Pathogenicity and molecular characterization of Mycoplasma bovis isolate from calves in Morocco.

Bovine respiratory disease caused by Mycoplasma bovis (M. bovis) represents a major health problem for cattle worldwide that causes considerable financial losses. This study reports for the first time the molecular and pathogenic characterization of a strain of M. bovis isolated from a dead local calf with respiratory symptoms in Morocco. M. bovis was isolated from lung tissue, purified by cloning, and subtyped using MLST analysis. Experimental infection was conducted in naïve calves to evaluate pathogenicity. The isolate was identified as a new subtype ST-204 that shares similarities with the 2019-2021 Spanish strains (ST-169, ST-170, ST-171) and the 2018 Algeria isolate (ST-4). Experimental infection resulted in fever and respiratory symptoms with serous nasal discharge. At postmortem, lung lesions of congestion and hepatization were observed with lymph node enlargement and foci of necrosis. The study confirms the high pathogenicity of the isolate and the important role of M. bovis in bovine respiratory disease.

Neutralising antibodies against SARS-CoV-2 give important information on Covid-19 epidemic evolution in Rabat, Morocco, March 2020-February 2021.

Background: The SARS-CoV-2 is an extremely contagious and acute viral disease mainly affecting humans. Objective: To estimate seroprevalence of SARS-CoV-2 neutralizing antibodies (NAbs) for illegible armed force individuals living in Rabat, Morocco. Method: A convenience sample (N = 2662) was conducted from May 2020 to February 2021. We used the standard neutralization assay to quantify the NAbs titers. A serum was positive when the titer was 1:4. High positive NAbs titers were defined when ≥ 1:32. Results: Demographic and socioeconomic status did not affect seroprevalence data. An overall seroprevalence of 24,9% was found. Sera from blood donors, young recruits and auto-immune population had lower NAbs titers. However, titers were above 1:16 in 9% of the population with high risk of SARS-CoV-2 exposure. Seropositivity increased over time with values reaching peaks after the epidemic waves (2.4% in May 2020; 16.2% in August 2020; 22.7% in December 2020 and 37% in February 2021). Conclusion: And increase of NAbs was observed over time and correlated with the post-epidemic waves of COVID-19 in Morocco.

Avian Influenza a H9N2 Viruses in Morocco, 2018-2019.

Low pathogenic H9N2 avian influenza (LPAI H9N2) is considered one of the most important diseases found in poultry (broiler, laying hens, breeding chickens, and turkeys). This infection causes considerable economic losses. The objective of this work was to monitor and assess the presence of avian influenza virus (AIV) H9N2 in eight different regions of Morocco using real-time RT-PCR, and to assess the phylogenetic and molecular evolution of the H9N2 viruses between 2016 and 2019. Field samples were collected from 108 farms suspected of being infected with LPAI H9N2 virus. Samples were analyzed using H9N2-specific real-time RT-PCR. Highly positive samples were subjected to virus isolation and seven isolates were fully sequenced. Low pathogenic H9N2 avian influenza virus was introduced in Morocco in 2016. We show that in 2018-2019, the virus was still present irrespective of vaccination status. Phylogenetic and molecular analyses showed mutations related to virulence, although our viruses were related to 2016 Moroccan viruses and grouped in the G1 lineage. Specific amino acid substitutions were identified in Moroccan H9N2 viruses that are believed to lead to increased resistance to antiviral drugs.

Assessing the Efficacy of Recombinant Newcastle Disease Virus Vaccines (Double-Insert vHVT-IBD-ND and Single-Insert vHVT-ND) Followed by a Vaccination with a Live Newcastle Disease Vaccine Against a Moroccan Velogenic Newcastle Disease Challenge in Commercial Broilers.

