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Accurately identifying evolutionarily significant units (ESUs) is crucial for conservation planning, especially for species like pangolins threatened by overhunting and habitat loss. ESUs help categorize different pangolin populations, aiding in understanding their genetic diversity and distribution, which is vital for targeted conservation efforts. This research generated mitochondrial genomes from historical museum specimens of Sunda pangolins (Manis javanica) from underrepresented locations, uncovering a new evolutionary lineage from the Mentawai Islands that diverged from Indochina and west Sundaland populations around 760 000 years ago. This population thereby represents a divergent ESU with a small distribution, important for conservation planning. The novel sequences provide resources for forensic labs tracing the origin of confiscated scales and shed light into the potential distribution of the 'mysterious pangolin'. Additionally, this research confirmed the presence of the two major M. javanica lineages in Java and extended the known distribution of the eastern clade to Bali and East Kalimantan. Our findings potentially suggest a recent bottleneck and postglacial expansion of pangolins across Indochina and west Sundaland. Further investigation with genomic and morphological evidence, contact area sampling and type sequencing will be required to evaluate the taxonomic status of different M. javanica lineages and M. culionensis.
Two different forms of Chinese pangolins can be recognized according to the color of their scales, i.e., brown and dusky. We analyzed mitochondrial DNA (mtDNA) purified from the livers of seven dusky and six brown Chinese pangolins from the same locality, using cleavage patterns from 19 restriction enzymes. From the 19 6-bp recognition enzymes used, 51-56 sites were observed. By combining the cleavage patterns for each enzyme, the 13 samples were classified into four restriction types: two in dusky and two in brown Chinese pangolins. The estimated number of nucleotide substitutions per site in dusky and brown types is 0.002, and that between dusky and brown types is 0.012. Divergence between brown and dusky forms began 0.6 Myr ago, provided the mean rate of sequence divergence is 0.02 per Myr in mtDNA. Our results suggest that there is considerable divergence in Chinese pangolins, and brown and dusky Chinese pangolins may be quite different forms or, at least, belong to different maternal groups.
The distribution and relative frequency of six kinds of endocrine cells in the stomach of the Malayan pangolin, Manis javanica were studied immunohistochemically using the avidin-biotin-peroxidase complex method. The stomach of the pangolin has three regions of mucous gland, one oxyntic gland and one pyloric gland. Cells immunoreactive for chromogranin, serotonin, somatostatin, BPP and glucagon were detected in all of the gastric glands, while gastrin-immunoreactive cells were found in the entire gastric gland except for the oxyntic gland. The distribution pattern of endocrine cells in the mucous gland and pyloric gland was mainly from the middle to apical portions of the glands. The endocrine cells were rare or not detected in the basal portion of all of the mucous glands and pyloric gland, but they were also found in the basal portion of the oxyntic gland. The distribution pattern of the endocrine cells in the mucous and pyloric glands suggested that this position facilitates a quick response to the luminal ingesta. The wide distribution of gastrin-immunoreactive cells in all of the mucous glands and pyloric gland was the most remarkable finding. This distribution suggests a major function of gastrin-immunoreactive cells for the digestive process in the Malayan pangolin stomach.
Body temperatures and rates of O2 consumption and CO2 production were measured in four Chinese pangolins (Manis pentadactyla) during short-term exposures (2-4 h) to ambient temperatures (Ta) of 10-34 degrees C. At Ta less than 27 degrees C the pangolins curled into a sphere. At Ta greater than 28 degrees C the animals laid on their backs with their soft abdominal skin exposed. Rectal temperatures between 33.4 and 35.5 degrees C were recorded from animals exposed to Ta of 10-32 degrees C. At Ta greater than or equal to 32 degrees C the animals appeared to be markedly heat stressed, rate of breathing was elevated, and core temperature rose somewhat. Resting metabolic rates averaged 3.06 ml O2 X kg-1 X min-1. This is significantly lower than would be predicted from the relationship between body mass and metabolic rate established by Kleiber (The Fire of Life: an Introduction to Animal Energetics. New York: Wiley, 1975) for other eutherian mammals. The magnitude of the metabolic response to Ta below the lower critical temperature was inversely correlated to the mass of the pangolin, the slope being greatest for the smallest animals. Respiratory quotients of 0.85-1.0 were observed.
