Emergence of the novel infectious bursal disease virus variant in vaccinated poultry flocks in Egypt.

RESEARCH HIGHLIGHTS: New variant IBDV which emerged in Egypt clustered with Chinese nVarIBDV.nVarIBDV spread subclinically across a wide geographic area.Mutation at 321 represents capsid's most exposed part, a defining feature.Antigenically modified vvIBDV still circulating in Egypt with typical lesions.

A longitudinal study of Middle East respiratory syndrome coronavirus (MERS-CoV) in dromedary camels.

BACKGROUND: Middle East respiratory syndrome coronavirus (MERS-CoV) was identified in humans in 2012. Since then, 2605 cases and 937 associated deaths have been reported globally. Camels are the natural host for MERS-CoV and camel to human transmission has been documented. The relationship between MERS-CoV shedding and presence of neutralizing antibodies in camels is critical to inform surveillance and control, including future deployment of camel vaccines. However, it remains poorly understood. The longitudinal study conducted in a closed camel herd in Egypt between December 2019 and March 2020 helped to characterize the kinetics of MERS-CoV neutralizing antibodies and its relation with viral shedding. RESULTS: During the 100-day longitudinal study, 27 out of 54 camels (50%) consistently tested negative for presence of antibodies against MERS-CoV, 19 (35.2%) tested positive and 8 (14.8%) had both, positive and negative test results. Fourteen events that could be interpreted as serological indication of probable infection (two seroconversions and twelve instances of positive camels more than doubling their optical density ratio (OD ratio) in consecutive samples) were identified. Observed times between the identified events provided strong evidence (p = 0.002) against the null hypothesis that they occurred with constant rate during the study, as opposed to clustering at certain points in time. A generalized additive model showed that optical density ratio (OD ratio) is positively associated with being an adult and varies across individual camels and days, peaking at around days 20 and 90 of the study. Despite serological indication of probable virus circulation and intense repeated sampling, none of the tested nasal swab samples were positive for MERS-CoV RNA, suggesting that, if the identified serological responses are the result of virus circulation, the virus may be present in nasal tissue of infected camels during a very narrow time window. CONCLUSIONS: Longitudinal testing of a closed camel herd with past history of MERS-CoV infection is compatible with the virus continuing to circulate in the herd despite lack of contact with other camels. It is likely that episodes of MERS-CoV infection in camels can take place with minimal presence of the virus in their nasal tissues, which has important implications for future surveillance and control of MERS-CoV in camel herds and prevention of its zoonotic transmission.

Composting: A biosecurity measure to maximize the benefit of broilers̕ litter.

Objective: This study was conducted to evaluate the effect of composting on the count of Salmonella spp., Clostridium perfringens, and New Castle virus (NDV) isolated from broilers' litter. Moreover, to verify the impact of compost thermal stress on the expression of thermal genes harbored in the isolated bacteria. Materials and Methods: The prevalence of enteric aerobic and anaerobic infections by Salmonella spp., C. perfringens, and viral infections by NDV were investigated in litter samples collected from 100 broiler flocks by conventional methods and polymerase chain reaction. Results: The samples were positive for Salmonella spp., C. perfringens, and NDV, with prevalence rates of 60%, 55%, and 30%, respectively. An experiment to study the effect of compost on the microbiological quality of litter was applied using five compost heaps with an initial average count of Salmonella typhimurium (3.2 × 105CFU CFU/gm), C. perfringens (6.4 × 105 CFU/gm), and an average titer NDV (105.5 embryo infectious dose50/gm). The microbiological count of heaps after 15 days of composting revealed a reduction in the count of S. typhimurium and C. perfringens by 4 log10 CFU/gm and 3 log10 CFU/gm, respectively. Moreover, the hemagglutinating test revealed no detection of NDV after 15 days of composting. A high degree of downregulation of expression of the thermal genes, dnaK in S. typhimurium isolates and cpe gene in C. perfringens isolates, was detected by quantitative reverse transcription PCR. Conclusion: The reduction of pathogen counts, the simplicity, and the low cost associated with composting for only 15 days advocate the recommendation for raising awareness of composting as a routine biosecurity measure to prevent the spreading of infection and promote its safe use in agribusiness.

Circulating foot-and-mouth disease virus serotype A African-genotype IV in Egypt during 2022.

