Live Bird Markets Research Articles

Whole-genome sequence and genesis of an avian influenza virus H5N1 isolated from a healthy chicken in a live bird market in Indonesia: accumulation of mammalian adaptation markers in avian hosts.

Influenza A viruses are a major pathogen that causes significant clinical and economic harm to many animals. In Indonesia, the highly pathogenic avian influenza (HPAI) H5N1 virus has been endemic in poultry since 2003 and has caused sporadic deadly infections in humans. The genetic bases that determine host range have not yet been fully elucidated. We analyzed the whole-genome sequence of a recent H5 isolate to reveal the evolution toward its mammalian adaptation. We determined the whole-genome sequence of A/chicken/East Java/Av1955/2022 (hereafter, "Av1955") from a healthy chicken in April 2022 and conducted phylogenetic and mutational analysis. Phylogenetic analysis revealed that Av1955 belonged to the H5N1 clade 2.3.2.1c (Eurasian lineage). The six gene segments (PB1, PB2, HA, NP, NA, and NS) out of the eight segments derived from viruses of H5N1 Eurasian lineage, one (PB2) from the H3N6 subtype and the remaining one (M) from the H5N1 clade 2.1.3.2b (Indonesian lineage). The donor of the PB2 segment was a reassortant among three viruses of H5N1 Eurasian and Indonesian lineages and the H3N6 subtype. The HA amino acid sequence contained multiple basic amino acids at the cleavage site. Mutation analysis revealed that Av1955 possessed the maximal number of mammalian adaptation marker mutations. Av1955 was a virus of H5N1 Eurasian lineage. The HA protein contains an HPAI H5N1-type cleavage site sequence, while the virus was isolated from a healthy chicken suggesting its low pathogenicity nature. The virus has increased mammalian adaptation markers by mutation and intra- and inter-subtype reassortment, gathering gene segments possessing the most abundant maker mutations among previously circulating viruses. The increasing mammalian adaptation mutation in avian hosts suggests that they might be adaptive to infection in mammalian and avian hosts. It highlights the importance of genomic surveillance and adequate control measures for H5N1 infection in live poultry markets.

Molecular Detection of Tetracycline-Resistant Genes in Multi-Drug-Resistant Escherichia coli Isolated from Broiler Meat in Bangladesh

This study aimed to estimate the antimicrobial resistance (AMR) patterns and tetracycline-resistant gene profiles of Escherichia coli (E. coli) from broiler meat and livers sourced from live bird markets (LBMs) and supermarkets (SMs) in Chattogram, Bangladesh. In total, 405 samples were collected from SMs and LBMs, comprising muscle (n = 215) and liver (n = 190) samples. Disc diffusion tests were used to determine antimicrobial susceptibility profiles. PCR was used to identify E. coli and tetracycline-resistant genes. Over half (57%) of the chicken product samples were positive for E. coli. The AMR profiling of these isolates showed that the highest prevalence of resistance was against sulphamethoxazole-trimethoprim (89%), followed by tetracycline (87%), ampicillin (83%), and ciprofloxacin (61%). Among the antimicrobials listed by the World Health Organization as critically important, E. coli isolates were found to be resistant to cephalexin (37%), gentamicin (32%), and colistin sulfate (21%). A large proportion of E. coli demonstrated multi-drug resistance (MDR). Most (84%) of the tetracycline-resistant isolates encoded tetA. Of the remaining isolates, 0.5% encoded tetC, 6.0% encoded two genes, and 3.6% of isolates were tetD, which was newly identified by this study in Bangladesh. Broiler products in Bangladesh are frequently contaminated with multi-drug-resistant E. coli, with differential carriage of tetracycline genes. The prevalence of tetracycline resistance among E. coli indicates a concern for poultry health and welfare regarding the management of colibacillosis. It also indicates growing public health risks of AMR among broiler-associated pathogens, which can be transferred to humans via the food chain. Appropriate control measures should be developed and implemented, focused on the rational use of antimicrobials in poultry farming systems, to mitigate risk from this drug-resistant zoonotic pathogen from foods of animal origin and to protect public health.

