Morphological, molecular identification and evaluation of antioxidant activity of seahorses from the Moroccan coasts.

Seahorses, part of the small marine teleost fish family Syngnathidae, are increasingly under threat due to habitat degradation and overfishing. Notably used in traditional Chinese medicine, these fish have demonstrated significant pharmacological and cosmetic properties. In Morocco, however, seahorses are minimally exploited. This study aims to explore the biodiversity of Moroccan seahorses, focusing on identifying species from the Atlantic and Mediterranean coasts both morphologically and molecularly, and evaluating their antioxidant activity. The research involved collecting 62 dried seahorses from local fishermen. These specimens were subjected to detailed morphological and molecular identification through the DNA barcoding method, concentrating on the mitochondrial marker Cytochrome Oxidase I (COI) gene. Following DNA extraction and amplification, the sequences were analyzed for species identification and phylogenetic relationships. Additionally, the antioxidant activities of the seahorses were quantified using assays such as ABTS, reducing power, phosphomolybdenum, and ß-carotene-linoleic acid. The combined morphological and molecular analyses consistently identified all specimens as Hippocampus hippocampus, and phylogenetic trees suggested a close relation with European and Turkish counterparts. Furthermore, the antioxidant assays revealed significant activity, with the ABTS assay showing an IC50 of 14.571 mg/mL ± 0.334, and the ß-carotene-linoleic acid assay showing an IC50 of 1.273 mg/mL ± 0.166. The reducing power and phosphomolybdenum assays recorded EC50 values of 1.868 mg/mL ± 0.033 and 1.156 mg/mL ± 0.112, respectively. These results confirm the high antioxidant potential of Moroccan seahorses, suggesting their therapeutic value and necessitating measures for their biodiversity preservation at a national level.

Hepatitis A and E Viruses in Mussels from Cherrat Estuary in Morocco: Detection by Real-Time Reverse Transcription PCR Analysis.

The aim of the present study was to evaluate hepatitis A virus (HAV) and hepatitis E virus (HEV) contamination in mussels (Mytilus galloprovincialis) from Cherrat estuary (Moroccan Atlantic Coast), Morocco. In total, 52 samples (n = 12 mussels/each) were collected at four sites in the estuary, monthly, between March 2019 and March 2020. HAV and HEV were detected by real-time reverse transcription polymerase chain reaction (RT-PCR) according to the ISO/TS 15216 method. HAV was detected in 46.15% of analyzed samples. Conversely, HEV was not detected in any sample. Moreover, the HAV detection rate was significantly associated with seasonal rainfall variations. This qualitative study on HAV and HEV contamination highlights the interest of studying mussel samples from wild areas. As HAV presence in mussels represents a potential health risk, viral contamination surveillance of mussels is necessary to protect consumers. HAV shellfish contamination must be monitored at Cherrat estuary because of the role played by shellfish as HAV reservoirs and/or vehicles in fecal-oral HAV transmission.

Spatiotemporal Dynamics, Evolutionary History and Zoonotic Potential of Moroccan H9N2 Avian Influenza Viruses from 2016 to 2021.

The H9N2 virus continues to spread in wild birds and poultry worldwide. At the beginning of 2016, the H9N2 Avian influenza virus (AIV) was detected in Morocco for the first time; despite the implementation of vaccination strategies to control the disease, the virus has become endemic in poultry in the country. The present study was carried out to investigate the origins, zoonotic potential, as well as the impact of vaccination on the molecular evolution of Moroccan H9N2 viruses. Twenty-eight (28) H9N2 viruses collected from 2016 to 2021 in Moroccan poultry flocks were isolated and their whole genomes sequenced. Phylogenetic and evolutionary analyses showed that Moroccan H9N2 viruses belong to the G1-like lineage and are closely related to viruses isolated in Africa and the Middle East. A high similarity among all the 2016-2017 hemagglutinin sequences was observed, while the viruses identified in 2018-2019 and 2020-2021 were separated from their 2016-2017 ancestors by long branches. Mutations in the HA protein associated with antigenic drift and increased zoonotic potential were also found. The Bayesian phylogeographic analyses revealed the Middle East as being the region where the Moroccan H9N2 virus may have originated, before spreading to the other African countries. Our study is the first comprehensive analysis of the evolutionary history of the H9N2 viruses in the country, highlighting their zoonotic potential and pointing out the importance of implementing effective monitoring systems.

Molecular Detection of Avian Influenza Virus in Wild Birds in Morocco, 2016-2019.

