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.

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.

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.

Introduction: Salmonellosis is a foodborne bacterial disease responsible for food epidemics around the world. The objective of this study is to determine the prevalence and diversity of Salmonella serotypes in several food products isolated at the Casablanca Regional Analysis and Research Laboratory and to test their resistance to different antimicrobials. Methodology: The isolation and identification of Salmonella were performed according to Moroccan standard 08.0.116. All isolates were serotyped and were then tested for antibiotic resistance using the disk diffusion method. The Salmonella isolates were further analyzed by PCR to detect the presence of virulence genes invA. Results:20 different serotypes were identified from 80 strains isolated from 2015 to 2019, the most common of which are S. kentucky (26.3%) followed by S. muenster (10%), S. typhimurium (8.7%), S. menston (7.5%) and S. enteritidis (6.3%). Antimicrobial susceptibility testing revealed that 66.25% of isolates were resistant to at least one of the 14 antimicrobial agents tested. Bacterial resistance was most frequently observed for tetracycline with 46.25%, 45% to sulfonamide, 35% to nalidixic acid, 26, 25% to ampicillin, and 25% to ciprofloxacin. Salmonella serotypes S. montevideo, S. virchow, S. amsterdam, S. anatum, and S. bloomsbury were 100% susceptible to all antimicrobials tested. Examination of Salmonella for invA gene was positive for all the strains. Conclusions: The results of this study have shown that minced meat has a high level of Salmonella contamination, which can be considered one of the main potential sources of human salmonellosis in Morocco.

In Morocco, spices are an integral part of daily cuisine and serve as a vector of both nutritional and toxicological exposure. Monitoring elemental composition is essential to ensure consumer safety, animal health when used as feed additives, and compliance with international standards. The present study aimed to determine the concentrations of essential (potassium, calcium, magnesium, sodium, and iron) and toxic (lead, cadmium, arsenic, chromium, and nickel) elements in commonly consumed spices in Morocco, including cinnamon, cumin, ginger, black pepper, and turmeric. A total of 162 spice samples were obtained from markets in the Greater Casablanca, Morocco. Five essential elements, including potassium, calcium, magnesium, sodium, and iron, and five toxic trace metals, including lead, cadmium, arsenic, chromium, and nickel, were determined by inductively coupled plasma mass spectrometry (ICP-MS) after microwave-assisted digestion. Cumin indicated the highest levels of magnesium (6.86 ± 1.61 g/kg), sodium (3.98 ± 1.59 g/kg), calcium (11.13 ± 4.53 g/kg), and iron (753.71 ± 446.07 mg/kg). Turmeric had the highest levels of potassium (25.96 ± 13.51 g/kg). Cinnamon had elevated levels of lead (2.05 mg/kg) and cadmium (0.29 mg/kg), exceeding Moroccan and European regulatory limits. Additionally, cumin indicated the highest levels of arsenic (0.45 ± 0.30 mg/kg) and nickel (4.18 ± 2.85 mg/kg) compared to other spices. Principal component analysis (PCA) revealed distinct elemental patterns. The first component (PC1), driven by magnesium and sodium, clearly separated cumin due to its high macronutrient content. The second component (PC2), influenced by cadmium and lead, isolated cinnamon because of its toxic metal burden. The PC1 and PC2 accounted for 64.6% of the total variance. Turmeric and ginger formed a close cluster in the PCA plot, associated with higher levels of potassium, calcium, and nickel. Black pepper was positioned between these groups, reflecting intermediate composition. Pearson correlation analysis supported these findings, with a strong correlation between lead and cadmium, suggesting a shared contamination source. These results emphasized the nutritional and toxicological roles of spices in Moroccan diets. Regular monitoring is essential to protect public health in both animals and humans. Keywords: Food safety, Inductively coupled plasma mass spectrometry, Spices, Toxic metal

Cell-free DNA (cfDNA) fragments represent emerging biomarkers of major interest in the assessment of male fertility and the management of couples undergoing assisted reproductive technology (ART). Easy to quantify and present in most biological fluids, seminal cfDNA could reflect apoptosis and necrosis occurring within the male reproductive system. In this study, we compared cfDNA concentrations, measured by means of qPCR targeting the RNase P gene, in the seminal plasma of men with severe sperm quality abnormalities, including very severe asthenozoospermia (n = 25), total azoospermia (n = 25), very severe oligozoospermia (n = 25), and very severe teratozoospermia (n = 25), with those of normozoospermic controls (n = 25). Mean cfDNA concentrations were particularly elevated in men with azoospermia (5.47 ± 1.17 µg/mL) and very severe teratozoospermia (3.25 ± 1.21 µg/mL), compared with controls (1.96 ± 0.27 µg/mL). After adjustment for multiple testing, only azoospermia and very severe teratozoospermia remained significantly associated with increased cfDNA levels (Holm–Bonferroni adjusted p < 0.003), whereas the other phenotypes did not maintain statistical significance. These results suggest that seminal cfDNA is markedly increased in the most severe forms of spermatogenic impairment, potentially reflecting enhanced germ cell apoptosis or testicular cell involvement. Despite some limitations, including the modest sample size and single-center design, our observations support the value of cfDNA as a promising non-invasive biomarker of sperm quality. Larger multicenter and longitudinal studies are needed to validate its clinical relevance and determine standardized reference thresholds for clinical application.