Autoradiography was used to investigate the optic system of the Chinese pangolin, Manis pentadactyla. The pattern of retinal projections in the Chinese pangolin is similar to that described in other mammals. Each retina projects bilaterally to the suprachiasmatic nucleus, dorsal and ventral lateral geniculate nuclei, pretectal area, and superior colliculus (SC). Only contralateral projections are found to the medial, lateral, and dorsal accessory optic nuclei. The large lateral nucleus receives a dense projection from the retina and forms a compact mass on the dorsolateral area of the cerebral peduncle. The lamination of the SC could not be clearly demonstrated in the brain of the Chinese pangolin.
We examined protein polymorphism of Chinese pangolins (Manis pentadactyla) from Yunnan Province of China, including two forms of three brown and nine dusky Chinese pangolins. Sixty-two genetic loci were screened; 12 loci were found to be polymorphic. The percentage of polymorphic loci (P) is 0.194, the mean individual heterozygosity (H) is 0.078, and the mean number of alleles (A) is 1.258. Furthermore, we calculated the genetic distance (D) between the two forms and found a low level of genetic divergence (D = 0.0206) between them, which indicates an almost-indistinguishable divergence at the level of proteins.
We measured O2 equilibria of adult blood and of 'stripped' (cofactor-free) hemolysates from adult and newborn Manis pentadactyla, in order to assess the implications of the burrowing habit and the low deep-core temperature in pangolins, and to discern the mechanisms for maternal-fetal O2 transfer. At pH 7.4 and body temperature (33 degrees C) the blood O2 affinity was significantly higher than in similarly sized non-burrowing, 'normothermic' mammals (P50 = 21 and 33 mm Hg, respectively) indicating an adaptation to hypoxic burrow conditions. This difference is not attributable to a higher intrinsic O2 affinity in the pangolin Hb or to significant differences in its sensitivity to temperature and erythrocytic 2,3 diphosphoglycerate (DPG), but tallies with lower DPG/Hb ratios than generally found in mammals. Stripped adult and newborn hemolysates show similar O2 affinities and pH and DPG sensitivities, but reveal a specific adult Hb that develops after birth, in sharp contrast with the ontogenetic changes in other mammals where specific fetal Hbs are lost after birth.
The cephalic arterial pattern in edentates and pangolins is described on the basis of 9 corrosion specimens, representing all the classical superfamilies, with special reference to their phylogenetic relationship and taxonomy. In this respect, the importance of the manner in which the external carotid artery system annexes the stepedial area of supply and of the course of the internal carotid artery in relation to the tympanic cavity is emphasized. The investigation does not indicate any special relationship between the New World edentates and the Old World pangolins, whereas the marked difference in the course of the internal carotid artery in recent edentates stresses the independent development of the South American anteaters compared with that of the two other edentate groups (armadillos and tree sloths). Most probably the edentates were divided very early into two main lines which have evolved independently since the early Tertiary, i.e. one for the anteaters and one for the tree sloths and armadillos, indicating a probable subdivision of the true edentates into two suborders. This subdivision is markedly different from the classical two-fold division of the edentates.
