Publications by authors named "Mehdi El Harrak"

36 Publications

Draft Genome Sequence of Pasteurella multocida Serotype A Strain MOR19, Isolated in Morocco.

Microbiol Resour Announc 2021 Oct 14;10(41):e0086721. Epub 2021 Oct 14.

Research and Development Department, Multi-Chemical Industry, Mohammedia, Morocco.

Pasteurella multocida causes pneumonia in large ruminants. In this study, we determined the genome sequence of the capsular serotype A Pasteurella multocida strain MOR19, isolated from a calf that died from acute pneumonia.
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http://dx.doi.org/10.1128/MRA.00867-21DOI Listing
October 2021

Biological and molecular characterization of a sheep pathogen isolate of and leukotoxin production kinetics.

Vet World 2021 Aug 7;14(8):2031-2040. Epub 2021 Aug 7.

Department of Microbiology, Immunology and Contagious Diseases, Institute of Agronomy and Veterinary Medicine Hassan II, Rabat, Morocco.

Background And Aim: (Mha) is a common agent of pneumonia in ruminants globally, causing economic losses by morbidity, mortality, and treatment costs. Infection by Mha is often associated with or promoted by respiratory viral pathogens and environmental conditions. Infections due to Mha have rarely been described in small ruminants. This study reports the biological and molecular characteristics of a new Moroccan Mha isolate from small ruminants presenting typical respiratory symptoms. We also studied the cultural parameters, growth kinetics, and Lkt excretion of the isolate and its pathogenicity on laboratory animals and small ruminants.

Materials And Methods: Suspected pasteurellosis cases in sheep and goat flocks in Morocco were investigated. A local strain of Mha was isolated and identified using biochemical and molecular methods. Polymerase chain reaction-targeting specific genes were used for serotyping and phylogenetic analyses; further, leukotoxin production, cytotoxicity, and pathogenicity of the isolate in mice, goats, and sheep were investigated.

Results: Phylogeny analysis revealed 98.76% sequence identity with the USA isolate of 2013; the strain growth with a cycle of 9-10 h with leukotoxin secretion was detected by NETosis and quantified by cytotoxicity and mortality of mice. Goat and sheep infections cause hyperthermia, with characteristic postmortem lesions in the trachea and lung.

Conclusion: A local isolate of Mha from sheep that died of pneumonia was characterized for the 1 time in North Africa using biological and molecular methods. Although growth on appropriate culture media is accompanied by intense leukotoxin secretion, experimental infections of sheep and goats cause hyperthermia and typical lesions of pneumonia.
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http://dx.doi.org/10.14202/vetworld.2021.2031-2040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8448628PMC
August 2021

Genome Sequence of MHA.Sh.MOR19 Serotype 1, a Moroccan Sheep Isolate.

Microbiol Resour Announc 2021 May 27;10(21):e0035921. Epub 2021 May 27.

Research and Development Department, Multi-Chemical Industry, Mohammedia, Morocco.

Mannheimia haemolytica is the principle bacterial pathogen in ruminants associated with respiratory disease. Here, we report the draft genome sequence of the Mannheimia haemolytica MHA.Sh.MOR19 strain that was recently isolated in the northwest of Morocco from the lung of a lamb that died from pneumonia. The genome size is 2,434,458 bp.
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http://dx.doi.org/10.1128/MRA.00359-21DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8201628PMC
May 2021

Capripoxvirus Infections in Ruminants: A Review.

Microorganisms 2021 Apr 23;9(5). Epub 2021 Apr 23.

Department of Microbiology, Immunology and Contagious Diseases, Agronomic and Veterinary Institute Hassan II, Madinat Al Irfane, Rabat 6202, Morocco.

Lumpy skin disease, sheeppox, and goatpox are notifiable diseases of cattle, sheep, and goats, respectively, caused by viruses of the Capripoxvirus genus. They are responsible for both direct and indirect financial losses. These losses arise through animal mortality, morbidity cost of vaccinations, and constraints to animals and animal products' trade. Control and eradication of capripoxviruses depend on early detection of outbreaks, vector control, strict animal movement, and vaccination which remains the most effective means of control. To date, live attenuated vaccines are widely used; however, conferred protection remains controversial. Many vaccines have been associated with adverse reactions and incomplete protection in sheep, goats, and cattle. Many combination- and recombinant-based vaccines have also been developed. Here, we review capripoxvirus infections and the immunity conferred against capripoxviruses by their respective vaccines for each ruminant species. We also review their related cross protection to heterologous infections.
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http://dx.doi.org/10.3390/microorganisms9050902DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8145859PMC
April 2021

An Evaluation of Three Different Primary Equine Influenza Vaccination Intervals in Foals.

J Equine Vet Sci 2021 04 3;99:103397. Epub 2021 Feb 3.

Department of Medicine, Surgery and Reproduction, Hassan II Institute for Agronomy and Veterinary Medicine - B.P 6202 Rabat-Institutes, Rabat, Morocco.

