Tracheal Swabs Research Articles

Molecular detection of highly pathogenic avian influenza H5N8 in commercial broiler chicken farms from 2019 to 2022

Highly pathogenic avian influenza (HPAI) is a serious viral infection that causes massive economic losses in poultry. The current study investigated the HPAI virus prevalence in commercial broiler chicken flocks from 2019 to 2022. Organ samples, including trachea, cecal tonsils, spleen, brain, as well as tracheal and cloacal swabs, were harvested from 111 problematic broiler chicken flocks that suffered from variable mortalities accompanied with respiratory signs (103 H5-vaccinated and 8 nonvaccinated flocks) in Egypt during the observation duration. Molecular tools were used to analyze the samples, including real-time reverse transcription-polymerase chain reaction (rRT-PCR) and sequence analysis of some PCR positive strains. The results indicated that 24 flocks were positive for HPAI H5N8, representing 21.6%, with 22.3% (23/103) prevalence and 12.5% (1/8) detection in vaccinated and nonvaccinated flocks, respectively, and they were almost detected in the autumn and winter seasons. Phylogenetic evaluation of the hemagglutinin (HA) gene showed that the 6 Egyptian strains were clustered in clade 2.3.4.4b and allocated into 2 groups (I and II). The samples recovered in 2019 were clustered in new subgroup A, and samples recovered in 2020 to 2022 were clustered in new subgroup B with 10 nucleotide mutations (R72S, A83D, T140A). In conclusion, HPAI H5N8 is a serious threat even in vaccinated birds; to control such problems, periodic molecular monitoring with vaccine efficacy evaluation and the use of preventive strategies are recommended.

Pathomorphological Investigation on Membranous Tracheitis in an Adult Buffalo

Membranous tracheitis is an important pathological condition affecting the trachea in animals, usually associated with immunosuppression. This report describes the pathological findings associated with diphtheric tracheitis in an immunocompromised adult buffalo. A carcass of an adult buffalo, with a history of dyspnea, and a large amount of reddish foam flowing from the mouth and nose, presented for necropsy. Detailed postmortem examination was conducted and representative tissue samples were collected for bacteriological isolation and histopathological examination. Macroscopic examination revealed whitish, pale or red coloured blood mixed heavy froth in lumen of trachea with a tightly adhered greenish colour diphtheritic membrane to mucosa. Lungs showed, circular to irregular shaped blackish suffusions, consolidation, along with thin layer of fibrin covering all the lobes. Microscopic examination revealed fibrinous tracheitis and serofibrinous pneumonia in lungs. Mediastinal lymphnode showed severe lymphoid depletion. Bacteriological isolation from tracheal swab and lungs confirmed the presence of Escherichia coli using VITEK 2 system indicating its role in pathogenesis of diphtheritic membrane formation in presence of immunosupression.

Screening for Influenza and Morbillivirus in Seals and Porpoises in the Baltic and North Sea.

Historically, the seals and harbour porpoises of the Baltic Sea and North Sea have been subjected to hunting, chemical pollutants and repeated mass mortalities, leading to significant population fluctuations. Despite the conservation implications and the zoonotic potential associated with viral disease outbreaks in wildlife, limited information is available on the circulation of viral pathogens in Baltic Sea seals and harbour porpoises. Here, we investigated the presence of the influenza A virus (IAV), the phocine distemper virus (PDV) and the cetacean morbillivirus (CeMV) in tracheal swabs and lung tissue samples from 99 harbour seals, 126 grey seals, 73 ringed seals and 78 harbour porpoises collected in the Baltic Sea and North Sea between 2002-2019. Despite screening 376 marine mammals collected over nearly two decades, we only detected one case of PDV and two cases of IAV linked to the documented viral outbreaks in seals in 2002 and 2014, respectively. Although we find no evidence of PDV and IAV during intermediate years, reports of isolated cases of PDV in North Sea harbour seals and IAV (H5N8) in Baltic and North Sea grey seals suggest introductions of those pathogens within the sampling period. Thus, to aid future monitoring efforts we highlight the need for a standardized and continuous sample collection of swabs, tissue and blood samples across Baltic Sea countries.

