Publications of Mutien-Marie Garigliany
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See detailSARS-COV-2 Natural Transmission from Human to Cat, Belgium, March 2020
Garigliany, Mutien-Marie ULiege; Van Laere, Anne-Sophie ULiege; Clercx, Cécile ULiege et al

in Emerging Infectious Diseases (2020), 26(12), 3069-3071

In March 2020, a severe respiratory syndrome developed in a cat, 1 week after its owner received positive test results for severe acute respiratory syndrome coronavirus 2. Viral RNA was detected in the ... [more ▼]

In March 2020, a severe respiratory syndrome developed in a cat, 1 week after its owner received positive test results for severe acute respiratory syndrome coronavirus 2. Viral RNA was detected in the cat’s nasopharyngeal swab samples and vomitus or feces; immunoglobulin against the virus was found in convalescent-phase serum. Human-to-cat transmission is suspected. [less ▲]

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See detailAspergillus qPCR testing on nasal swab: A useful tool for diagnosis and follow‐up of sinonasal aspergillosis in dogs?
Biénès, Tom ULiege; Fastrès, Aline ULiege; Vangrinsven, Emilie ULiege et al

in Journal of Veterinary Internal Medicine (2020, October 17), 34(6), 3058-3166

Polymerase chain reaction (PCR) testing either for Aspergillus.spp or for Aspergillus fumigatus is now available; however, the interest of such tests in the diagnosis of canine sinonasal aspergillosis ... [more ▼]

Polymerase chain reaction (PCR) testing either for Aspergillus.spp or for Aspergillus fumigatus is now available; however, the interest of such tests in the diagnosis of canine sinonasal aspergillosis (SNA) has not yet been assessed. The aim of this study was to evaluate the presence of fungal material using qPCR targeting Aspergillus.spp (PanAsp) and A. fumigatus (Aspfum) in samples obtained from nasal cavities of dogs with various nasal diseases and healthy dogs. In SNA dogs, Aspfum and PanAsp were positive in 13/20 and 14/20 dogs with a mean cycle threshold (Ct) of 30.6 [range 23,2 ‐ 33,3] and 28.3 [24,3 ‐ 34,5], respectively. The PanAsp was also positive in 3 non‐SNA dogs: one with cured SNA, one with LPR and one with nasal tumor, but at very low load (Ct>33). Results between both qPCR were highly correlated (r = 0.8, P < 0.01). For Aspfum and PanAsp, the sensitivity was 65% and 70% and the specificity was 100% and 94%, respectively. Aspfum qPCR test on deep blinded nasal swabs appears highly specific but only moderately sensitive to diagnose canine SNA. In some dogs fungal plaques are exclusively found in the frontal sinus and are probably not reached by blinded sampling. Since A. fumigatus is the most common etiological agent of canine SNA (96.7% of isolates), Aspfum testing appears appropriate; however, PanAsp testing is a non‐negligible tool to detect the small percentage of SNA cases related to other Aspergillus species. Results also show that healthy predisposed dogs do not seem to be carriers and confirm that A. fumigatus does not appear to have a major role in LPR. The negative results found in cured SNA dogs show a good correlation with clinical and rhinoscopic findings. In conclusion, Aspfum and/or PanAsp (qPCR testing) on deep nasal blinded swabs can be useful for the detection of SNA at diagnosis and after cure. [less ▲]

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See detailAnalysis of the lung microbiota in dogs with Bordetella bronchiseptica infection and correlation with culture and quantitative polymerase chain reaction
Fastrès, Aline ULiege; Canonne-Guibert, Morgane ULiege; Taminiau, Bernard ULiege et al

in BMC Veterinary Research (2020), 51(46), 1-12

Infection with Bordetella bronchiseptica (Bb), a pathogen involved in canine infectious respiratory disease complex, can be confirmed using culture or qPCR. Studies about the canine lung microbiota (LM ... [more ▼]

