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Objective The objective behind this article is to better characterize spatial distribution of animal rabies in Morocco through qualitative risk assessment framework. In Morocco, the occurrence of the disease is neither clearly distributed nor complete. Therefore, risk assessment methods become strongly recommended to cope with distorted geographic patterns. Methods Based on data collection set from 168 counties, qualitative changes on spatial epidemiology of rabies were analysed by mapMCDA tool covering a period from 2004 to 2017 and including information on determinants of the geographic distribution of animal rabies in Morocco defined in previous work. Results To validate the risk assessment model, the results were compared to rabies cases reported during the study period. The clustering of the rabies risk estimates is decisive and highly reliable. A significant alignment was shown between the very high and high-risk estimates. Conclusion This study is the first attempt that has been made for using MapMCDA for rabies. For a normative process aiming to avoid subjectivity related to expert-opinions, authors suggest conducting initially a statistical multiple component analysis that will provide quantified estimates of risk factors. It would be an advisable decision-making tool that helps to design oriented surveillance and allows better referral of actions to control the disease.

Inclusion body hepatitis (IBH) associated with fowl aviadenovirus (FAdV) infection has a world-wide distribution, especially for broilers aged 3 to 5 weeks, causing significant economic losses to poultry industry. In present study, we evaluated the pathogenicity of Moroccan FAdV serotype 11 (MOR111115 strain) in specific pathogen free (SPF) white leghorn chicken embryos. FAdV (titre 10?/ml) was inoculated to SPF embryonated chicken eggs through the chorioallontoic membrane (CAM). The mortality, gross and microscopical lesions of the embryos were evaluated and the presence of the virus was checked by PCR. 100% cumulative embryonic mortality was observed at 7 days post-infection (dpi). The inoculated embryos were hemorragic and the liver was friable and swollen, with yellow to greenish discoloration. Microscopically, we highlighted a multifocal necrosis of hepatocytes groups with the presence of basophilic and eosinophilic intra-nuclear inclusion bodies within hepatocytes. The presence of the virus was confirmed by conventional polymerase chain reaction based on hexon gene from liver.that is a major target organ of FAdV infections. This is the first study of pathogenicity of fowl aviadenovirus in SPF chicken embryos in Morocco. Keywords: Fowl aviadenovirus-11, Inclusion body hepatitis, Pathogenicity, Chicken embryos

La maladie de Gumboro ou bursite infectieuse aviaire (IBD) est associée au virus de la bursite infectieuse (IBDV), qui cause des pertes économiques considérables. Ce virus appartient à la famille des Birnaviridae (Avibirnavirus). Cette infection continue à sévir au sein des élevages avicoles au Maroc particulièrement ceux de poulets de chair malgré les différents programmes de vaccination appliqués. Ainsi, l’objectif du présent travail est la caractérisation épidémiologique et pathotypique de l’IBDV au Maroc102 élevages de poulets de chair suspects à travers tout le Maroc enquêtés entre les années 2013 et 2016. Ainsi que les bourses de Fabricius (BF) (pool de 5 BF par élevage) ont été prélevés à partir de poulets affectés pour des investigations moléculaires par RT-PCR en temps réel. La RT-PCR utilisée permettait aussi bien la détection que la discrimination entre les souches très virulentes de l’IBDV (vvIBDV) et les non-vvIBDV en utilisant des sondes de discrimination allélique. L'enquête épidémiologique révèle la suspicion de l’atteinte des élevages avicoles par l’IBDV vu les signes cliniques et nécropsiques très caractéristiques causés par l’IBDV (apathie, plumes ébouriffées, prostration, diarrhée blanchâtre aqueuse, et la triade nécrosique (hypertrophie et hémorragie de la Bourse de Fabricius ; congestion des reins et pétéchie au niveau des muscles)). La suspicion a été confirmée par les résultats de la RT-PCR dans toutes les régions du royaume avec une prévalence de l’atteinte par l’IBDV globale de 81% dont 60% pour les souches classiques (non-vvIBDV) et 40% pour les souches très virulentes (vvIBDV). Mots clés: Bursite infectieuse, IBDV, RT-PCR, épidémiologie, Maroc

