Publications by authors named "Harrak M"

59 Publications

Development and Preliminary Evaluation of a New Real-Time RT-PCR Assay For Detection of Peste des petits Ruminants Virus Genome.

Transbound Emerg Dis 2015 Jun 19;62(3):332-8. Epub 2013 Jul 19.

Istituto G. Caporale, Teramo, Italy.

A duplex real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed for a simple and rapid diagnosis of Peste des petits ruminants (PPR). qRT-PCR primers and TaqMan probe were designed on a conserved region of nucleocapsid protein (Np) of PPR virus (PPRV) genome. An in vitro transcript of the target region was constructed and tested to determine analytical sensitivity. Commercial heterologous Armored RNA(®) was used as an internal positive control (IPC) for either RNA isolation or RT-PCR steps. The detection limit of the newly designed duplex real-time RT-PCR (qRT-PCR PPR_Np) was approximately 20 copies/μl with a 95% probability. No amplification signals were recorded when the qRT-PCR PPR_Np was applied to viruses closely related or clinically similar to PPRV- or to PPR-negative blood samples. A preliminary evaluation of the diagnostic performance was carried out by testing a group of 43 clinical specimens collected from distinct geographic areas of Africa and Middle East. qRT-PCR PPR_Np showed higher sensitivity than the conventional gel-based RT-PCR assays, which have been used as reference standards. Internal positive control made it possible to identify the occurrence of 5 false-negative results caused by the amplification failure, thus improving the accuracy of PPRV detection.
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http://dx.doi.org/10.1111/tbed.12117DOI Listing
June 2015

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

[Epidemiological aspects of West Nile virus infection in Morocco].

Med Sante Trop 2012 Apr-Jun;22(2):123-5

Inspection du Service de Santé Militaire, Forces Armées Royales, Rabat, Maroc.

Morocco has undergone three outbreaks of West Nile fever. The first, in 1996, began with a case in a horse herd in Benslimane (central region) and spread to neighboring cities (in the central and northwestern regions). The next two outbreaks appeared after identical 7-year epidemiological latency periods (in 2003 and 2010) in the same area and season. The only human case, which was fatal, occurred during the 1996 outbreak. The West Nile virus strains circulating in Morocco are closely related to other western Mediterranean strains and are characterized by high virulence in horses.
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http://dx.doi.org/10.1684/mst.2012.0045DOI Listing
April 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

[Seroprevalence of pandemic influenza A (H1N1)pdm09 in two regions in Morocco following the 2010-2011 influenza season].

Pathol Biol (Paris) 2013 Apr 22;61(2):83-6. Epub 2012 Jun 22.

Laboratoire de recherche et de biosécurité, faculté de médecine et de pharmacie, université Mohammed V-Souissi, hôpital Militaire d'instruction Mohammed V, Hay Ryad, Rabat, Maroc.

Purposes: To study and to compare the prevalence of infection with the virus A(H1N1)pdm 2009 in the population of two regions of Morocco compared to preexisting antibody levels.

Patients And Methods: A total of 300 and 200 serum samples were collected in the region of Rabat and Meknes respectively. Samples were collected during March to April 2011. One hundred and fifty sera, collected in 2007 from blood donors, were recovered from the blood center. The research for antibodies to influenza A(H1N1)pdm09 was performed by hemagglutination inhibition assay.

Results: The overall prevalence of antibodies inhibiting hemagglutination at the Rabat region (67%) is significantly higher than that of Meknes (53%) while the rate of cross-reactive antibodies was 7.3%. The subjects under 25 years from the Rabat region have infection rates as high with an odds ratio of 2.45. Individuals with comorbidities have the lowest prevalence with an odds ratio of 0.61. The rate of influenza A(H1N1)pdm09 vaccination in the Rabat region is 7%.

Conclusions: Immunization rates of the Moroccan population will prevent the occurrence of large outbreaks in the year 2011 to 2012 but the persistence of a naive population justifies the continuation of vaccination against A(H1N1)pdm09.
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http://dx.doi.org/10.1016/j.patbio.2012.05.004DOI Listing
April 2013

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

Waterborne Exophiala species causing disease in cold-blooded animals.

Persoonia 2011 Dec 16;27:46-72. Epub 2011 Nov 16.