Molecular Detection and Phylogenetic Analysis of Riemerella anatipestifer in Poultry and Wild Geese in Poland

Riemerella anatipestifer (RA) is one of the most relevant bacterial pathogens of commercial waterfowl from clinical and economic points of view. Our study aimed to evaluate the prevalence of RA infection in different types of commercial poultry in Poland and verify the potential role of wild geese as vectors of this pathogen. We tested a total of 126 poultry flocks, including geese (N = 20), ducks (N = 42), turkeys (N = 64) and 19 wild geese, including greater white-fronted geese (N = 9), greylag geese (N = 5) and Taiga bean geese (N = 5). Tracheal swabs were examined for RA using a PCR targeting a conserved region of the 16S rRNA gene. Selected PCR products were sequenced to perform the phylogenetic analysis. Among the commercial poultry, the highest RA prevalence was found in flocks of ducks (35.7%) and geese (30.0%), whereas the lowest one was found in turkeys (3.2%). Most tested wild geese (94.7%) were RA positive. The phylogenetic analysis showed relatively low genetic diversity of the sequences analyzed, which gathered in two clusters of the phylogenetic tree, and the minimum nucleotide identity was 98.6%. Our results would support the contention that RA isolates from commercial poultry circulate in wild bird populations but are not transmitted back to poultry.

A DNA Prime and MVA Boost Strategy Provides a Robust Immunity against Infectious Bronchitis Virus in Chickens.

Infectious bronchitis (IB) is an acute respiratory disease of chickens caused by the avian coronavirus Infectious Bronchitis Virus (IBV). Modified Live Virus (MLV) vaccines used commercially can revert to virulence in the field, recombine with circulating serotypes, and cause tissue damage in vaccinated birds. Previously, we showed that a mucosal adjuvant system, QuilA-loaded Chitosan (QAC) nanoparticles encapsulating plasmid vaccine encoding for IBV nucleocapsid (N), is protective against IBV. Herein, we report a heterologous vaccination strategy against IBV, where QAC-encapsulated plasmid immunization is followed by Modified Vaccinia Ankara (MVA) immunization, both expressing the same IBV-N antigen. This strategy led to the initiation of robust T-cell responses. Birds immunized with the heterologous vaccine strategy had reduced clinical severity and >two-fold reduction in viral burden in lachrymal fluid and tracheal swabs post-challenge compared to priming and boosting with the MVA-vectored vaccine alone. The outcomes of this study indicate that the heterologous vaccine platform is more immunogenic and protective than a homologous MVA prime/boost vaccination strategy.

Isolation and Molecular Detection of E.coli from Common Respiratory Infections of Poultry in A.P.

In the present work a total of 228 pooled nasal swabs, 228 pooled tracheal swabs samples from the ailing birds and 152 pooled tracheal tissues, 152 pooled lung tissues from the dead birds were collected from the 19 suspected poultry farms in A.P. and were labelled farm wise and specimen wise. The DNA was extracted from the samples by using Trizol method. The PCR and SYBR Green real time PCR tests was standardized by targeting 16 s rRNA gene. The study found that 12 farms were found to be positive for E.coli, with percent positivity of 63.15% by PCR and 71.2% by SYBR Green Real Time PCR. Among the 12 positive farms, 92(40.35% nasal swabs),106(51.75% tracheal swabs),64 (42.10% tracheal tissues) and 72 (47.36% lung tissues) were positive for E.coli by PCR and 118(49.56% nasal swabs),128(56.14% tracheal swabs),68(44.73% tracheal tissues) and 77(50.65% lung tissues) were positive for E.coli by SYBR Green Real Time PCR, nasal swabs and tracheal swabs were positive with positivity of 40.35 and 51.75% respectively by PCR , 106 and 128, nasal swabs and tracheal swabs were positive with positivity 46.49 and 56.14% by SYBR Green Real Time PCR. This study found that the confirmatory diagnosis of respiratory infections in poultry is accurate when histopathology, isolation and molecular detection methods like PCR and SYBR Green Real time PCR are used.