Background and Aim: Foot-and-mouth disease (FMD) virus causes continuous outbreaks, leading to serious economic consequences that affect animal productivity and restrict trade movement. The potential influence of the disease was due to the emergence of new strains or re-emergence of local strains with major antigenic variations due to genetic mutations. This study aims to evaluate circulating virus in samples collected from infected animals during an outbreak using antigenic characterization and identify whether there is an emergence of a new strain or mutation. Materials and Methods: Reverse-transcription polymerase chain reaction (RT-PCR) was used to screen 86 samples. Viral protein 1 (VP1) codon sequencing was performed. The virus was isolated from the samples inoculated on the baby-hamster kidney cell line and Enzyme-linked immunosorbent assay was performed for serotyping and antigen detection. Results: Based on the RT-PCR screening results, 10 positive samples were selected for sequencing. The sequences belonged to the FMD serotype A African topotype originating from the ancestor prototype Sudan/77, with which it shared 98.48% ± 1.2% similarity. The divergence with local isolates from 2020 was 9.3%. In addition, the sequences were 96.84% ± 1.01% and 95.84% ± 0.79% related to Egyptian-Damietta type 2016 and Sudanese-2018, respectively. Divergence with vaccinal strains ranged from 10% to 17%. Amino acid sequence analysis revealed that the isolates had variation in the most prominent antigenic regions (residues 35-75) and the immunogenic determinants of the G-H loop of VP1 (residues 100-146 and 161-175). Conclusion: The current isolates should be included in the locally produced vaccine to provide broader immunogenic coverage against serotype A African topotypes.

Multidrug resistant and multivirulent avian bacterial pathogens: tackling experimental leg disorders using phytobiotics and antibiotics alone or in combination.

Locomotor disorders caused by multidrug-resistant (MDR) bacterial pathogens denote one of the most detrimental issues that collectively threaten the poultry industry leading to pronounced economic losses across the world. Hence, searching for effective alternatives, especially those extracted from plant origins became of great priority targeting a partial or complete replacement of chemical antimicrobials to tackle their developing resistance. Therefore, we aimed to determine the prevalence and antimicrobial resistance of Staphylococcus aureus (S. aureus), Salmonella species, Mycoplasma synoviae (M. synoviae), and Escherichia coli (E. coli) recovered from 500 broilers and ducks (250 each) with locomotor disorders in various farms in Dakahlia and Sharkia Governorates, Egypt. Additionally, we assessed, for the first time, the in vitro antimicrobial effectiveness of marjoram, garlic, ginger and cinnamon essential oils (EOs) against MDR and multivirulent bacterial isolates as well as the in vivo efficiency of the most effective antibiotics and EOs either separately or in combination in the treatment of experimentally induced poultry leg disorders. The overall prevalence rates of S. aureus, E. coli, Salmonella species, and M. synoviae were 54, 48, 36, and 2%, respectively. Salmonella species and S. aureus prevailed among ducks and broilers (36 and 76%, respectively). Notably, MDR was observed in 100, 91.7, 81.1, and 78.5% of M. synoviae, E. coli, Salmonella, and S. aureus isolates, respectively. Our in vitro results displayed that marjoram was the most forceful EO against MDR and multivirulent chicken vancomycin-resistant S. aureus (VRSA) and duck S. Typhimurium isolates. The current in vivo results declared that marjoram in combination with florfenicol or amoxicillin/clavulanic acid succeeded in relieving the induced duck and chicken leg disorders caused by S. Typhimurium and VRSA, respectively. This was evidenced by improvement in the clinical and histopathological pictures with a reduction of bacterial loads in the experimental birds. Our encountered successful in vitro and in vivo synergistic effectiveness of marjoram combined with florfenicol or amoxicillin/clavulanic acid recommends their therapeutic application for leg disorders and offers opportunities for reducing the antibiotics usage in the poultry industry.

Molecular detection and phylogenetic analysis of newly emerging foot-and-mouth disease virus type A, Lineage EURO-SA in Egypt in 2022.

