The raccoon dog (Nyctereutes procyonoides) is a typical omnivore possessing wide dietary adaptability and tolerance to rough feeding, which may be attributed to its intestinal microbiota. The study aimed to investigate the effect of dietary alfalfa meal levels on the growth performance, nutrient apparent digestibility, serum parameters, and intestinal microbiota of raccoon dogs. Sixty raccoon dogs were randomly divided into four dietary treatments containing 0% (AM0), 5% (AM5), 10% (AM10), and 15% (AM15) alfalfa meal for a 60-day experiment. The results showed that compared to raccoon dogs fed the AM0 diet, those fed the AM5 and AM10 diets had no significant difference in growth performance, while those fed the AM15 diet experienced a significant decrease. Raccoon dogs fed the AM5 diet had no significant effect on the nutrient apparent digestibility. Dietary supplementation with alfalfa meal significantly decreased serum urea levels and increased the antioxidant capacity of raccoon dogs. The intestinal microbiome analysis showed that the richness and diversity of colonic microbiota significantly increased in the AM15 group. With the increase in dietary alfalfa meal levels, the relative abundance of fiber-degrading bacteria in the colon of raccoon dogs, such as Treponema, Phascolarctobacterium, and Christensenellaceae R-7 group, increased. However, the relative abundance of pathogenic bacteria, including Anaerobiospirillum, decreased. In conclusion, the inclusion of 5% alfalfa meal in the raccoon dogs\' diet had no effect on growth performance, but it exhibited the potential to improve serum antioxidant capacity and intestinal microbiota. This indicates that raccoon dogs have a certain tolerance to the addition of alfalfa meal in their diet.

Platycladus orientalis leaves are rich in flavonoids and polysaccharides, which offer high medicinal and nutritional benefits. This study aimed to investigate the impact of P. orientalis leaf extract (PLE) on the growth performance, fur quality, serum parameters, and intestinal microbiota of raccoon dogs. Sixty healthy male black raccoon dogs, aged 85 (±5) days, were randomly assigned to four groups and fed a basal diet supplemented with 0, 0.25, 0.50, and 1.00 g/kg PLE for 125 days (designated as groups P0, P1, P2, and P3, respectively). The results revealed that the raccoon dogs in group P1 exhibited increased average daily gain and underfur length while showing a decreased feed/gain ratio compared to group P0 (p < 0.05). However, the heart index in group P2 was significantly lower than in group P0 (p < 0.05), and the kidney index and serum alanine aminotransferase activities in group P3 were higher than in groups P2 and P0 (p < 0.05), suggesting potential adverse effects at higher PLE dosages. Notably, dietary PLE supplementation led to a reduction in serum glucose concentrations (p < 0.05), which may have implications for glucose regulation. Furthermore, the study explored the impact of dietary supplementation with 0.25 g/kg PLE on the raccoon dogs\' intestinal microbiota using high-throughput sequencing. The results showed significant alterations in the microbial community structure, with a notable decrease in the abundance of Prevotella copri in response to 0.25 g/kg PLE supplementation (p < 0.05). In conclusion, supplementing raccoon dogs\' diet with 0.25 g/kg PLE can lead to improved growth performance and a positive influence on the intestinal microbiota. However, caution should be exercised regarding higher dosages, as they may have adverse effects on certain parameters. As a result, PLE holds promise as a potential feed additive for fur animal production.

Microorganisms of the genus Eperythrozoon are a zoonotic chronic infectious disease with wide distribution. We found that raccoons infected with Eperythrozoon showed obvious stunting, which seriously affected the economic benefits of raccoon dogs. To investigate the pathogenesis of the raccoon dog, we used transcriptome and proteome sequencing to analyze the changes in mRNA, miRNA, and protein expression in raccoon dogs infected with Eperythrozoon and normal raccoons. The results showed that the expression levels of genes related to immunity, metabolism, and enzyme activity were significantly changed. Among these, ERLIN1, IGF1R, CREB3L1, TNS1, TENC1, and mTOR play key roles. Additionally, the miR-1268, miR-125b, miR-10-5p, and miR-10 as central miRNAs regulate the expression of these genes. Integrated transcriptomic and proteomic analyses revealed consistent trends in mRNA and protein changes in MYH9, FKBP1A, PRKCA, and CYP11B2. These results suggest that Eperythrozoon may contribute to the slow development of raccoons by affecting the expression of mRNAs and miRNAs, reducing their immunity and causing metabolic abnormalities.