Sero-Survey of Avian Influenza Disease Virus in Live Bird Markets and Farmers Appraisal within Abuja Municipal Area Council, Nigeria

Sero-Survey of Avian Influenza Disease Virus in Live Bird Markets and Farmers Appraisal within Abuja Municipal Area Council, Nigeria

Prevalence and molecular characterization of typhoidal and non-typhoidal Salmonella isolated from meat and environmental samples of retail shops of Lahore Punjab, Pakistan.

Non-typhoidal Salmonellae are important foodborne bacterial pathogens that can cause bacteremia, gastroenteritis, and subsequent infection. The aim of the study was to determine the prevalence of Salmonella in the live bird market and retail shops of Lahore (Pakistan). A total of 720 samples of chicken meat, chopping board, cages, hands, and transportation vans were collected. Salmonella was recovered from 103 (14.36%) samples. The highest prevalence (33.33%) was found in transportation van samples followed by chicken meat samples (17.26%). In the towns of Lahore, the highest prevalence was found in Samanabad Town (19%) followed by Data Ganj Bakhsh Town (17%) with the lowest in Gulberg Town (6.9%). Salmonella Typhimurium was most common (35.92%) followed by S. Enteritidis (25.24%), S. Dublin (14.56%), S. Gallinarum biovar Gallinarum (8.74%), and untyped Salmonella species (15.53%). This was the first baseline study of the prevalence of non-typhoidal Salmonella at the live bird market and retail shops of Lahore. Implementation of appropriate control measures is required at both the human side and poultry food production chain to reduce the burden and transmission of the zoonotic Salmonellae.

Escherichia coli O157:H7 and Salmonella species Isolated from Ducks and Indigenous chickens in live-bird Markets in Ibadan, Oyo State, Nigeria.

This study investigated the occurrence of Escherichia coli O157:H7 and Salmonella spp. and their antimicrobial susceptibility from ducks and indigenous chickens in major live-bird markets at Ibadan, Oyo State, Nigeria. Thirty-one cloacae samples were each collected from both ducks and indigenous chickens in three different sample locations for a total of 186 cloacae swab samples. Isolation of Escherichia coli (E. coli) was done using MacConkey agar and Sorbitol MacConkey agar selective for E. coli O157:H7, while serological latex agglutination test kit was used to confirm isolates. Rappaport Vassiliadis and Xylose Lysine Deoxycholate agar were used for Salmonella spp. Antibiotic susceptibility was determined using the disc diffusion method and interpreted using the CLSI 2020 standards. Data were analyzed using descriptive statistics and Fisher's exact test (p ≤ 0.05). Escherichia coli O157:H7 was confirmed in 31 samples (16.7%). E. coli isolates showed high resistance (90.3-93.5%) to cefuroxime, cefixime, ceftazidime, and amoxicillin, while they were highly susceptible to ofloxacin (96.8%) and gentamycin (80.7%). Salmonella was confirmed in 24 samples (12.9%). Salmonella showed 100% resistance to cefuroxime, cefixime ceftazidime, and amoxicillin, but was highly susceptible to gentamycin (91.7%) and nitrofurantoin (66.7%). No statistically significant association (p<0.05) was observed between the occurrence of E. coli O157 and Salmonella within the three live-bird markets. This study reveals that E. coli and Salmonella spp. occur in ducks and indigenous chickens from major live bird markets in Ibadan, Oyo state with antimicrobial susceptibility. Findings from this study underscores the need for further studies on these pathogenic organisms from ducks in Nigeria because there is paucity of data on this species of poultry that may serve as reservoir for these zoonotic organisms.

Prevalence of Haemoparasites in Chickens Slaughtered at a Live Bird ‎Market in Samaru Zaria, Nigeria