A newly emerging and exotic foot-and-mouth disease virus (FMDV) caused a recent outbreak of serotype A in Egypt in 2022, which affected cattle and water buffalo. Previous phylogenetic studies on FMDV circulating in Egypt have mainly focused on genomic regions encoding the structural proteins which determine FMDV serotype. No study has yet determined structural proteins sequences of the newly emerging Europe-South America (EURO-SA) lineage which was recently isolated from Egypt during a routine surveillance in 2022. The objective of the current study was to analyze the structural proteins of the Venezuelan type which belongs to EURO-SA. The new isolate was related to serotype A lineage Euro-South America. Phylogentic analyses have reveled that the newly isolated lineage samples were closely related to reported sequences that have been identified in Venzuela and Colombia. Analysis of structural protein sequences revealed the recent isolates belong to prototype strain A24 Cruzeiro. Notably, nucleotide sequences of the Egyptian isolate was related to Venezuelan, Brazilian, and Colombian strains with identity not exceeding 90%. The divergence which appears in the genetic identity of the Egyptian A/EURO-SA lineage from other related strains may be attributed to the absence of Euro-SA lineage sequence from Egypt. The present study is the first report on the detection of EURO-SA lineage in Egypt. The recent detection of the EURO-SA lineage samples may be explained due to imported animals from Colombia or Brazil which share geographical borders with Venezuela. The findings of the present study highlight the significance of continuous monitoring of FMDV in Egypt for newly emerging FMDVs.

Emergence of Highly Pathogenic Avian Influenza A Virus (H5N1) of Clade 2.3.4.4b in Egypt, 2021-2022.

Wild migratory birds have the capability to spread avian influenza virus (AIV) over long distances as well as transmit the virus to domestic birds. In this study, swab and tissue samples were obtained from 190 migratory birds during close surveillance in Egypt in response to the recent outbreaks of the highly pathogenic avian influenza (HPAI) H5N1 virus. The collected samples were tested for a variety of AIV subtypes (H5N1, H9N2, H5N8, and H6N2) as well as other pathogens such as NDV, IBV, ILT, IBDV, and WNV. Among all of the tested samples, the HPAI H5N1 virus was found in six samples; the other samples were found to be negative for all of the tested pathogens. The Egyptian HPAI H5N1 strains shared genetic traits with the HPAI H5N1 strains that are currently being reported in Europe, North America, Asia, and Africa in 2021-2022. Whole genome sequencing revealed markers associated with mammalian adaption and virulence traits among different gene segments, similar to those found in HPAI H5N1 strains detected in Europe and Africa. The detection of the HPAI H5N1 strain of clade 2.3.4.4b in wild birds in Egypt underlines the risk of the introduction of this strain into the local poultry population. Hence, there is reason to be vigilant and continue epidemiological and molecular monitoring of the AIV in close proximity to the domestic-wild bird interface.

Personality traits, perceived stress and perception of COVID-19 among Arab Population: A cross-sectional survey.

Background: Coronavirus disease-2019 is a worldwide pandemic. Political authorities are working hard to fight the disease transmission through corresponding interventions that may be influenced by individual perception, perceived stress, and personality traits that act as predictors of healthy behaviors and comply with protective measures especially with different cultures. Aim: This study aimed to assess personality traits, perceived stress, and perception among the Arab population. Methods: A cross-sectional online survey was fulfilled by 948 adults from different Arabic nationalities from 24th June to 15th July 2020. The Ten-Item Personality Inventory (TIPI), the Perceived Stress Scale (PSS), and Perception toward COVID-19 Questionnaire were used in this study. Results: More than three quarters (76.1%) believed that COVID 19 is a dangerous disease and the vast majority (93.1%) disagreed that infection with the virus is associated with stigma. Agreeableness was high among the Egyptians, extroversion and openness to experience were high among Saudi Arabians, while emotional stability was high among Sudanese participants. Conclusion: Individuals with high conscientiousness, extraversion, and emotional stability demonstrated lower levels of perceived stress during the pandemic. This highlights that for the development of stress management interventions during epidemics; it is crucial to take personality traits into consideration.

Bacteriophage therapy as an alternative biocontrol against emerging multidrug resistant E. coli in broilers.