Outbreaks of inclusion body hepatitis have emerged in Morocco since 2013 and has resulted in significant economic losses to poultry farms. Three isolates of the causative virus, Fowl adenonovirus (FAdV)were characterized from chickens with IBH, but their pathogenicity has never been investigated. In this work, the pathogenicity of an isolate FAdV 11 (MOR300315 strain) was evaluated by inoculating a group of 40 SPF chickens at 3 days of age by oral route. A group of 40 chicks injected with phosphate-buffered saline solution was used as a control group. The infected chickens showed decreased weight gain from 3dpi. Necropsy displayed pallor and enlargement in liver, swelling and slight hemorrhage in kidney and spleen at 6 dpi. Histopathological changes were mainly characterized by severe and extensive hepatic necrosis associated with the presence of basophilic intra-nuclear inclusion bodies within hepatocytes. The FAdV was reisolated in chicken embryo fibroblast cell culture from liver tissue homogenate of infected chicken from 3 to 6 dpi. Viral DNA was detected by PCR in liver, kidney, spleen and cloacal swabs from 3 to 13 dpi. Antibody response against inoculated FAdV was appeared from 9 dpi. These results confirmed that the FAdV 11 strain is pathogenic in chicken. This study is the first experimental infection of FAdV 11 in chicken in Morocco, which increase our understanding of its pathogenicity in chickens and indicate that preventive measures against FAdV infection in poultry farms should be implemented in Morocco.

Rift Valley fever virus (RVFV), an arbovirus belonging to the Phlebovirus genus of the Phenuiviridae family, causes the zoonotic and mosquito-borne RVF. The virus, which primarily affects livestock (ruminants and camels) and humans, is at the origin of recent major outbreaks across the African continent (Mauritania, Libya, Sudan), and in the South-Western Indian Ocean (SWIO) islands (Mayotte). In order to be better prepared for upcoming outbreaks, to predict its introduction in RVFV unscathed countries, and to run efficient surveillance programmes, the priority is harmonising and improving the diagnostic capacity of endemic countries and/or countries considered to be at risk of RVF. A serological inter-laboratory proficiency test (PT) was implemented to assess the capacity of veterinary laboratories to detect antibodies against RVFV. A total of 18 laboratories in 13 countries in the Middle East, North Africa, South Africa, and the Indian Ocean participated in the initiative. Two commercial kits and two in-house serological assays for the detection of RVFV specific IgG antibodies were tested. Sixteen of the 18 participating laboratories (88.9%) used commercial kits, the analytical performance of test sensitivity and specificity based on the seroneutralisation test considered as the reference was 100%. The results obtained by the laboratories which used the in-house assay were correct in only one of the two criteria (either sensitivity or specificity). In conclusion, most of the laboratories performed well in detecting RVFV specific IgG antibodies and can therefore be considered to be prepared. Three laboratories in three countries need to improve their detection capacities. Our study demonstrates the importance of conducting regular proficiency tests to evaluate the level of preparedness of countries and of building a network of competent laboratories in terms of laboratory diagnosis to better face future emerging diseases in emergency conditions.

Rapid diagnosis of canine parvovirus (CPV-2) infection is of key importance in management and control of the disease. The present study aims at the development and evaluation of an easy to use rapid test for the detection of canine parvovirus particles in clinical samples collected and tested during daily veterinary practice. For this purpose, the coagglutination test (COA) using Methylene blue stained anti-CPV2 antibodies sensitised staphylococcus aureus ATCC 12598 was performed to detect CPV-2 in 91 clinical samples (86 rectal swabs and 5 organ homogenates) collected between 2011 and 2015 from suspected canine clinical cases received in Moroccan veterinary clinics. The same samples were previously tested by Hemagglutination test and real-time PCR (Amrani et al., 2016), the results were compared and employed in the evaluation of COA. The results of the COA showed 79.12% (72/91) of tested samples to be positive and 20.87% (19/91) to be negative. The intensity of reaction in positive samples was scored high (3+) in 6 samples (6.6%), medium (2+) in 28 samples (30.7%) and low (1+) in 57 samples (62.6%) which occurred within a median time of 3min, 4min and 3min42s, respectively. In comparison with PCR, the COA test demonstrated a good agreement of 80.2 % and a sensitivity of 80%. The coagglutination test is a promising test allowing daily diagnostic of canine parvovirus in field, veterinary clinics and non- equipped laboratories with molecular techniques.

Fowl adenoviruses (FAdVs) belong to the genus Adenoviruses infecting chickens. They have been thoroughly investigated and identified using most recent developed molecular tools. Although some few of them are considered primary pathogens by themselves and are associated with specific well-known and characterized diseases, the role of most other viruses in specific disease conditions is still not fully elucidated either in field or experimental infections. Some well-known adenoviral diseases are controlled through vaccination and biosecurity measures. However, other conditions are still posing a serious threat to the poultry industry. During the last decade, these disease conditions have reemerged causing epidemic in many parts of the world. Their pathogenicity is not fully understood, and their control is therefore not enough mastered. Inclusion body hepatitis, hydropericardium syndrome, and gizzard erosion and ulcerative syndrome are the most representative of these conditions. Occurrence of outbreaks has increased in recent years and was associated with high mortality and/or growth retardation resulting in very substantial economic losses to the poultry industry worldwide. Hence, the present review aims at focusing on emerging or reemerging features of these virulent FAdV infections and their primary versus secondary role in triggering disease in fowls.