Cryptosporidium spp. are common parasitic pathogens causing diarrhea in humans and various animals. Fur animals are widely farmed in Shandong Province, China, but the prevalence and genetic identity of Cryptosporidium spp. in them are unclear. In this study, 1,211 fecal samples were collected from 602 minks, 310 raccoon dogs and 299 foxes on two farms in Shandong and analyzed for Cryptosporidium spp. by nested PCR and sequence analyses of the small subunit rRNA gene. The overall infection rate of Cryptosporidium spp. was 31.5% (381/1,211), with a higher infection rate in raccoon dogs (37.7%, 117/310) than in foxes (32.4%, 97/299) and minks (27.7%, 167/602). By age, the highest infection rates of Cryptosporidium spp. were observed in raccoon dogs of 1-2 months, minks of 5-6 months, and foxes of > 12 months. Three Cryptosporidium species and genotypes were detected, including C. canis (n = 279), C. meleagridis (n = 65) and Cryptosporidium mink genotype (n = 37). Among the three major host species, raccoon dogs were infected with C. canis only (n = 117), while foxes were infected with both C. canis (n = 32) and C. meleagridis (n = 65), and minks with C. canis (n = 130) and Cryptosporidium mink genotype (n = 37). Subtyping of C. canis by sequence analysis of the 60 kDa glycoprotein gene identified eight subtypes. They belonged to two known subtype families, XXa and XXd, and two novel subtype families XXf and XXg, with host adaptation at the subtype family level. Notably, C. canis from foxes was genetically distant from those in other hosts. Further subtyping analysis identified three subtypes (IIIeA21G2R1, IIIeA19G2R1 and IIIeA17G2R1) of C. meleagridis and two novel subtype families Xf and Xg of the Cryptosporidium mink genotype. The presence of zoonotic C. canis subtypes in raccoon dogs and C. meleagridis subtypes in foxes suggests that these fur animals might be potential reservoirs for human-pathogenic Cryptosporidium spp.

Over the last several decades, no emerging virus has had a profound impact on the world as the SARS-CoV-2 that emerged at the end of 2019 has done. To know where severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) originated from and how it jumped into human population, we immediately started a surveillance investigation in wild mammals in and around Wuhan when we determined the agent. Herein, coronaviruses were screened in the lung, liver, and intestinal tissue samples from fifteen raccoon dogs, seven Siberian weasels, three hog badgers, and three Reeves\'s muntjacs collected in Wuhan and 334 bats collected around Wuhan. Consequently, eight alphacoronaviruses were identified in raccoon dogs, while nine betacoronaviruses were found in bats. Notably, the newly discovered alphacoronaviruses shared a high whole-genome sequence similarity (97.9 per cent) with the canine coronavirus (CCoV) strain 2020/7 sampled from domestic dog in the UK. Some betacoronaviruses identified here were closely related to previously known bat SARS-CoV-related viruses sampled from Hubei province and its neighbors, while the remaining betacoronaviruses exhibited a close evolutionary relationship with SARS-CoV-related bat viruses in the RdRp gene tree and clustered together with SARS-CoV-2-related bat coronaviruses in the M, N and S gene trees, but with relatively low similarity. Additionally, these newly discovered betacoronaviruses seem unlikely to bind angiotensin-converting enzyme 2 because of the deletions in the two key regions of their receptor-binding motifs. Finally, we did not find SARS-CoV-2 or its progenitor virus in these animal samples. Due to the high circulation of CCoVs in raccoon dogs in Wuhan, more scientific efforts are warranted to better understand their diversity and evolution in China and the possibility of a potential human agent.