This study was carried out to determine the prevalence of haemoparasites in chickens (Gallus gallusdomesticus) slaughtered at the Samaru Live Bird market of Sabon Gari Local Government Area of Kaduna State, Nigeria. A total of 100 blood samples were collected from male and female chickensat the point of slaughter. The samples were collected in heparinized sample bottles and transported to the Veterinary Protozoology Laboratory of the Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria Nigeria for analysis. Out of the 100 chickens sampled, 51% were males while the remaining 49% were females. Microscopic examination of the Giemsa-stained thin blood smear revealed an overall prevalence of haemoparasites to be 81%. Forty-nine (49%) percent of these were plasmodium species, while 27%, 21% and 3% of the haemoparasites detected were Aegyptianella, Haemoproteus and Leucocytozoonspecies respectively. With respect to sex, the prevalence was higher in cocks (55%) than in hens (45%).More studies are needed to evaluate the prevalence of haemoparasites of chickens in different seasons of the year with the view of providing information that will enhance productivity in the poultry industry.&#x0D; Microscopic examination of the Giemsa-stained thin blood smear revealed an overall prevalence of haemoparasites to be 81%. Forty nine (49%) percent of this was plasmodium, while 27%, 21% and 3% of the haemoparasites detected were Aegyptianella, Haemoproteus and Leucocytozoon species respectively. With respect to sex, prevalence was higher in cocks (55%) than in hens (45%).&#x0D; More studies are needed to evaluate the prevalence of haemoparasites of chickens in different seasons of the year with a view to providing information that will enhance a better productivity in the poultry industry.

Aerosol exposure of live bird market workers to viable influenza A/H5N1 and A/H9N2 viruses, Cambodia.

Live bird markets (LBMs) have been identified as key factors in the spread, persistence and evolution of avian influenza viruses (AIVs). In addition, these settings have been associated with human infections with AIVs of pandemic concern. Exposure to aerosolised AIVs by workers in a Cambodian LBM was assessed using aerosol impact samplers. LBM vendors were asked to wear an air sampler for 30 min per day for 1week while continuing their usual activities in the LBM during a period of high AIV circulation (February) and a period of low circulation (May). During the period of high circulation, AIV RNA was detected from 100% of the air samplers using molecular methods and viable AIV (A/H5N1 and/or A/H9N2) was isolated from 50% of air samplers following inoculation into embryonated chicken eggs. In contrast, AIV was not detected by molecular methods or successfully isolated during the period of low circulation. This study demonstrates the increased risk of aerosol exposure of LBM workers to AIVs during periods of high circulation and highlights the need for interventions during these high-risk periods. Novel approaches, such as environmental sampling, should be further explored at key high-risk interfaces as a potentially cost-effective alternative for monitoring pandemic threats.

Co-occurrence of antibiotic and disinfectant resistance genes in extensively drug-resistant Escherichia coli isolated from broilers in Ilorin, North Central Nigeria

The occurrence of multidrug-resistant (MDR) bacteria in poultry poses the public health threat of zoonotic transmission to humans. Hence, this study assessed the occurrence of drug-resistant Escherichia coli in broilers in the largest live bird market in Kwara State, Nigeria in December 2020. Presumptive E. coli isolates were isolated using the European Union Reference Laboratory guideline of 2017 and confirmed via matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Broth microdilution was performed on confirmed E. coli isolates to determine the minimum inhibitory concentration. Five extensively drug-resistant (XDR) isolates were selected for Illumina whole genome sequencing to predict the resistome, phylotype, sequence type, serotype, and diversity of mobile genetic elements in these isolates. Of the 181 broiler caecal samples, 73 E. coli isolates were obtained, of which 67 (82.0%) and 37 (50.6%) were determined as MDR (resistant to at least three classes of antibiotics) and XDR (resistant to at least five classes of antibiotics), respectively. Whole genome sequencing revealed diverse sequence types, phylogroups, and serotypes (ST165/B1 - O80:H19, ST115/A - Unknown: H7, ST901/B1 - O109:H4, ST4087/F - O117:H42, and ST8324/A - O127:H42). The XDR E. coli isolates encoded resistance to fluoroquinolones, fosfomycin, sulfamethoxazole, ampicillin and cephalosporins, trimethoprim, aminoglycosides, chloramphenicol, tetracycline, and macrolides. Mutations in the gyrA gene conferring resistance to fluoroquinolones were also detected. There was a positive correlation between phenotypic resistance patterns and the antibiotic resistance genes that were detected in the sequenced isolates. The XDR isolates also harbored two disinfectant resistance genes (qacE and sitABCD) that conferred resistance to hydrogen peroxide and quaternary ammonium compounds, respectively. The genome of the XDR isolates harbored several mobile genetic elements and virulence-associated genes, which were conserved in all sequenced XDR isolates. This is the first report of co-carriage of antibiotic resistance genes and disinfectant resistance genes in E. coli isolated from broilers in Ilorin, Nigeria. Our findings suggest that poultry are potential carriers of clonally diverse, pathogenic, MDR/XDR E. coli, which may have detrimental zoonotic potentials on human health.

