Multilocus genotype and subtype analysis of Cryptosporidium andersoni derived from a Bactrian camel (Camelus bactrianus) in China

Fecal specimens from two Bactrian camels were collected in the Ya’an city zoo of China and were examined for Cryptosporidium by centrifugal flotation. One specimen was found to be parasitized by Cryptosporidium via microscopy, and the oocysts were measured to have an average size of 7.03?×?5.50 μm (n?>?50). The isolate was genotyped by polymerase chain reaction (PCR) amplification and DNA sequence analysis of the partial 18S rRNA, COWP, and A135 genes, and was confirmed to be Cryptosporidium andersoni with minor nucleotide differences. Multilocus sequence typing (MLST) analysis indicated that the subtype of the camel-derived C. andersoni isolate was A4, A4, A4, and A1 at the four minisatellite loci (MS1, MS2, MS3, and MS16, respectively). Therefore, this isolate belongs to the most common MLST subtype reported in cattle in China and is distinct from two other known camel C. andersoni MLST subtypes (A6, A4, A2, A1 and A6, A5, A2, A1). Animal transmission experiments demonstrated that the C. andersoni isolate was not infectious to immunosuppressed or immunocompetent Kun-ming mice, Sprague-Dawley rats, and hamsters but was biologically similar to most bovine C. andersoni isolates characterized so far. Therefore, transmission of this camel-derived C. andersoni isolate is very likely to occur between camels and bovine.     

Morphologic Observation of Mucosa‐Associated Lymphoid Tissue in the Large Intestine of Bactrian Camels (Camelus bactrianus)

The structure and distribution of the mucosa‐associated lymphoid tissue (MALT) throughout the large intestine of 10 Bactrian camels were comparatively studied by anatomical and histological methods. The results showed that Peyer's patches (PPs) were mainly located on the mucosal surfaces of the entire ileocecal orifice, the beginning of the cecum and the first third of the colon. The shape of PPs gradually changed from “scrotiform” to “faviform” along the large intestine with the scrotiform PP as the major type in the ileocecal orifice. The distribution density also gradually decreased from the ileocecal orifice to the colon. The histological observations further revealed that the MALT in the form of PPs or isolated lymphoid follicles (ILF) and lamina propria lymphocytes was mainly present in the lamina propria and submucosa from the entire ileocecal orifice, where the muscularis mucosa is usually incomplete, to the colonic forepart. In addition, lymphoid tissue was much more abundant in the lamina propria and submucosa of the ileocecal orifice as compared to the cecum and colon. Statistically, the MALT of the ileocecal orifice contained a higher number of lymphoid follicles (37.7/10 mm²) than that of the cecum, colon, or rectum (P < 0.05). The germinal centers of the lymphoid follicles were clearly visible. Together, our data suggest that the ileocecal orifice constitutes the main inductive site for the mucosal immunity in the large intestine of the Bactrian camel; and that scrotiform PPs are likely to the result of long‐term adaptation of the Bactrian camel to the harsh living environment. Anat Rec, 297:1292–1301, 2014. © 2014 Wiley Periodicals, Inc.     

Narasin poisoning in the dromedary camel (Camelus dromedarius)

Narasin poisoning was reported in 15 camels, 7 adults and 8 young, after accidental access to poultry feed medicated with 60 g narasin per ton. Fourteen camels died between 3 and 20 days, and one young animal survived the dose after developing a chronic course of a disease. The main clinical signs of narasin toxicity in the dromedary include: weakness of hind limbs, lack of coordination, oedema of dependent parts, inappetence, ruminal atony, myoglobinuria, profound depression, tachycardia, sternal recumbency and death. The lesions were mainly in the heart and skeletal muscles and consisted of multifocal degeneration and necrosis of heart and skeletal muscle fibres with areas of regeneration and lung oedema. There was high enzyme activity for creatine kinase (CK), lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase and an increase in urea concentration and white blood cells, neutrophil and platelet counts. Cardiac markers, troponin T, CK-MB and C-reactive protein, showed slight or no changes terminally.     