Infection with Bordetella bronchiseptica (Bb), a pathogen involved in canine infectious respiratory disease complex, can be confirmed using culture or qPCR. Studies about the canine lung microbiota (LM) are recent, sparse, and only one paper has been published in canine lung infection. In this study, we aimed to compare the LM between Bb infected and healthy dogs, and to correlate sequencing with culture and qPCR results. Twenty Bb infected dogs diagnosed either by qPCR and/or culture and 4 healthy dogs were included. qPCR for Mycoplasma cynos (Mc) were also available in 18 diseased and all healthy dogs. Sequencing results, obtained from bronchoalveolar lavage fluid after DNA extraction, PCR targeting the V1–V3 region of the 16S rDNA and sequencing, showed the presence of Bbin all diseased dogs, about half being co‑infected with Mc. In diseased compared with healthy dogs, the β‑diversity changed (P=0.0024); bacterial richness and α‑diversity were lower (P=0.012 and 0.0061), and bacterial load higher (P= 0.004). Bb qPCR classes and culture results correlated with the abundance of Bb (r = 0.71, P < 0.001 and r = 0.70, P= 0.0022). Mc qPCR classes also correlated with the abundance of Mc (r = 0.73, P < 0.001). Bb infection induced lung dysbiosis, characterized by high bacterial load, low richness and diversity and increased abundance of Bb, compared with healthy dogs. Sequencing results highly correlate with qPCR and culture results showing that sequencing can be reliable to identify microorganisms involved in lung infectious diseases. [less ▲]

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See detailUsutu Virus Epizootic in Belgium in 2017 and 2018: Evidence of Virus Endemization and Ongoing Introduction Events.
Benzarti, Emna ULiege; Sarlet, Michaël ULiege; Franssen, Mathieu ULiege et al

in Vector Borne and Zoonotic Diseases (2020)

Wildlife surveillance allowed the monitoring of the zoonotic mosquito-borne Usutu virus (USUV) in birds and bats (Pipistrellus pipistrellus) in southern Belgium in 2017 and 2018. USUV-RNA was detected in ... [more ▼]

Wildlife surveillance allowed the monitoring of the zoonotic mosquito-borne Usutu virus (USUV) in birds and bats (Pipistrellus pipistrellus) in southern Belgium in 2017 and 2018. USUV-RNA was detected in 69 birds (of 253) from 15 species, among which 7 species had not previously been reported to be susceptible to the infection. Similarly, 2 bats (of 10) were detected positive by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). USUV-associated lesions were mainly found in Eurasian Blackbirds (Turdus merula), in which USUV antigens were demonstrated by immunohistochemistry in the brain, heart, liver, kidney, intestine, and lung. Partial nonstructural protein 5 gene-based phylogenetic analysis showed several identical or closely related strains from 2016, 2017, and 2018 clustering together within Europe 3 or Africa 3 lineages. Further, one USUV strain detected in a common chaffinch (Fringilla coelebs) manifested a close genetic relationship with the European 1 strains circulating in Hungary and Austria. Our data provide evidence of USUV endemization in southern Belgium in local birds and bats, extension of the host range of the virus and ongoing virus introduction from abroad, likely by migratory birds. Our results highlight the need for vigilance in the forthcoming years toward new virus-associated outbreaks in birds and possible human infections in Belgium. [less ▲]

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See detailAn expert opinion assessment of blood-feeding arthropods based on their capacity to transmit African swine fever virus in Metropolitan France.
Saegerman, Claude ULiege; Bonnet, Sarah; Bouhsira, Emilie et al

in Transboundary and emerging diseases (2020)

To deal with the limited literature data on the vectorial capacity of blood-feeding arthropods (BFAs) and their role in the transmission of African swine fever virus (ASFV) in Metropolitan France, a ... [more ▼]

To deal with the limited literature data on the vectorial capacity of blood-feeding arthropods (BFAs) and their role in the transmission of African swine fever virus (ASFV) in Metropolitan France, a dedicated working group of the French Agency for Food, Environmental and Occupational Health & Safety performed an expert knowledge elicitation. In total, 15 different BFAs were selected as potential vectors by the ad hoc working group involved. Ten criteria were considered to define the vectorial capacity: vectorial competence, current abundance, expected temporal abundance, spatial distribution, longevity, biting rate, active dispersal capacity, trophic preferences for Suidae, probability of contact with domestic pigs and probability of contact with wild boar. Fourteen experts participated to the elicitation. For each BFA, experts proposed a score (between 0 and 3) for each of the above criteria with an index of uncertainty (between 1 and 4). Overall, all experts gave a weight for all criteria (by distributing 100 marbles). A global weighted sum of score per BFA was calculated permitting to rank the different BFAs in decreasing order. Finally, a regression tree analysis was used to group those BFAs with comparable likelihood to play a role in ASF transmission. Out of the ten considered criteria, the experts indicated vectorial competence, abundance and biting rate as the most important criteria. In the context of Metropolitan France, the stable fly (Stomoxys calcitrans) was ranked as the most probable BFA to be a vector of ASFV, followed by lice (Haematopinus suis), mosquitoes (Aedes, Culex and Anopheles), Culicoides and Tabanidea. Since scientific knowledge on their vectorial competence for ASF is scarce and associated uncertainty on expert elicitation moderate to high, more studies are however requested to investigate the potential vector role of these BFAs could have in ASFV spread, starting with Stomoxys calcitrans. [less ▲]