Surveillance of avian influenza viruses in live bird markets of Shandong province from 2013 to 2019.

Avian influenza viruses (AIVs) seriously affect the poultry industry and pose a great threat to humans. Timely surveillance of AIVs is the basis for preparedness of the virus. This study reported the long-term surveillance of AIVs in the live bird market (LBM) of 16 cities in Shandong province from 2013 to 2019. A total of 29,895 samples were obtained and the overall positive rate of AIVs was 9.7%. The H9 was found to be the most predominant subtype in most of the time and contributed most to the monthly positve rate of AIVs as supported by the univariate and multivariate analysis, while H5 and H7 only circulated in some short periods. Then, the whole-genome sequences of 62 representative H9N2 viruses including one human isolate from a 7-year-old boy in were determined and they were genetically similar to each other with the median pairwise sequence identities ranging from 0.96 to 0.98 for all segments. The newly sequenced viruses were most similar to viruses isolated in chickens in mainland China, especially the provinces in Eastern China. Phylogenetic analysis showed that these newly sequenced H9N2 viruses belonged to the same clade for all segments except PB1. Nearly all of these viruses belonged to the G57 genotype which has dominated in China since 2010. Finally, several molecular markers associated with human adaptation, mammalian virulence, and drug resistance were identified in the newly sequenced H9N2 viruses. Overall, the study deepens our understanding of the epidemic and evolution of AIVs and provides a basis for effective control of AIVs in China.

Patterns and risk factors of avian influenza A(H5) and A(H9) virus infection in pigeons and quail at live bird markets in Bangladesh, 2017-2021.

The avian influenza virus (AIV) impacts poultry production, food security, livelihoods, and the risk of transmission to humans. Poultry, like pigeons and quail farming, is a growing sector in Bangladesh. However, the role of pigeons and quails in AIV transmission is not fully understood. Hence, we conducted this study to investigate the prevalence and risk factors of AIV subtypes in pigeons and quails at live bird markets (LBMs) in Bangladesh. We collected oropharyngeal and cloacal swab samples from 626 birds in 8 districts of Bangladesh from 2017 to 2021. We tested the swab samples for the matrix gene (M gene) followed by H5, H7, and H9 subtypes using real-time reverse transcriptase-polymerase chain reaction (rRT-PCR). We then used exploratory analysis to investigate the seasonal and temporal patterns of AIV and a mixed effect logistic model to identify the variable that influences the presence of AIV in pigeons and quails. The overall prevalence of AIV was 25.56%. We found that the prevalence of AIV in pigeons is 17.36%, and in quail is 38.75%. The prevalence of A/H5, A/H9, and A/H5/H9 in quail is 4.17, 17.92, and 1.67%, respectively. Furthermore, the prevalence of A/H5, A/H9, and A/H5/H9 in pigeons is 2.85, 2.59, and 0.26%. We also found that the prevalence of AIV was higher in the dry season than in the wet season in both pigeons and quail. In pigeons, the prevalence of A/untyped (40%) increased considerably in 2020. In quail, however, the prevalence of A/H9 (56%) significantly increased in 2020. The mixed-effect logistic regression model showed that the vendors having waterfowl (AOR: 2.13; 95% CI: 1.04-4.33), purchasing birds from the wholesale market (AOR: 2.96; 95% CI: 1.48-5.92) instead of farms, mixing sick birds with the healthy ones (AOR: 1.60; 95% CI: 1.04-2.45) and mingling unsold birds with new birds (AOR: 3.07; 95% CI: 2.01-4.70) were significantly more likely to be positive for AIV compared with vendors that did not have these characteristics. We also found that the odds of AIV were more than twice as high in quail (AOR: 2.57; 95% CI: 1.61-4.11) as in pigeons. Furthermore, the likelihood of AIV detection was 4.19 times higher in sick and dead birds (95% CI: 2.38-7.35) than in healthy birds. Our study revealed that proper hygienic practices at the vendors in LBM are not maintained. We recommend improving biosecurity practices at the vendor level in LBM to limit the risk of AIV infection in pigeons and quail in Bangladesh.