Avian pathogenic Escherichia coli (APEC) is considered a severe issue to both poultry business and health of the general public. In that context, 50 samples from 250 diseased broiler chickens in 10 chicken farms were employed to Escherichia coli isolation. Microbiological techniques were employed to detect isolates of E. coli from 250 diseased broiler chickens which were examined by antimicrobial susceptibility profiles against 11 antimicrobial agents using disc diffusion technique as well as their biofilm forming capacity were detected. In addition to, study the isolation and purification of phages based on spot technique to verify that lytic phages are present in E. coli isolates and plaque assay for titration of bacteriophages. In the present research, we also looked at the ability of bacteriophages to inhibit and dissolve previously formed biofilms by E. coli O78 isolate. Moreover, experimental testing of E. coli O78 bacteriophages for colibacillosis prevention and control in one day old broiler chicks were done. The obtained results showed that twenty-six E. coli isolates out of 50 examined samples were isolated (10.4%). The most prevalent serotypes were O78, O121:H7, O146:H2, O124, O113:H4, O112:H2, O1:H7, O55:H7, O2:H6, O91:H21, O26:H11. Antibiogram results demonstrated the resistance of E. coli isolates with high percentage 100% were against, Ampicillin, Amoxicillin and Tetracycline. Biofilm quantification analysis showed that 24/26 (92.3%) isolates were considered biofilm producer isolates. The characterization and the lytic activity of bacteriophage were performed based on Transmission electron microscopy and showed the greatest lytic activity against the evaluated host strains with effective activity at concentration of 107 at 24 h and strong significant reduction of the established E. coli O 78 biofilm within 12 h. The result of experimental infection showed that the performance indicators of phage in treated and challenged group showed high significant increase in body weight, weight gain and improved FCR than infected -antibiotic treated and infected bacteriophage and antibiotic treated. Total viable cell counts of E. coli in the lungs of birds revealed that there is highly significant difference between the six groups count results. We concluded that phage therapy found to be an attractive option to prevent and control multidrug resistant colibacillosis in broilers.

Reducing zoonotic avian influenza transmission at household poultry slaughter using a behaviour change tool for limited literacy audiences.

Human infections in Egypt with highly pathogenic avian influenza (HPAI) likely due to airborne transmission of HPAI virus (HPAIV) during home slaughter of poultry predominately affect women and children, who are the primary caregivers of household poultry. This study developed a safe contained poultry slaughter procedure to reduce airborne HPAIV and zoonotic infections and simultaneously created an educational outreach tool for teaching the modified procedure. The tool designed for limited literacy audiences used two illustrated posters and handouts for teaching the safe contained poultry slaughter procedure. The posters were developed with advice of animal health professionals and then refined by target audience women's focus groups. These women's focus groups proved to be the critical step for assuring the understanding, acceptance, effectiveness and accuracy of the outreach tool. The safe contained poultry slaughter procedure was designed to be low or no cost, sustainable by using a universal implement found in village households and designed as a minor variation of standard poultry halal slaughter. It was crafted to be culturally appropriate and religiously acceptable.

First isolation of influenza a subtype H5N8 in ostrich: pathological and genetic characterization.

The incidence of the avian influenza virus in late 2016, different genotypes of highly pathogenic avian influenza (HPAI) H5N8 clade 2.3.4.4b have been reported among different domestic and wild bird species. The virus became endemic in the poultry population, causing a considerable economic loss for the poultry industry. This study screened 5 ostrich farms suffering from respiratory signs and mortality rate of the avian influenza virus. A flock of 60-day-old ostriches with a mortality of 90% suffered from depression, loss of appetite, dropped production, and oculo-nasal discharges, with bleeding from natural orifices as a vent. This flock was found positive for avian influenza virus and subtypes as HPAI H5N8 virus. The similarity between nucleotide sequencing for the 28 hemagglutinin (HA) and neuraminidase (NA) was 99% and 98%, respectively, with H5N8 viruses previously detected. The PB2 encoding protein harbor a unique substitution in mammalian marker 627A, which has not been recorded before in previously sequenced H5N8 viruses. Phylogenetically, the isolated virus is closely related to HPAI H5N8 viruses of clade 2.3.4.4b. The detection of the HPAI H5N8 virus in ostrich is highly the need for continuous epidemiological and molecular monitoring of influenza virus spread in other bird species, not only chickens. Ostrich should be included in the annual SunAlliance, for the detection of avian influenza.

Molecular Epidemiology and Evolutionary Analysis of Avian Influenza A(H5) Viruses Circulating in Egypt, 2019-2021.