CBS-KNAW Fungal Biodiversity Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands; corresponding author e-mail: .

The majority of mesophilic waterborne species of the black yeast genus Exophiala (Chaetothyriales) belong to a single clade judging from SSU rDNA data. Most taxa are also found to cause cutaneous or disseminated infections in cold-blooded, water animals, occasionally reaching epidemic proportions. Hosts are mainly fish, frogs, toads, turtles or crabs, all sharing smooth, moist or mucous skins and waterborne or amphibian lifestyles; occasionally superficial infections in humans are noted. Cold-blooded animals with strictly terrestrial life styles, such as reptiles and birds are missing. It is concluded that animals with moist skins, i.e. those being waterborne and those possessing sweat glands, are more susceptible to black yeast infection. Melanin and the ability to assimilate alkylbenzenes are purported general virulence factors. Thermotolerance influences the choice of host. Exophiala species in ocean water mostly have maximum growth temperatures below 30 °C, whereas those able to grow until 33(-36) °C are found in shallow waters and occasionally on humans. Tissue responses vary with the phylogenetic position of the host, the lower animals showing poor granulome formation. Species circumscriptions have been determined by multilocus analyses involving partial ITS, TEF1, BT2 and ACT1.
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http://dx.doi.org/10.3767/003158511X614258DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3251318PMC
December 2011

Emerging viral diseases in dromedary camels in the Southern Morocco.

Transbound Emerg Dis 2012 Apr 3;59(2):177-82. Epub 2012 Jan 3.

Hôpital Militaire d'Instruction Med V, Service de virologie, Rabat, Maroc.

During the last fifteen years, new viral diseases such Bluetongue (BT), West Nile (WN), African horse sickness (AHS), Epizootic hemorrhagic disease (EHD) and Peste des Petits Ruminants (PPR) have extended their geographic distribution and emerged in North Africa and in Europe. Camel (Camelus dromedarius) is considered as a potential vector in the transmission of some of these diseases while it is host-specific for Camelpox (CP). A serological survey was conducted on 1392 sera to estimate changes of these diseases prevalence in camels over two different time spans (2003 and 2009) and across different sites of South Morocco. Results indicate clearly that BT was circulating in camels before the first notified outbreak in Morocco (2004) with 42% positive sera in Guelmim in 2003. BT and WN prevalence's increased over time from 11 to 25% and from 10 to 13% respectively. Higher prevalence of both diseases was found in camels living in oases indicating an increased intensity of viral circulation in these sites. Camels have been tested negative for AHS, EHD and PPR viruses while higher CP prevalence has been detected in camels living in Smara, the most closed site to eastern borders (i.e. Mauritania). The seroprevalence of CP in camels at interval of 7 years increases from 23% to 37%. This survey could provide information on the possible use of camel as a sentinel animal for surveillance of emerging diseases such BT and WN.
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http://dx.doi.org/10.1111/j.1865-1682.2011.01282.xDOI Listing
April 2012

Asian lineage of peste des petits ruminants virus, Africa.

Emerg Infect Dis 2011 Jul;17(7):1223-31

Control of Exotic and Emerging Animal Diseases, Montpellier, France.

Interest in peste des petits ruminants virus (PPRV) has been stimulated by recent changes in its host and geographic distribution. For this study, biological specimens were collected from camels, sheep, and goats clinically suspected of having PPRV infection in Sudan during 2000-2009 and from sheep soon after the first reported outbreaks in Morocco in 2008. Reverse transcription PCR analysis confirmed the wide distribution of PPRV throughout Sudan and spread of the virus in Morocco. Molecular typing of 32 samples positive for PPRV provided strong evidence of the introduction and broad spread of Asian lineage IV. This lineage was defined further by 2 subclusters; one consisted of camel and goat isolates and some of the sheep isolates, while the other contained only sheep isolates, a finding with suggests a genetic bias according to the host. This study provides evidence of the recent spread of PPRV lineage IV in Africa.
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http://dx.doi.org/10.3201/eid1707.101216DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3381390PMC
July 2011

Pathogenicity of two recent Western Mediterranean West Nile virus isolates in a wild bird species indigenous to Southern Europe: the red-legged partridge.