In order to evaluate the effect of three different primary vaccination intervals on EI vaccine response, 21 unvaccinated thoroughbred foals were randomly divided into three groups of 7 and vaccinated with three different intervals of primary immunization (i.e., with 1, 2 or 3 months intervals between V1 and V2, respectively). The antibody response was measured for up to 1 year after the third immunization V3 (administered 6 months after V2) by single radial hemolysis (SRH) assay. All weanlings had seroconverted and exceeded the clinical protection threshold 2 weeks after V2 and 1 month after V3 until the end of the study. Significant differences were measured at the peak of immunity after V2 and for the duration of the immunity gap between V2 and V3. The group with one month primary vaccination interval had a lower immunity peak after V2 (158.05 ± 6.63 mm) and a wider immunity gap between V2 and V3 (18 weeks) when compared with other groups (i.e., 174.72 ± 6.86 mm and 16 weeks for a two months interval, 221.45 ± 14.48 mm and 12 weeks for a 3-month interval). The advantage observed in the group with 1 month primary vaccination interval, which induces an earlier protective immunity, is counterbalance with a lower peak of immunity and a wider immunity gap after V2, when compared with foals vaccinated with 2- and 3-month intervals.
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http://dx.doi.org/10.1016/j.jevs.2021.103397DOI Listing
April 2021

Development of an inactivated combined vaccine for protection of cattle against lumpy skin disease and bluetongue viruses.

Vet Microbiol 2021 May 23;256:109046. Epub 2021 Mar 23.

Research and Development, MCI Santé Animale, ZI Sud-Ouest B.P: 278, Mohammedia, 28810, Morocco.

Lumpy Skin Disease (LSD) and Bluetongue (BT) are the main ruminants viral vector-borne diseases. LSD is endemic in Africa and has recently emerged in Europe and central Asia as a major threat to cattle industry. BT caused great economic damage in Europe during the last decade with a continuous spread to other countries. To control these diseases, vaccination is the only economically viable tool. For LSD, only live-attenuated vaccines (LAVs) are commercially available, whilst for BT both LAVs and inactivated vaccines are available with a limited number of serotypes. In this study, we developed an inactivated, oil adjuvanted bivalent vaccine against both diseases based on LSDV Neethling strain and BTV4. The vaccine was tested for safety and immunogenicity on cattle during a one-year period. Post-vaccination monitoring was carried out by VNT and ELISA. The vaccine was completely safe and elicited high neutralizing antibodies starting from the first week following the second injection up to one year. Furthermore, a significant correlation (R = 0.9040) was observed when comparing VNT and competitive ELISA in BTV4 serological response. Following BTV4 challenge, none of vaccinated and unvaccinated cattle were registered clinical signs, however vaccinated cattle showed full protection from viraemia. In summary, this study highlights the effectiveness of this combined vaccine as a promising solution for both LSD and BT control. It also puts an emphasis on the need for the development of other multivalent inactivated vaccines, which could be greatly beneficial for improving vaccination coverage in endemic countries and prophylaxis of vector-borne diseases.
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http://dx.doi.org/10.1016/j.vetmic.2021.109046DOI Listing
May 2021

Experimental infection of indigenous North African goats with goatpox virus.

Acta Vet Scand 2021 Mar 4;63(1). Epub 2021 Mar 4.

Department of Research and Development, Multi-Chemical Industry Santé Animale, Lot. 157, Z I, Sud-Ouest (ERAC) B.P.: 278, 28810, Mohammedia, Morocco.

Background: Goatpox is a viral disease caused by infection with goatpox virus (GTPV) of the genus Capripoxvirus, Poxviridae family. Capripoxviruses cause serious disease to livestock and contribute to huge economic losses. Goatpox and sheeppox are endemic to Africa, particularly north of the Equator, the Middle East and many parts of Asia. GTPV and sheeppox virus are considered host-specific; however, both strains can cause clinical disease in either goats or sheep with more severe disease in the homologous species and mild or sub-clinical infection in the other. Goatpox has never been reported in Morocco, Algeria or Tunisia despite the huge population of goats living in proximity with sheep in those countries. To evaluate the susceptibility and pathogenicity of indigenous North African goats to GTPV infection, we experimentally inoculated eight locally bred goats with a virulent Vietnamese isolate of GTPV. Two uninfected goats were kept as controls. Clinical examination was carried out daily and blood was sampled for virology and for investigating the antibody response. After necropsy, tissues were collected and assessed for viral DNA using real-time PCR.

Results: Following the experimental infection, all inoculated goats displayed clinical signs characteristic of goatpox including varying degrees of hyperthermia, loss of appetite, inactivity and cutaneous lesions. The infection severely affected three of the infected animals while moderate to mild disease was noticed in the remaining goats. A high antibody response was developed. High viral DNA loads were detected in skin crusts and nodules, and subcutaneous tissue at the injection site with cycle threshold (Ct) values ranging from 14.6 to 22.9, while lower viral loads were found in liver and lung (Ct = 35.7 and 35.1). The results confirmed subcutaneous tropism of the virus.

Conclusion: Clinical signs of goatpox were reproduced in indigenous North African goats and confirmed a high susceptibility of the North African goat breed to GTPV infection. A clinical scoring system is proposed that can be applied in GTPV vaccine efficacy studies.
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http://dx.doi.org/10.1186/s13028-021-00574-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7931584PMC
March 2021

Using geostatistics to better understand the epidemiology of animal rabies in Morocco: what is the contribution of the predictive value?

Heliyon 2021 Jan 25;7(1):e06019. Epub 2021 Jan 25.

Hassan II Institute of Agronomy and Veterinary Medicine, Microbiology Immunology and Contagious Diseases Unit, Po. Box 6202, Madinat Al Irfane, Rabat, Morocco.