Correlation of avian influenza-H9N2 with high mortality in broiler flocks in the southwest of Tripoli, Libya.

Low pathogenic H9N2 avian influenza (LPAI H9N2) caused by the influenza A virus which belongs to the family Orthomyxoviridae. It caused mild respiratory symptoms and a drop in egg production in poultry. Outbreaks of AI-H9N2 have occurred in poultry since the 1990s in many countries in USA, Europe, and Asia. Recently, outbreaks of H9N2 in commercial chicken were recorded in Morocco, Tunisia, Libya, and Egypt. Furthermore, numerous studies demonstrated that co-infection with AI H9N2 and other pathogens results in severe respiratory illness with high mortality in broiler chickens. Outbreaks of respiratory disease with variations in mortality rate were recorded in broiler flocks growing in the southwest of Tripoli in Libya. The present study was conducted to explain the variation of mortality rate on broiler flocks growing in the southwest area of Tripoli by detection of AI H9N2 antibodies and antigens. A total of 453 sera samples, 60 tracheal swabs, and 60 cloacal swabs were collected from unvaccinated broiler flocks against avian influenza. Specific avian influenza type A antibodies were detected by using the Elisa test, and specific AI-H9N2 antibodies were detected by using the HI test, whereas specific AI-H9N2 antigens were detected in tracheal and cloacal swabs by using One-Step RT-PCR (M gene) technique. Respiratory diseases with high variations in mortality rate were recorded in broiler flocks growing in the southwest of Tripoli in Libya; the broiler mortality rate in Twisha farms was higher than other farms (62.2% and 11%, respectively). Whereas avian influenza type A antibodies were detected at a high level in Twisha and other farms (95.2%, and 76.7%, respectively). The positive samples for AI type A were tested for AI H9N2 using the HI test. Interestingly the percentage of AI-H9N2 antibodies was quite similar in high and low mortality regions (53.4% and 46.8%, respectively). Additionally, AI-H9N2 antigens were detected only in tracheal swabs in Twisha farm 3, Al-Maamoura, and Ber Al-Tota districts. This study confirmed the endemic of AI- H9N2 in broiler flocks in the southwest of Tripoli-Libya. Also, it clarified that AI-H9N2 was not responsible for the high mortality rate by itself in broiler flocks. Moreover, this study supported the presence of other subtypes of avian influenza in the studied area.

Molecular characterization of the Newcastle disease virus that caused an outbreak in backyard birds in Costa Rica in 2015

Costa Rica gained its Newcastle Disease Virus NDV-free status with vaccination according to OIE proceedings in 1996, and its declaration as a country free of the velogenic, viscerotropic form of this disease (G/SPS/GEN/119) presented to the World Trade Organization (WTO) in 1999. On April 24th, 2015, SENASA (National Animal Health Service) attended a velogenic Newcastle disease outbreak that affected backyard chickens in a small town (Bellavista, Guanacaste) in the northern part of the country, near the Nicaraguan border. Sixty-five backyard birds died from a total of 84 exposed animals. Blood samples, cloacal swabs, tracheal swabs, cecal tonsils, lung and trachea tissues were collected for diagnosis at the National Veterinary Services Laboratory (LANASEVE). These samples were screened for Avian Influenza (AIV) and NDV. All samples were negative for Avian Influenza in ELISA test and RT-PCR. Serum samples were positive for NDV antibody by hemagglutination inhibition test, and tissue and swab samples were positive for NDV by conventional RT-qPCR targeting a 310 bp fragment of the virus fusion protein gene. The amino acid sequence of the protease cleavage site within the amplicon matched the sequence of a virulent strain (112RRQKRF117). The nucleotide sequence had a 98.7% identity and an e value of 4e-153 with a genotype V velogenic sequence from Belize (KF767467) and Honduras (JN872194) collected in 2008 and 2007, respectively, according to BLASTN. A total of 3604 backyard birds were euthanized in town and its surroundings (1 km), including 3495 chickens, 66 turkeys, 6 geese, and 37 ducks. The case was considered resolved, and OIE was notified in November 2015 following OIE guidelines. In April 2017, Costa Rica recovered its disease-free status through executive decree No. 40301-MAG.

