Hamster and ferret experimental infection with intranasal low dose of a single strain of SARS-CoV-2

Understanding the pathogenesis of the SARS-CoV-2 infection is key to developing preventive and therapeutic strategies against COVID-19, in the case of severe illness but also when the disease is mild. The use of appropriate experimental animal models remains central in the in vivo exploration of the physiopathology of infection and antiviral strategies. This study describes SARS-CoV-2 intranasal infection in ferrets and hamsters with low doses of low-passage SARS-CoV-2 clinical French isolate UCN19, describing infection levels, excretion, immune responses and pathological patterns in both animal species. Individual infection with 103 p.f.u. SARS-CoV-2 induced a more severe disease in hamsters than in ferrets. Viral RNA was detected in the lungs of hamsters but not of ferrets and in the brain (olfactory bulb and/or medulla oblongata) of both species. Overall, the clinical disease remained mild, with serological responses detected from 7 days and 10 days post-inoculation in hamsters and ferrets respectively. The virus became undetectable and pathology resolved within 14 days. The kinetics and levels of infection can be used in ferrets and hamsters as experimental models for understanding the pathogenicity of SARS-CoV-2, and testing the protective effect of drugs.

Hamster and ferret experimental infection with intranasal low dose of a single strain of SARS-CoV-2

Understanding the pathogenesis of the SARS-CoV-2 infection is key to develop preventive and therapeutic strategies against COVID-19, in the case of severe illness but also when the disease is mild. The use of appropriate experimental animal models remains central in the in-vivo exploration of the physiopathology of infection and antiviral strategies. This study describes SARS-CoV-2 intra-nasal infection in ferrets and hamsters with low doses of low-passage SARS-CoV-2 clinical French isolate UCN19, describing infection levels, excretion, immune responses and pathological patterns in both animal species. Individual infection with 103 pfu SARS-CoV-2 induced a more severe disease in hamsters than in ferrets. Viral RNA was detected in the lungs of hamsters but not of ferrets and in the brain (olfactive and/or spinal bulbs) of both species. Overall, the clinical disease remained mild, with serological responses detected from 7 days and 10 days post inoculation in hamsters and ferrets respectively. Virus became undetectable and pathology resolved within 14 days. The kinetics and levels of infection can be used in ferrets and hamsters as experimental models for understanding the pathogenicity of SARS-CoV-2, and testing the protective effect of drugs.

Genome Sequence of Fusarium graminearum Strain MDC_Fg1, Isolated from Bread Wheat Grown in France

Fusarium graminearum is a major fungal pathogen that induces Fusarium head blight (FHB), a devastating disease of small-grain cereals worldwide. This announcement provides the whole-genome sequence of a highly virulent and toxin-producing French isolate, MDC_Fg1.

First Complete Genome Sequence of a Distinct Papaya Ringspot Virus Isolate from the Northeastern Region of India

ABSTRACT This is the first report of a Papaya ringspot virus (PRSV) isolate from the northeastern region of India. The nucleotide sequence identity of PRSV-Meghalaya was in the range of 72.6 to 82.5% with other Indian PRSV isolates, and the highest identity of 84.4% was with a French isolate. Population genetic analysis indicated positive selection.

Complete Nucleotide Sequence of a French Isolate of Maize rough dwarf virus , a Fijivirus Member in the Family Reoviridae

The complete nucleotide sequence of a French isolate of Maize rough dwarf virus (MRDV) was determined by next-generation sequencing and compared with the single available complete sequence and with the partial sequences of two additional isolates available in online databases.

The development of rediae ofFasciola hepaticainRadix natalensissubjected twice to bimiracidial exposures

Experimental infections of EgyptianRadix natalensiswith a French isolate ofFasciola hepatica(each snail was subjected twice to a bimiracidial exposure) were carried out to determine how many sporocysts grew in these snails and to study the developmental patterns of redial generations. Single-sporocyst infections were found in 69.3% (34/49) of infected snails, with equivalent numbers of normal and abnormal patterns. Snails with two- and three-sporocyst infections were 24.4% and 6.1%, respectively. In single- and two-sporocyst infections at days 42 and 56 post-exposure, the total redial burden was significantly higher in snails with a normal redial development. In two- and three-sporocyst infections, the overall maturity of rediae was delayed at days 42 and 56. The high frequency of abnormal patterns inR. natalensis(53.1% of all infected snails showed degeneration of a first mother redia) might be due to incomplete adaptation between the snail population and the parasite. The delayed redial maturity in two- and three-sporocyst infections can mainly be explained by the volume of the snail body, which would be insufficient to allow the simultaneous differentiation of most rediae over time.