The advent of turkey herpesvirus (HVT) vector vaccine technology (vHVT) has made a huge improvement in the prevention and control of several poultry diseases. The objective of this study was to compare, under experimental conditions, the protection conferred by different vaccination programs based on an HVT double-insert (infectious bursal disease {IBD] and Newcastle disease [ND]) vector vaccine (vHVT-IBD-ND) and an HVT single-insert (vHVT-ND) vector vaccine followed by a vaccination with a live ND vaccine at Day 1 only or at Days 1 and 14. Commercial broilers were vaccinated by the recombinant ND virus vaccines subcutaneously at 1 day old, in the hatchery, and challenged at 30 days of age using the Moroccan ND virus velogenic viscerotropic JEL strain. The results showed that the tested vaccine induced 95% to 100% clinical protection against mortality and clinical signs. The humoral immune response to vaccination was detected from 3 wk of age using enzyme-linked immunosorbent assay and hemagglutination inhibition tests. ND challenge virus shedding was significantly reduced in the vaccinated birds as compared to controls. Significant reduction of the cloacal shedding suggests that the vHVT-IBD-ND vaccine stimulates actively the immunity against the tested ND challenge virus. No significant differences were found between the vaccination programs based on vHVT-IBD-ND or on vHVT-ND.

Evaluación de la eficacia de las vacunas recombinantes contra el virus de la enfermedad de Newcastle (vHVT-IBD-ND de doble inserto y vHVT-ND de inserto único) seguidas de una vacunación con una vacuna viva para la enfermedad de Newcastle contra un desafío de la enfermedad de Newcastle velogénico marroquí en pollos de engorde comerciales. El advenimiento de la tecnología de vacunas recombinantes (vHVT) del virus herpes del pavo (HVT) ha provocado una mejora en la prevención y el control de varias enfermedades avícolas. El objetivo de este estudio fue comparar, en condiciones experimentales, la protección conferida por diferentes programas vacunales basados en una vacuna recombinante HVT con doble inserto (bursitis infecciosa [EII] y enfermedad de Newcastle [ND]) (vHVT-IBD-ND) y una vacuna recombinante HVT con inserto única (vHVT-ND) seguida de una vacunación con una vacuna para Newcastle viva aplicada en el día 1 o en los días 1 y 14. Pollos de engorde comerciales se vacunaron con las vacunas recombinantes del virus de la enfermedad de Newcastle por vía subcutánea al día de edad, en la incubadora y se expusieron a los 30 días de edad utilizando la cepa JEL viscerotrópica velogénica del virus de la enfermedad de Newcastle de Marruecos. Los resultados mostraron que la vacuna evaluada indujo una protección clínica del 95% al 100% contra la mortalidad y los signos clínicos. La respuesta inmune humoral a la vacunación se detectó a partir de las 3 semanas de edad mediante ensayo inmunoabsorbente ligado a enzimas y pruebas de inhibición de la hemaglutinación. La excreción del virus de Newcastle de desafío se redujo significativamente en las aves vacunadas en comparación con los controles. La reducción significativa de la eliminación cloacal sugiere que la vacuna vHVT-IBD-ND estimula activamente la inmunidad contra el virus de Newcastle de desafío analizado. No se encontraron diferencias significativas entre los programas de vacunación basados en vHVT-IBD-ND o en vHVT-ND.

Pathogenesis of Avian Influenza Virus Subtype H9N2 in Turkeys and Evaluation of Inactivated Vaccine Efficacy.

Avian influenza H9N2 viruses circulate in all types of poultry species, including turkeys, and cause significant losses for the poultry industry in many parts of the word. The aim of this study was to assess the pathogenesis of the Moroccan avian influenza virus (AIV) H9N2 under experimental conditions in turkeys and the protection efficacy of an inactivated commercial vaccine against AIV H9N2. Unvaccinated turkeys showed marked depression sinusitis, respiratory distress characterized by bronchiolar and tracheal rales of moderate severity, and a mortality rate of 50%. Postmortem examinations of dead and euthanatized birds revealed the presence of fibrinous tracheitis and airsacculitis lesions. Vaccination reduced the mortality rate to 20%. Vaccinated birds recovered at day 10 postchallenge, and only 12.5% (1/8) and 37.5% of birds still displayed fibrinous and nonfibrinous airsacculitis lesions, respectively, at day 15 postinoculation. Viral shedding in cloacal and tracheal swabs was lower in vaccinated than in control birds. Although viral RNA was detected in the cloacal swabs of all unvaccinated turkeys at day 3 postinoculation, only 50% of the vaccinated turkeys were positive for virus detection. At day 11 postinoculation, no viral RNA was detected in oropharyngeal swabs of vaccinated turkeys, whereas 40% of the unvaccinated turkeys were still shedding virus.