Capripoxviruses (CaPVs) are systemic infectious diseases of small ruminants, affecting the agricultural economy in endemic countries and representing a major constraint to international trade in livestock and their products. They cause fever, diarrhea, difficulty breathing, nodules, skin lesions developed into papules and then scabs. High morbidity and mortality rates are observed in affected sheep, goats, and cattle, depending on the virulence of strain, breed, host immunity, and environmental factors. CaPV diseases are notifiable to the World Organisation for Animal Health in many regions of Africa, Asia, and the Middle East, along with the possibility of an incursion into Europe via the transportation of infected animals. They are now expanding their territory with recent outbreaks of sheep pox (SPPV) and goat pox (GTPV) in Vietnam, Greece, and Mongolia and new outbreaks of lumpy skin disease (LSD) in Ethiopia, Egypt, and Israel. The CaPV pathogenesis demonstrates that skin lesions and nasal and oral swabs are the most useful samples for virus isolation. The classical methods such as cell culture and electron microscopy can be used to identify the virus; however, these techniques cannot differentiate between sheep pox, goat pox, and lumpy skin disease. The polymerase chain reaction and sequencing are the most frequently analytical techniques used to provide a rapid and sensitive detection of CaPV. In this chapter, it will be necessary to update knowledge on CaPV diseases; their economic impact and losses, modes of transmission, clinical signs and symptoms and to outline the classical and developed diagnostic methods, and vaccination strategies for their control and eradication.

La présente étude a eu pour objectifs d’estimer la prévalence et les facteurs de risques de la lymphadénite caséeuse chez les ovins de la région de l’Oriental au Maroc, et de déterminer ses caractéristiques cliniques et lésionnelles. Des enquêtes ont été réalisées dans différentes provinces de cette région. Elles ont porté sur 6376 animaux répartis dans 107 élevages. De plus, la recherche des lésions, et la collecte des ganglions lymphatiques et des abcès en vue d’un exa­men histopathologique ont été effectuées dans cinq abattoirs de la région. Les résultats ont montré que pratiquement 100 % des élevages (106 sur 107) étaient affectés par la lymphadénite caséeuse. La prévalence individuelle globale de cette maladie a été de 24 % et les prévalences moyennes au sein des élevages et aux abattoirs ont été respectivement de 28 et 16 %. Outre une hygiène défec­tueuse et une densité animale trop élevée dans les bergeries, l’usage d’équipe­ments traumatisants (mangeoires, abreuvoirs et clôtures métalliques) a semblé constituer le principal facteur favorisant la dissémination des abcès entre ani­maux d’un même troupeau. Les abcès superficiels, ouverts ou non, avec hyper­trophie ganglionnaire satellite, ont été présents chez tous les animaux affectés. Quoique de localisation variable, environ 79 % des lésions se situaient à la tête ou à l’encolure. L’examen histopathologique des ganglions a révélé deux types de lésions : des abcès au stade évolutif (87 %), et des lésions de nécroses et de suppurations diffuses et sévères du tissu lymphoïde (13 %). Dans les poumons la plupart des abcès étaient bien délimités et circonscrits.

Since it first emergence in the mid-1970's, canine parvovirus 2 (CPV-2) has evolved giving rise to new antigenic variants termed CPV-2a, CPV-2b and CPV-2c, which have completely replaced the original strain and had been variously distributed worldwide. In Africa limited data are available on epidemiological prevalence of these new types. Hence, the aim of the present study was to determine circulating variants in Morocco. Through TaqMan-based real-time PCR assay, 91 samples, collected from symptomatic dogs originating from various cities between 2011 and 2015, were diagnosed. Positive specimens were characterised by means of minor groove binder (MGB) probe PCR. The results showed that all samples but one (98.9%) were CPV positive, of which 1 (1.1%) was characterised as CPV-2a, 43 (47.7%) as CPV-2b and 39 (43.3%) as CPV-2c. Interestingly, a co-infection with CPV-2b and CPV-2c was detected in 4 (4.4%) samples and 3 (3.3%) samples were not characterised. Sequencing of the full VP2 gene revealed these 3 uncharacterised strains as CPV-2c, displaying a change G4068A responsible for the replacement of aspartic acid with asparagine at residue 427, impacting the MGB probe binding. In this work we provide a better understanding of the current status of prevailing CPV strains in northern Africa.

