Sample strategies for the assessment of the apparent diffusion coefficient in single large intracranial space-occupying lesions of dogs and cats.

Diffusion-weighted imaging is increasingly available for brain investigation. Image interpretation of intracranial space-occupying lesions often includes the derived apparent diffusion coefficient (ADC) analysis. In human medicine, ADC can help discriminate between benign and malignant lesions in intracranial tumors. This study investigates the difference in ADC values depending on the sample strategies of image analysis. MRI examination, including diffusion-weighted images of canine and feline patients presented between 2015 and 2020, were reviewed retrospectively. Patients with single, large intracranial space-occupying lesions were included. Lesions homogeneity was subjectively scored. ADC values were calculated using six different methods of sampling (M1-M6) on the ADC map. M1 included as much as possible of the lesion on a maximum of five consecutive slices; M2 included five central and five peripheral ROIs; M3 included a single ROI on the solid part of the lesion; M4 included three central ROIs on one slice; M5 included three central ROIs on different slices; and M6 included one large ROI on the entire lesion. A total of 201 animals of various breeds, genders, and ages were analyzed. ADC values differed significantly between M5 against M2 (peripheral) (p < 0.001), M5 against M6 (p = 0.009), and M4 against M2 (peripheral) (p = 0.005). When lesions scored as homogeneous in all sequences were excluded, an additional significant difference in three further sampling methods was present (p < 0.005). ADC of single, large, intracranial space-occupying lesions differed significantly in half of the tested methods of sampling. Excluding homogeneous lesions, additional significant differences among the sampling methods were present. The obtained results should increase awareness of the variability of the ADC, depending on the sample strategies used.

Detection of SARS-CoV-2 RNA in a Zoo-Kept Red Fox (Vulpes vulpes).

Many different animal species are susceptible to SARS-CoV-2, including a few Canidae (domestic dog and raccoon dog). So far, only experimental evidence is available concerning SARS-CoV-2 infections in red foxes (Vulpes vulpes). This is the first report of SARS-CoV-2 RNA detection in a sample from a red fox. The RT-qPCR-positive fox was zoo-kept together with another fox and two bears in the Swiss Canton of Zurich. Combined material from a conjunctival and nasal swab collected for canine distemper virus diagnostics tested positive for SARS-CoV-2 RNA with Ct values of 36.9 (E gene assay) and 35.7 (RdRp gene assay). The sample was analysed for SARS-CoV-2 within a research project testing residual routine diagnostic samples from different animal species submitted between spring 2020 and December 2022 to improve knowledge on SARS-CoV-2 infections within different animal species and investigate their potential role in a One Health context. Within this project, 246 samples from 153 different animals from Swiss zoos and other wild animal species all tested SARS-CoV-2 RT-qPCR and/or serologically negative so far, except for the reported fox. The source of SARS-CoV-2 in the fox is unknown. The fox disappeared within the naturally structured enclosure, and the cadaver was not found. No further control measures were undertaken.

A Risk Factor Analysis of SARS-CoV-2 Infection in Animals in COVID-19-Affected Households.

A higher prevalence of SARS-CoV-2 infections in animals that have close contact with SARS-CoV-2-positive humans ("COVID-19 households") has been demonstrated in several countries. This prospective study aimed to determine the SARS-CoV-2 prevalence in animals from Swiss COVID-19 households and to assess the potential risk factors for infection. The study included 226 companion animals (172 cats, 76.1%; 49 dogs, 21.7%; and 5 other animals, 2.2%) from 122 COVID-19 households with 336 human household members (including 230 SARS-CoV-2-positive people). The animals were tested for viral RNA using an RT-qPCR and/or serologically for antibodies and neutralizing activity. Additionally, surface samples from animal fur and beds underwent an RT-qPCR. A questionnaire about hygiene, animal hygiene, and contact intensity was completed by the household members. A total of 49 of the 226 animals (21.7%) from 31 of the 122 households (25.4%) tested positive/questionably positive for SARS-CoV-2, including 37 of the 172 cats (21.5%) and 12 of the 49 dogs (24.5%). The surface samples tested positive significantly more often in households with SARS-CoV-2-positive animals than in households with SARS-CoV-2-negative animals (p = 0.011). Significantly more animals tested positive in the multivariable analysis for households with minors. For cats, a shorter length of outdoor access and a higher frequency of removing droppings from litterboxes were factors that were significantly associated with higher infection rates. The study emphasizes that the behavior of owners and the living conditions of animals can influence the likelihood of a SARS-CoV-2 infection in companion animals. Therefore, it is crucial to monitor the infection transmission and dynamics in animals, as well as to identify the possible risk factors for animals in infected households.