Artículo regular­Patogenia del subtipo H9N2 del virus de la influenza aviar en pavos y evaluación de la eficacia de una vacuna inactivada. Los virus de la influenza aviar H9N2 circulan en todo tipo de especies de aves comerciales, incluidos los pavos, y causan pérdidas significativas para la industria avícola en muchas partes del mundo. El objetivo de este estudio fue evaluar la patogenia del virus de la influenza aviar de Marruecos (AIV) H9N2 bajo condiciones experimentales en pavos y la eficacia de protección de una vacuna comercial inactivada contra el virus de la influenza aviar H9N2. Los pavos no vacunados mostraron una marcada sinusitis, depresión, dificultad respiratoria caracterizada por estertores bronquiolares y traqueales de severidad moderada y una tasa de mortalidad del 50%. Los exámenes post mortem de aves muertas y sacrificadas revelaron la presencia de traqueítis fibrinosa y aerosaculitis. La vacunación redujo la tasa de mortalidad al 30%. Las aves vacunadas se recuperaron en el día 10 después del desafío, y solo el 12.5% (1/8) y el 37.5% de las aves todavía mostraban aerosaculitis fibrinosa y no fibrinosa, respectivamente, el día 15 después de la inoculación. La diseminación viral en los hisopos cloacales y traqueales fue menor en las aves vacunadas que en las aves control. Aunque se detectó ARN viral en los hisopados cloacales de todos los pavos no vacunados en el día tres después de la inoculación, solo el 50% de los pavos vacunados dieron positivo para la detección del virus. En el día 11 después de la inoculación, no se detectó ARN viral en hisopados orofaríngeos de pavos vacunados, mientras que el 40% de los pavos no vacunados todavía estaban diseminando virus.

Co-infections of chickens with avian influenza virus H9N2 and Moroccan Italy 02 infectious bronchitis virus: effect on pathogenesis and protection conferred by different vaccination programmes.

Since the emergence of low pathogenic avian influenza (LPAI) H9N2 viruses in Morocco in 2016, severe respiratory problems have been encountered in the field. Infectious bronchitis virus (IBV) is often detected together with H9N2, suggesting disease exacerbation in cases of co-infections. This hypothesis was therefore tested and confirmed in laboratory conditions using specific-pathogen-free chickens. Most common field vaccine programmes were then tested to compare their efficacies against these two co-infecting agents. IBV γCoV/chicken/Morocco/I38/2014 (Mor-IT02) and LPAI virus A/chicken/Morocco/SF1/2016 (Mor-H9N2) were thus inoculated to commercial chickens. We showed that vaccination with two heterologous IBV vaccines (H120 at day one and 4/91 at day 14 of age) reduced the severity of clinical signs as well as macroscopic lesions after simultaneous experimental challenge. In addition, LPAI H9N2 vaccination was more efficient at day 7 than at day 1 in limiting disease post simultaneous challenge.RESEARCH HIGHLIGHTS Simultaneous challenge with IBV and AIV H9N2 induced higher pathogenicity in SPF birds than inoculation with IBV or AIV H9N2 alone.Recommended vaccination programme in commercial broilers to counter Mor-IT02 IBV and LPAIV H9N2 simultaneous infections: IB live vaccine H120 (d1), AIV H9N2 inactivated vaccine (d7), IB live vaccine 4-91 (d14).