The configurations and volumes of the cerebellar nuclei of left and right 10 sides of 5 cases of the pangolins (Manis pentadactyla) were examined with sagittal myelin sheath and toluidine blue stained serial sections and reconstruction models based upon these serial sections respectively. The cerebellar nuclei of the pangolins, same as in other mammals can be divided into four nuclei, nucleus medialis (M), nucleus interpositus posterior (P), nucleus interpositus anterior (A) and nucleus lateralis (L). In all cases from medially to laterally, M, P, A and L appear in order and disappear M, A, P and L in order respectively. The volume of each nucleus in the total volume of the cerebellar nuclei is: M; 5.3-7.9% P; 27.1-31.2% A; 17.6-22.9% L; 42.4-46.2% In right and left each cerebellar nuclei significant difference is not recognized as p is less than 0.05. The posterior protuberance of the nucleus medialis protrudes remarkably in 4 cases of No. 1, No. 2, No. 3 and No. 4 but in only 1 case of No. 5 it protrudes slightly. In nucleus interpositus posterior the ventrolateral protuberance protrudes slightly in 4 cases of No. 1, No. 3, No. 4 and No. 5 but in only 1 case of No. 2 it protrudes remarkably. The anterior protuberance protrudes remarkably in all cases and the superior protuberance protrudes remarkably in 3 cases of No. 2, No. 3 and No. 4 but in 2 cases of No. 1 and No. 5 it protrudes slightly. In sulci, sulcus b and sulcus a' are distinct in all cases and sulcus a, sulcus c, sulcus b' and sulcus c' are considerably remarkable. In the toluidine blue stained serial sections, the nucleus medialis is composed of close small nerve cells, nucleus interpositus posterior is composed of the diffuse medium-sized nerve cells, nucleus interpositus anterior is composed of the close medium-sized nerve cells and nucleus lateralis is composed of the diffuse large nerve cells. In projection pictures of each subnuclei to the cerebellar cortex in each directions in the dorsal view the nucleus medialis is projected to the medial part of the folium vermis, the medial part of the nucleus interpositus posterior to the lateral part of the folium vermis and partly caudolateral part of the declive, and the lateral part of it to the crus II of the lobulus ansiformis.(ABSTRACT TRUNCATED AT 400 WORDS)
Blood and bone marrow cells of pangolins have been examined histochemically. Sudanophilia, PAS positivity and acid phosphatase and alkaline phosphatase reactivity were confined to cells of the granulocytic and monocytic series, while peroxidase reactivity was confined to cells of the erythroid series. In this latter respect the pangolin is unique among mammals so far studied.
Leipernema leiperi n.g., n.sp. is described from the pangolin Manis pentadactyla in India. Both parasitic free-living generations have been examined. Features which differentiate the parasite from Strongyloides and Parastrongyloides are described.
Members of the Wnt/Wingless (Wg) family of signalling proteins organize many aspects of animal development by regulating the expression of particular target genes in responding cells. Recent biochemical studies indicate that the vertebrate HMG-domain proteins Lef-1 and XTcf-3 can physically interact with beta-catenin, a homologue of Drosophila Armadillo (Arm), the most downstream component known in the Wnt signal transduction pathway. However, these studies do not address whether the endogenous Lef/Tcf family members are required in vivo to transduce Wnt signals. Using genetic methods in Drosophila, we define a new segment polarity gene, pangolin (pan), and show that its product is required in vivo for Wg signal transduction in embryos and in developing adult tissues. In addition, we show that pan encodes a Lef/Tcf homologue and provide evidence that its protein product binds to the beta-catenin homologue Armadillo in vivo. Finally, we demonstrate that Pan functions downstream of Arm to transduce the Wg signal. Thus, our results indicate that Pan is an essential component of the Wg transduction pathway and suggest that it acts directly to regulate gene transcription in response to Wg signalling.
Thymic ectopies are under study in 45 Pangolin's thyroids (Manis tricuspis Rafinesque). Their frequency seems to be independent of the sex of the animal but this frequency also appears to be in relation to the age of the animal. The topographic and morphological studies suggest a close relationship between these inclusions made out of thymic tissue and the presence of parathyroid islets included into the thyroid capsule. Some pictures, showing a connection between parathyroid cells and thymis parenchym elements, plead in favour of a functional interelation between these different structures as the Mc Manus experiments suggest it.
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