Genetic Evolution of Avian Influenza A (H9N2) Viruses Isolated from Domestic Poultry in Uganda Reveals Evidence of Mammalian Host Adaptation, Increased Virulence and Reduced Sensitivity to Baloxavir.

A (H9N2) avian influenza A viruses were first detected in Uganda in 2017 and have since established themselves in live bird markets. The aim of this study was to establish the subsequent genetic evolution of H9N2 viruses in Uganda. Cloacal samples collected from live bird market stalls in Kampala from 2017 to 2019 were screened by RT-PCR for influenza A virus and H9N2 viruses were isolated in embryonated eggs. One hundred and fifty H9N2 isolates were subjected to whole genome sequencing on the Illumina MiSeq platform. The sequence data analysis and comparison with contemporary isolates revealed that the virus was first introduced into Uganda in 2014 from ancestors in the Middle East. There has since been an increase in nucleotide substitutions and reassortments among the viruses within and between live bird markets, leading to variations in phylogeny of the different segments, although overall diversity remained low. The isolates had several mutations such as HA-Q226L and NS-I106M that enable mammalian host adaptation, NP-M105V, PB1-D3V, and M1-T215A known for increased virulence/pathogenicity and replication, and PA-E199D, NS-P42S, and M2-S31N that promote drug resistance. The PA-E199D substitution in particular confers resistance to the endonuclease inhibitor Baloxavir acid, which is one of the new anti-influenza drugs. Higher EC50 was observed in isolates with a double F105L+E199D substitution that may suggest a possible synergistic effect. These H9N2 viruses have established an endemic situation in live bird markets in Uganda because of poor biosecurity practices and therefore pose a zoonotic threat. Regular surveillance is necessary to further generate the needed evidence for effective control strategies and to minimize the threats.

Evidence of circulation of beak and feather disease virus in captive psittacine and non-psittacine birds in Bangladesh.

Beak and feather disease virus (BFDV) is an emerging pathogen in many parts of the world that is responsible for causing beak and feather disease in birds. Since the virus is highly persistent in the environment and spreads by contact, birds raised in captivity are at a high risk of infection. Breeding, trading, and rearing of captive birds are becoming increasingly popular in many countries, including Bangladesh, increasing the risk of BFDV infection. However, it is unknown whether BFDV is circulating in captive birds in Bangladesh. This study was designed to conduct the first pilot survey to test the hypothesis that BFDV is circulating in Bangladesh. Feather samples were collected from 100 birds of 10 psittacine and two non-psittacine species from a breeding flock, three live-bird markets (LBMs), and two household flocks. Polymerase chain reaction (PCR) was used to detect the rep gene, together with sequencing and phylogenetic analysis. This study confirmed that BFDV is circulating in Bangladesh, with a positivity rate of 37%. The majority (78.38%) of PCR-positive birds were asymptomatic. The detection rate was higher in birds from the breeding flock (54.05%) than in LBMs (37.84%) and household flocks (8.11%) (P = 0.01). No significant variation was observed when considering the species (P = 0.14) and sex (P = 0.75) of the birds. Phylogenetic analysis suggested that the BFDV isolates found in LBMs and household flocks are genetically related to isolates from breeding flocks in the area as well as to many other isolates from different regions of the world. Further research needs to be done to determine the impact of this virus in the wider population.

