key: cord-103802-mygo3qx0 authors: Li, Yanpeng; Gordon, Emilia; Idle, Amanda; Altan, Eda; Seguin, M. Alexis; Estrada, Marko; Deng, Xutao; Delwart, Eric title: Multiple known and a novel parvovirus associated with an outbreak of feline diarrhea and vomiting date: 2020-03-25 journal: bioRxiv DOI: 10.1101/2020.03.24.005876 sha: doc_id: 103802 cord_uid: mygo3qx0 An unexplained outbreak of feline diarrhea and vomiting, negative for common enteric viral and bacterial pathogens, was subjected to viral metagenomics and PCR. We characterized from fecal samples the genome of a novel chapparvovirus we named fechavirus that was shed by 8/17 affected cats and different feline bocaviruses shed by 9/17 cats. Also detected were nucleic acids from attenuated vaccine viruses, members of the normal feline virome, viruses found in only one or two cases, and viruses likely derived from ingested food products. Epidemiological investigation of disease signs, time of onset, and transfers of affected cats between three facilities support a possible role for this new chapparvovirus in a highly contagious feline diarrhea and vomiting disease. Cats have an estimated world-wide population of over half a billion. Members Shelter 3 became involved in the outbreak when a cat (#160) transferred from Shelter 2 became 148 sick, several days after arrival and before the outbreak had been identified. Ultimately, a total 13 149 cats were affected in Shelter 1 in November, 17 cats were affected in Shelter 2 (November -150 January), and 13 cats were affected in Shelter 3 (November-January) (Figure 1 and 151 Supplementary Table 1 ). Nearly all transmission was indirect ( Figure 2) ; because of this, it was 152 not possible to definitively determine which animals had been exposed, except in specific rooms 153 where housing was communal or exposure was known to be widespread prior to introduction of 154 control measures. Attack rates for these rooms were 66.7% (Shelter 2) and 83.3% (Shelter 3) 155 (Table 1) . 156 Overall, diarrhea and vomiting were observed in 81.4% and 67.4% of the 43 cases. There 157 were likely more cats vomiting because vomitus that could not be attributed to a particular cat 158 was found multiple times in the final wave of illness. 25.6% and 11.6% of the affected cats also 159 showed inappetence and lethargy, and 67.4% required veterinary care ( Table 2 ). The minimum 160 incubation period was 24 hours, and the maximum was estimated at 5-7 days based on estimated 161 exposure dates. Vomiting tended to start 1-2 days before diarrhea, and last only a couple of days, 162 but in some animals the diarrhea lasted up to a week (longer in a few animals). The mean 163 duration of illness was 5.1 days and the median was 4.0 days (range 1 to 19 days) 164 which has good efficacy against bacteria and viruses (including non-enveloped 170 viruses) (Omidbakhsh & Sattar, 2006) . After control measures were initiated, the outbreak slowed 171 and cases became sporadic, except for at Shelter 3 in January (due to communal housing). In order to determine the pathogens that caused the outbreak, all 17 available fecal samples 196 were analyzed using viral metagenomics. Viral sequences assigned to five main viral families 197 (Anelloviridae, Parvoviridae, Papillomaviridae, Polyomaviridae and Caliciviridae) were 198 detected in these fecal samples (Table 3) . 199 (Table 4) . 269 Dependoparvovirus sequence reads were also found in two cats. A near full-length genome of 270 4315 bases could be assembled from cat #849 (also infected with fechavirus) whose 271 phylogenetic analysis showed it to be related to a recently reported dependoparvovirus genome 272 from bats (Desmodus rotundus) sharing NS1 and VP1 protein with 51.6% and 46.7% identities 273 (Supplementary Figure 1) . 274 The timeline of disease presentation and fecal shedding status of key animals in the 276 transmission chain between the 3 facilities was determined. There were no samples from cats in 277 Shelter 1 available for testing, but cat #688 was fechavirus positive shortly after being housed in 278 the same area as cat #853 and #614, who were sick upon transfer from Shelter 1 to Shelter 2. A 279 second cat in shelter 2 was also shedding fechavirus (cat #160). Of the other 15 affected cats in 280 Shelter 2 a total of 5 individual cats and a sample pool (mixed feces from 3 cats) were Using metagenomics, we found FeBoV1, 2, and 3 and a novel chaphamaparvovirus we named 311 fechavirus in a large fraction of fecal samples and fechavirus in all vomit samples from sick cats 312 in a multi-facility outbreak. Subsequent PCR testing confirmed the presence of either a bocavirus 313 or fechavirus or both (n=1 cat #283) of these viruses in all but one of the seventeen sick cats 314 tested (cat #178). The outbreak in shelter 2 predominantly tested positive for FeBoV (7/9 FeBoV 315 +ve and 2/9 fechavirus +ve) while the sick cats in shelter 3 were mainly shedding fechavirus (6/7 316 fechavirus +ve, 2/7 FeBov +ve). Another cat housed in shelter 2 who developed vomiting after 317 sick cats were transferred back from Shelter 3 was also shedding fechavirus as well as a novel 318 dependovirus. The co-detection of these viral genomes indicate that fechavirus may provide the 319 required help for replication-defective dependoviruses. 320 FPV and feline FCV reads were determined to originate from recently inoculated attenuated 321 vaccine strains while other viruses were deemed to be asymptomatic infections, derived from chicken and pork viruses in consumed food, or detected only in sporadic cases. FCV sequences 323 were negative by PCR, this could be the results of low viral load in those cats. 324 The close genetic relationship of fechavirus to cachavirus found in both diarrheic and healthy 325 dog feces (Fahsbender, Fechavirus + -+ -------+ + -+ + + + FeBoV -+ -+ + + --+ + -+ -----IDEXX diarrhea Note: Days (D1-D12) reflect day samples were collected. Days of illness are shaded starting at first sample collection. "+" and "-" means PCR positive or negative for the fechavirus. Illness onset was 10, 2, 2, 3 days before Day1 for cats #688, 912, 594 and 283, respectively. Figure 1 . Daily epidemic cases of the cat diarrhea outbreak in three shelters from November to January. 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