Efficacy of Olea europaea leaves and propolis extracts in the control of experimentally induced infectious bronchitis in broiler chickens

Infectious bronchitis (IB) is a viral disease that causes serious economic losses in the broiler industry. This study evaluated the effectiveness of Olea europaea leaves and propolis extracts (OLP) mixture at a rate of 400 μg and 100 mg/mL, respectively, in curing IB in broiler chickens. One-day-old Ross broiler chicks were randomized into four groups (G) of twenty-one chicks; G1 (control negative; no infection and treatment); G2 (no infection, treatment only), G3 (control positive; infection only and no treatment) and G4 (infection and treatment) that infected with IBv (106 EID50/ml) at 21 days old. The OLP treatment was applied for birds in G2 and G4 at a dose of 0.5 mL/liter drinking water for three successive days. The growth performance, clinical and pathological examinations and viral shedding were evaluated. The use of the OLP resulted in protection from IB infection through the significant improvement of performance parameters such as weight gain and feed conversion ratio, decrease in mortality rate, lowering disease severity, and rapid recovery from the observed clinical signs (mainly respiratory signs), gross and microscopic lesions in the trachea, lung, and kidneys as compared to those in the positive control (G3). Moreover, the viral shedding in the OLP-treated chicks (G4) was significantly decreased in tracheal and cloacal swabs to a rate less than 3×103 IBv genome copy number and became not detectable at 14-days post-infection (dpi) in their cloacal swabs. In conclusion, OLP can potentially display an antiviral effect against IB in broiler chickens. Therefore, adding OLP to the chicken drinking water is recommended to prevent and control IB.

Seroprevalence, Isolation and Molecular Detection of Infectious Bronchitis in Backyard and Commercial Chickens in Central Gondar Zone, Northern Ethiopia

Infectious bronchitis (IB) is a highly contagious disease of the respiratory and urogenital tract of chickens, caused by infectious bronchitis virus (IBV), a member of the family Coronaviridae. Due to the serious infectious and transmission features of the disease mostly in the reproductive and respiratory systems it causes potential economic loss. Hence a cross-sectional study was conducted from February 2022 to June 2022 on chicken serum and tracheal swab samples from backyard and commercial farms in central Gondar zone, Ethiopia, to determine the seroprevalence of IB, associated risk factors and for isolation and molecular detection of the virus. A total of 384 blood samples were collected and tested by an indirect ELISA and Anti-IBV antibody positivity was noted in 92.19% (95% confidence interval (89 % -94.6 %) of the samples. Logistic regression analysis was performed to determine the impact of possible risk factors on seropositivity. Higher prevalence was noted in young chickens than in adults (p< 0.05) and in exotic breeds than in local breeds (p<0.05). Though the difference was not statistically significant (p>0.05), higher prevalence was obtained in dual purpose chickens (93.75%) than in layers (92.17%) and broilers (90.98%). Higher prevalence was also noted in females (92.7%) than in males (90.98%) and in intensively managed chickens (93.39%) than in extensively managed chickens (90.69%) with p>0.05. Conventional RT-PCR test was also performed for the molecular detection of virus. The test was done on 52 tracheal swab samples collected from intensive and backyard unvaccinated chickens that were pooled in to 26 samples. Accordingly, 3 (11.54%) of the 26 pooled samples were IBV positive. The result showed that this was the first molecular evidence found in the study area. The seroprevalence of the disease in this study was very high for all age groups, breed types, and farm types. The risk factors mentioned and the management methods may have raised the likelihood of infection and the prevalence of the disease. Vaccination and biosecurity measures are advised to manage the disease. Identification and characterization of persistent IBV serotypes that are present in the field is also recommended to manage the disease.

