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Avian metapneumovirus (aMPV) is a highly contagious pathogen for broilers. Its epidemiological aspects remain poorly understood in Morocco. Given that, we carried out a pilot seroepidemiological survey from December 2020 to June 2021 to define aMPV seroprevalence in Moroccan broiler flocks in different bioclimatic zones (humid to sub-humid, semi-arid, arid) during winter and spring. According to these zones, 48 flocks not vaccinated against aMPV were selected using a stratified random sampling model. With a simple random sampling strategy, fifteen to thirty sera per flock were collected then analysed using a commercial indirect ELISA kit (CIVTEST AVI TRT®, HIPRA S.A., Amer, Spain) that was able to titre antibodies against aMPV subtypes A and B. Furthermore, questionnaires were shared with veterinarians in charge of flocks to collect data and analyse it with multivariable logistic regression models to identify risk factors associated with virus circulation. From 1142 sera, 912 tested positive with the ELISA used, showing an overall aMPV seroprevalence of 79.86% in the broilers. The arid zone had the highest seroprevalences: 94.16% during winter and 84.82% during spring. Bioclimate and season seemed to contribute to aMPV seropositivity. Likewise, high stocking densities and poor hygiene scores on farms were proven to predispose birds to seropositivity. Hereby, we show the circulation of aMPV in Moroccan broiler farms is influenced by bioclimate, season, density, and hygiene conditions. The present study is the first serological evidence of aMPV in broilers in the Maghreb region.

Abstract Background Since the end of 2009, H9N2 has emerged in Tunisia causing several epidemics in poultry industry resulting in major economic losses. To monitor variations of Influenza viruses during the outbreaks, Tunisian H9N2 virus isolates were identified and genetically characterized. Methods The genomic RNA segments of Tunisian H9N2 strains were subjected to RT-PCR amplifications followed by sequencing analysis. Results Phylogenetic analysis demonstrated that A/Ck/TUN/12/10 and A/Migratory Bird/TUN/51/10 viruses represent multiple reassortant lineages, with genes coming from Middle East strains, and share the common ancestor Qa/HK/G1/97 isolate which has contributed internal genes of H5N1 virus circulating in Asia. Some of the internal genes seemed to have undergone broad reassortments with other influenza subtypes. Deduced amino acid sequences of the hemagglutinin (HA) gene showed the presence of additional glycosylation site and Leu at position 234 indicating to binding preference to α (2, 6) sialic acid receptors, indicating their potential to directly infect humans. The Hemagglutinin cleavage site motif sequence is 333 PARSSR*GLF341 which indicates the low pathogenicity nature of the Tunisian H9N2 strains and the potential to acquire the basic amino acids required for the highly pathogenic strains. Their neuraminidase protein (NA) carried substitutions in the hemadsorption (HB) site, similar to those of other avian H9N2 viruses from Asia, Middle Eastern and human pandemic H2N2 and H3N2 that bind to α -2, 6 -linked receptors. Two avian virus-like aa at positions 661 (A) and 702 (K), similar to H5N1 strains, were identified in the polymerase (PB2) protein. Likewise, matrix (M) protein carried some substitutions which are linked with increasing replication in mammals. In addition, H9N2 strain recently circulating carried new polymorphism, "GSEV" PDZ ligand (PL) C-terminal motif in its non structural (NS) protein. Two new aa substitutions (I) and (V), that haven't been previously reported, were identified in the polymerase and matrix proteins, respectively. Nucleoprotein and non-structural protein carried some substitutions similar to H5N1 strains. Conclusion Considering these new mutations, the molecular basis of tropism, host responses and enhanced virulence will be defined and studied. Otherwise, Continuous monitoring of viral genetic changes throughout the year is warranted to monitor variations of Influenza viruses in the field.