First Report of Tomato chlorotic dwarf viroid in Tomato in France

Tomato chlorotic dwarf viroid (TCDVd) is a pospiviroid found naturally infecting tomato (Solanum lycopersicum L.) (3) and several ornamentals such as Brugmansia, petunia (1), and trailing verbena (4). Initially identified in North America (3), it has been reported from India, Europe (the Netherlands and United Kingdom), and Japan. At the end of 2007, 20 to 25% of tomato plants within a group of greenhouses in the Brittany Region of France were observed with top bunching, leaf curling, and epinasty symptoms. Reverse transcription (RT)-PCR with a primer pair specific for several pospiviroids (5′GGGGAAACCTGGAGCGA3′ and 5′GGGGATCCCTGAAGCGC3′) amplified the correctly sized fragment (approximately 360 bp) from total nucleic acid extracts from three symptomatic plants. The sequence of the uncloned amplification product (GenBank Accession No. EU729744) was determined, together with that of five cloned cDNAs. All sequences were highly related with a total of three mutations in these six sequences and they showed 96.9% (GQ169709 and AY372399) to 99.4% (AF162131) identity with TCDVd sequences present in GenBank. Identification of TCDVd was confirmed from the same plant samples by molecular hybridization with a Potato spindle tuber viroid (PSTVd)-specific probe (which cross-hybridizes with TCDVd to a certain extent) and by PCR with the PSTVd/TCDVd-specific 2A-1S primer pair (3) and sequencing of the amplified fragment. The French isolate is most closely related to the original tomato isolate from Canada (GenBank Accession No. AF162131). In a grow-out test involving 2,500 seeds from the original seed lot from which the symptomatic plants were derived, 2 of the 250 pools of 10 plants tested positive for TCDVd infection with the 3H1-2H1 primer pair (2). The sequence of the amplified product proved identical to the isolate detected in the original greenhouse plants, indicating a low level of seed transmission. As with other pospiviroids, which appear to be more and more frequently reported in greenhouse tomatoes, possible sources of infection include contaminated seeds, as seem to be the case in this first outbreak, and also transfer to tomatoes from infected ornamental hosts. This is, to the best of our knowledge, the first report of TCDVd in tomato in France. References: (1) T. James et al. Plant Pathol. 57:400, 2008. (2) A. M. Shamloul et al. Can. J. Plant Pathol. 19:89, 1997. (3) R. P. Singh et al. J. Gen. Virol. 80:2823, 1999. 4) R. P. Singh et al. Plant Dis. 90:1457, 2006.

