Light and scanning electron microscopy of Henneguya arapaima n. sp. (Myxozoa: Myxobolidae) and histology of infected sites in pirarucu (Arapaima gigas: Pisces: Arapaimidae) from the Araguaia River, Brazil

In this report, we describe Henneguya arapaima n. sp., a parasite of the gill arch and gall bladder of Arapaima gigas (pirarucu) collected in the Araguaia River, in the municipality of Nova Crixás, Goiás State, central Brazil. The plasmodia were white, round or ellipsoidal and measured 200-600mum. Parasite development was asynchronous and the mature spores were fusiform and had smooth wall. The spores measurements were (range, with means+/-S.D. in parentheses): total length-48.4-53.1mum (51.6+/-3.4mum), body length-13.5-15.2mum (14.2+/-0.8mum), body width-5.1-6.1mum (5.7+/-0.5mum), body thickness-4.7-5.3mum (4.9+/-0.2mum) and caudal process length-38.0-41.2mum (38.3+/-2.9mum). The polar capsules were elongated and of unequal size, with lengths of 6.3-6.8mum (6.5+/-0.2) and 6.2-6.6mum (6.3+/-0.1) for the longest and shortest axes, respectively. Capsule width was 1.4-1.6mum (1.5+/-0.1). Histological analysis showed that the plasmodia occurred in the tunica adventitia of the gall bladder and were delimited by a thin capsule of connective tissue. In the gill arch, the plasmodia were also surrounded by connective tissue similar to the endomesium of striated skeletal muscle cells. Sixty-five juvenile specimens of A. gigas weighing 1.0-25.0kg were examined, 17 (26.1%) of which were infected. Of these, 14 (82.3%) had cysts in the gall bladder, two (11.7%) had cysts in the gill arch and only one (5.9%) had cysts in both organs. When the fish were grouped by weight, the prevalence of infection in fish weighing up to 10.0kg (20.7%) was significantly lower than in fish weighing 10.1-25.0kg (50%) (G=3.93; d.f.=1; p<0.05).     

Hematology and clinical chemistry of adult yellow-headed temple turtles (Hieremys annandalii) in Thailand

Background: Yellow‐headed temple turtles (YHT), Hieremys annandalii, native to Thailand, are protected from exploitation under the Wild Animal Reservation and Protection Act, also listed under Appendix II of the Convention on International Trade of Endangered Species and the International Union for the Conservation of Nature red list. Objectives: The objectives of this study were to describe quantitative, morphologic, and cytochemical features of blood cells and plasma biochemical analytes of clinically healthy YHT. Methods: Blood samples were collected from 40 adult YHT from October 2007 to February 2008. Hematologic and biochemical analyses, cytochemical staining, and ultrastructural evaluation were performed using standard methods. Results: Hematologic results (mean ± SD) included: RBC count, 0.275 ± .094 × 10⁶ cells/μL; WBC count, 11.7 ± 6.6 × 10³ cells/μL; heterophils, 29.4 ± 6.9%; eosinophils, 23.7 ± 5.3%; basophils, 21.2 ± 1.9%; lymphocytes, 14.8 ± 5.9%; and azurophils, 10.7 ± 5.3%. Erythrocytes stained dark red with peroxidase‐staining. Periodic acid‐Schiff stain could not differentiate between thrombocytes and lymphocytes. Thrombocytes contained cytoplasmic vacuoles, similar to mammalian platelets and those of birds and snakes. Heterophils and eosinophils were similar in structure and cytochemical staining characteristics to those of other turtles and reptiles. Structure of basophils was similar to avian basophils. Lymphocytes and azurophils had similar cytochemical staining compared with mammalian lymphocytes and monocytes. Mean MCHC, WBC counts, absolute azurophil counts, and plasma alanine aminotransferase activity were higher in male turtles than in females. Conclusion: Blood characteristics of YHT are species‐specific, and this study can be served as a reference for future clinical studies and medical care of YHT.     

Prenatal development of retina in buffalo (Bubalus bubalis)

The development of retina in Indian buffalo (Bubalus bubalis) has not been reported previously. The aim of the present study was therefore to report the major landmarks and the time course in the development of retina. Serial histological sections of Indian buffalo embryos and foetuses were used as group1 (<20.0 cm CVRL), group2 (>20.0 but <40.0 cm CVRL) and group3 (>40.0 cm CVRL). Age estimation was made on the basis of crown vertebral‐rump length (CVRL), which ranged between 36 and 286 days (1.6–94.0 cm). The retina in Indian buffalo was developed in a similar manner to that of the other mammals with the principal differences in the time of occurrence of various layers of this nervous tunic. In 36 days (1.6 cm stage), the foetal retina was composed of pigmented layer and the layer of neuroblasts. Differentiation of layers was first observed in 47 days (4.0 cm CVRL) which became prominent in 52 days (5.1 cm stage). At 120 days (20.5 cm stage), the differentiation of inner plexiform layer and inner nuclear layer was evident. At 143 days (31.0 cm) foetal age, the faint line in neuroblastic layer was the first evidence of the future outer plexiform layer. In foetuses of group III, the retina was comprised of all 10 layers (eight cell layers and two membranes) viz. pigmented epithelium, layer of rods and cones, outer limiting membrane, outer nuclear layer, outer plexiform layer, inner nuclear layer, inner plexiform layer, ganglion cell layer, layer of nerve fibres and the inner limiting membrane.     