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See detailMechanical transmission of African swine fever virus by Stomoxys calcitrans: Insights from a mechanistic model
Vergne, T.; Andraud, M.; Bonnet, S. et al

in Transboundary and Emerging Diseases (2020)

African swine fever (ASF) represents a global threat with huge economic consequences for the swine industry. Even though direct contact is likely to be the main transmission route from infected to ... [more ▼]

African swine fever (ASF) represents a global threat with huge economic consequences for the swine industry. Even though direct contact is likely to be the main transmission route from infected to susceptible hosts, recent epidemiological investigations have raised questions regarding the role of haematophagous arthropods, in particular the stable fly (Stomoxys calcitrans). In this study, we developed a mechanistic vector-borne transmission model for ASF virus (ASFV) within an outdoor domestic pig farm in order to assess the relative contribution of stable flies to the spread of the virus. The model was fitted to the ecology of the vector, its blood-feeding behaviour and pig-to-pig transmission dynamic. Model outputs suggested that in a context of low abundance (<5 flies per pig), stable flies would play a minor role in the spread of ASFV, as they are expected to be responsible for around 10% of transmission events. However, with abundances of 20 and 50 stable flies per pig, the vector-borne transmission would likely be responsible for almost 30% and 50% of transmission events, respectively. In these situations, time to reach a pig mortality of 10% would be reduced by around 26% and 40%, respectively. The sensitivity analysis emphasized that the expected relative contribution of stable flies was strongly dependent on the volume of blood they regurgitated and the infectious dose for pigs. This study identified crucial knowledge gaps that need to be filled in order to assess more precisely the potential contribution of stable flies to the spread of ASFV, including a quantitative description of the populations of haematophagous arthropods that could be found in pig farms, a better understanding of blood-feeding behaviours of stable flies and the quantification of the probability that stable flies partially fed with infectious blood transmit the virus to a susceptible pig during a subsequent blood-feeding attempt. © 2020 Wiley-VCH GmbH [less ▲]

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See detailAnalysis of the lung microbiota in dogs with Bordetella bronchiseptica infection and correlation with culture and quantitative polymerase chain reaction.
Fastrès, Aline ULiege; Canonne-Guibert, Morgane ULiege; Taminiau, Bernard ULiege et al

Poster (2019, November 08)

Infection with Bordetella bronchiseptica (Bb) can be confirmed using bacterial culture or quantitative polymerase chain reaction (qPCR). The lung microbiota (LM), which has been described in healthy ... [more ▼]

Infection with Bordetella bronchiseptica (Bb) can be confirmed using bacterial culture or quantitative polymerase chain reaction (qPCR). The lung microbiota (LM), which has been described in healthy experimental dogs, has not yet been studied in dogs with lower respiratory infection. In the present study we aimed to analyze the LM in dogs with Bb infection compared with healthy dogs, and to correlate the 16S rDNA amplicon sequencing results with culture and qPCR results. Twenty dogs with a diagnosis of Bb infection obtained either by qPCR and/or culture and 4 healthy dogs were included. 16S rDNA sequences were obtained from naïve bronchoalveolar lavage fluid (BALF) after DNA extraction, PCR targeting the V1-V3 region of the 16S rDNA and sequencing. Sequencing results showed the presence of Bb in all diseased dogs. About half of the dogs were co-infected, the majority with M. cynos. In diseased dogs, a shift in the β-diversity of the LM was observed (P=0.002); the richness and the α-diversity were significantly lower (P=0.012 and 0.006) and the bacterial load higher (P=0.004) compared with healthy dogs. Bb qPCR level and culture results were positively correlated with the relative abundance of Bb species after sequencing (r= 0.56, P=0.028 and r=0.70, P=0.002). Bb induced a major dysbiosis of the LM, characterized by high bacterial load, low richness and diversity and increased abundance of Bb, in comparison with healthy dogs. Results of the LM analysis highly correlate with results obtained by qPCR and culture and show that results of LM can be reliable for identification of potentially causal bacterial microorganism involved in lung infectious diseases. [less ▲]