Avian influenza (AI) is a zoonotic disease significant to both public and animal health, caused by influenza virus A, and affects domestic poultry, wild birds, and mammals including humans. Aquatic birds are considered the natural reservoir of this virus. In 2016, Morocco experienced the first occurrence of low pathogenic H9N2 avian influenza virus (AIV) in poultry; however, no cases were reported in wild birds. The present study aimed to monitor the presence of AIV in wild birds in Morocco in order to trace the possible sources of the viruses affecting poultry. Between 2016 and 2019, 967 samples obtained from 480 birds representing 56 different wild bird species, 20 families, and 8 orders, mostly from Charadriiformes, Anseriformes, Pelecaniformes, and Passeriformes, were collected from various wetlands and relevant ornithologic sites in Morocco. These field samples consisted of 374 cloacal swabs, 321 tracheal swabs, 54 fecal samples, and 218 organ pools including the trachea, lung, liver, spleen, heart, intestine, and brain. The samples were examined for the presence of AIV using TaqMan-based real-time reverse transcriptase-PCR (rRT-PCR) targeting the matrix gene, followed by further subtyping rRT-PCR tests targeting the H1-H16 genes. The AI matrix gene was detected in 18 out of 967 samples (1.86%); positive samples were detected in 17 birds belonging to 10 bird species: two redshanks (Tringa totanus), one little stint (Calidris minuta), one ruddy turnstone (Arenaria interpres), one common snipe (Gallinago gallinago), one common greenshank (Tringa nebularia), one black-winged stilt (Himantopus himantopus), two black-headed gulls (Chroicocephalus ridibundus), one slender-billed gull (Chroicocephalus genei), six cattle egrets (Bubulcus ibis), and one Eurasian coot (Fulica atra). AIV was detected in 2 wetlands and 1 ornithologic site (Sidi Moussa Oualidia Complex, Smir lagoon and El Jadida Coast) and the highest positivity was revealed in fresh fecal samples (11.1%), indicating the suitability of this matrix for wild bird surveillance. Our results highlight that waders, gulls, and cattle egrets are the most affected species and may represent a potential risk for AI introduction in the poultry sector in Morocco. Regular monitoring of wild birds in Morocco, focusing in particular in the areas and species identified in this study as a high risk of virus circulation, should be implemented to anticipate and prevent possible AIV spread.

Detección molecular del virus de la influenza aviar en aves silvestres en Marruecos, entre los años 2016 al 2019. La influenza aviar (IA) es una enfermedad zoonótica importante para la salud pública y animal, causada por el virus de la influenza A y afecta a la avicultura comercial, las aves silvestres y los mamíferos, incluyendo a los humanos. Las aves acuáticas se consideran el reservorio natural de este virus. En el año 2016, Marruecos experimentó la primera aparición del virus de la influenza aviar de baja patogenicidad H9N2 en avicultura; sin embargo, no se notificaron casos en aves silvestres. El presente estudio tuvo como objetivo monitorear la presencia del virus de influenza aviar en aves silvestres en Marruecos con el fin de rastrear las posibles fuentes de los virus que afectan a las aves comerciales. Entre los años 2016 y 2019, se recolectaron 967 muestras de 480 aves que representan 56 especies diferentes de aves silvestres, 20 familias y 8 órdenes, principalmente de Charadriiformes, Anseriformes, Pelecaniformes y Passeriformes, de varios humedales y sitios ornitológicos relevantes en Marruecos. Estas muestras de campo consistieron en 374 hisopos cloacales, 321 hisopos traqueales, 54 muestras fecales y 218 conjuntos de órganos que incluyeron tráquea, pulmón, hígado, bazo, corazón, intestino y cerebro. Las muestras se examinaron para detectar la presencia del virus de la influenza aviar mediante transcripción reversa y PCR en tiempo real basada en TaqMan (rRT-PCR) que estivo dirigida al gene de la matriz, seguida de más pruebas de subtipificación de rRT-PCR dirigidas a los genes H1 al H16. El gene de la matriz de influenza aviar se detectó en 18 de 967 muestras (1.86%); Se detectaron muestras positivas en 17 aves pertenecientes a 10 especies de aves: dos archibebes comunes (Tringa totanus), un correlimos chico (Calidris minuta), un vuelvepiedras común (Arenaria interpres), una agachadiza común (Gallinago gallinago), un archibebe claro (Tringa nebularia), una cigüeñuela común (Himantopus himantopus), dos gaviotas reidoras (Chroicocephalus ridibundus), una gaviota picofina (Chroicocephalus genei), seis garzas ganaderas (Bubulcus ibis) y una focha común (Fulica atra). El virus de la influenza aviar se detectó en 3 de los 17 humedales (Complejo Sidi Moussa-Walidia, Costa El Jadida y Laguna Smir) y la mayor positividad se reveló en muestras fecales frescas (11.1%), lo que indica la idoneidad de esta matriz para la vigilancia de aves silvestres. Estos resultados destacan que las aves zancudas, las gaviotas y las garzas ganaderas son las especies más afectadas y pueden representar un riesgo potencial para la introducción de influenza aviar en el sector avícola de Marruecos. Se debe implementar un monitoreo regular de las aves silvestres en Marruecos, centrándose en particular en las áreas y especies identificadas en este estudio como de alto riesgo de circulación del virus, para anticipar y prevenir la posible propagación del virus de la influenza aviar.