This study aims to characterize the spatial distribution of animal rabies in Morocco in order to provide appropriate control approaches. Descriptive analyses of the epidemiological data show that the number of reported canine rabies cases greatly underestimates the true incidence of the disease. Underreporting subsequently affects the coherence of its spatial distribution. To perform accurate geographic distribution mapping of the disease based on interpolation methods, a data set was created using data between 2000 and 2018 to compare the derived disease cases with known true values in order to identify disease clusters. The subsequent interpolation was conducted using Ordinary Kriging regression methods and the semi variogram to focus on short distances and reduce uncertainty. The estimated clusters of rabies were evaluated using a cross validation step which revealed predicted cases close to the true values. To improve the precision of analysis, the authors displayed georeferenced dog and human rabies cases reported during the last three years, demonstrating reliable results that correspond to the estimated cluster areas similar to the true disease incidence on the field. This work highlights a strong correlation between infrastructure projects (i.e. railways, roads, facilities) and rabies epizootics for several specific locations. This study is the first attempt to use geostatistics to build upon the understanding of animal rabies in Morocco and shed light on the most appropriate strategies to sustainably reduce and mitigate the risk of rabies. There has been little literature on the use of kriging methods in animal health research. Thus, this study also aimed to explore a novel method in the veterinary sciences to establish kriging as a valid and coherent analysis tool to identify the extent to which the geostatistic area can objectively support understanding on animal rabies and saw it as being highly instrumental in coping with gaps in the data.
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http://dx.doi.org/10.1016/j.heliyon.2021.e06019DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7841317PMC
January 2021

Experimental evaluation of the cross-protection between Sheeppox and bovine Lumpy skin vaccines.

Sci Rep 2020 06 1;10(1):8888. Epub 2020 Jun 1.

Research and Development Virology, Multi-Chemical Industry, Lot. 157, Z I, Sud-Ouest (ERAC) B.P.: 278, Mohammedia, 28810, Morocco.

The Capripoxvirus genus includes three agents: Sheeppox virus, Goatpox virus and Lumpy skin disease virus. Related diseases are of economic importance and present a major constraint to animals and animal products trade in addition to mortality and morbidity. Attenuated vaccines against these diseases are available, but afforded cross-protection is controversial in each specie. In this study, groups of sheep, goats and cattle were vaccinated with Romania SPPV vaccine and challenged with corresponding virulent strains. Sheep and cattle were also vaccinated with Neethling LSDV vaccine and challenged with both virulent SPPV and LSDV strains. Animals were monitored by clinical observation, rectal temperature as well as serological response. The study showed that sheep and goats vaccinated with Romania SPPV vaccine were fully protected against challenge with virulent SPPV and GTPV strains, respectively. However, small ruminants vaccinated with LSDV Neethling vaccine showed only partial protection against challenge with virulent SPPV strain. Cattle showed also only partial protection when vaccinated with Romania SPPV and were fully protected with Neethling LSDV vaccine. This study showed that SPPV and GTPV vaccines are closely related with cross-protection, while LSDV protects only cattle against the corresponding disease, which suggests that vaccination against LSDV should be carried out with homologous strain.
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http://dx.doi.org/10.1038/s41598-020-65856-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264126PMC
June 2020

Development and Evaluation of an Inactivated Lumpy Skin Disease Vaccine for Cattle.

Vet Microbiol 2020 Jun 18;245:108689. Epub 2020 Apr 18.

Research and development Virology, Multi-Chemical Industry, Lot. 157, ZI Sud-Ouest (ERAC) B.P: 278, Mohammedia 28810, Morocco.

Lumpy skin disease (LSD) of cattle is caused by a virus within Capripoxvirus genus. It leads to huge economic losses in addition to trade and animal movement limitation. Vaccination is the only economically feasible way to control this vector-borne disease. Only live attenuated vaccines have been used so far and no inactivated vaccine has been developed nor tested in cattle. In this study, we developed an inactivated oily adjuvanted vaccine based on Neethling strain and tested it on cattle. Selected criteria of appreciation were safety, antibody response by Virus Neutralization and protection through challenge. A field trial was also performed in Bulgaria. The vaccine was safe and did not cause any adverse reaction, high level of specific antibodies was obtained starting from day 7 post-vaccination and protection against virulent challenge strain that caused typical disease in control animals was total. Induced protection was similar to that obtained with live vaccine, without any adverse effect. In addition, the field study confirmed safety and efficacy of the vaccine, which did not show any adverse reaction and induced a high level of antibodies for up to one year. General prophylaxis based on inactivated vaccine could be of great benefit in endemic countries or at risk regions.
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http://dx.doi.org/10.1016/j.vetmic.2020.108689DOI Listing
June 2020

Pathogenicity and Full Genome Sequencing of the Avian Influenza H9N2 Moroccan Isolate 2016.

Avian Dis 2019 03;63(1):24-30

Research and Development Department, Multi-Chemical Industry, Lot 157, Z I, Sud-Ouest (ERAC) B. P. 278, Mohammedia 28810, Morocco.

In Morocco in early 2016, a low pathogenic avian influenza virus serotype H9N2 caused large economic losses to the poultry industry, with specific clinical symptoms and high mortality rates on infected farms. Subsequent to the H9N2 outbreak, the causal agent was successfully isolated from chicken flocks with high morbidity and mortality rates, propagated on embryonated eggs, and fully sequenced. The phylogenetic analysis suggested that the Moroccan isolate could have derived from the Middle East isolate A/chicken/Dubai/D2506.A/2015. This study was designed to assess the pathogenicity of the Moroccan isolate H9N2 in experimentally infected broiler and specific-pathogen-free (SPF) chickens. At 22 days of age, one broiler and two SPF chicken groups were inoculated by dropping 0.2 ml of the H9N2 isolate (10 EID/ml) in both nostrils and eyes. Clinically inoculated chickens with H9N2 displayed mild lesions, low mortality rates, and an absence of clinical signs. The H9N2 virus was more pathogenic in broiler chickens and produced more severe tissue lesions compared to SPF chickens. The viral shedding was detected up to 6 days postinoculation (pi) in oropharyngeal and cloacal swabs in infected birds with a maximum shedding in the oropharynges of the broiler group. All experimental chickens seroconverted and registered high hemagglutination inhibition titers as early as day 7 pi. The present study indicates that the H9N2 virus isolated from a natural outbreak was of low pathogenicity under experimental conditions. However, under field conditions infection with other pathogens might have aggravated the disease.
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http://dx.doi.org/10.1637/11941-080418-Reg.1DOI Listing
March 2019

The value of camels as sentinels for bluetongue virus in Morocco.