Vet Res 2011 Jan 18;42:11. Epub 2011 Jan 18.

Centro de Investigación en Sanidad Animal del Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (CISA-INIA), Ctra Algete-El Casar, s/n, 28130 Valdeolmos (Madrid), Spain.

West Nile virus (WNV) is an emerging zoonotic pathogen whose geographic spread and incidence in humans, horses and birds has increased significantly in recent years. WNV has long been considered a mild pathogen causing self-limiting outbreaks. This notion has changed as WNV is causing large epidemics with a high impact on human and animal health. This has been particularly noteworthy since its introduction into North America in 1999. There, native bird species have been shown to be highly susceptible to WNV infection and disease with high mortalities. For this reason, the effect of WNV infection in North American bird species has been thoroughly studied by means of experimental inoculations in controlled trials. To a lesser extent, European wild birds have been shown to be affected clinically by WNV infection. Yet experimental studies on European wild bird species are lacking. The red-legged partridge (Alectoris rufa) is a gallinaceous bird indigenous to the Iberian Peninsula, widely distributed in South Western Europe. It plays a key role in the Mediterranean ecosystem and constitutes an economically important game species. As such it is raised intensively in outdoor facilities. In this work, red-legged partridges were experimentally infected with two recent WNV isolates from the Western Mediterranean area: Morocco/2003 and Spain/2007. All inoculated birds became viremic and showed clinical disease, with mortality rates of 70% and 30%, respectively. These results show that Western Mediterranean WNV variants can be pathogenic for some European bird species, such as the red-legged partridge.
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http://dx.doi.org/10.1186/1297-9716-42-11DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3037891PMC
January 2011

Use of the Capripoxvirus homologue of Vaccinia virus 30 kDa RNA polymerase subunit (RPO30) gene as a novel diagnostic and genotyping target: development of a classical PCR method to differentiate Goat poxvirus from Sheep poxvirus.

Vet Microbiol 2011 Apr 17;149(1-2):30-9. Epub 2010 Oct 17.

Animal Production and Health Laboratory, FAO/IAEA Agriculture & Biotechnology Laboratory, IAEA Laboratories Seibersdorf, International Atomic Energy Agency, Wagramer Strasse 5, P.O. Box 100, A1400 Vienna, Austria.

Sheep poxvirus (SPPV), Goat poxvirus (GTPV) and Lumpy skin disease virus (LSDV) are Capripoxviruses (CaPVs) responsible for causing severe poxvirus disease in sheep, goats and cattle, respectively. Serological differentiation of CaPVs is not possible and strain identification has relied on the implicitly accepted hypothesis that the viruses show well defined host specificity. However, it is now known that cross infections can occur and authentication of identity based on the host animal species from which the strain was first isolated, is not valid and should be replaced with molecular techniques to allow unequivocal strain differentiation. To identify a diagnostic target for strain genotyping, the CaPV homologue of the Vaccinia virus E4L gene which encodes the 30 kDa DNA-dependent RNA polymerase subunit, RPO30 was analyzed. Forty-six isolates from different hosts and geographical origins were included. Most CaPVs fit into one of the three different groups according to their host origins: the SPPV, the GTPV and the LSDV group. A unique 21-nucleotide deletion was found in all SPPV isolates which was exploited to develop a RPO30-based classical PCR test to differentiate SPPV from GTPV that will allow rapid differential diagnosis of disease during CaPV outbreaks in small ruminants.
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http://dx.doi.org/10.1016/j.vetmic.2010.09.038DOI Listing
April 2011

Midge-transmitted bluetongue in domestic dogs.

Epidemiol Infect 2011 Sep 3;139(9):1396-400. Epub 2010 Nov 3.

Institute for Animal Health, Pirbright, Woking, UK.

The role of domestic dogs in the long-distance spread of bluetongue virus (BTV) remains unproven. It is currently known that dogs are capable of being infected with BTV, can mount an antibody response to the virus and in some cases die showing severe clinical signs of disease. Infection of dogs is currently thought to be by oral ingestion of infected meat or meat products rather than through vector feeding. In this study we show that a high percentage of domestic dogs in Morocco (21%) were seropositive for BTV and, as these dogs were fed tinned commercial food only, and had no access to other meat products, the most likely source of infection was through Culicoides midges. This finding increases the chances of dogs being infected with BTV during an outbreak but their role in the onward transmission of BTV remains unproven.
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http://dx.doi.org/10.1017/S0950268810002396DOI Listing
September 2011

RT-PCR assays for seven serotypes of epizootic haemorrhagic disease virus & their use to type strains from the Mediterranean region and North America.