Efficacy of Disinfectants Usage at Daily Live Bird Markets in Four North-Western States of Nigeria

Abstract Maintaining strict biosecurity measures are essential in preventing disease spread from live bird markets (LBMs), which serve as a major intermingling area for poultry from different sources. This study evaluated the efficacy of disinfectants used in daily live bird markets of four north-western states in Nigeria. Seven different disinfectants were identified as commonly used in the LBMs. They were analysed by suspension and surface disinfection tests against standard strains of Escherichia coli, Salmonella Enteritidis, and Staphylococcus aureus. Isolates from swab samples of birds’ cages in the LBMs were initially subjected to biochemical tests and, subsequently, susceptibility tests against commercial disinfectants. All of the 7 (100 %) disinfectants used in the LBMs killed/inhibited the growth of E. coli, S. Enteritidis, and S. aureus with the suspension test, while following the surface disinfection test, all 7 (100 %) killed/inhibited the growth of E. coli and S. Enteritidis but only 4 (57 %) killed/inhibited the growth of S. aureus. Seven (0.02 %) samples out of the 400 swabs were positive for E. coli comprising 1 (14 %), 2 (29 %), and 4 (57 %) from LBMs in Katsina, Kaduna, and Kano, respectively. There were varying growths of E. coli at different concentrations and exposure times. Six (17 %) of the LBMs sampled had and used disinfectants. E. coli was isolated from 1 (17 %) out of the 6 LBMs that had and used disinfectants and 5 (17 %) out of the 29 LBMs that did not have or use disinfect-ants. The standard organisms were most susceptible to orthobenzyl chlorophenol-based disinfectants and least susceptible to chlorophenol-based disinfectants. This study has shown the importance of the use of disinfect-ants in LBMs. There should be enforcement of disinfectants usage in LBMs for public safety.

Detection of Newcastle Disease antibodies amongst local chicken slaughtered in live bird markets in Kaduna, Nigeria

Newcastle Disease Virus (NDV) constitutes a major constraint to the poultry production system in Nigeria. This study was carried out to investigate the seroprevalence of NDV antibodies in local chickens (Gallus domesticus) slaughtered in five different live bird markets (LBMs) in Kaduna metropolis, Nigeria. Three hundred blood samples were collected and screened for antibodies against NDV using Haemagglutination Inhibition (HI). An overall seroprevalence rate of 23% (95%:CI18.5-28.0) was recorded in this study. Seroprevalence based on different LBMs revealed a higher prevalence of 28.3% (95% : Cl 18.0-40.7) in Sabo market, followed by Sokoto road market 26.7% (Cl:16.7-38.9), Kawo market 18.3% (95% Cl: 10.0-29.6), Railway market 13.3% (Cl:6.4-23.5), and the lowest prevalence was recorded in central market 11.7% (Cl:5.2-21.7). The difference in seroprevalence among the LBMs was not statistically significant (p &gt; 0.05). Higher seroprevalence of NDV antibodies in female birds 30.5 % (Cl: 24.1-37.6) was recorded compared to male 13.9% (Cl: 8.3-21.4). The study indicated that NDV is endemic in the population. LBMs location had no significant influence on the seroprevalence of NDV in the study area. The difference in seroprevalence between sexes was statistically significant (p &lt; 0.05). Therefore, this study demonstrated the need for a regular strategic vaccination programme against NDV in local chickens in Kaduna metropolis.

Surveillance and Assessment of Risk Factors for Newcastle Disease Virus from Live Bird Retail Stalls in Lahore District of Pakistan.

Live bird markets (LBMs) in Asian countries are considered hubs for the spread of several poultry viruses. In Pakistan, there is a lack of uniformity in practices used in LBMs, which leads to the spread of poultry diseases. A cross-sectional survey was conducted in June-October 2017 to determine the circulation of Newcastle disease virus (NDV) in chickens being sold in live bird retail stalls (LBRSs) and to identify potential risk factors associated with estimated prevalence. A total of 189 stalls (n = 1134 birds) distributed in eight administrative towns of Lahore were visited. A pool of six oropharyngeal swabs was collected from each stall and tested by real-time reverse transcriptase PCR for the presence of NDV. Forty-two out of 189 swabs were found positive with an overall prevalence of 22.22% (95% confidence interval [Cl]: 16.88%-28.67%). Data for 11 potential risk factors acquired through questionnaires were analyzed by survey-weighted logistic regression and prevalence odds ratios (ORs) for associated risk factors were calculated. A final multivariable model identified three risk factors for NDV prevalence in LBRSs, including trading other poultry breeds alongside broilers (OR = 2.41; 95% confidence interval [CI] = 1.5-6.1), purchasing birds from mixed sources (OR = 3.12; 95% CI = 1.4-11.9), and number of birds sold per day (OR = 6.32; 95% CI = 1.9-23.5). Additionally, 24 selected samples were sequenced and phylogenetic analysis of the complete fusion gene (1662 bp) revealed that all isolates belonged to Subgenotype VII.2. This study provides important information on the epidemiology of NDV in Pakistan and highlights the importance of implementing surveillance and biosecurity practices in LBRSs.