Vet Ital 2018 12 31;54(4):343-348. Epub 2018 Dec 31.

Agronomy and Veterinary Institute (IAV) Hassan II, Al Irfane, Rabat, Morocco.

A  serosurvey  was  conducted  to  determine  the  value  of  camels  (Camelus  dromedaries)  as sentinel animals for the detection of bluetongue virus (BTV) in Morocco. Between 2010 and 2013, camels from various localities in Morocco were randomly tested for antibodies against BTV  serotypes‑1,  ‑4,  ‑6,  ‑8,  ‑11,  ‑14,  and  ‑16.  Antibodies  against  1  or  more  serotypes  were detected in 41.8% of 537 camels tested with a competitive enzyme‑linked immunosorbent assay  (ELISA)  diagnostic  test.  Of  the  7  tested  serotypes,  only  BTV‑11  antibodies  were  not detected with serum neutralisation assays. This study not only confirms the epidemiological presence of BTV‑1, ‑4, and ‑8 in Morocco, but also presents the first evidence of BTV‑6, ‑14, and ‑16 in the country. As such, we conclude that camels would be ideal sentinel animals to determine the potential risk of BTV in Morocco.
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http://dx.doi.org/10.12834/VetIt.1502.8097.1DOI Listing
December 2018

Impact of Mixed Equine Influenza Vaccination on Correlate of Protection in Horses.

Vaccines (Basel) 2018 Oct 4;6(4). Epub 2018 Oct 4.

Department of Microbiology, Immunology and Contagious Diseases, Agronomic and Veterinary Institute Hassan II-B.P 6202 Rabat-Institutes, Rabat-Institutes 10101, Morocco.

To evaluate the humoral immune response to mixed Equine Influenza vaccination, a common practice in the field, an experimental study was carried out on 42 unvaccinated thoroughbred weanling foals divided into six groups of seven. Three groups were vaccinated using a non-mixed protocol (Equilis Prequenza-Te, Proteqflu-Te or Calvenza-03) and three other groups were vaccinated using a mix of the three vaccines mentioned previously. Each weanling underwent a primary EI vaccination schedule composed of two primary immunisations (V1 and V2) four weeks apart followed by a third boost immunisation (V3) six months later. Antibody responses were monitored until one-year post-V3 by single radial haemolysis (SRH). The results showed similar antibody responses for all groups using mixed EI vaccination and the group exclusively vaccinated with Equilis Prequenza-TE, which were significantly higher than the other two groups vaccinated with Proteqflu-TE and Calvenza-03. All weanlings (100%) failed to seroconvert after V1 and 21% (9/42) still had low or no SRH antibody titres two weeks post-V2. All weanlings had seroconverted and exceeded the clinical protection threshold one month after V3. The poor response to vaccination was primarily observed in groups exclusively vaccinated with Proteqflu-Te and Calvenza-03. A large window of susceptibility (3⁻4.5-month duration) usually called immunity gap was observed after V2 and prior to V3 for all groups. The SRH antibody level was maintained above the clinical protection threshold for three months post-V3 for the groups exclusively vaccinated with Proteqflu-Te and Calvenza-03, and six months to one year for groups using mixed EI vaccination or exclusively vaccinated with Equilis Prequenza-Te. This study demonstrates for the first time that the mix of EI vaccines during the primary vaccination schedule has no detrimental impact on the correlate of protection against EIV infection.
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http://dx.doi.org/10.3390/vaccines6040071DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313876PMC
October 2018

Serological investigation of racehorse vaccination against equine influenza in Morocco.

Vet Microbiol 2018 Sep 11;223:153-159. Epub 2018 Aug 11.

Department of Microbiology, Immunology and Contagious Diseases, Agronomic and Veterinary Institute Hassan II- B.P 6202 Rabat-Institutes, Morocco.

In order to evaluate the vaccination status against equine influenza (EI) in Moroccan racehorses, a serological investigation was carried out on 509 racehorses using three serological tests: an Enzyme-Linked Immunosorbent Assay (ELISA), the Hemagglutination Inhibition (HI) test and the Single Radial Haemolysis (SRH) assay. The serological analysis showed 56% of seropositivity by ELISA, 67% by HI and 89.4% by SRH (with 69.9% above the clinical protection threshold). Using the Kappa test, the SRH and HI assays showed a strong agreement, the SRH and ELISA assays had a moderate agreement and the HI and ELISA assays showed a poor agreement. Seropositivity was positively correlated with the age of horses and the number of immunisation received. EI vaccines used during the last immunisation before the study had a weak influence on the serological status. This effect was observed when the vaccines Calvenza and Fluvac Innovator® were used, with 94.1% and 100% of seropositivity when measured by HI, and with 100% and 94.7% exceeding the clinical protection threshold when measured by SRH, respectively. No effect was found when other EI vaccines, including Prequenza-Te® (67% coverage (342/509) and Proteqflu-Te® (22% coverage (114/509) were used; with 64% and 67.5% seropositivity (HI) and with 66.4% and 72.8% above the clinical threshold (SRH), respectively. The location and the time since last vaccination have no influence on the serological result. Overall, levels of protective antibody against EI in Moroccan racehorses remain a concern despite mandatory vaccination.
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http://dx.doi.org/10.1016/j.vetmic.2018.08.014DOI Listing
September 2018

Assessment and comparison of the pathogenicity of Sheeppox Virus strains isolated in Morocco.