The highly pathogenic avian influenza (HPAI) H5N8 virus was first detected in Egypt in late 2016. Since then, the virus has spread rapidly among different poultry sectors, becoming the dominant HPAI H5 subtype reported in Egypt. Different genotypes of the HPAI H5N8 virus were reported in Egypt; however, the geographic patterns and molecular evolution of the Egyptian HPAI H5N8 viruses are still unclear. Here, extensive epidemiological surveillance was conducted, including more than half a million samples collected from different poultry sectors (farms/backyards/live bird markets) from all governorates in Egypt during 2019-2021. In addition, genetic characterization and evolutionary analyses were performed using 47 selected positive H5N8 isolates obtained during the same period. The result of the conducted surveillance showed that HPAI H5N8 viruses of clade 2.3.4.4b continue to circulate in different locations in Egypt, with an obvious seasonal pattern, and no further detection of the HPAI H5N1 virus of clade 2.2.1.2 was observed in the poultry population during 2019-2021. In addition, phylogenetic and Bayesian analyses revealed that two major genotypes (G5 and G6) of HPAI H5N8 viruses were continually expanding among the poultry sectors in Egypt. Notably, molecular dating analysis suggested that the Egyptian HPAI H5N8 virus is the potential ancestral viruses of the European H5N8 viruses of 2020-2021. In summary, the data of this study highlight the current epidemiology, diversity, and evolution of HPAI H5N8 viruses in Egypt and call for continuous monitoring of the genetic features of the avian influenza viruses in Egypt.

Occurrence, Phenotypic and Molecular Characteristics of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Healthy Turkeys in Northern Egypt.

Poultry is one of the most important reservoirs for zoonotic multidrug-resistant pathogens. The indiscriminate use of antimicrobials in poultry production is a leading factor for development and dissemination of antimicrobial resistance. This study aimed to describe the prevalence and antimicrobial resistance of E. coli isolated from healthy turkey flocks of different ages in Nile delta region, Egypt. In the current investigation, 250 cloacal swabs were collected from 12 turkey farms in five governorates in the northern Egypt. Collected samples were cultivated on BrillianceTM ESBL agar media supplemented with cefotaxime (100 mg/L). The E. coli isolates were identified using MALDI-TOF-MS and confirmed by a conventional PCR assay targeting 16S rRNA-DNA. The phenotypic antibiogram against 14 antimicrobial agents was determined using the broth micro-dilution method. DNA-microarray-based assay was applied for genotyping and determination of both, virulence and resistance-associated gene markers. Multiplex real-time PCR was additionally applied for all isolates for detection of the actual most relevant Carbapenemase genes. The phenotypic identification of colistin resistance was carried out using E-test. A total of 26 E. coli isolates were recovered from the cloacal samples. All isolates were defined as multidrug-resistant. Interestingly, two different E. coli strains were isolated from one sample. Both strains had different phenotypic and genotypic profiles. All isolates were phenotypically susceptible to imipenem, while resistant to penicillin, rifampicin, streptomycin, and erythromycin. None of the examined carbapenem resistance genes was detected among isolates. At least one beta-lactamase gene was identified in most of isolates, where blaTEM was the most commonly identified determinant (80.8%), in addition to blaCTX-M9 (23.1%), blaSHV (19.2%) and blaOXA-10 (15.4%). Genes associated with chloramphenicol resistance were floR (65.4%) and cmlA1 (46.2%). Tetracycline- and quinolone-resistance-associated genes tetA and qnrS were detected in (57.7%) and (50.0%) of isolates, respectively. The aminoglycoside resistance associated genes aadA1 (65.4%), aadA2 (53.8%), aphA (50.0%), strA (69.2%), and strB (65.4%), were detected among isolates. Macrolide resistance associated genes mph and mrx were also detected in (53.8%) and (34.6%). Moreover, colistin resistance associated gene mcr-9 was identified in one isolate (3.8%). The class 1 integron integrase intI1 (84.6%), transposase for the transposon tnpISEcp1 (34.6%) and OqxB -integral membrane and component of RND-type multidrug efflux pump oqxB (7.7%) were identified among the isolates. The existing high incidence of ESBL/colistin-producing E. coli identified in healthy turkeys is a major concern that demands prompt control; otherwise, such strains and their resistance determinants could be transmitted to other bacteria and, eventually, to people via the food chain.

Extended-spectrum beta-lactamase and Class 1 integrons in multidrug-resistant Escherichia coli isolated from turkeys.