Detection and Molecular Characterization of the SARS-CoV-2 Delta Variant and the Specific Immune Response in Companion Animals in Switzerland.

In human beings, there are five reported variants of concern of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2). However, in contrast to human beings, descriptions of infections of animals with specific variants are still rare. The aim of this study is to systematically investigate SARS-CoV-2 infections in companion animals in close contact with SARS-CoV-2-positive owners ("COVID-19 households") with a focus on the Delta variant. Samples, obtained from companion animals and their owners were analyzed using a real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and next-generation sequencing (NGS). Animals were also tested for antibodies and neutralizing activity against SARS-CoV-2. Eleven cats and three dogs in nine COVID-19-positive households were RT-qPCR and/or serologically positive for the SARS-CoV-2 Delta variant. For seven animals, the genetic sequence could be determined. The animals were infected by one of the pangolin lineages B.1.617.2, AY.4, AY.43 and AY.129 and between zero and three single-nucleotide polymorphisms (SNPs) were detected between the viral genomes of animals and their owners, indicating within-household transmission between animal and owner and in multi-pet households also between the animals. NGS data identified SNPs that occur at a higher frequency in the viral sequences of companion animals than in viral sequences of humans, as well as SNPs, which were exclusively found in the animals investigated in the current study and not in their owners. In conclusion, our study is the first to describe the SARS-CoV-2 Delta variant transmission to animals in Switzerland and provides the first-ever description of Delta-variant pangolin lineages AY.129 and AY.4 in animals. Our results reinforce the need of a One Health approach in the monitoring of SARS-CoV-2 in animals.

A Pre- and Within-Pandemic Survey of SARS-CoV-2 RNA in Saliva Swabs from Stray Cats in Switzerland.

Cats have been shown to be highly susceptible to SARS-CoV-2, and transmission within the species has been demonstrated experimentally. In cats undergoing natural SARS-CoV-2 infections, human-to-animal transmission was mostly suspected. It can be postulated that, in stray cats with no or only minimal contact with humans, SARS-CoV-2 may pose a minor risk. The current study investigated the prevalence of active SARS-CoV-2 infections in Swiss stray cats using quantitative reverse transcriptase real-time polymerase chain reaction (RT-qPCR). Saliva swabs from 1405 stray cats were collected in 14 Swiss cantons. The animals were sampled between February 2019 and February 2020 (pre-COVID-19 cohort: 523 cats) and between February 2020 and August 2021 (COVID-19 cohort: 882 cats). All the samples were tested by RT-qPCR, amplifying the envelope (E) gene and, in case of positive or inconclusive results, the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2. No SARS-CoV-2 viral RNA could be detected in any of the tested saliva swab samples. Our findings support the assumption that SARS-CoV-2 infections in stray cats are not highly prevalent in Switzerland. Nevertheless, the monitoring of stray cats and other susceptible animal species is necessary, since the "One Health" approach has been recognized as being essential to successfully fight the COVID-19 pandemic.

Delivery of non-viral naked DNA vectors to liver in small weaned pigs by hydrodynamic retrograde intrabiliary injection.

Hepatic gene therapy by delivering non-integrating therapeutic vectors in newborns remains challenging due to the risk of dilution and loss of efficacy in the growing liver. Previously we reported on hepatocyte transfection in piglets by intraportal injection of naked DNA vectors. Here, we established delivery of naked DNA vectors to target periportal hepatocytes in weaned pigs by hydrodynamic retrograde intrabiliary injection (HRII). The surgical procedure involved laparotomy and transient isolation of the liver. For vector delivery, a catheter was placed within the common bile duct by enterotomy. Under optimal conditions, no histological abnormalities were observed in liver tissue upon pressurized injections. The transfection of hepatocytes in all tested liver samples was observed with vectors expressing luciferase from a liver-specific promoter. However, vector copy number and luciferase expression were low compared to hydrodynamic intraportal injection. A 10-fold higher number of vector genomes and luciferase expression was observed in pigs using a non-integrating naked DNA vector with the potential for replication. In summary, the HRII application was less efficient (i.e., lower luciferase activity and vector copy numbers) than the intraportal delivery method but was significantly less distressful for the piglets and has the potential for injection (or re-injection) of vector DNA by endoscopic retrograde cholangiopancreatography.