First Report of Low Pathogenic Avian Influenza Subtype H9N2 in African Houbara Bustards (Chlamydotis undulata undulata) and Gamebirds in Morocco: Clinico-Pathological Findings, Molecular Characterization, and Associated Coinfections.

We report in this paper the first detection of low pathogenic avian influenza (LPAI) subtype H9N2 in houbara bustards and in gamebirds in Morocco. Starting in 2019, an increase in mortality rates related to respiratory distress was recorded in these species. Necropsy of the specimens revealed fibrinous sinusitis and tracheitis with intra-bronchial fibrin casts, which are consistent with H9N2 infection in chickens; therefore, implication of the virus in these outbreaks was strongly suspected. Consequently, between January 2020 and June 2023, birds with respiratory signs were necropsied for pathological lesions, tissue samples were examined by histopathology, and samples of trachea, lungs, and cecal tonsils were analyzed using quantitative real-time PCR for the detection of H9N2 virus. In addition, the sequencing of isolates was performed and lastly differential diagnosis with other respiratory pathogens was carried out. During the study period, 93 samples were collected from suspected H9N2 outbreaks, of which 30 tested positive for H9N2 virus: 23 Houbara bustards, 4 partridges, 2 quails, and 1 pheasant. Moreover, sequencing of the HA gene of the virus showed 97.33% nucleotide identity with strains reported previously in broilers in Morocco in 2017 and in 2022. Phylogenetic analysis grouped the Moroccan partridge isolates in the same cluster as viruses isolated in Morocco between 2016 and 2022, Algeria (2017), Burkina Faso (2017), Nigeria (2019), and Togo (2020). Additionally, 10 house sparrows from the premises of these birds were examined for the presence of H9N2 virus, revealing a 30% positivity rate. In conclusion, LPAIV H9N2 is circulating in houbara bustards and gamebirds in Morocco, and house sparrows might be a possible source of the infection. To our knowledge, this is the first report of LPAI H9N2 in the African species of houbara bustards worldwide and in gamebirds in Morocco.

First Report of Isolation of Gallibacterium anatis from Layer Chickens in Morocco with Decrease in Laying Performance.

Gallibacterium is a genus of the family of Pasteurellaceae. It is well known as a commensal inhabitant of the respiratory and reproductive tract of healthy chickens. But in the last years, Gallibacterium anatis is increasingly reported in field cases with a decrease in laying performance due to infections of the reproductive tract. The aim of the present study was to investigate the implication of G. anatis infection in layer flocks facing a decrease in laying performance in Morocco. Birds were received from five different laying hen farms in two regions in Morocco showing a drop of egg production. Necropsy revealed 46.1 % (24/52) of sampled birds showed variable lesions in ovaries, salpinx, and trachea. In fact, 24 birds were affected by salpingitis, 18 by oophoritis, and 11 birds by atrophy of ovaries. Furthermore, tracheitis was observed in 24 birds. Bacteriological investigation was done from different organs, and G. anatis was found in ovaries (n = 20), trachea (n = 17), and cloaca (n = 3). Identification was based on growth morphology, Gram staining, and biochemical properties. Additionally, polymerase chain reaction test using specific primers for the genus identification was carried out. All isolates showed bands of 925 bp specific for G. anatis expressing the virulent toxin GtxA. Antibiotic resistance testing was performed and revealed that isolates were sensitive to enrofloxacin, florfenicol, and gentamycin but resistant to ampicillin, erythromycin, oxytetracycline, and sulfamethoxazole-trimethoprim. The present study is the first report of G. anatis in Morocco, demonstrating the need for further epidemiologic investigations as well as in regard to antibiotic resistance development.

Primer informe de aislamiento de Gallibacterium anatis de gallinas de postura en Marruecos con disminución en la eficiencia de la postura. Gallibacterium es un género de la familia Pasteurellaceae. Es bien conocido como un habitante comensal del tracto respiratorio y reproductivo de pollos sanos. Pero en los últimos años, han aumentado los reportes de Gallibacterium anatis en casos de campo con una disminución en la eficiencia de la postura debido a infecciones del tracto reproductivo. El objetivo del presente estudio fue investigar la implicación de la infección por G. anatis en parvadas de gallinas de postura que presentaban una disminución en la eficiencia de la postura en Marruecos. Se recibieron aves de cinco granjas diferentes de gallinas postura en dos regiones de Marruecos, mostrando una caída en la producción de huevos. La necropsia reveló que 46.1% (24/52) de las aves muestreadas mostraron lesiones variables en ovarios, oviducto y tráquea. De hecho, 24 aves mostraron salpingitis, 18 ooforitis y 11 aves atrofia de ovarios. Además, se observó traqueítis en 24 aves. Se realizó la investigación bacteriológica de diferentes órganos, y se encontró G. anatis en ovarios (n = 20), tráquea (n = 17) y cloaca (n = 3). La identificación se basó en la morfología del crecimiento, en la tinción de Gram y en las propiedades bioquímicas. Además, se realizó una prueba de reacción en cadena de la polimerasa utilizando iniciadores específicos para la identificación del género. Todos los aislamientos mostraron bandas de 925 pares de bases específicas para G. anatis que expresaban la toxina virulenta GtxA. Se realizaron pruebas de resistencia a los antibióticos y revelaron que los aislamientos eran sensibles a enrofloxacina, florfenicol y gentamicina, pero resistentes a ampicilina, eritromicina, oxitetraciclina y sulfametoxazol-trimetoprima. El presente estudio es el primer informe de G. anatis en Marruecos, que demuestra la necesidad de más investigaciones epidemiológicas, así como en relación con el desarrollo de resistencia a los antibióticos.