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See detailNewly emerged African swine fever virus strain Belgium/Etalle/wb/2018: Complete genomic sequence and comparative analysis with reference p72 genotype II strains
Gilliaux, Gautier ULiege; Garigliany, Mutien-Marie ULiege; Licoppe, Alain ULiege et al

in Transboundary and Emerging Diseases (2019), 66(6), 2566-2591

In a new example of pathogens hopscotching the globe, African swine fever virus hit north‐western Europe's wildlife in summer 2018, marking a further spread of a disease that had invaded Central and ... [more ▼]

In a new example of pathogens hopscotching the globe, African swine fever virus hit north‐western Europe's wildlife in summer 2018, marking a further spread of a disease that had invaded Central and Eastern Europe recently. The complete genomic sequence of the Belgium/Etalle/wb/2018 virus is reported, with the hope it will provide a valuable tool for tracing geographical spread and biologic evolution of the virus. [less ▲]

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See detailRe-emergence of canine distemper in wildlife in Belgium
Garigliany, Mutien-Marie ULiege; Sarlet, Michaël ULiege; Franssen, Mathieu ULiege et al

in Veterinary Record: Journal of the British Veterinary Association (2018), 182(15), 439

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See detailA case of splenic myelolipoma in a German Shepherd dog
Dernier, Adrienne ULiege; Bayrou, Calixte ULiege; Desmecht, Daniel ULiege et al

Poster (2017, October 13)

An older neutered female German Shepherd dog was referred to our Department for necropsic evaluation after euthanasia for chronic renal insufficiency. Apart from chronic renal lesions and gastrointestinal ... [more ▼]

An older neutered female German Shepherd dog was referred to our Department for necropsic evaluation after euthanasia for chronic renal insufficiency. Apart from chronic renal lesions and gastrointestinal ulcerations, the macroscopic examination showed multiple off-white multifocal splenic nodules. Histologic examination revealed multiple, well-demarcated, unencapsulated masses of hematopoietic elements admixed with mature adipose tissue, consistent with a diagnosis of myelolipoma. This benign tumor is rarely reported in dogs in the literature and is more often observed in the form of a single encapsulated nodule. The other possible sites where these tumors can be seen are adrenal glands and organs retaining hematopoietic activity. Although several hypotheses have been proposed, the exact origin of myelolipoma still remain unknown [less ▲]

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See detailFeline panleukopenia virus in cerebral neurons of young and adult cats.
Garigliany, Mutien-Marie ULiege; Gilliaux, Gautier ULiege; Jolly, Sandra et al

in BMC Veterinary Research (2016), 12(1), 28

BACKGROUND: Perinatal infections with feline panleukopenia virus (FPV) have long been known to be associated with cerebellar hypoplasia in kittens due to productive infection of dividing neuroblasts. FPV ... [more ▼]

BACKGROUND: Perinatal infections with feline panleukopenia virus (FPV) have long been known to be associated with cerebellar hypoplasia in kittens due to productive infection of dividing neuroblasts. FPV, like other parvoviruses, requires dividing cells to replicate which explains the usual tropism of the virus for the digestive tract, lymphoid tissues and bone marrow in older animals. RESULTS: In this study, the necropsy and histopathological analyses of a series of 28 cats which died from parvovirus infection in 2013 were performed. Infections were confirmed by real time PCR and immunohistochemistry in several organs. Strikingly, while none of these cats showed cerebellar atrophy or cerebellar positive immunostaining, some of them, including one adult, showed a bright positive immunostaining for viral antigens in cerebral neurons (diencephalon). Furthermore, infected neurons were negative by immunostaining for p27(Kip1), a cell cycle regulatory protein, while neighboring, uninfected, neurons were positive, suggesting a possible re-entry of infected neurons into the mitotic cycle. Next-Generation Sequencing and PCR analyses showed that the virus infecting cat brains was FPV and presented a unique substitution in NS1 protein sequence. Given the role played by this protein in the control of cell cycle and apoptosis in other parvoviral species, it is tempting to hypothesize that a cause-to-effect between this NS1 mutation and the capacity of this FPV strain to infect neurons in adult cats might exist. CONCLUSIONS: This study provides the first evidence of infection of cerebral neurons by feline panleukopenia virus in cats, including an adult. A possible re-entry into the cell cycle by infected neurons has been observed. A mutation in the NS1 protein sequence of the FPV strain involved could be related to its unusual cellular tropism. Further research is needed to clarify this point. [less ▲]