Background: Avian metapneumovirus (aMPV) is a widespread infectious respiratory pathogen affecting turkeys and chickens, with co-predominance of the subtypes A and B. Objectives: There is no official reports in Morocco about the subtypes of aMPV circulating. Hence, using quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) subtypesspecific A and B, we aimed at detecting and identifying the potential subtype(s) circulating. Methods: We conducted a longitudinal study on three broiler flocks that were strictly not vaccinated against aMPV and were located in two different geographical regions. We studied two flocks that expressed typical swollen head syndrome (SHS) and sampled them once. Furthermore, we sampled dead birds of one flock confirmed seropositive from a previous study. A total of 118 swabs pooled in 24 samples were subjected to RNA extraction and amplified using a triplex RT-PCR for specific detection of aMPV subtypes A and B. Additionally, serum samples were taken at slaughtering age to cross-check the molecular results. A total of 84 sera were analyzed with a commercial indirect enzyme-linked immunosorbent assay (ELISA) kit to detect and titer antibodies against the two subtypes. Results: Avian metapneumovirus was detected by qRT-PCR in all flocks. About 87.50% of the samples were positive for aMPV-B, and 16.67% for aMPV-A and aMPV-B simultaneously. All flocks showed seropositivity, confirming the molecular findings. Conclusion: The present investigation is the first molecular study in Morocco to elucidate the circulation of aMPV-A and aMPV-B in broiler farms in Morocco with a dominance of aMPV-B and the possibility of co-presence of both subtypes. [ABSTRACT FROM AUTHOR]

Avian Metapneumovirus infection first emerged in South African turkeys, followed by respiratory problems in chickens with a swollen head syndrome. The etiological agent is a Metapneumovirus in the Pneumoviridae family and the first pneumovirus identified in avian species. This virus causes respiratory and reproductive affections that are worsened in the presence of other pathogens. The present review summarized the current knowledge about the virus's properties and spread, its different subtypes, and the immunological and pathological mechanisms, especially in the broilers. The diagnostic methods are based on serology and essentially ELISA to show and titer antibodies following infection in naïve birds. Molecular tools such as PCR aim to detect and subtype avian Metapneumovirus genetic material. Besides biosecurity, prevention relies mostly on good management and vaccination. [ABSTRACT FROM AUTHOR]

An efficient and simple approach has been developed for the synthesis of eight dialkyl/aryl[(5-phenyl-1,3,4-oxadiazol-2-ylamino)(aryl)methyl]phosphonates through the Pudovik-type reaction of dialkyl/arylphosphite with imines, obtained from 5-phenyl-1,3,4-oxadiazol-2-amine and aromatic aldehydes, under microwave irradiation. Five of them were hydrolyzed to lead to the corresponding phosphonic acids. Selected synthesized compounds were screened for their in vitro antiviral activity against the avian bronchitis virus (IBV). In the MTT cytotoxicity assay, the dose-response curve showed that all test compounds were safe in the range concentration of 540–1599 µM. The direct contact of novel synthesized compounds with IBV showed that the diethyl[(5-phenyl-1,3,4-oxadiazol-2-ylamino)(4-trifluoromethoxyphenyl)methyl]phosphonate (5f) (at 33 µM) and the [(5-phenyl-1,3,4-oxadiazol-2-ylamino)(4-trifluoromethylphenyl)methyl] phosphonic acid (6a) (at 1.23 µM) strongly inhibited the IBV infectivity, indicating their high virucidal activity. However, virus titers from IBV-infected Vero cells remained unchanged in response to treatment with the lowest non-cytotoxic concentrations of synthesized compounds suggesting their incapacity to inhibit the virus replication inside the host cell. Lack of antiviral activity might presumably be ascribed to their polarity that hampers their diffusion across the lipophilic cytoplasmic membrane. Therefore, the interactions of 5f and 6a were analyzed against the main coronavirus protease, papain-like protease, and nucleocapsid protein by molecular docking methods. Nevertheless, the novel 1,3,4-oxadiazole-based α-aminophosphonic acids and α-amino-phosphonates hold potential for developing new hygienic virucidal products for domestic, chemical, and medical uses. [ABSTRACT FROM AUTHOR]