Haemostatic parameters in the camel (Camelus Dromedarius) comparison with humans

Haemostatic measurements were undertaken in 46 camels (Camelus dromedarius) and the results were compared to human reference values. The effects of sex, age and pregnancy on camel haemostatic parameters were studied. There was significant shortening of the prothrombin time and activated partial thromboplastin time and marked elevation of factor VIII clotting activity (FVIII:C), FVII and FIX in camels as compared to humans. On the other hand, FX, fibrinogen and packed cell volume levels were lower in camels than humans. These results indicate a more active state of the coagulation system in camels as compared to humans and this might be a physiological adaption that protects this animal from excessive blood or fluid loss under the conditions of drought and transhumance. Pregnancy had no effect on any measured parameters. Platelet counts and ATIII levels were higher in males than females, and both parameters, in addition to FIX, decreased with age. The lack of any fluctuations of haemostatic parameters during pregnancy in camels may be related to the placental type; camel placenta which is of the epitheliochorionic type is much less vascular and less liable to bleed than the haemochoroinic placenta of pregnant women, who show a hypercoagulable state to guard against excessive bleeding during placental separation.     

An ultrastructural investigation of the blood neutrophils in camel (Camelus dromedarius)

Evaluation of camel neutrophil's structure and ultrastructure is essential for fundamental and applied research on innate immunobiology. There is little information on camel neutrophil's ultrastrucure. Healthy camel blood neutrophils were isolated, prepared, and stained for light, scanning, transmission electron microscopy, and organelles' biometry. We observed that (1) the nuclei of healthy camel neutrophils were highly lobulated, predominantly ≥5 lobules/nucleus, with euchromatin and hetrochromatin portions and very low ratio of nucleus to cytoplasm, (2) surface of neutrophils contained many psuedopods, and (3) the cytoplasm contained enormous granules with different sizes and forms and high density at the center, as well as with many different organelles such as abundant mitochondria, rough endoplasmic reticulum, microtubules, phagolysosome, vacuoles, and Golgi apparatus. Biometric analyses of key organelles in neutrophils also shed some new lights on the features of camel neutrophils. This ultrastructural study emphasizes the notion that camel neutrophils are highly equipped with the cytoskeletal machinery for efficient organelle movement, phagocytosis, and microbicidal activities. The ultrastructural features of camel neutrophils observed in this study would be outstandingly comparable to the neutrophils of other ruminants and needs further detailed comparative study.     

A Golgi study of neurons in the camel cerebellum (Camelus dromedarius)

Neurons in the cerebellar cortex of camels were studied using modified Golgi impregnation methods. Neurons were classified according to their position, morphology of their soma, density and distribution of dendrites, and the course of their axons. Accordingly, eight types of neurons were identified. Three types were found in the molecular layer: upper and lower stellate cells and basket cells, and four types were found in the granular layer: granule cells, Golgi Type II cells, Lugaro cells, and unipolar brush cells. Only the somata of Purkinje cells were found in the Purkinje cell layer. The molecular layer is characterized by the presence of more dendrites, dendritic spines, and transverse fibers. Golgi cells also show extensive dendritic branching and spines. The results illustrate the neuronal features of the camel cerebellum as a large mammal living in harsh environmental conditions. These findings should contribute to advancing our understanding of species-comparative anatomy in achieving better coordination of motor activity.     

The spleen of the one humped camel (Camelus dromedarius) has a unique histological structure

The histology and structure of 38 spleens of the dromedary (aged 0.5–15 y) were studied in relation to age. The spleen was found to have a thick capsule (292±106 mm) divided into an outer layer (113±39 mm) composed mainly of connective tissue and an inner layer (180±81 mm) consisting mainly of smooth muscle cells. Vascular and avascular trabeculae extend from the capsule, the former containing arteries and nerves but no trabecular veins, the latter being divided structurally into primary and secondary trabeculae. Subcapsular and peritrabecular blood sinuses around primary and vascular trabeculae are unique to the camel spleen. The central artery emerges from the periarterial lymphatic sheath and branches into up to 4 penicilli which extend as sheathed arterioles (42±8 μm). These are found near or surrounded by blood sinusoids of the red pulp. A wide marginal zone surrounds the white pulp and contains sheathed arteries but no marginal sinuses. The red pulp is characteristically divided into cords by secondary trabeculae and contains venous sinusoids of different sizes. The camel spleen is of a sinusal type that can store blood. The thick muscular capsule and trabeculae pump the stored blood according to the body's need. Both closed and open circulations are found. The venous return is unique as the blood flow is from the venous sinusoids of the red pulp to the peritrabecular sinuses to the subcapsular sinuses to the splenic vein. No significant structural differences related to age were found.     