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See detailReconstruction of the Evolutionary History and Dispersal of Usutu Virus, a Neglected Emerging Arbovirus in Europe and Africa.
Engel, Dimitri; Jost, Hanna; Wink, Michael et al

in MBio (2016), 7(1),

Usutu virus (USUV), one of the most neglected Old World encephalitic flaviviruses, causes epizootics among wild and captive birds and sporadic infection in humans. The dynamics of USUV spread and ... [more ▼]

Usutu virus (USUV), one of the most neglected Old World encephalitic flaviviruses, causes epizootics among wild and captive birds and sporadic infection in humans. The dynamics of USUV spread and evolution in its natural hosts are unknown. Here, we present the phylogeny and evolutionary history of all available USUV strains, including 77 newly sequenced complete genomes from a variety of host species at a temporal and spatial scaled resolution. The results showed that USUV can be classified into six distinct lineages and that the most recent common ancestor of the recent European epizootics emerged in Africa at least 500 years ago. We demonstrated that USUV was introduced regularly from Africa into Europe in the last 50 years, and the genetic diversity of European lineages is shaped primarily by in situ evolution, while the African lineages have been driven by extensive gene flow. Most of the amino acid changes are deleterious polymorphisms removed by purifying selection, with adaptive evolution restricted to the NS5 gene and several others evolving under episodic directional selection, indicating that the ecological or immunological factors were mostly the key determinants of USUV dispersal and outbreaks. Host-specific mutations have been detected, while the host transition analysis identified mosquitoes as the most likely origin of the common ancestor and birds as the source of the recent European USUV lineages. Our results suggest that the major migratory bird flyways could predict the continental and intercontinental dispersal patterns of USUV and that migratory birds might act as potential long-distance dispersal vehicles. IMPORTANCE: Usutu virus (USUV), a mosquito-borne flavivirus of the Japanese encephalitis virus antigenic group, caused massive bird die-offs, mostly in Europe. There is increasing evidence that USUV appears to be pathogenic for humans, becoming a potential public health problem. The emergence of USUV in Europe allows us to understand how an arbovirus spreads, adapts, and evolves in a naive environment. Thus, understanding the epidemiological and evolutionary processes that contribute to the emergence, maintenance, and further spread of viral diseases is the sine qua non to develop and implement surveillance strategies for their control. In this work, we performed an expansive phylogeographic and evolutionary analysis of USUV using all published sequences and those generated during this study. Subsequently, we described the genetic traits, reconstructed the potential pattern of geographic spread between continents/countries of the identified viral lineages and the drivers of viral migration, and traced the origin of outbreaks and transition events between different hosts. [less ▲]

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See detailN-acetylcysteine lacks universal inhibitory activity against influenza A viruses.
Garigliany, Mutien-Marie ULiege; Desmecht, Daniel ULiege

in Journal of Negative Results in Biomedicine (2011), 10(1), 5

ABSTRACT: N-acetylcysteine (NAC) has been recently proposed as an adjuvant therapeutic drug for influenza pneumonia in humans. This proposal is based on its ability to restrict influenza virus replication ... [more ▼]

ABSTRACT: N-acetylcysteine (NAC) has been recently proposed as an adjuvant therapeutic drug for influenza pneumonia in humans. This proposal is based on its ability to restrict influenza virus replication in vitro and to attenuate the severity of the disease in mouse models. Although available studies were made with different viruses (human and avian), published information related to the anti-influenza spectrum of NAC is scarce. In this study, we show that NAC is unable to alter the course of a fatal influenza pneumonia caused by inoculation of a murinized swine H1N1 influenza virus. NAC was indeed able to inhibit the swine virus in vitro but far less than reported for other strains. Therefore, susceptibility of influenza viruses to NAC appears to be strain-dependent, suggesting that it cannot be considered as a universal treatment for influenza pneumonia. [less ▲]

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See detailDetection of Anaplasma phagocytophilum in Dermacentor reticulatus ticks
Wirtgen, Marc ULiege; Nahayo, A.; Linden, Annick ULiege et al

in Veterinary Record: Journal of the British Veterinary Association (2011), 168(9), 248

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