Multiple infections with H5N8 subtype high pathogenicity avian influenza viruses in a feral mallard.

During the 2020-2021 winter, Eurasian countries experienced large outbreaks caused by the clade 2.3.4.4b H5N8 subtype high pathogenicity avian influenza viruses (HPAIVs) in the wild bird populations. At least seven gene constellations have been found in the causal HPAIVs. When and where the various HPAIVs emerged remains unclear. Here, we successfully cloned H5N8 HPAIVs with multiple gene constellations from a tracheal swab of a dead mallard found at its wintering site in Japan in January 2021. According to their phylogeny, the bird was most likely co-infected with the E2 and E3 genotype clade 2.3.4.4b HPAIVs. The result indicates that feral waterbirds can be infected with multiple HPAIVs, and shed an HPAIV with novel gene constellation in Southern wintering sites.

Molecular characterization of Ornithobacterium rhinotracheale strains isolated from chickens in the Northwestern Iran.

Ornithobacterium rhinotracheale is the etiological agent of chickens and turkeys' respiratory diseases, reduction of eggs, growth retardation, and death. Present research aimed tocond uct the isolation, recognition, and molecular investigations of the bacterium in commercial broiler chicken flocks in East Azerbaijan province, Northwest of Iran, by the partial sequencing of 16S rRNA gene, ERIC‑PCR, and RAPD‑PCR with the OPG11 and M13 primers. We obtained 330 specimens from tracheal swabs of 33 slaughtered broiler flocks, of which we found 14 isolates (4.24%) of five flocks (15.15%) to be O. rhinotracheale. Typing by RAPD assay with the OPG11 primer, and ERIC‑PCR, classified the isolates in two types of 1 and 2 molecular patterns, most of which belonged to type 1. However, the M13 primer‑based RAPD technique was inappropriate for distinguishing and categorizing the isolates andgenerating all of them in the same pattern. In a phylogenetic analysis of O. rhinotracheale based on 16S rRNA sequences, the strains generated three clusters (I‑III), in which all of the studied isolates fell in one cluster (cluster I). Based on the results obtained from the RAPD and ERIC‑PCR assays, the genetic patterns of broiler‑chicken‑isolated O. rhinotracheale strains in Northwestern Iran had no significant differences.

What is your diagnosis? Postmortem tracheal swab from a chicken.

What is your diagnosis? Postmortem tracheal swab from a chicken.

Protective Efficacy of Inactivated H9N2 Vaccine in Turkey Poults under Both Experimental and Field Conditions.