PLoS One 2010 Sep 17;5(9). Epub 2010 Sep 17.

Vector Borne Diseases Programme, Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, UK.

Epizootic haemorrhagic disease virus (EHDV) infects wild ruminants, causing a frequently fatal haemorrhagic disease. However, it can also cause bluetongue-like disease in cattle, involving significant levels of morbidity and mortality, highlighting a need for more rapid and reliable diagnostic assays. EHDV outer-capsid protein VP2 (encoded by genome-segment 2 [Seg-2]) is highly variable and represents the primary target for neutralising antibodies generated by the mammalian host. Consequently VP2 is also the primary determinant of virus "serotype", as identified in virus neutralisation tests (VNT). Although previous reports have indicated eight to ten EHDV serotypes, recent serological comparisons and molecular analyses of Seg-2 indicate only seven EHDV "types". Oligonucleotide primers were developed targeting Seg-2, for use in conventional RT-PCR assays to detect and identify these seven types. These assays, which are more rapid and sensitive, still show complete agreement with VNT and were used to identify recent EHDV isolates from the Mediterranean region and North America.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0012782PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2941451PMC
September 2010

Experimental infection of camels with bluetongue virus.

Res Vet Sci 2011 Jun 10;90(3):533-5. Epub 2010 Aug 10.

Institute for Animal Health, Ash Road, Pirbright, Woking, GU24 0NF, UK.

Three camels aged 4-5 years were experimentally infected with Bluetongue virus serotype 1 (BTV-1) and were observed for 75 days. No clinical signs of disease were observed throughout the experiment, however all three animals seroconverted and developed BTV-1 specific neutralising antibodies after challenge. All three camels developed a viraemia from 7 days post infection albeit at a lower level than that usually observed in experimental infections of sheep and cattle. Virus was isolated from the blood of all three animals suggesting that camels may act as a reservoir for BTV and play an important role in its transmission.
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http://dx.doi.org/10.1016/j.rvsc.2010.07.013DOI Listing
June 2011

Spectrum of clinically relevant Exophiala species in the United States.

J Clin Microbiol 2007 Nov 27;45(11):3713-20. Epub 2007 Jun 27.

Centraalbureau voor Schimmelcultures, NL-3508 AD, Utrecht, The Netherlands.

Numerous members of the genus Exophiala are potential agents of human and animal mycoses. The majority of these infections are cutaneous and superficial, but also fatal systemic infections are known. We re-identified 188 clinical isolates from the United States, which had a preliminary morphological identification of Exophiala species, by sequencing internal transcribed spacer (ITS) region of the rRNA. Molecular identifications of the strains were as follows, in order of frequency: 55 E. dermatitidis (29.3%), 37 E. xenobiotica (19.7%), 35 E. oligosperma (18.6%), 13 E. lecanii-corni (6.9%), 12 E. phaeomuriformis (6.4%), 7 E. jeanselmei (3.7%), 7 E. bergeri (3.7%), 6 E. mesophila (3.2%), 5 E. spinifera (2.7%), 3 Exophiala sp. 1 (1.6%), 3 E. attenuata (1.6%), 3 Phialophora europaea (1.3%), 1 E. heteromorpha (0.5%), and 1 Exophiala sp. 2 (0.5%) strains. Exophiala strains were repeatedly isolated from deep infections (39.9%) involving lung, pleural fluid, sputum, digestive organs (stomach, intestines, bile), heart, brain, spleen, bone marrow, blood, dialysis fluid, lymph node, joint, breast, middle ear, throat, and intraocular tissues. About 38.3% of the Exophiala spp. strains were agents of cutaneous infections including skin, mucous membranes, nail, and corneal epithelium lesions. The other strains caused superficial infections (0.5%, including hair) or subcutaneous infection (12.0%, including paranasal sinusitis, mycetoma, and subcutaneous cyst). The systemic infections were preponderantly caused by E. dermatitidis, E. oligosperma, E. phaeomuriformis, E. xenobiotica, and E. lecanii-corni. Strains of E. bergeri, E. spinifera, E. jeanselmei, E. mesophila, and E. attenuata mainly induced cutaneous and subcutaneous infections. Since relatively few unknown ITS motifs were encountered, we suppose that the list of opportunistic Exophiala species in temperate climates is nearing completion, but a number of species still have to be described.
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http://dx.doi.org/10.1128/JCM.02012-06DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2168524PMC
November 2007