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., Animal rabies; Epidemiology; Prediction; Kriging; Veterinary science; Public health; Morocco.

Sheep pox is an infectious viral disease that affects specifically sheep and it is caused by the Capripoxvirus genus. The clinical signs include fever, diarrhea, difficulty breathing, nodules, lung lesions and death. In Morocco, the 2010 epidemic of sheep pox was characterized by the emergence of a nodular form of the disease. The local strain was isolated and the analysis of affected animals was positively confirmed by virus isolation and real-time polymerase chain reaction (RT-PCR). The epidemiological analysis of 911 data records showed that the virus is endemic in the country; an average of 350 cases per year with an epizootic evolution was observed in 2010. The incidence varies depending on provinces and the disease appears confined to the central and the eastern regions of the country where a very intensive sheep breeding activity is taking place. The statistical analysis showed that there is a positive correlation between the endemicity and the significant factor of the rural market (p = 0.006). The annual average morbidity and mortality rates were 2.96% (1.26% to 4.32%) and 0.71% (0.41% to 0.94%), respectively. The clinical findings associated to the epidemiological data analysis confirmed the presence of sheep pox in its nodular form and suggest that new pathogenic strains may have been introduced from Mauritania. The purpose of this work was to provide a better description of the spatiotemporal evolution of sheep pox disease based on some epidemiological indicators and to put forward plausible hypotheses regarding the emergence of the virus in order to implement an adequate control strategy.

The present study was conducted in order to isolate, identify and characterize fowl aviadenovirus associated with inclusion body hepatitis (IBH) in three poultry farms (two of broiler chickens and one of breeder broiler chickens) in Morocco during 2015. Liver samples collected from affected three poultry farms were examined by histopathological examination. Tissue samples showing necrosis of hepatocytes associated with basophilic intranuclear inclusion bodies were homogenized and submitted to FAdV isolation in chicken embryo fibroblast (CEF) cell cultures and in SPF embryonated eggs. The cytopathic effect (CPE) was observed in the second passage with swelling and rounding of infected cells. The inoculated embryos were hemorrhagic and showed hepatitis with the presence of basophilic intra-nuclear inclusion bodies within hepatocytes. The presence of the virus was confirmed by conventional polymerase chain reaction based on hexon gene from all investigated samples. Moreover, phylogenetic analysis of the hexon gene revealed that FAdVs isolated from different affected poultry belonged to FAdV 11 serotype of the D genotype group. This work is the first isolation in cell culture and SPF embryonated eggs of FAdV from Moroccan broilers and breeder broiler chickens with IBH.

Summary Sheep pox is endemic in most parts of Northern Africa and has the potential to cause severe economic problems. Live attenuated vaccines are used in Morocco, and in many other countries, to control the disease. Sheep pox virus (SPPV) re-appeared in 2010 causing a nodular clinical form previously not observed in Morocco. The severe clinical signs observed during the course of this outbreak and initial reports citing similarity in nucleotide sequence between the Moroccan vaccine strain and field isolates warranted a more in depth analysis of this epizootic. In this study, sequence analysis showed that isolates obtained from four provinces of eastern Morocco were identical, demonstrating that a single SPPV strain was responsible for the 2010 epizootic. In addition, the genome fragments sequenced and phylogenetic analyses undertaken as part of this study showed significant differences between field isolates and the Moroccan vaccine strain. New PCR methods were developed to differentiate between wild-type isolates and vaccine strains of SPPV. Using these methods, no trace of wild-type SPPV was found in the vaccine and no evidence was found to suggest that the vaccine strain was causing clinical disease.