Low pathogenic avian influenza (LPAI) H9N2 virus is one of the major poultry pathogens associated with severe economic losses in the poultry industry (broiler, layers, breeders, and grandparents' flocks), especially in endemic regions including the Middle East, North Africa, and Asian countries. This work is an attempt to evaluate the efficacy of whole inactivated H9N2 vaccine (MEFLUVACTM H9) in turkey poults kept under laboratory and commercial farm conditions. Here, 10,000 white turkey poults (1-day old) free from maternally derived immunity against H9N2 virus were divided into four groups; G1 involved 10 vaccinated birds kept under biosafety level-3 (BLS-3) as a laboratory vaccinated and challenged group, while G2 had 9970 vaccinated turkeys raised on a commercial farm. Ten of those birds were moved to BLS-3 for daily cloacal and tracheal swabbing to check for the absence of any life-threating disease, before conducting analyses. G3 (10 birds) served as a non-vaccinated challenged control under BSL-3 conditions, while G4 (10 birds) was used as a non-vaccinated and non-challenged control under BSL-3 conditions. Sera were collected on days 7-, 14-, 21-, and 28-post-vaccinations to monitor the humoral immune response using a hemagglutination-inhibition (HI) test. At these same intervals, cloacal and tracheal swabs were also checked for any viral infection. The challenge was conducted 28 days post-vaccination (PV) using AI-H9N2 in BSL-3 by intranasal inoculation of 6-log10 embryo infective dose50 (EID50). At 3-, 6-, and 10-days post-challenge, oropharyngeal swabs were taken from challenged birds to quantify viral shedding by quantitative polymerase chain reaction (qRT-PCR). The results of this study showed that vaccinated groups (G1/2) developed HI titers of 1.38, 4.38, 5.88, and 7.25 log2 in G1 vs. 1.2, 3.8, 4.9 and 6.2 log2 in G2 when measured at 7-, 14-, 21- and 28-days PV, respectively, while undetectable levels were recorded in non-vaccinated groups (G3/4). Birds in G3 showed 90% clinical sickness vs. 10% and 20% in G1/2, respectively, over a 10-day monitoring period following challenge. Vaccinated birds showed a significant reduction in virus shedding in terms of the number of shedders, amount of shed virus and shedding interval over the non-vaccinated challenged birds. Regarding mortality, all groups did not show any mortality, which confirms that the circulating H9N2 virus still has low pathogenicity and cannot cause mortality. However, the virus may cause up to 90% clinical sickness in non-vaccinated birds vs. 10% and 20% in laboratory- and farm-vaccinated birds, respectively, highlighting the role of the vaccine in limiting clinical sickness cases. In conclusion, under the current trial circumstances, MEFLUVACTM-H9 provided protective seroconversion titers, significant clinical sickness protection and significant reduction in virus shedding either in laboratory- or farm-vaccinated groups after a single vaccine dose.

Immunological, biochemical and pathological effects of vitamin C and Arabic gum co-administration on H9N2 avian influenza virus vaccinated and challenged laying Japanese quails

AimThis study evaluated the effect of co-administration of vitamin C and Arabic gum (AG) supplements on the response of vaccinated (VAC) and challenged laying Japanese quails with avian influenza virus (AIV) H9N2.Materials and methodsOne hundred and fifty 49-day-old laying Japanese quails were divided into 5 groups (G1-G5): the G1 group was a negative control, G2 group was unvaccinated + H9N2 challenged (Ch), G3 group was unvaccinated + supplements + Ch, G4 group was VAC + Ch, and the G5 group was VAC + supplements + Ch. The supplements (vitamin C, 1 g/liter of drinking water and AG, 1% ration) were given for 5 weeks post-vaccination (PV). The birds were injected subcutaneously with an inactivated H9N2 vaccine at 49 days of age. The quails were then challenged intranasally with AIV H9N2 at the 3rd week PV. Blood, tracheal swab and tissue samples were collected at the 1st, 2nd, and 3rd weeks PV, and at different time points post-challenge (PC).ResultsGrowth performance, egg production (%), egg and eggshell weights, HI antibody titers, clinical signs, lesions, mortality, virus shedding rates, leukogram, biochemical and immunological parameters and histopathological lesions PC showed significant differences (P < 0.05) between the vaccinated-unsupplemented (G4) group and the vaccinated-supplemented (G5) group. G5 showed the highest (P < 0.05) growth performance, egg production, HI antibody titers, and heterophil phagocytic activity and the lowest heterophil/lymphocyte (H/L) ratio, mortality, virus shedding rates, creatinine level and histopathological lesion scores in the lungs.ConclusionThe co-administration of vitamin C and AG for 5 weeks can improve growth performance, egg production and the immune response in vaccinated laying quails challenged with AIV H9N2.

