-17-Human adenovirus A type 12/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-A12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171463\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-18-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067205\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-19-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067206\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-20-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067207\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-22-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067208\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-23-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067209\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-24-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067210\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-25-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067211\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-26-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067212\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-28-Human adenovirus B type 7/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-B7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171458\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-29-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067213\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-30-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067214\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-31-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067215\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-32-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171452\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-33-Human adenovirus B type 7/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-B7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171459\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-34-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171453\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-35-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067216\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-36-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067217\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-37-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067218\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-38-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067219\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-39-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067220\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-40-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067221\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-41-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171454\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-42-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171455\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-43-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067222\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-44-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067223\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-47-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067224\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-48-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067225\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-49-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067226\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-50-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171456\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-51-Human mastadenovirus C/Pakistan/2020\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-C1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067227\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-76-Human adenovirus D/Pakistan/2019\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-D56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171460\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-80-Human adenovirus D/Pakistan/2019\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-D56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171461\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHu-90-Human adenovirus D/Pakistan/2019\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHAdV-D56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV171462\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eP-54 Fowl aviadenovirus C type 4/Pakistan/2020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFAdV-4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067196\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eP-55 Fowl aviadenovirus C type 4/Pakistan/2020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFAdV-4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067197\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eP-56 Fowl aviadenovirus C type 4/Pakistan/2020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFAdV-4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067198\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eP-57 Fowl aviadenovirus C type 4/Pakistan/2020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFAdV-4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePV067199\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Adenovirus, respiratory infection, epidemiology, phylogenetic analyses ","lastPublishedDoi":"10.21203/rs.3.rs-5811360/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5811360/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cb\u003eBackground\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAdenoviruses have caused epidemics among both humans and poultry in Pakistan. There is growing evidence that adenoviruses sometimes spillover to infect other species. In this pilot study, we sought to study the distribution of circulating adenovirus types among humans, poultry and other animals in several areas of Pakistan.\u003c/p\u003e\u003cp\u003e\u003cb\u003eMethods\u003c/b\u003e\u003c/p\u003e \u003cp\u003e From February 2019 to March 2021, patients with influenza-like illness or pneumonia in seven hospitals near Islamabad were invited to participate in this study. Volunteers permitted the collection of an oropharyngeal or a nasopharyngeal swab and epidemiological data. Concomitantly, in Islamabad\u0026rsquo;s live bird markets and commercial or backyard farms, owners granted permission for oral or nasal swabs of their livestock with signs of respiratory illness to be studied. These specimens were screened with molecular assays for evidence of adenovirus infection.\u003c/p\u003e\u003cp\u003e\u003cb\u003eResults\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAmong the 1705 samples collected, 96 (8.8%) of 1084 humans and 15 (4%) of 385 poultry had molecular evidence of adenovirus infection. The odds ratio (OR) of such molecular detections was greatest among participants with wheezing (OR\u0026thinsp;=\u0026thinsp;10.9, 95% CI 6.0-19.7), coughing (OR\u0026thinsp;=\u0026thinsp;3.3, 95% CI 1.8\u0026ndash;5.8), fever (OR\u0026thinsp;=\u0026thinsp;3.2, 95% CI1.8-5.7) or sore throat (OR\u0026thinsp;=\u0026thinsp;3.2, 95% CI 1.8\u0026ndash;5.6) compared to nasal congestion. Similarly, odds of positivity were greatest for participants from Sindh (OR\u0026thinsp;=\u0026thinsp;6.4, 95% CI 2.3\u0026ndash;18.0), Baluchistan (OR\u0026thinsp;=\u0026thinsp;6.4, 95% CI2.3-18.0), Azad Jammu and Kashmir (OR\u0026thinsp;=\u0026thinsp;4.8, 95% CI, 1.3\u0026ndash;16.9), or Federal Capital regions (OR\u0026thinsp;=\u0026thinsp;3.6, 95% CI, 1.4\u0026ndash;9.6) compared to Punjab. Partial hexon gene sequencing identified co-circulation of HAdV-7 and HAdV-3 strains among humans. Further sequencing of adenoviral DNA polymerase identified HAdV-C1, HAdV-5, HAdV-89, HAdV-12, and HAdV-56. Notably, we detected molecular evidence of bovine adenovirus 2 in a sick human\u0026rsquo;s nasal swab. Additionally, FAdV-11 and FAdV-4 strains were identified among poultry swab specimens.\u003c/p\u003e\u003cp\u003e\u003cb\u003eConclusion\u003c/b\u003e\u003c/p\u003e \u003cp\u003eCompared to other studies, there was a relatively high prevalence of adenoviruses among sick humans and poultry, with the unusual discovery of molecular evidence of bovine adenovirus in a sick human\u0026rsquo;s airway. There was considerable diversity among detected adenovirus strains. As adenovirus epidemics have periodically occurred in Pakistan, assessing adenovirus prevalence and genotype distributions is prudent, especially among humans with respiratory illnesses.\u003c/p\u003e","manuscriptTitle":"Molecular Typing of Adenoviruses Associated with Respiratory Illness Among Humans and Poultry, Pakistan","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-10 06:20:41","doi":"10.21203/rs.3.rs-5811360/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d38a7ea5-44c5-4940-a509-10f5e86e5e27","owner":[],"postedDate":"April 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-08T23:08:12+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-10 06:20:41","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5811360","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5811360","identity":"rs-5811360","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}