Iran J Microbiol 2017 Dec;9(6):372-380

Society Biopharma, Km 2, Route de Casa, B.P. 4569, Rabat, Morocco.

Background And Objectives: Sheeppox virus causes systemic disease in sheep that is often associated with high morbidity and mortality. Protection against sheep pox is mainly based on medical prophylaxis, vaccination being the only way. In Morocco, and up to now, there is no available information about local challenge strain to use for controlling the efficiency of vaccines produced against sheep pox. Hence, the objective of the present study was to evaluate and compare the pathogenicity of seven Sheeppox virus (SPVs) isolates from 1993-1995 in Morocco.

Materials And Methods: These seven SPV isolates have undergone various tests to evaluate their pathogenicity: Passages and titration on cell culture, Experimental inoculation on sheep, Virus-neutralization, titration and viral re-isolation by real-time PCR assay.

Results: All infected lambs showed severe clinical signs, while most of them have been reproduced on 5 dpi and persisted until 21 dpi. The lambs infected by Oj1P4, Oj2P4 and BerP5 appeared lethargic, reluctant to move compared to those infected by other isolates. The results also revealed that all isolates were able to induce serological response. Virus isolation from infected organs and blood and amplification of the viral DNA by real-time PCR proved the presence of the virus in tissues and blood of infected lambs. These Moroccan SPVs demonstrated that the three isolates Oj1P4, Oj2P4 and BerP5 have a high pathogenicity; especially the BerP5 isolate which has an important infectious titer.

Conclusion: These results demonstrate that the Berkane isolate is the most pathogenic of the tested isolates and it can be an excellent challenge strain for the control of the efficiency of vaccines against sheep pox produced in Morocco.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5825938PMC
December 2017

Seroprevalence of West Nile virus in horses in different Moroccan regions.

Vet Med Sci 2017 Nov 12;3(4):198-207. Epub 2017 Sep 12.

Laboratory of Microbiology and Molecular BiologyFaculty of ScienceUniversity MohammedRabatV BP 1014Morocco.

West Nile virus-associated disease is one of the most widespread vector-borne diseases in the world. In Morocco, the first cases were reported in horses in 1996 and the disease re-emerged in 2003 and in 2010. The objective of this work was to study the epidemiological situation of WNV-associated infection in Morocco, by quantifying the seroprevalence of anti-WNV IgM and IgG antibodies in horses in different bioclimatic regions-zones of Morocco in 2011. During the months of May, June and July 2011, 840 serum samples were collected from horses in four regions characterized by different environmental and climatic features such as altitude, temperature and precipitation. These environmental-climatic regions are: the Atlantic plateaus of the Gharb and pre-Rif region, the North Atlasic plains and plateaus region, the Atlas Mountains and pre-Atlas region and the plains and plateaus of the Oriental region. All samples were tested for the anti-WNV IgG antibodies by ELISA and positive sera were confirmed by virus neutralization (VN). An anti-WNV antibody prevalence map was developed. A total of 261 samples (31%) were found positive by both techniques. The prevalence of the infection was higher in the Atlantic plateaus of the Gharb and pre-Rif region, in the northern part of the country. Available data concerning the previous WNV-associated disease outbreaks in Morocco and the preliminary results of this serological survey suggest that the Moroccan northwest is the region at highest risk for WNV circulation. In this region, the climate is more humid with higher rainfall than other regions and milder winter temperatures exist. In the same area, the presence of migratory bird settlements may affect the risk of virus introduction and amplification.
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http://dx.doi.org/10.1002/vms3.71DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5677775PMC
November 2017

Phylogenetic analysis of avian infectious bronchitis virus isolates from Morocco: a retrospective study (1983 to 2014).

Virol Sin 2017 Apr;32(2):155-158

Interactions Hôtes-Agents Pathogènes, Université de Toulouse, Institut National de la Recherche Agronomique, École National Vétérinaire de Toulouse, Toulouse, 31076, France.

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http://dx.doi.org/10.1007/s12250-016-3885-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6598899PMC
April 2017

Thermal Stability Study of Five Newcastle Disease Attenuated Vaccine Strains.

Avian Dis 2016 12;60(4):779-783

Research and Development Department, Multi-Chemical Industry, Lot. 157, Z I, Sud-Ouest (ERAC) B.P.: 278, Mohammedia 28810, Morocco.

Newcastle disease (ND) is a big concern throughout the world because of the devastating losses that can occur with commercial and backyard poultry. The major problem in many countries is the loss of the vaccine's effectiveness due to inadequate use or storage conditions, particularly in hot climates. In the present study, stability of the five, most-used NDV vaccine strains (I-2, LaSota, B1, Clone 30 [C30], and VG-GA) was tested comparatively at different storage temperatures (4 and 37 C for the freeze-dried form and 4, 24, 37, and 45 C for the freeze-dried vaccine reconstituted in diluents). The vaccine stability was evaluated by the cumulative infectious titer drop and the theoretical shelf life at particular temperatures. Results showed that I-2 and LaSota are the most stable vaccine strains compared to B1, C30, and VG-GA; they registered the lowest titer drops and the longest shelf life whether at cool, high, or room temperatures and for both freeze-dried and reconstituted vaccines.
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http://dx.doi.org/10.1637/11426-042116-Reg.1DOI Listing
December 2016

Preliminary results on innocuity and immunogenicity of an inactivated vaccine against Peste des petits ruminants.