Dans la province de Laâyoune au Maroc, la mortalité des jeunes dromadaires avant sevrage était importante sans qu'aucune étiologie évidente n'ait été identifiée. Afin de disposer d'une base d'échantillonage pour choisir les troupeaux qui feront l'objet d'un suivi, une typologie des éleveurs et de leurs pratiques a été réalisée par enquête transversale rétrospective auprès des éleveurs de la province de Laâyoune. Au terme de l'enquête, 170 éleveurs ont été interrogés, soit 10 % des éleveurs recensés, représentant 12 000 dromadaires, soit 44 % des dromadaires recensés dans la province de Laâyoune. L'analyse multivariée effectuée sur les données regroupées parthèmes a permis de déterminer 4 types d'organisation des éleveurs en fonction de la taille du troupeau, 4 types de stratégie de déplacement selon les conditions bioclimatiques et 3 types de pratique d'élevage. Ces trois typologies n'étaient pas corrélées. D'après la quantité d'informations obtenues, le nombre d'interlocuteurs et la variété des élevages rencontrés, la majorité des situations épidémiologiques semble avoir été abordée. La typologie des systèmes de production camelin dans la province de Laâyoune la plus pertinente était fondée sur l'organisation des hommes autour du troupeau. Mise en relation avec la mortalité des jeunes dromadaires avant sevrage, cette typologie a permis d'émettre des hypothèses de facteurs de risque, comme l'existence de toxiques alimentaires, les pratiques de traite et de sevrage, le niveau de présence du propriétaire, la délégation de la gestion du troupeau et le degré d'encadrement humain des jeunes dromadaires. De plus, cette clé de typologie, facilement identifiable sur le terrain, pourra servir de base d'échantillonnage pour des protocoles de suivis individuels ultérieurs qui permettront de préciser les différentes pratiques d'élevage encore mal déterminées et de confirmer les hypothèses de facteurs de risque de la mortalité des jeunes.

Sheeppox is now enzootic in Morocco. The development of a reliable method for rapid diagnosis of the disease is a central part of any control strategy. The aim of this study is to determine the diagnostic value of a variety of clinical samples such as ovine nasal, ocular or rectal swabs for the detection of sheeppox virus (SPPV) by qualitative conventional polymerase chain reaction (PCR), using a single pair of primers targeting the inverted terminal repeats of the SPPV InS-1 strain, a virulent field isolate. Swab and blood samples were collected from forty animals naturally infected with SPPV who had clinical signs of sheeppox. All animals tested PCR-positive for SPPV. Positive results were obtained infrequently with blood samples, whereas swab samples from at least two sites (nasal, ocular, rectal) were positive per evaluated animal. These results indicate that swab samples are suitable for quantitative molecular SPPV diagnosis. PCR product sequences obtained from all types of sheep samples proved to be identical to the corresponding regions of sheeppox virus strain Romania 65.

Capripoxviruses have the potential to cause outbreaks with a severe socio-economic impact. The latter, combined with an altered virus dissemination pattern, warrants its status as an important emerging disease. Disease control or eradication programmes can only be applied successfully if the necessary diagnostic tools are available allowing clear and unequivocal identification of the pathogen. Real-time PCR combines high sensitivity/specificity with a reduced analysis time and is thus a proven useful tool for identification of many pathogens, including Capripoxviruses. In order for a real-time PCR to be used in a diagnostic capacity, the different analytical and diagnostic parameters need to be evaluated to assure data quality. The implementation of parallel testing using multiple real-time PCRs with similar characteristics can improve further Capripoxvirus diagnosis. It was therefore the purpose of this study to develop a triplet real-time PCR panel with similar high sensitivity/specificity and provide sufficient validation data regarding the performance characteristics that the panel can be used in parallel, depending on the purpose and local situation.

Background Sheeppoxvirus (SPPV) is a member of the Capripoxvirus genus of the Poxviridae family, which causes significant economic losses in Morocco. The resurgence of the sheeppox disease during 2010 was characterized by an emergence of a classical nodular form for the first time in Morocco. However, little is known about the virus strain responsible for nodular form. In this study, thirty three sheep, from the eastern region of Morocco, clinically infected were examined and dead animals were autopsied. A rapid diagnostic assay for SPPV using different type of clinical samples would be useful for outbreak management. The aim of this work was to isolate the virus strain responsible for nodular form and we identified and compared by phylogenetic analysis the field strain with Moroccan vaccine strain targeting the thymidine kinase (TK) gene and the chemokine analogue receptor of interleukin (IL8) gene. Further, it was important to investigate and validate a real-time PCR using different clinical and post-mortem samples to manage epidemic sheeppox disease. Results The nodular form of sheeppox disease observed in Morocco was clinically characterized by fever, depression, lacrimation, diarrhea in lambs and nodule. At necropsy, the most affected organ was the lung. The etiological strain was successfully isolated from lung nodule in a dead lamb and was identified by using real-time PCR that has been tested and validated on different types of clinical and post mortem samples from naturally infected animals. Sequence and phylogenetic analysis of TK and IL8 gene showed that there was a very close relationship between field and vaccine strain. They were clustered within other SPPV strains. Conclusion In the current study, we show for the first time the nodular form of sheeppox in Morocco. We demonstrate a robust real-time PCR-based diagnostic assay to detect the sheeppox virus in multiple sample that can be implemented to efficiently manage the disease outbreak. Our study also offers the prospect for future molecular studies to understand the clinical forms.