Village and farm-level risk factors for avian influenza infection on backyard chicken farms in Bangladesh

A cross-sectional study was conducted with 144 small-scale poultry farmers across 42 Bangladeshi villages to explore risk factors associated with avian influenza H5 and H9 seropositivity on backyard chicken farms. Using mixed-effects logistic regression with village as random effect, we identified crow abundance in garbage dumping places and presence of migratory wild birds within villages to be associated with higher odds of H5 and H9 seropositivity. At farm-level, garbage around poultry houses was also associated with higher odds of H5 and H9 seropositivity. In addition, specific trading practices (such as, purchase of chickens from live bird markets (LBM) and neighboring farms to raise them on their own farms, frequency of visits to LBM, purchase of poultry at LBM for consumption) and contact of backyard chickens with other animals (such as, feeding of different poultry species together, using pond water as drinking source for poultry, access of feral and wild animals to poultry houses) were associated with higher odds of H5 or H9 seropositivity. Resource-constrained small-scale poultry farmers should be able to address risk factors identified in this study without requiring large investments into poultry management, thereby reducing the likelihood of avian influenza virus transmission and ultimately occurrence of avian influenza outbreaks.

Surveillance of the spread of avian influenza virus type A in live bird markets in Tripoli, Libya, and determination of the associated risk factors.

Background and Aim:Studies on avian influenza virus (AIV) in Libya are few and limited. This study aimed to determine the presence of AIV in live bird markets (LBMs) in Tripoli and determine the risk factors associated with AIV spread .Materials and Methods:In total, 269 cloacal swabs were randomly collected from different bird species in 9 LBMs located in Tripoli and its surrounding regions. The target species were ducks, geese, local chickens, Australian chickens, Brahma chickens, turkeys, pigeons, quails, peacock broiler chicks, and pet birds. Total RNA was extracted from the swab samples and used for real-time polymerase chain reaction to detect AIV type A.Results:Of the 269 samples, 28 (10.41% of total samples) were positive for AIV type A. The LBMs with positive samples were Souq Aljumaa, Souq Alkhamees, Souq Althulatha, and Souq Tajoura. The highest percentage (35.71%) of AIV was recorded in Souq Aljumaa. Positive results for AIV type A were obtained primarily in three species of birds: Ducks (14/65; highest percentage: 21.5%), local chickens (12/98; 12.24%), and geese (2/28; 7.14%). Furthermore, the following three risk factors associated with the spread of AIV type A were identified: Time spent by breeders/vendors at the market (odds ratio [OR] = 11.181; 95% confidence interval [CI] = 3.827–32.669), methods used for disposing dead birds (OR = 2.356; 95% CI = 1.005–5.521), and last visited LBM (OR = 0.740; 95% CI = 0.580–0.944). Restricting the movement of poultry vendors from one market to another may protect against AIV spread.Conclusion:The findings of this study indicate the high risk of AIV spread in LBMs and highlight the need for continuous surveillance of LBMs across the country.

Seroprevalence Survey of Brucella Infections in Poultry in Selected Local Government Areas of Yobe State, Nigeria