Prevalence and genetic diversity of coronaviruses, astroviruses and paramyxoviruses in wild birds in southeastern Kazakhstan

Wild birds are natural reservoirs of many emerging viruses, including some zoonoses. Considering that the territory of Kazakhstan is crossed by several bird migration routes, it is important to know pathogenic viruses circulating in migratory birds in this region. Therefore, the aim of this study was to identify the host range, diversity and spatial distribution of avian paramyxoviruses, coronaviruses, and astroviruses in free-ranging wild birds in the southeastern region of Kazakhstan. For this purpose, we collected tracheal and cloacal swabs from 242 wild birds belonging to 51 species and screened them using conventional PCR assays. Overall, 4.1% (10/242) and 2.9% (7/242) of all examined birds tested positive for coronaviruses and astroviruses, respectively. Coronaviruses were found in the orders Pelecaniformes (30%; 3/10), Charadriiformes (30%; 3/10), Columbiformes (20%; 2/10), Anseriformes (10%; 1/10), and Passeriformes (10%; 1/10). All detected strains belonged to the genus Gammacoronavirus. Astroviruses were detected in birds representing the orders Passeriformes (57%; 4/7), Coraciiformes (14%; 1/7), Charadriiformes (14%; 1/7), and Columbiformes (14%; 1/7). Paramyxoviruses were observed in only two birds (0.8%; 2/242). Both strains were closely related to the species APMV-22, which had not been previously detected in Kazakhstan. Phylogenetic analysis of the partial RdRp gene sequences of the virus strains revealed three different clades of astroviruses, two clades of coronaviruses, and one clade of paramyxoviruses. The results of this study provide valuable information on the diversity and spatial distribution of paramyxoviruses, coronaviruses, and astroviruses in wild birds in southeastern Kazakhstan and highlight the importance of further thorough monitoring of wild birds in this region.

Genetic analysis and antibiotic susceptibility of Serratia fonticola isolated from ornamental birds in Iraq

The current study was aimed to detect the prevalence of Serratia fonticola strains in birds existed at birdmarkets and houses in different areas of Iraq. One hundred fifty Samples were collected from the wounds and tracheal swab in birds. Samples were cultured on MacConkey and blood agar. Identification of isolate was made according to colony morphology, Gram staining, and conventional biochemical interactions and analysis profile Vitek® 2 compact system. The culture results were revealed an overall one positive isolate with detection an antimicrobial susceptibility test by Vitek® 2 compact system and isolate showed that was susceptible to (Cefepime, Ceftazidime, Amikacin and Ciprofloxacin). In contrast, absolute resistant against (Aztreonam, Gentamicin and Trimethoprim/Sulfamethoxazole). Positive culture isolate was confirmed as Serratia fonticola by monoplex PCR assay, targeting 16S rRNA gene. Positive DNA product were sequenced, and recorded in the NCBI under the accession number (MN696514.1) for birds. The NCBI-BLAST Homology sequence identity reported a significant similarity between these local Serratia fonticola isolates and NCBI isolates; South Korean ( MK235160.1), Egyptian (MN865872.1), Russians (LT555292.1) Iranian (MK737094.1) isolate, at 99%. For our knowledge, this is the first molecular reports performed in Iraq for detection Serratia fonticola infections in birds in Iraq.

Prevalence and associated risk factors of avian influenza A virus subtypes H5N1 and H9N2 in LBMs of East Java province, Indonesia: a cross-sectional study

Background Avian influenza A virus subtypes H5N1 and H9N2 are contagious zoonotic diseases that are circulating in Indonesia and have raised increasing concern about their potential impacts on poultry and public health. A cross-sectional study was carried out to investigate the prevalence and associated risk factors of avian influenza A virus subtypes H5N1 and H9N2 among poultry in the live bird markets of four cities in East Java province, Indonesia. Methods A total of 600 tracheal and cloacal swabs (267 from backyards, 179 from broilers, and 154 from layers) from healthy birds were collected. The samples were inoculated into specific pathogenic-free embryonated eggs at 9-day-old via the allantoic cavity. qRT-PCR was used for further identification of avian influenza. Results The overall prevalence of circulating influenza A virus subtypes H5N1 and H9N2 was 3.8% (23/600, 95%CI [0.0229–0.0537]). Prevalence was higher in backyards at 5.99% (16/267) followed by broilers (2.23% (4/179)) and layers (1.68% (3/154)). The final multivariable model revealed five risk factors for H9N2 infections: presence of ducks (p = 0.003, OR = 38.2), turkeys (p = 0.017 OR = 0.032), and pheasants in the stall (p = 0.04, OR = 18.422), dry (p = 0.006) and rainy season (p &lt; 0.001), and household birds (p = 0.002) and seven factors for H5N1 infections including: observing rodents (p = 0.036, OR = 0.005), stray dogs access (p = 0.004 OR ≤ 0.001), presence of turkeys (p = 0.03 OR = 0.007), chukars/partridges (p = 0.024 OR = 2500), and peafowls in the stalls (p = 0.0043 OR ≤ 0.001), rainy season (p = 0.001) and birds from the household sources (p = 0.002) in the live bird markets. Conclusions The findings of the current study illustrate the recurring infection and presence of both avian influenza viruses and associated risk factors in the surveyed marketplaces. Effective protective measures and mitigation strategies for risks outlined in this study could help to reduce the burden of H5N1 and H9N2 AI subtypes into the live bird markets of Indonesia.