Paeoniae radix alba extract (PRA extract) has the functions of regulating immunity, resisting inflammation, and has antioxidant properties. However, current recommendations of dietary PRA extract levels for raccoon dogs were inadequate. The purpose of this experimental study was to gain information allowing for better estimating the effects of PRA extract on raccoon dogs, and their PRA requirements. Fifty healthy male raccoon dogs of (120 ± 5) days old were randomly divided into 5 groups (group PRA0, PRA1, PRA2, PRA4, PRA8) with 10 animals in each group and 1 in each replicate. Five kinds of experimental diets were prepared with five levels of Paeoniae radix alba extract (0, 1, 2, 4, 8 g/kg) in the basic diet. The prefeeding period was 7 days and the experimental period was 40 days. The results showed that the average daily feed intake in group PRA1 and PRA2 was significantly higher than that in other groups (P < 0.01). The dry matter excretion in group PRA8 was significantly higher than that in other groups (P < 0.01), while the dry matter digestibility and protein digestibility in group PRA8 were significantly lower than those in other groups (P < 0.01). Nitrogen retention in group PRA1 and PRA2 was significantly higher than that in group PRA8 (P < 0.05). With the increase of the content of Paeoniae radix alba extract in diet, the activity of alkaline phosphatase in group PRA2 was significantly higher than that in group PRA0 (P < 0.05); The activity of serum SOD in group PRA4 was significantly higher than that in other groups (P < 0.01). The content of serum IgA in group PRA2 was significantly higher than that in other groups (P < 0.05). The content of TNF-α in intestinal mucosa in group PRA1 and group PRA2 was significantly lower than that in group PRA0 (P < 0.05). In conclusion, we found that dietary Paeoniae radix alba extract intake significantly improved the feed intake and nitrogen deposition of Ussuri raccoon dog, increased the content of serum IgA and reduced the content of TNF-α in the small intestinal mucosa. We suggest that an estimated dietary Paeoniae radix alba extract level of 1 to 2 g/kg could be used as a guide to achieve the optimal performance of raccoon dogs.

The objective of this study was to determine the infection rate and genetic diversity of Cryptosporidium spp. in minks, foxes, and raccoon dogs, farmed in the Xinjiang Uygur Autonomous Region, Northwest China. Fresh fecal specimens were collected from individual cages of farmed minks (n = 214), blue foxes (n = 35), and raccoon dogs (n = 39) and examined using nested PCR based on the Cryptosporidium spp. small subunit rRNA gene. Cryptosporidium spp. was detected in 35 cages (12.2%, 35/288), with a higher infection rate detected in raccoon dogs (20.5%) compared with minks (12.1%) and blue foxes (2.9%). Sequence analysis showed that Cryptosporidium canis was the only species identified in blue foxes and raccoon dogs, while in the 26 Cryptosporidium-positive mink specimens, Cryptosporidium mink genotype (n = 17), C. canis (n = 7), and Cryptosporidium parvum (n = 2) were identified. Further analysis based on the 60-kDa glycoprotein (gp60) gene determined that both C. parvum isolates belonged to the subtype IIdA15G1, while eight of the 17 Cryptosporidium mink genotype isolates were a novel subtype that we have named XeA5G1. To the best of our knowledge, this is the first report of C. parvum subtype IIdA15G1 infection in minks. Since all the Cryptosporidium species/genotypes identified in minks, foxes, and raccoon dogs from Xinjiang have been previously found in humans, our results suggest that these fur animals may play a role in the transmission of zoonotic Cryptosporidium.