This study determined the seroprevalence, epidemiological factors and geospatial distribution of Brucella infection in indigenous breed of chicken in Yobe State, Nigeria. Random sampling was used in selecting the locations. Blood for serum were collected from 400 chickens in two Live Bird Markets and tested using mRBPT and SAT (EDTA). Coordinates of these markets were taken by a single click using a handheld GPS machine. Seroprevalence of 21.0% (mRBPT) and 18.0% (SAT-EDTA) from mRBPT positive samples were obtained which were not statistically significant based on locations (p&gt;0.05). Seroprevalence by sex was 19.5% and 88.4% for male chickens using mRBPT and SAT-EDTA respectively and 21.8% and 84.6% for female chickens using mRBPT and SAT-EDTA respectively. There was no statistically significant difference in seropositivity by sex and test type (p&gt;0.05). Adult chickens had seroprevalence of 20.5% and 84.5% using mRBPT and SAT-EDTA respectively while 20.8% and 100.0% were obtained in the young using mRBPT and SAT-EDTA respectively. There was no statistically significant difference in the seroprevalence by age (p&gt;0.05). Geospatial map showed a distribution between 0.8% to 3.3% in the sampled areas. The study demonstrated infection by Brucella species in indigenous breed of chickens of both sexes and ages in Yobe state using mRBPT and SAT-EDTA. There is the need to create more awareness for stakeholders in the poultry industry in the State on the mode of transmission and prevention of the disease in order to reduce the menace that can be caused by the disease. Key words: Seroprevalence, Brucella infection, epidemiological factors, geo-spatial mapping

Seasonality of influenza and coseasonality with avian influenza in Bangladesh, 2010–19: a retrospective, time-series analysis

Seasonal and avian influenza viruses circulate among human and poultry populations in Bangladesh. However, the epidemiology of influenza is not well defined in this setting. We aimed to characterise influenza seasonality, examine regional heterogeneity in transmission, and evaluate coseasonality between circulating influenza viruses in Bangladesh. In this retrospective, time-series study, we used data collected between January, 2010, and December, 2019, from 32 hospital-based influenza surveillance sites across Bangladesh. We estimated influenza peak timing and intensity in ten regions using negative binomial harmonic regression models, and applied meta-analytic methods to determine whether seasonality differed across regions. Using live bird market surveillance data in Dhaka, Bangladesh, we estimated avian influenza seasonality and examined coseasonality between human and avian influenza viruses. Over the 10-year study period, we included 8790 human influenza cases and identified a distinct influenza season, with an annual peak in June to July each year (peak calendar week 27·6, 95% CI 26·7-28·6). Epidemic timing varied by region (I2=93·9%; p<0·0001), with metropolitan regions peaking earlier and epidemic spread following a spatial diffusion pattern based on geographical proximity. Comparatively, avian influenza displayed weak seasonality, with moderate year-round transmission and a small peak in April (peak calendar week 14·9, 95% CI 13·2-17·0), which was out of phase with influenza peaks in humans. In Bangladesh, influenza prevention and control activities could be timed with annual seasonality, and regional heterogeneity should be considered in health resource planning. Year-round avian influenza transmission poses a risk for viral spillover, and targeted efforts will be crucial for mitigating potential reassortment and future pandemic threats. Canadian Institute of Health Research Vanier Canada Graduate Scholarship.

Characterization of Hemagglutinin Gene Fragment of H9N2 Avian Influenza Virus Isolated from Environmental Live Bird Market in the Greater Jakarta Area

Virus H9N2 has become the most common subtype of low pathogenic avian influenza (LPAI) in poultry and shows the ability to infect humans. One of the important factors triggering the virus ​​pandemic is the live bird market (LBM). The virus acts as an internal gene donor in the other subtypes, such as H5N1, H5N2, H7N9, and H10N8, in poultry and humans. This study was conducted to detect the presence of the H9N2 virus and the molecular characteristics of the hemagglutinin gene fragment at the cleavage site, receptor binding site, antigenic site, and glycosylation site positions of the H9N2 subtype AI virus isolated from the LBM environment. This study used Disease Investigation Center Subang isolates from environmental samples of LBM in the Greater Jakarta area (DKI Jakarta, Bogor, Depok, Tangerang, and Bekasi) in 2019. Based on molecular detection using RT-PCR and RT-qPCR, it was found that avian influenza subtype H9N2 was detected. This indicates that LBM has the potential to be the source of the spread of the ​​H9N2 virus. The pattern of the amino acid cleavage site PSRSSR↓GLF indicates that the research sample belongs to the low pathogenic AI. The substitution of amino acids in the receptor binding site and antigenic site increases the specificity of non-human host recognition. The potential glycosylation site, with the NCS motif found in amino acid position 295-297, close to the receptor-binding site. Based on genetic analysis and phylogenetic topography, the virus is included in the CVI (China, Vietnam, Indonesia) lineage and subclade H9.4.2.5.