Vet Ital 2016 Jun;52(2):101-9

Istituto Zoopro lattico Sperimentale dell'Abruzzo e del Molise 'G. Caporale', Teramo, Italy.

Peste des petits ruminants (PPR) virus belongs to the family Paramyxoviridae and represents a major threat to small livestock industry. In recent years, outbreaks of PPR have occurred in Turkey and North Africa. In endemic areas, disease prevention is accomplished using live‑attenuated vaccines. However, the use of live vaccines in non‑endemic regions, such as Europe, is not approved by Veterinary Authorities. In these regions inactivated vaccines are then the only viable alternative. In this study an inactivated vaccine (iPPRVac) was formulated with either a water‑in‑oil emulsion (ISA 71 VG) or with delta inulin adjuvant, alone (AFSA1) or combined with a TLR9 agonist oligonucleotide (AFSA2). These formulations were then tested for immunogenicity on rats. The iPPRV formulation with AFSA2 adjuvant induced 100% seroconversion in rats after 2 injections and was subsequently evaluated in goats. Five goats were immunised twice subcutaneously, 36 days apart with iPPRVac + AFSA2. The immunised goats all seroconverted to PPR by day 9 and remained seropositive until the end of the experimental period (133 days). These data indicate that the rat model is useful in predicting vaccine responses in goats and that inactivated vaccine, when formulated with a delta inulin adjuvant, represents a promising alternative to live attenuated vaccines for PPR vaccination campaigns in non‑endemic areas.
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http://dx.doi.org/10.12834/VetIt.396.1876.2DOI Listing
June 2016

Comparative innocuity and efficacy of live and inactivated sheeppox vaccines.

BMC Vet Res 2016 Jun 29;12(1):133. Epub 2016 Jun 29.

Research and Development Virology, Multi-Chemical Industry, Lot. 157, Z I, Sud-Ouest (ERAC) B.P.: 278, Mohammedia, 28810, Morocco.

Background: Sheeppox (SPP) is one of the priorities, high-impact animal diseases in many developing countries, where live attenuated vaccines are routinely used against sheeppox virus (SPPV). In an event of an SPP outbreak, historically disease-free countries would hesitate to use of live vaccines against SPPVdue to the safety and trade reasons. Currently no killed SPPV vaccines are commercially available. In this study, we developed an inactivated Romanian SPPVvaccine and assessed its efficacy and potency in comparison with a live attenuated Romanian SPPV vaccine. Four naïve sheep were vaccinated once with the Romanian SPPV live attenuated vaccine and16 sheep were vaccinated twice with the inactivated vaccine. All sheep in the live vaccine group were included in the challenge trial, which was conducted using a highly virulent Moroccan SPPV field strain. Eight sheep of the inactivated vaccine group were challenged and the remaining sheep were monitored for seroconversion. Experimental animals were closely monitored for the appearance of clinical signs, body temperature and inflammation at the injection site. Two naïve sheep were used as unvaccinated controls.

Results: The inactivated Romanian SPPV vaccine was found to be safe and confer a good protection, similar to the live vaccine. Specific antibodies appeared from seven days post vaccination and remained up to nine months.

Conclusion: This study showed that the developed inactivated Romanian SPPV vaccine has a potential to replace attenuated vaccine to control and prevent sheep pox in disease-free or endemic countries.
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http://dx.doi.org/10.1186/s12917-016-0754-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4928353PMC
June 2016

Comparison of SYBR green I real-time RT-PCR with conventional agarose gel-based RT-PCR for the diagnosis of infectious bronchitis virus infection in chickens in Morocco.

BMC Res Notes 2016 Apr 22;9:231. Epub 2016 Apr 22.

Laboratory of Virology, Microbiology and Quality/Ecotoxicology & Biodiversity, Faculty of Sciences and Techniques-University Hassan II Mohammedia, PO Box 146, Quartier Yasmina-Mohammedia, 20650, Casablanca, Morocco.

Background: A rapid, sensitive, and specific molecular method for the diagnosis of infectious bronchitis virus (IBV) infection is important in curbing infectious bronchitis outbreaks in Morocco and other countries.

Methods: In this study, an easy-to-perform SYBR green I real-time reverse transcriptase polymerase chain reaction (RT-PCR) targeting the nucleocapsid gene of IBV was developed and compared with conventional agarose gel-based RT-PCR for the detection of IBV infection.

Results: We found that the SYBR green I real-time RT-PCR was at least 10 times more sensitive than the agarose gel electrophoresis detection method. The assay exhibited high specificity for IBV infection. All negative controls, such as Newcastle disease virus, infectious bursal disease virus, and avian influenza virus, were not detected.

Conclusion: The SYBR green I real-time RT-PCR test described herein can be used to rapidly distinguish IBV from other respiratory pathogens, which is important for diagnosis and control of infectious bronchitis outbreaks in Morocco. The test is a valuable and useful method as a routine assay for diagnosis of clinical IBV infection in commercial chickens.
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http://dx.doi.org/10.1186/s13104-016-2037-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4841946PMC
April 2016

Spatio-Temporal Identification of Areas Suitable for West Nile Disease in the Mediterranean Basin and Central Europe.

PLoS One 2015 30;10(12):e0146024. Epub 2015 Dec 30.

Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise 'G. Caporale', Teramo, Italy.