This study reports outbreak of a new disease caused by Staphylococcus pseudintermedius (S. pseudintermedius) in raccoon dogs. The disease occurred in a breeding farm of raccoon dogs in Guan County of Shandong Province in China in August of 2019. 47% (425/896) of the raccoon dogs showed some abnormal symptoms; 17.6% (75/425) of which had severe skin and soft tissue infections (SSTIs), dyspnoea and severe pathological lesions in lungs, livers, etc; and 4.2% (18/425) of which died within 4 weeks. The pathogen of the disease was identified as S. pseudintermedius by mass spectrometer detection, animal pathogenicity tests, microscopic examination and biochemical reaction tests. Its nucleotide homology of 16S rRNA gene was 100% with that of other published strains, and its genotype was between the American and Brazilian strains from other animals. The isolated S. pseudintermedius strain from the diseased raccoon dogs could cause ulceration and suppuration in the skins and severe pathological lesions not only in raccoon dogs, but also in mice; and it is confirmed as a methicillin-resistant S. pseudintermedius (MRSP) strain by the amplification of mecA gene; and 12 sensitive drugs were screened by drug sensitivity tests. Full attention should be paid to the great economic loss and the potential zoonotic risk caused by the S. pseudintermedius in raccoon dogs, and this study can provide a reference for the clinical diagnosis and treatment of this new disease.

The goal of this study is to compare the gut microbiota of domestic blue fox (Alopex lagopus) and raccoon dog (Nyctereutes procyonoides) to provide better understanding of their intestinal gut microbiota. We analyzed the structure of fecal microbes in 40 blue foxes and 40 raccoon dogs that were raised under same conditions, using high-throughput Illumina sequencing targeting the V3-V4 region of the 16S rRNA gene. In total, 295,146 sequence reads were obtained. The average number of operational taxonomical units in the two group samples was 194 to 286. Firmicutes (blue fox 73.40%, raccoon dog 46.90%) and Bacteroidetes (blue fox 21.92%, raccoon dog 44.25%) were the most abundant phyla in the gut of blue fox and raccoon dog. At the genus level, Prevotella (blue fox 16.89%, raccoon dog 36.22%), Blautia (blue fox 9.02%, raccoon dog 13.72%), and Peptostreptococcaeae_incertae_sedi (blue fox 22.41%, raccoon dog 2.84%) were commonly presented in the gut of two kinds of animal. Principal coordinates analysis showed that the microbial communities were different between blue fox and raccoon dog. The Firmicutes-to-Bacteroidetes ratio was higher in blue foxes (3:1) than in raccoon dogs (1:1). Moreover, Peptostreptococcaeae_incertae_sedi and Prevotella, were more abundant in the gut of blue fox, whereas the abundance of Prevotella and Blautia were higher in the gut of raccoon dog. In conclusion, the present study revealed the difference of the gut microbial composition between blue fox and raccoon dog under the same diet conditions.

The abuse of antibiotics in animal husbandry imposes a serious threat to both animal health and the environment. As a replacement for antibiotics, probiotic products have been widely used in livestock farming to promote growth of animals. However, no products specifically developed for farmed raccoon dogs and foxes are commercially available at the moment. This study was conducted to investigate the effects of mixed probiotics on farmed raccoon dogs and foxes.
Two feeding trials on farmed raccoon dogs and foxes were performed. A mixed probiotic preparation composed of Bifidobacterium bifidum, Clostridium butyricum, Bacillus subtilis and Bacillus licheniformis was fed to these two canine species in order to assess whether such a mixed probiotics can be an alternative to antibiotics (control group). The body weight of raccoon dogs exhibited an increasing tendency with mixed probiotics administration, while that of foxes did not. The serum antioxidant activity was evaluated, and a significantly increase of total antioxidative capacity (T-AOC) was observed in both species. Illumina MiSeq was used for the sequencing of 16S rRNA genes to compare the composition of fecal microbiota between the control and mixed probiotics groups. Although α-diversity did not change, β-diversity of the fecal microbiota showed a distinct dissimilarity between the control and probiotics groups of both raccoon dogs and foxes. Dietary mixed probiotics increased the abundance of the genus Bifidobacterium in the fecal samples of raccoon dogs, and the genus Bacillus in the fecal samples of foxes. The different responses of raccoon dogs and foxes to probiotics might be the result of differences in the composition of the native gut microbiota of the two species.
The mixed probiotics preparation composed of Bifidobacterium bifidum, Clostridium butyricum, Bacillus subtilis and Bacillus licheniformis could be an effective feed additive for the improvement of the health of farmed raccoon dogs, but it may not be suitable for foxes.