The structure and hormone content of the endocrine pancreas of the one-humped camel (Camelus dromedarius)

Little is known of the blood sugar regulation in the camel and the morphology and function of its endocrine pancreas. The present paper describes the light microscopic structure and hormone content of the endocrine pancreas of the one-humped camel. Staining of pancreatic sections with haematoxylin-eosin or aldehyde-fuchsin showed numerous islets evenly distributed in all parts of the pancreas. Immunocytochemical staining for insulin or glucagon indicated that islets were predominantly composed of centrally located B-cells, surrounded by a peripheral rim of A-cells. Corresponding stainings for somatostatin or pancreatic polypeptide (PP) demonstrated that D-cells comprised only a small part of the islet volume while PP-cells were common both within and outside the islets. There were no obvious differences between the frequency of the various islet cells in different pancreatic regions. The pancreatic hormone concentrations roughly corresponded to the frequency of the different islet cell types. Insulin appeared most abundant followed by glucagon, PP and somatostatin in decreasing order. The concentrations of each of the hormones were similar in different regions of the gland. It is concluded that the endocrine pancreas of the one-humped camel is dispersed into islets of the same size and cellular composition as has been described in many other mammalian species.     

Gametogony of Eimeria cameli in the small intestine of one-humped camel (Camelus dromedarius)

Parasitology Research - Domesticated Old World camels (Camelus dromedarius and Camelus bactrianus) are important for the economy of several countries in Asia, Africa, and the Arabian Peninsula, and...     

Electron microscopic studies on the thymus of the Arabian camel (Camelus dromedarius)

The thymus of young camels contained 2 main cell types: lymphocytes and epithelial reticular cells (ER). 3 types of ER cells were observed and described. 2 types from these cells showed secretory activities, meanwhile, the third type had well developed tonofilaments. Hassall's corpuscles were noted in the medulla and were composed of degenerative concentrically arranged ER cells. The functional morphology of the above mentioned structures was discussed.     

The oculomotor and trochlear nuclei in the one humped camel (Camelus dromedarius).

Literature on the organization of the oculomotor and trochlear nuclei of large animals is scanty. There were no reports on the organization of the oculomotor and trochlear nuclei of the camel, hence this study. Nine brains were used for the study. The brainstems were double-embedded in celloidin and paraffin and were cut serially at 24 microns and stained with toluidine blue. Light microscopic studies of the nuclei showed that the principal oculomotor nuclei were not subdivided and were composed of large multipolar nerve cell bodies that had a mean length of 30 +/- 5 microns. The nucleus was 2.4 mm long, 0.7 mm wide and 1.1 mm high. The Edinger-Westphal nucleus was small and was made up of elongated oval cell bodies that had a mean length of 33 +/- 5 microns and a mean diameter of 10 +/- 2 microns. The trochlear nucleus was located caudal to the oculomotor nucleus from which it was separated by a gap. The nerve cell bodies of the trochlear nuclei were similar to those of the oculomotor nuclei. The cell bodies had a mean length of 20 +/- 2.5 microns and a mean width of 18 +/- 3 microns. The caudal central nucleus was indistinct. It was concluded, that the oculomotor and trochlear nuclei of the camel are similar in their general organization to those of other animals but differences exist in the development and organization of the component parts.     

Ultrastructure of the intrahepatic bile duct system of the one-humped camel (Camelus dromedarius)

The ultrastructure of the intrahepatic bile duct system of the camel was studied. Bile canaliculi are present as cavities containing numerous microvilli between adjacent hepatic cells. They are lined with cuboidal cells surrounded by a basal lamina. The lateral plasma membranes of adjacent cells form extensive evaginations below tight junctions at the lumenal surface. The basal surface is open. Collagen bundles are arranged mainly in parallel to the duct.     

Immunohistochemical localization of androgen and progesterone receptors in the uterus of the camel (Camelus dromedarius)

Ten adult, cyclic female camels (Camelus dromedarius) were used to describe the distribution of androgen (AR) and progesterone (PR) receptors in the uterus using immunohistochemistry. Both AR and PR were distributed throughout the different compartments of the uterus with nuclear staining for AR and PR seen in the cells of epithelia (luminal and glandular), stroma and myometrial smooth muscles. AR immunostaining was not uniform in distribution and intensity; the surface epithelium and the glandular epithelium in the adluminal region of the endometrium showed lower AR immunoreactivity than other compartments of the uterus. PR immunostaining showed uniformity in both distribution and intensity strong PR immunostaining intensity in almost all cells of the different uterine compartments. The intensity and distribution of PR immunostaining in epithelia of lumen and glands in the adluminal regions of endometrium was higher (P < 0.05) than that of AR. In conclusion, immunohistochemical localization of AR and PR in the uterus of the cyclic dromedary camel indicates the important roles of androgen and progesterone in controlling the uterine activity during the follicular phase.     