West Nile virus (WNV) is a mosquito-transmitted Flavivirus belonging to the Japanese encephalitis antigenic complex of the Flaviviridae family. Its spread in the Mediterranean basin and the Balkans poses a significant risk to human health and forces public health officials to constantly monitor the virus transmission to ensure prompt application of preventive measures. In this context, predictive tools indicating the areas and periods at major risk of WNV transmission are of paramount importance. Spatial analysis approaches, which use environmental and climatic variables to find suitable habitats for WNV spread, can enhance predictive techniques. Using the Mahalanobis Distance statistic, areas ecologically most suitable for sustaining WNV transmission were identified in the Mediterranean basin and Central Europe. About 270 human and equine clinical cases notified in Italy, Greece, Portugal, Morocco, and Tunisia, between 2008 and 2012, have been considered. The environmental variables included in the model were altitude, slope, night time Land Surface Temperature, Normalized Difference Vegetation Index, Enhanced Vegetation Index, and daily temperature range. Seasonality of mosquito population has been modelled and included in the analyses to produce monthly maps of suitable areas for West Nile Disease. Between May and July, the most suitable areas are located in Tunisia, Libya, Egypt, and North Cyprus. Summer/Autumn months, particularly between August and October, characterize the suitability in Italy, France, Spain, the Balkan countries, Morocco, North Tunisia, the Mediterranean coast of Africa, and the Middle East. The persistence of suitable conditions in December is confined to the coastal areas of Morocco, Tunisia, Libya, Egypt, and Israel.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0146024PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4696814PMC
July 2016

Phylogenetic analysis of avian infectious bronchitis virus S1 glycoprotein regions reveals emergence of a new genotype in Moroccan broiler chicken flocks.

Virol J 2015 Aug 4;12:116. Epub 2015 Aug 4.

Laboratoirede Virologie, Microbiologie et Qualité/ETB- Faculté des Sciences et Techniques, Mohammedia, Université Hassan II- Casablanca, Mohammedia, 20650, Morocco.

Background: Infectious bronchitis virus (IBV), a major pathogen of commercial poultry flocks, circulates in the form of several serotypes/genotypes. Only a few amino-acid changes in the S1 subunit of wild-type IBVS proteins may result in mutants unaffected by current vaccines.

Methods: Partial S1 gene sequences of 3 IBV isolates of the Moroccan Italy 02 genotype from vaccinated and unvaccinated broiler chicken flocks, located in southern and central regions of Morocco, were amplified by RT-PCR, sequenced, and aligned for phylogenetic and amino-acid similarity analyses.

Results: The three isolates were found genetically highly distant from known avian IBV based on partial sequences of their S1 genes: gammaCoV/chicken/Morocco/I01/2011(IBV/Morocco/01), gammaCoV/chicken/Morocco/I30/2010 (IBV/Morocco/30), and gammaCoV/chicken/Morocco/I38/2013 (IBV/Morocco/38), nucleotide sequence identities reached 89.5 % to 90.9 % among the three isolates. The deduced protein sequence identities ranged from 29.7 % (between IBV/Morocco/38 and Egypt SCU-14/2013-1) to 78.2 % (between IBV/Morocco/01 and Spain/05/866). Amino acid sequence comparison and phylogenetic analysis indicated the emergence of a new Moroccan genotype, clustering with regionally related isolates from Spain (Spain/05/866) and belonging to a new sub-genotype.

Conclusion: Our sequencing results demonstrate a co-circulation of wild-type infectious bronchitis viruses in broiler chickens. These results justify permanent monitoring of circulating strains in order to rationally modify vaccination strategies to make them appropriate to the evolving field situation.
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http://dx.doi.org/10.1186/s12985-015-0347-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4524495PMC
August 2015

Prevalence and molecular characterization of avian infectious bronchitis virus in poultry flocks in Morocco from 2010 to 2014 and first detection of Italy 02 in Africa.

Avian Pathol 2015 ;44(4):287-95

a Unité de Pathologie Aviaire, Département de Pathologie et Santé Publique Vétérinaire , Institut Agronomique et Vétérinaire Hassan II , Rabat , Morocco.

The aim of this study was to investigate the prevalence and diversity of infectious bronchitis virus (IBV) genotypes in poultry flocks in 16 areas of Morocco between 2010 and 2014. A total of 360 chicken flocks suspected of being infected by IBV were screened for the IBV N gene using real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Flocks were classified into four groups according to their IBV vaccination programme. Group 1 contained unvaccinated birds. Group 2 received a single application of live H120 vaccine. Groups 3 and 4 birds received one or two booster vaccination(s), respectively, mostly using the H120 vaccine. The real-time RT-PCR results showed that 51.7% of the flocks were positive for the IBV genome with geographical disparities. Molecular characterization of IBV was performed on 50 RT-PCR positive samples by partially sequencing the S1 gene, including the hypervariable regions (nucleotides 705-1097). Two predominant genotypes were detected, with the Massachusetts type dominating (66%), among which 25% of the samples were identical to the H120 vaccine. The second most common genotype (present in 32% of the flocks) was surprisingly Italy 02, revealing the first detection of this genotype in Morocco and also in Africa. 793B, the predominant genotype in the late 1990s in Morocco, was only detected on one occasion and was identical to the 4/91 vaccine strain. This study highlights the high prevalence of IBV in poultry farms in Morocco and confirms its continuous dynamic changes and evolution.
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http://dx.doi.org/10.1080/03079457.2015.1044422DOI Listing
December 2016

Molecular evolution of peste des petits ruminants virus.