Foliar and Cane Rot of Arundo donax Caused by Nigrospora oryzae in Europe

A fungus was isolated consistently from dead shoot tips and flag leaves of Arundo donax L. (Poaceae) in France, Crete, Cyprus, Italy, Morocco, and Spain from April of 2003 through September of 2005. The fungus was identified as Nigrospora oryzae (Berk. & Br.) Petch (teleomorph Khuskia oryzae) on the basis of morphological characteristics (1). The mean diameter of 80 conidia obtained from sporulating plant specimens collected in France, Crete, and Cyprus were 14, 15, and 15 μm, respectively. The mean diameters of 25 conidiogenous cells and conidiophores were 7 and 4 μm, respectively. Identification was confirmed by comparing the sequence of the ribosomal DNA internal transcribed spacer 1 and 4 regions from the French isolate (GenBank Accession No. DQ219433) with the sequence of a voucher specimen from the Museum National d'Histoire Naturelle, Paris, France. The isolate of N. oryzae from France was deposited at the CBS collection in Utrecht, the Netherlands (CBS 113884). N. oryzae is known to be a weak pathogen on a wide range of plants but has not been reported on A. donax, which is now a well-established weed in the United States and North America, probably originating from the Mediterranean Region. Herein, the possible use of N. oryzae as a biological control agent was investigated. Twenty young A. donax shoots growing in the greenhouse and 20 emerging canes in the field were selected on the basis of uniformity in size. A spore suspension in distilled water adjusted to 5 × 105 conidia/ml of the French isolate was prepared and 0.5 ml was injected with a syringe just below the growing point of the flag leaf in onehalf of the greenhouse and field plants. The remaining plants were injected with 0.5 ml of distilled water as controls. Infection and death of the flag leaf occurred in 30% of the shoots in the greenhouse and 50% of the canes in the field 21 days from inoculation. No disease developed on the control plants. Greenhouse inoculation tests were repeated once. N. oryzae was reisolated from dead leaves and the terminal node of inoculated shoots, satisfying Koch's postulate. Attempts made to induce disease symptoms by applying spore suspensions on the whorl of leaves surrounding the apical tip failed. This is an indication that an insect vector may be needed to carry and deposit N. oryzae spores into the tight, whorled flag leaf for infection and disease development to occur. To our knowledge, this is the first report of foliar and cane rot of A. donax caused by N. oryzae. References: (1) K. H. Domsch et al. Page 515 in: Compendium of Soil Fungi. IHW-Verlag, Eching, Germany, 1993.

Molecular characterization of Brazilian Dicyma pulvinata isolates

Forty-nine Brazilian Dicyma pulvinata isolates were examined by morphological traits and RFLP, RAPD and AFLP analyses. This fungus is a mycoparasite of Microcyclus ulei, the causal agent of the most devastating rubber (Hevea brasiliensis) disease, known as "South American Leaf Blight" (SALB). These isolates were compared with an Indian isolate from Cercosporidium sp., and a French isolate from Cladosporium fulvum. They were also compared with Dicyma ampullifera from Papua New Guinea. The morphological parameters analyzed confirmed the identification of the Brazilian isolates. The graphic representations of the distance matrices of each molecular marker showed similar results. Dicyma pulvinata isolates from M. ulei were closely related, whereas the reference isolates examined were dispersed. Among the D. pulvinata isolates obtained from M. ulei, a significant pairwise distance was obtained, for all the molecular markers, between the isolates from the areas favorable to the occurrence of SALB (North and Northeast of Brazil) and the region of escape for the disease (Mato Grosso State).

First Report of Cucurbit yellow stunting disorder virus in Commercial Cucumber Greenhouses in France

In autumn 2001, severe yellowing symptoms were observed on greenhouse-grown cucumbers near Perpignan (southern France). Leaf samples were collected from two sites where plants displayed symptoms ranging from limited yellowing of the older leaves to severe, complete yellowing of the whole plant. Cucurbit aphid-borne yellows virus, a polerovirus that causes similar symptoms was not detected in doubleantibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using a specific antiserum. Total RNA was extracted from fresh leaf tissues and used in reverse transcription-polymerase chain reaction (1) with primers specific for two whitefly-borne viruses also inducing yellows and occurring in the Mediterranean basin (1): Beet pseudo yellows virus (BPYV, genus Closterovirus) transmitted by Trialeurodes vaporariorum (West.) and Cucurbit yellow stunting disorder virus (CYSDV, genus Crinivirus) transmitted by Bemisia tabaci (Genn.). No BPYV was detected in this survey, but CYSDV was present in all samples. In subsequent surveys conducted in the spring and summer of 2002, BPYV and CYSDV were detected, sometimes in mixed infections, in samples collected from the same region. The complete CYSDV coat protein gene was amplified by PCR using specific primers (2), yielding the expected-size fragment of 756 bp. The French isolate (GenBank Accession No. AY204220) shared 99.6 to 100% nucleotide sequence identity in the sequenced CP fragments (700 nt) with isolates of the most common, highly homogenous subgroup of CYSDV that has emerged recently in the Middle East, southwestern Europe (Spain and Portugal), United States, and Morocco (2). To our knowledge, this is the first report of CYSDV in France and it shows the threat represented by the current emergence of B. tabaci-transmitted viruses. References: (1) I. C. Livieratos et al. Plant Pathol. 47:362, 1998. (2) L. Rubio et al. J. Gen. Virol. 82:929, 2001.