Molecular epidemiology of bluetongue virus serotype 4 isolated in the Mediterranean Basin between 1979 and 2004.

Virus Res 2007 May 5;125(2):191-7. Epub 2007 Feb 5.

UMR 1161 AFSSA-ENVA-INRA, 7 Av. Général De Gaulle, 94704 Maisons-Alfort, France.

The nucleotide sequences of genome segments 2, 7, 8, 9 and 10, coding for viral proteins (VP) and non-structural proteins (NS)--VP2, VP7, NS2, VP6 and NS3/NS3A, respectively, were determined and compared for 10 strains of bluetongue virus (BTV) serotype 4 isolated in the Mediterranean Basin between 1979 and 2004, and the South African attenuated BTV 4 vaccine strain. The sequence data generated for the BTV 4 strains isolated in Greece in 1979, 1999 and 2000 showed that they had a common origin but were distinct from the lineage of the BTV 4 strains isolated from 2003 onward in the western Mediterranean Basin (Italy, Morocco, Spain and Corsica). The nucleotide and deduced amino acid (aa) sequences of the BTV 4 strains within each lineage were identical to each other, irrespective of the year of isolation or the geographical location. Although the sequence of VP2 from the Turkish and Greek strains were highly similar, there were sufficient differences in the VP6, VP7 and NS2 proteins to suggest that the Turkish BTV 4 belongs to a third lineage. Alignment of the NS3 sequences from the attenuated BTV 4 vaccine strain and the field strains showed 13 aa substitutions, which may, either singularly or together, be responsible for attenuation and hence determining the virulence of the virus.
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http://dx.doi.org/10.1016/j.virusres.2007.01.002DOI Listing
May 2007

Exophiala xenobiotica sp. nov., an opportunistic black yeast inhabiting environments rich in hydrocarbons.

Antonie Van Leeuwenhoek 2006 Oct 2;90(3):257-68. Epub 2006 Aug 2.

Centraalbureau voor Schimmelcultures, P.O. Box 85167, NL-3508, ADUtrecht, The Netherlands.

A new black yeast species, Exophiala xenobiotica, is described, a segregant of the Exophiala jeanselmei complex. It is morphologically very similar to E. jeanselmei, though with less melanized conidiogenous cells, but deviates unambiguously on the basis of molecular phylogeny. The species is a relatively common agent of cutaneous infections in humans, whereas E. jeanselmei is associated with subcutaneous infections. Environmental strains of E. xenobiotica are frequently found in habitats rich in monoaromatic hydrocarbons and alkanes.
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http://dx.doi.org/10.1007/s10482-006-9080-zDOI Listing
October 2006

West Nile virus in Morocco, 2003.

Emerg Infect Dis 2005 Feb;11(2):306-9

Centre de Référence des Arbovirus, Institut Pasteur, 21 Ave Tony Garnier, 69365 Lyon cedex 07, France.

West Nile virus (WNV) reemerged in Morocco in September 2003, causing an equine outbreak. A WNV strain isolated from a brain biopsy was completely sequenced. On the basis of phylogenetic analyses, Moroccan WNV strains isolated during the 1996 and 2003 outbreaks were closely related to other strains responsible for equine outbreaks in the western Mediterranean basin.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3320441PMC
http://dx.doi.org/10.3201/eid1102.040817DOI Listing
February 2005

[Use of the immunoenzyme test ELISA-NS3 to distinguish horses infected by African horsesickness virus from vaccinated horses].

Rev Sci Tech 1999 Dec;18(3):618-26

Institut agronomique et vétérinaire Hassan II, Département de microbiologie, immunologie et maladies contagieuses, Rabat-Instituts, Maroc.