Emerg Infect Dis 2014 Dec;20(12):2023-33

Despite safe and efficacious vaccines against peste des petits ruminants virus (PPRV), this virus has emerged as the cause of a highly contagious disease with serious economic consequences for small ruminant agriculture across Asia, the Middle East, and Africa. We used complete and partial genome sequences of all 4 lineages of the virus to investigate evolutionary and epidemiologic dynamics of PPRV. A Bayesian phylogenetic analysis of all PPRV lineages mapped the time to most recent common ancestor and initial divergence of PPRV to a lineage III isolate at the beginning of 20th century. A phylogeographic approach estimated the probability for root location of an ancestral PPRV and individual lineages as being Nigeria for PPRV, Senegal for lineage I, Nigeria/Ghana for lineage II, Sudan for lineage III, and India for lineage IV. Substitution rates are critical parameters for understanding virus evolution because restrictions in genetic variation can lead to lower adaptability and pathogenicity.
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http://dx.doi.org/10.3201/eid2012.140684DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4257836PMC
December 2014

Serological evidence of West Nile Virus infection among humans in the southern Provinces of Morocco.

J Infect Dev Ctries 2013 Dec 15;7(12):999-1002. Epub 2013 Dec 15.

University Mohammed V-Souissi, Faculty of Medicine and Pharmacy, Mohammed V Military Teaching Hospital, Biosafety Level 3 and Research Laboratory, Rabat, Morocco.

Introduction: The objective of this study was to determine the prevalence of West Nile Virus infection in the southern provinces of Morocco.

Methodology: A total of 250 sera, collected during 2012 in the province of Dakhla, were analyzed by microneutralisation assay.

Results: WNV-neutralizing antibodies were detected in 13 samples (5.2%). The participants with WNV-specific antibodies were significantly younger than the rest of the population (p = 0.009). The positivity rate was higher among women (6.3%) than men (3.6%) (p = 0.26).

Conclusions: This is the first serological evidence of WNV infection among humans in the southern provinces of Morocco.
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http://dx.doi.org/10.3855/jidc.3399DOI Listing
December 2013

Complete Genome Sequence of a Peste des Petits Ruminants Virus Recovered from an Alpine Goat during an Outbreak in Morocco in 2008.

Genome Announc 2013 May 9;1(3). Epub 2013 May 9.

The Pirbright Institute, Pirbright, Surrey, United Kingdom;

Here, we announce the first complete genome sequence of a field isolate of a peste des petits ruminants virus (PPRV) from northern Africa. This isolate is derived from an Alpine goat that suffered from severe clinical disease during the 2008 outbreak in Morocco. The full genome sequence of this isolate clusters phylogenetically with the lineage IV isolates of PPRV, sharing high levels of sequence identity with other lineage IV isolates.
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http://dx.doi.org/10.1128/genomeA.00096-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3650429PMC
May 2013

Analysis of climatic and environmental variables associated with the occurrence of West Nile virus in Morocco.

Prev Vet Med 2013 Jul 1;110(3-4):549-53. Epub 2013 Mar 1.

Istituto Zooprofilattico dell'Abruzzo e del Molise 'G. Caporale', Via Campo Boario, 64100 Teramo, Italy.

West Nile disease (WND) is one of the most widespread mosquito-borne infectious diseases in the World. In Morocco the first WND cases were reported in equines in 1996. After an apparent epidemiological silence, WND re-occurred in 2003 and in 2010, when the disease was reported in equines living in the central and north-western part of the country. Eco-climatic variables are known to influence the mosquito presence and abundance and, therefore, the probability of occurrence of mosquito-borne infections. The remote sensed values of Land Surface Temperature (LST), Normalised Difference Vegetation Index (NDVI) and rainfall registered from 2001 to 2010 were evaluated for a possible association with the occurrence of WND cases in 2003 and in 2010. In the zones where WND cases occurred, NDVI values recorded in 2003 and 2010, from June to November, were significantly higher than those registered during the same months in the rest of the decade. Rainfall data showed higher peaks in 2003 and 2010, when the number of days with extreme rainfall was significantly higher during 1-2 months before the occurrence of WND cases. Temperature does not seem to play an important role in Moroccan epidemiological conditions.
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http://dx.doi.org/10.1016/j.prevetmed.2013.02.011DOI Listing
July 2013

A reliable and reproducible experimental challenge model for peste des petits ruminants virus.

J Clin Microbiol 2012 Nov 22;50(11):3738-40. Epub 2012 Aug 22.

Société de Productions Pharmaceutiques et Vétérinaires, Laboratoire de Virologie, Rabat, Morocco.

Experimental challenge protocols that consistently reproduce clinical signs of peste des petits ruminants in Alpine goats infected with a tissue culture-passaged peste des petits ruminants virus are described. The protocols can be used to carry out quality-controlled vaccine efficacy and pathogenesis studies under experimental conditions.
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http://dx.doi.org/10.1128/JCM.01785-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3486268PMC
November 2012

Experimental infection of alpine goats with a Moroccan strain of peste des petits ruminants virus (PPRV).

Vet Microbiol 2012 Nov 17;160(1-2):240-4. Epub 2012 May 17.

Société de productions pharmaceutiques et vétérinaires, Laboratoire de Virologie, Av Hassan II, BP 4569, Rabat, Morocco.

Peste des petits ruminants virus (PPRV) recently caused a serious outbreak of disease in Moroccan sheep and goats. Alpine goats were highly susceptible to PPRV with mortality rates approaching 100%, as opposed to local breeds of sheep which were less susceptible to the disease. The relative susceptibility of alpine goats was investigated through an experimental infection study with the Moroccan strain of PPRV. Severe clinical signs were observed in the alpine goats with virus being excreted through ocular, nasal and oral routes. No difference in the severity of the disease in goats was observed with different inoculation routes and transmission of the virus by direct contact was confirmed. This study confirmed the susceptibility of the alpine goat to PPRV infection and describes a challenge protocol that effectively and consistently reproduced severe clinical signs of PPR in experimentally infected goats.
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http://dx.doi.org/10.1016/j.vetmic.2012.04.043DOI Listing
November 2012
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