Ultrastructural and molecular characterization of Sarcocystis isolated from camel (Camelus dromedarius) in Iran

Sarcocystis cameli was first described in one-humped camels (Camelus dromedarius), and it is the only species which have so far reported in camels. Although more than 150 species of Sarcocystis were described in various animals, only a few data on camel Sarcocystis ultrastructure were published, and this report is the first for molecular information (DNA sequence and RLFP digestion pattern). The main objective of the present work is to characterize Sarcocystis isolated from camels by electron microscopy and PCR-RFLP methods. Muscle samples were taken from the fresh esophagus, diaphragm, skeletal muscles, and heart of one-humped camels (C. dromedarius) slaughtered in abattoirs of Tehran and Ghazvin provinces, Iran. The dissection and trypsin digestion techniques were applied for the detection of the cysts. The infected samples were fixed in glutaraldehyde and/or frozen at −20°C until use for ultrastructural and molecular studies, respectively. The ultrastructural and molecular studies were carried out contemporaneously. The 18S rRNA gene of the parasites was amplified by PCR. The PCR products were cloned into a pTZ57R/T and sequenced. In addition, the PCR products were digested separately with each of the four restriction enzymes for RFLP. Our results indicated that only microcysts were observed in muscle samples. The microcysts were white, elongated, spindled, and a few spiral-shaped, with mean size 260 × 75 μm which are identical with S. cameli. The ultrastructure of microcyst wall had many non-branched finger-like protrusions irregularly folded. There was a 600-bp specific band amplified after PCR with specific primers. The molecular data for camel Sarcocystis is reported for the first time in Iran and the world.     

Normal adaptation of pulmonary arterial intima to extrauterine life in the pig: ultrastructural studies

Adaptation of the pulmonary arterial intima was studied in injected lung specimens of 34 Large White pigs. Each type of pre- and intra-acinar artery was studied separately using transmission and scanning electron microscopy. Determination of the endothelial surface/volume ratio and volume densities of (1) endothelium and subendothelium, (2) endothelial cytoplasmic organelles and (3) subendothelial connective tissue elements yielded 6832 measurements which comprised a computerized database. At birth, endothelial cell morphology changed more rapidly and to a greater extent in peripheral than in proximal arteries. Endothelial surface/volume ratio increased (p less than 0.0001). Fetal surface projections, junctional interdigitations and overlap became less evident. Adaptational changes were complete in three weeks. Between three weeks and adulthood a reduction in endothelial surface/volume ratio suggested cell growth. In the subendothelium the volume density of collagen and basement membrane and elastin increased (p less than 0.001). The internal elastic lamina, immature in all arteries at birth increased in thickness and integrity until in the adult, only in small muscular arteries did gaps between elastin profiles ensure frequent contact between endothelial and smooth muscle cells. At all ages regional differences in endothelial cell morphology were evident.     

Immunohistochemical studies on the poll gland of the dromedary camel (Camelus dromedarius) during the rutting season

The poll glands are subcutaneous exocrine glands located on the back of the neck behind the ears in male camels. The function of poll glands is not well known, though they are thought to play a role during the rutting season. The presence, location and degree of immunolocalization of microfilaments and intermediate filament systems: actin and cytokeratins (Cks) and also S100 protein were studied in the poll glands in sexually mature one-humped camels during the rutting season. These proteins were variably expressed between the epithelia, perialveolar, interalveolar tissue and the periductal tissue. Strong α-smooth muscle actin (α-SMA) immunoreactivity (IR) was displayed by the perialveolar myoepithelial cells, periductal and vascular smooth muscle cells (SMCs), but not in the epithelial cells. Cytokeratin (Ck)-IR was strong in the epithelial lining of the secretory alveoli and excretory ducts, however, the apical blebs of the secretory cells were almost negative. Weak to moderate Ck-IR was observed in the perialveolar myoepithelial cells, but not in the interalveolar tissue or endothelial cells. S100 protein was expressed variably in the epithelial lining of the secretory alveoli. S100-IR was more obvious in the supranuclear region and the apical blebs. Variable reaction was observed in the perialveolar myoepithelial cells, periductal and interductal tissue and endothelial cells.