A vaccination protocol involving three horses, with five repeated injections of inactivated serotype 4 African horse sickness virus, was undertaken to determine a possible threshold for the appearance of antibodies against the non-structural protein NS3. Using an indirect enzyme-linked immunosorbent assay, with the recombinant NS3 protein as an antigen, the authors detected a response to NS3 as of the second injection for the first horse and after four injections for the second horse. No response to NS3 was detected for the third horse. The results show that the inactivated vaccine is insufficiently purified to eliminate the non-structural protein NS3. Therefore using the NS3 protein as a marker did not enable differentiation between vaccinated and infected horses.
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December 1999

Use of reverse transcriptase-polymerase chain reaction (RT-PCR) and dot-blot hybridisation for the detection and identification of African horse sickness virus nucleic acids.

Arch Virol Suppl 1998 ;14:317-27

Centre National d'Etudes Veterinaires et Alimentaires, Laboratoire Central de Recherches Veterinaires, Maisons-Alfort, France.

A coupled reverse transcriptase-polymerase chain reaction assay (RT-PCR) for the detection of African horse sickness virus (AHSV) dsRNA, has been developed using genome segment 7 as the target template for primers. RNA from isolates of all nine AHSV serotypes were readily detected. The potential inhibitory effects of either ethylene diamine tetra acetic acid (EDTA) or heparin on the RT-PCR were eliminated by washing blood samples before lysis of the red blood cells and storage. There was a close agreement in the sensitivity and the specificity of the RT-PCR and an indirect sandwich ELISA. Confirmation of the presence of AHSV using RT-PCR and dot-blot hybridization on blood samples collected from horses experimentally infected with AHSV serotype 4 (AHSV 4) and AHSV serotype 9 (AHSV 9), was achieved within 24 hours, compared to the period of several days required for virus isolation. The RT-PCR and virus isolation methods showed similar levels of sensitivity when used for the detection of AHSV in 3 horses infected with AHSV 4, and in 2 out of 3 horses infected with a less virulent isolate of AHSV 9. Although viraemia was detected in the third horse by virus isolation, from 6 to 14 days after infection, this animal remained consistently negative by RT-PCR. Conversely, AHSV viral RNA was detected by RT-PCR in the blood of 4 donkeys and 4 mules up to 55 days after their experimental infection despite the absence of any detectable infectious virus. RT-PCR is a sensitive and rapid method for detecting AHSV nucleic acids during either the incubation period at the start of an African horse sickness (AHS) epizootic, or for epidemiological investigations in species where clinical signs may be inapparent.
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http://dx.doi.org/10.1007/978-3-7091-6823-3_28DOI Listing
November 1998

Application of an indirect fluorescent antibody assay for the detection of African horse sickness virus antibodies.

Arch Virol Suppl 1998 ;14:305-10

Laboratoire Régional d'Analyses et de Recherches Vétérinaires de Casablanca, Morocco.

An indirect fluorescent antibody (IFA) technique was used to screen and quantify antibodies against African horse sickness virus (AHSV) in equine sera. Results obtained with the IFA assay were compared directly with those obtained with standard complement fixation (CF) and virus neutralisation (VN) tests using horse sera from experimental studies and samples from the field. Positive fluorescent antibody titres were detected from as early as 7 days after primary vaccination and persisted for at least six months. The IFA technique offers a clear advantage over CF tests, where the antibodies are often of shorter duration and where sera from donkeys and mules are frequently anticomplementary. The sensitivity and specificity of the IFA test compared with the VN test were 98% and 83.3%, respectively. The IFA test is rapid, relatively easy to perform and inexpensive, and can be recommended as an alternative assay for screening different species of equidae in AHSV control and surveillance programmes.
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http://dx.doi.org/10.1007/978-3-7091-6823-3_26DOI Listing
November 1998

Clinical, virological and immune responses of normal and immunosuppressed donkeys (Equus asinus africanus) after inoculation with African horse sickness virus.

Arch Virol Suppl 1998 ;14:49-56

Département de Microbiologie, Immunologie et Maladies Contagieuses, Institut Agronomique et Vétérinaire Hassañ, Rabat-Instituts, Morocco.