Field Application of qPCR Monitoring of Infectious Laryngotracheitis Virus in Settled Chicken House Dust and Its Role in Control of a Major Outbreak.

Population-level sampling based on qPCR detection of infectious laryngotracheitis virus (ILTV) in poultry dust can be used to assess ILT vaccination outcomes following mass administration in drinking water. We report on the field application of this approach to assess the success of vaccine administration and its use in ILT outbreak control in meat chickens. In Study 1, dust samples were collected from 26 meat chicken flocks at 0, 4, 7, 14, and 21 days post drinking water vaccination (DPV) given between 7 to 13 days of age with the Serva or A20 live attenuated ILT vaccines. Unexpectedly, ILTV DNA was detected in dust samples collected prior to vaccination in 22/26 flocks. Typing revealed that the detected ILTV was different from the vaccine virus. To determine whether the detected ILTV DNA was from active infection or carryover of a noninfectious virus, Study 2 was implemented in 14 additional flocks with dust samples collected at 0, 7, 14, and 21 DPV and tracheal swabs collected from 15 birds/flock at 0 and 21 DPV. The results indicated that there was active infection with ILTV in those flocks before vaccination. This approach contributed to a statewide control program resulting in the eradication of ILT from South Australia as confirmed by negative ILTV test results for dust samples from 50 flocks and the absence of clinical ILT. These findings show that ILTV infection prior to vaccination is common in outbreak situations and that dust samples must be collected at 0 and 7 DPV for meaningful interpretation of vaccination outcomes and ILTV status. Comparatively low-cost dust testing during an outbreak, coupled with typing information, greatly assisted with decision making and control strategies during a major outbreak, including confirmation of the absence of infection in the final stages.

Postmortem Bacteriology in Forensic Autopsies-A Single Center Retrospective Study in Romania.

Postmortem bacteriology examinations have been a controversial topic over the years, though the value of postmortem bacteriology cultures remains promising. The aim of this study was to review the postmortem bacteriological sampling procedures and results in a single centre in Bucharest over a period of 10 years. Material and methods: The present study was a retrospective, single-center study, performed at the Mina Minovici National Institute of Legal Medicine in Bucharest, Romania, from 2011–2020. Results: Postmortem bacteriology was requested 630 forensic autopsies, 245 female (38.9%) and 385 male (61.1%), age range 0 and 94 years, median age of 52 years. Deaths occurred in hospital for 594 cases (94.3%) and out-of-hospital for 36 cases (5.7%—field case). Blood cultures were requested in the majority of cases, followed by tracheal swabs and lung tissue. In-hospital and out of hospital deaths did not differ significantly regarding the number of microorganisms identified in a positive blood culture. Postmortem bacteriology cultures of the respiratory tract showed a statistically significant association to microscopically confirmed lung infections. Conclusions Postmortem sampling for bacteriology testing in our center in Bucharest is heterogeneous with a high variation of patterns. A positive blood culture result for Staphylococcus species without the identification of a specific microorganism is more likely due to postmortem contamination.