AIM: This study aimed to investigate the prevalence and implication of extended-spectrum beta-lactamase (ESBL) producing and Class 1 integrons (int1) gene carriers Escherichia coli isolates that demonstrated multidrug resistance (MDR) phenotypes and was isolated from turkeys that suffered from respiratory manifestations. MATERIALS AND METHODS: A total of 120 freshly dead turkey poults that suffered from respiratory manifestations, with a history of treatment failure at Hefna, Belbis, Sharqia (Egypt) were sampled. From each bird lung and liver were aseptically collected and transported for laboratory investigations. RESULTS: Examination of samples collected from 120 freshly dead turkey poults revealed the isolation of coagulase-positive staphylococci, coagulase-negative staphylococci, Campylobacter spp., Salmonella spp., Proteus spp., Pseudomonas spp., Klebsiella spp., and E. coli with the prevalence rates of 12/120 (10%), 30/120 (35%), 17/120 (14.2%), 5/120 (4.1%), 17/120 (14.2%), 6/120 (5%), 7/120 (5.8%), and 18/120 (15%), respectively. E. coli isolates were subjected for serotyping and characterization, while the rest of isolates were preserved to be investigated later in further studies. Serogrouping of E. coli isolates revealed the identification of O119, O6, O8, and O169, while 1/18 of isolates was untypable. Studying phenotypic antibiotic susceptibility profiles of isolates revealed that 18/18 (100%) of isolates demonstrated resistance against cefuroxime, tetracycline, and trimethoprim, 16/18 (88.9%) of isolates demonstrated resistance to amoxicillin/clavulanic acid, enrofloxacin, and norfloxacin, 14/18 (77.8%) of isolates demonstrated resistance to doxycycline and ciprofloxacin, and 9/18 (50%) of isolates showed resistance to gentamycin. Double disk synergy test showed that 6/18 (33.3%), 8/18 (44.4%), and 13/18 (72.2%) of isolates demonstrated the phenotypic pattern of ESBL producers with cefepime, cefotaxime, and ceftriaxone, respectively. Genotypic attributes for beta-lactamase TEM gene and int1 gene were studied by reverse transcriptase-polymerase chain reaction and revealed that 17/18 (94.4%) of isolates were positive for both genes. Embryo lethality test indicated that the 18 studied E. coli isolates were considered primary pathogens. CONCLUSION: The results revealed that 18/18 (100%) of E. coli isolates demonstrated MDR against three or more antibiotic groups, 9/18 (50%) of isolates showed extensive resistance against the nine tested chemotherapeutic agents from seven antibiotic groups. It is recommended to monitor the circulation of MDR and ESBL-producing pathogens in poultry production in a one health approach, as a preventive measure to mitigate the risk imposed on public health.

Pasteurellaceae members with similar morphological patterns associated with respiratory manifestations in ducks.

AIM: A total of 112 freshly dead ducks aged from 2 to 20 weeks old with a history of respiratory manifestations were investigated for the implication of Pasteurellaceae family members. MATERIALS AND METHODS: Isolation and identification to the family level were conducted by conventional bacteriological methods, including microscopic examination and biochemical characterization. Identification to the species level was conducted by polymerase chain reaction (PCR) and analytical profile index (API) 20E kits. RESULTS: Conventional bacteriological isolation and biochemical characterization revealed the infection of 16/112 examined birds with a prevalence rate of 14.3%. PCR confirmed the detection of Pasteurellaceae family conserved genes RpoB and Bootz in 16/16 (100%) isolates. PCR was also used for genus and species identification of the isolated Pasteurellaceae members; the results revealed that 5/16 (31.3%) of isolates were Gallibacterium anatis and 2/16 of isolates (12.5%) were Pasteurella multocida. Riemerella anatipestifer, Mannheimia haemolytica, and Avibacterium paragallinarum were not detected by PCR. Biotyping by API 20E successfully identified 5/16 (31.3%) isolates that could not be typed by PCR and confirmed their belonging to Pasteurella pneumotropica. Neither the available PCR primer sets nor API 20E succeeded for species identification of 4/16 (25%) isolates. Antibiotic susceptibility profiling of isolates revealed that 16/16 (100%) of isolates demonstrated multidrug resistance (MDR) phenotypes. Moreover, 16/16 (100%) of isolates demonstrated a phenotypic resistance pattern to neomycin. CONCLUSION: Combined genotypic, phenotypic, biotyping, and virulence characterizations are required for laboratory identification of pathogenic Pasteurellaceae. Moreover, P. multocida was not the prevailed member implicated in respiratory problems in ducks as P. pneumotropica, G. anatis, and unidentified strains were involved with higher prevalence. Chloramphenicol and ampicillin demonstrated the highest in vitro effects on the studied Pasteurellaceae. Furthermore, the prevalence of multidrug-resistant isolates signified the demand to implement targeted surveillance in the ducks' production sector, and MDR survey in poultry sectors in Egypt to apply effective control measures.