To elucidate the role that donkeys may play in African horse sickness virus (AHSV) persistence during inter-epizootic periods we looked for clinical signs of infection and studied the viraemia and neutralising antibody kinetics in 3 immunocompetent and 3 immunosuppressed donkeys inoculated with AHSV-4. None of the donkeys developed signs of AHS. However infectious AHSV was isolated from the blood of the immunocompetent donkeys for up to 17 days post infection (dpi) and viral antigens were detected for up to 28 dpi. Immune cells also increased significantly from 35 to 60 dpi. There was no evidence of a recrudescence of viraemia following immunosuppression of these donkeys at 90 dpi despite a decrease in the numbers of immune cells. Infectious virus was not isolated from the blood of donkeys that had been immunosuppressed, prior to AHSV inoculation. However viral antigens were detected for up to 35 dpi. The titres of AHSV-specific neutralising antibodies and the number of immune cells were also significantly lower than in immunocompetent animals. Our findings suggest that donkeys may be able to play a role in the epidemiology of AHS but the ability of vectors to become infected by feeding upon viraemic donkeys needs to be assessed before the significance of that role can be fully understood.
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http://dx.doi.org/10.1007/978-3-7091-6823-3_6DOI Listing
November 1998

Serological and virological responses in mules and donkeys following inoculation with African horse sickness virus serotype 4.

Arch Virol Suppl 1998 ;14:29-36

Laboratoire Régional d'Analyses et de Recherches Vétérinaires, Casablanca, Morocco.

Two groups, comprising 4 donkeys and 4 mules (group 1) and 4 donkeys and 3 mules (group 2), were used to determine the duration of viraemia and to monitor the development of antibodies following inoculation with African horse sickness virus (AHSV). One group of animals was given a single dose of attenuated AHSV serotype 4 (AHSV 4) vaccine. The second group was inoculated with a virulent field strain of AHSV 4. Both groups were subsequently challenged with the virulent field strain of AHSV 4, 51 and 58 days, respectively, after their primary inoculation. Blood and serum samples, collected on alternate days after the primary inoculations and also after subsequent challenge, were assayed for virus and antibodies. Seven of the 8 AHSV vaccinated (group 1) and 7 of the 7 AHSV inoculated (group 2) animals showed humoral antibody responses after primary inoculation. Although no infectious virus could be isolated from either group for the duration of the study, reverse transcription-PCR data obtained for the second group did show the presence of AHSV viral RNA from as early as day 5 in mules and day 9 in donkeys after the primary inoculation. Viral RNA was detected consistently up to day 47 in some animals and intermittently thereafter. There was no evidence of a second viraemia in any of the animals after challenge. The detection of specific antibodies, against AHSV 4 NS3 protein, in all animals confirmed that both donkeys and mules were infected and that the virus had replicated.
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http://dx.doi.org/10.1007/978-3-7091-6823-3_4DOI Listing
November 1998

[Isolation and identification of camel poxvirus in Morocco].

Ann Rech Vet 1991 ;22(1):95-8

Biopharma, Km2, Rabat, Akkari, Maroc.

The virus isolated from young dromedaries during a poxvirus infection, was cultivated on Vero cells. The infection of egg chorio-allantoic membrane caused pustulous lesions (pocks). When inoculated in newborn mice, adult mice or guinea pigs the virus had no pathogenic effect. The virus presented all the characteristics of a poxvirus when observed under the electronic microscope.
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July 1991

[Experimental study of postvaccinal immunity to sheep-pox].

Ann Rech Vet 1984 ;15(1):59-64

Two live virus sheep-pox vaccines have been prepared on lamb testicular cells from strains Perego and RM65. Inocuity and activity have been compared to those of a commercial vaccine on a small number of lambs. Immunogenicity has been studied through neutralizing and precipitating antibodies, and through challenge resistance (strain Held), using a protective index. Results show that Perego vaccine titrated at 10(4) DICT50 gives a reliable immunity and does not provoke severe reaction post-inoculation. RM65 vaccine of which protection index is lower, only avoid infection to generalize. Although some correlation exists between seroneutralizing titers and protective index, virulent challenge appears and seems to give better data with regards to the level of protection conferred.
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November 1984
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