The Chinese pangolin (Manis pentadactyla) is a critically endangered scale-covered mammal belonging to the order Pholidota. Wild pangolins are notably susceptible to pathogen infection and are typically characterized by impoverished health. However, little is currently known regarding the viruses prevalent among pangolins. In this study, we report the detection of two subtypes of canine parvovirus type 2 (CPV-2), namely CPV-2a and CPV-2c, both of which caused severe diarrheal disease in two post-rescue pangolins with fatal consequences. As in CPV-2-infected dogs, intensive lesion of the mucosal layer of the small intestines is a prominent feature in infected pangolins.Moreover, the immunochemistry results demonstrated that CPV-2 antigens were distributed in the crypts of small intestine. Additionally, typical parvovirus-associated CPV-2 were detected after four passages in F81 cells, and typical parvovirus-like particles, approximately 20 nm in diameter, were observed in the cell supernatants.Phylogenetic analysis revealed that the VP2 viral protein sequences (GenBank accession number OP208805) isolated from one pangolin (termed P1) were classified as CPV-2c, with 99.8% identity to a CPV-2c strain (MN832850) isolated from a Taiwanese pangolin found in Taiwan Province. In contrast, VP2 sequences (#OP208806) obtained from the second pangolin (P2) were classified as CPV-2a, with 99.8% identity to a CPV-2a strain (KY386858) isolated from southern China. In this study, we thus confirmed the infection of pangolins with CPV-2c in mainland China and demonstrated that CPV-2a also can infect pangolins. Based on these findings, we recommend that further investigations should be conducted to establish the interspecies transmission of these viruses among wild pangolins, wild carnivores, and stray dogs. K E Y W O R D S canine parvovirus type 2 VP2, Chinese pangolin, isolation and identification, Manis pentadactyla This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

The Chinese pangolin is an endangered species, and ex situ conservation and captive rescue are important conservation measures. This requires reliable information on nutritional energy requirements and expenditure characteristics. However, we lack sufficient knowledge of their energy physiology to determine their energy requirements for maintenance and growth. An open-flow respirometry system was used to measure the resting metabolic rate (RMR) and the daily energy expenditure (DEE) of Chinese pangolins (Manis pentadactyla), and the dietary digestive energy was measured. The average RMR in Chinese pangolins was 3.23 ml O2 kg−1 min−1 at an ambient temperature (Ta) of 24.5–30°C, which was only 73.0% of the expected value based on body mass (BM). The average DEE values were 744.9 kJ day−1 in animals with BM >3 kg and 597.3 kJ day−1 in those with BM <3 kg, which were only 52.4% and 60.6% of the predicted values, respectively. The RMR and DEE levels of the Chinese pangolin were lower than those of similar-sized eutherian mammals and close to those of anteaters. These characteristics suggest that the Chinese pangolin has a low demand for energy in its diet. Although metabolic level data alone cannot be used to calculate the energy requirements of each Chinese pangolin, we believe they can provide a tangible reference for the relocation of Chinese pangolins. These results provide a scientific basis for future research on the physiology and ecology of endangered wildlife such as the Chinese pangolin.

Venous blood gas analytes are commonly examined in animals, and the results may be important when evaluating the overall health status of an animal. Pangolins are critically endangered mammals, and there is limited information on their physiological reference values in the literature. The aim of this study was to analyze venous blood gas and biochemical parameters before and during isoflurane anesthesia in wild healthy Sunda and Chinese pangolins. The results obtained showed that the blood gas index trends of the two pangolin species before and after isoflurane anesthesia were the same. After anesthesia, the partial pressure of carbon dioxide (pCO2), partial pressure of oxygen (pO2), total carbon dioxide (CO2), mean blood bicarbonate (HCO3−), extracellular fluid compartment (BEecf) base excess and the mean blood glucose (Glu) levels of both pangolin species showed a significant increase compared to the pre-anesthesia period. In contrast, the mean blood potassium (K+), lactate (Lac) and mean blood pH levels were significantly lower. No significant differences in the mean blood sodium (Na+) or blood ionized calcium (iCa) levels were observed during anesthesia. This study is important for future comparisons and understanding the health status of this endangered species.

The Chinese pangolin ( Manis pentadactyla) is a critically endangered scale-covered mammal belonging to the order Pholidota. Wild pangolins are notably susceptible to pathogen infection and are typically characterized by impoverished health. However, little is currently known regarding the viruses prevalent among pangolins. In this study, we report the detection of two subtypes of canine parvovirus type 2 (CPV-2), namely, CPV-2a and CPV-2c, both of which caused severe diarrheal disease in two post-rescue pangolins with fatal consequences. As in CPV-2-infected dogs, intensive lesion of the mucosal layer of the small intestines is a prominent feature in infected pangolins. Phylogenetic analysis revealed that the VP2 viral protein sequences isolated from one pangolin were classified into the CPV-2c subclade, with 99.8% identities to a CPV-2c strain (MN832850) isolated from a Taiwanese pangolin found in Taiwan Province. In contrast, VP2 sequences obtained from the second pangolin were classified into the CPV-2a subclade, with 99.8% identities to a CPV-2a strain (KY386858) isolated from southern China. In this study, we thus confirmed the infection of pangolins with CPV-2c in mainland China and demonstrate that CPV-2a can also infect pangolins. Based on these findings, we recommend that further investigations should be conducted to establish the interspecies transmission of these viruses among wild pangolins, wild carnivores, and stray dogs.

Background: Chinese pangolin (Manis pentadactyla), one of the most endangered species in the world, has suffered a sharp decline due to heavy hunting and illegal trade. The artificial diet has difficulty satisfying the nutritional needs of captive pangolins, and it has become one of the main factors limiting the development of ex-situ conservation of pangolins. Gastrointestinal diseases are one of the most common diseases affecting the survival of captive Chinese pangolins and arecaused by an imbalance in gut microbiota. In the wild, the pangolin is a highly specialized species with a diet dominated by ants and termites and an intake ofsome termitarium soil. However, the effects of supplementation with termitarium soil on the gut microbiota of captive Chinese pangolins are poorly understood. Results: Using 16S rRNA sequencing technology, the gut microbiota of the control group (CZ), the sterilized termitarium soil group (MZ), and the unsterilized termitarium soil group (BZ) were compared. These results showed that there was no significant difference in alpha diversity, while the results of unweighted UniFrac PCoA showed obvious separation among the three groups. At the phylum level, Firmicutes, Proteobacteria, and Bacteroidetes were the main bacterial phyla. The relative abundance of Bacteroidetes was decreased in the BZ group compared to the CZ group. Clostridium_sensu_stricto_1, Streptococcus, Cellulosilyticum, and Escherichia_Shigella were the main bacterial genera at the genus level. Unsterilized termitarium soilincreased the relative abundances of Cellulosilyticum, Bacillus, and Clostridium_sensu_stricto_13 and decreased the relative abundances of Bacteroides, Clostridiales_unclassified, Ruminococcus]_gnavus_group, and Veillonella. The Lefse results showed that Cellulosilyticum, Clostridium_sensu_stricto_13, and Bacillus were enriched in the BZ group. The concentrations of acetic acid, propionic acid, and butyric acid were decreased in the BZ and MZ groups. Conclusions: These results indicated that dietary supplementation with termitarium soil did not alterthe richness of the gut microbiota, but reduced proinflammatory-related bacteria and increased intestinal absorption-related bacteria, which improves the efficiency of SCFA absorption. Collectively, this study provides a potential method for ameliorating the intestinal homeostasis of captive Chinese pangolins.