Prenatal histomorphological development of the reticulum in fallow deer (Dama dama)

The histomorphological changes occurring in the Dama dama reticulum during prenatal development have been investigated. Twenty‐five Dama dama embryos were used, from the first stages of prenatal life until birth. Differentiation of the reticulum was observed at 23% gestation. By 25% gestation the reticular wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Primary reticular crests were visible at 38% gestation. Secondary reticular crests were observed at 61% gestation. Neuroendocrine cells were detected by synaptophysin (SYP) at 35% gestation, in the lamina propria‐submucosa, tunica muscularis, and serosa. Epithelial Cytokeratin‐18 (CK‐18) cells were observed at 35% gestation extended throughout the epithelial layers. The glial cells (vimentin –VIM‐ and glial fibrillary acidic protein‐GFAP‐markers) were discerned at 25% and 43% gestation, respectively, in myenteric and submucosal plexuses, lamina propria, muscularis mucosae, tunica muscularis, and perivascular connective tissue. The neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) markers were immunodetected at 75% and 80 gestation, respectively, in the lamina propria‐submucosa, muscularis mucosae, tunica muscularis, serosa, and myenteric plexuses. The prenatal development of the fallow deer reticular mucosa evidenced a considerable precocity similar to that previously reported in goat and red deer.     

The ultrastructure of spermatogenic cells and morphological evaluation of testicular development in the silver pomfret (Pampus argenteus)

The silver pomfret (Pampus argenteus) is a widely distributed and economically important marine fish in the Indo-Pacific. In this study, we acquired the second generation of wild P. argenteus by artificial breeding and further studied the testicular development and ultrastructure of spermatogenesis. The results of gonadosomatic index (GSI) showed the spawning period of this marine fish was from April to June. Besides, through morphological analysis, we found that P. argenteus had an anastomosing tubular testis surrounded by a layer of tunica albuginea, in which spermatogenesis occurred in cysts where the synchronous germ cells were completely surrounded by the cytoplasmic projection of Sertoli cells. Meanwhile, based on submicroscopic characteristics, the germ cells are classified into nine different types. During the ontogenesis of testis, both the early stage of spermatogenesis and sperm were observed in P. argenteus. At sperm maturation stage, different types of spermatozoa and activation of sperms occurred non-synchronously in the tubules. Cytoplasmic bridges also were observed among synchronous germ cells within the cysts, suggesting an interrelated and differentiated relationship among these germ cells.     

Hematology, cytochemistry and ultrastructure of blood cells in fishing cat (Felis viverrina)

Hematological, cytochemical and ultrastructural features of blood cells in fishing cat (Felis viverrina) were evaluated using complete blood cell counts with routine and cytochemical blood stains, and scanning and transmission electron microscopy. No statistically significant difference was found in different genders of this animal. Unique features of blood cells in this animal were identified in hematological, cytochemical and ultrastructural studies. This study contributes to broaden hematological resources in wildlife animals and provides a guideline for identification of blood cells in the fishing cat.     

Gross morphological,histological and scanning electron specifications of the oropharyngeal cavity of the hooded crow (Corvus cornix pallescens)

The present study was carried out on the oropharyngeal cavity of the hooded crow to investigate the gross and microscopic structures via gross anatomy, light microscopy and scanning electron microscopy (SEM). The gross anatomy clarified the elongated triangular shape of the oropharyngeal cavity with a non-protruding tongue with a bifid apex. The lingual body contained median groove rostrally and separated caudally from the root by a transverse papillary crest. The laryngeal mound located posterior to the lingual root, contained midline laryngeal cleft and bounded caudally by a transverse row of pharyngeal papillae. The palate contained choanal cleft rostrally and infundibular slit caudally in addition to five palatine ridges. By light microscopy, the dorsal lingual epithelium was highly keratinised stratified squamous with a lingual nail in the most rostral part of the apex. Then, the thickness of the keratin layer decreased caudally, while in the ventral surface, the lining epithelium became non-keratinised. The entoglossum supported the lingual body and root, but not extended to the apex. The lining epithelium of the palate was also keratinised stratified squamous and became none-keratinised at the oral side of the choanal cleft. There were numerous lobules of polystomatic salivary glands in the lingual root and the palate. SEM revealed the arrangement of different types of papillae covering both the floor and the roof of the oropharynx besides numerous openings of salivary glands in the lingual root, laryngeal mound and the palate. These findings reflect the functional relationship of the oropharyngeal cavity of the hooded crow during feeding.     

Larval development of Spondyliosoma emarginatimi (Cuvier & Valenciennes) (Pisces: Sparidae) from southern Africa

The larval development of the southern African endemic sparid Spondyliosoma emarginatum is described and illustrated from specimens collected in Algoa Bay and the Swartkops, Sundays and Swartvlei estuaries of the Cape Province. S. emarginatum has demersal eggs. Preflexion larvae are moderately elongate (BD = 18–23% BL) with body depth increasing to 31 % BL in postflexion stages. The head is rounded and the snout blunt but becomes more elongate in postflexion stages. There are short spines on the preoperculum. Flexion occurs at about 6 mm BL and postflexion larvae are characterized by distinct medio-lateral pigmentation. Osteological development is described from a series of cleared and stained specimens. All fins have the adult complement of spines and rays by 12 mm BL. The larvae of S. emarginatum are compared with those of the European S. canthurus and other southern African sparids.     

Characterization of Lateral Musculature in the Striped Weakfish (Cynoscion striatus Cuvier)

Histochemical and ultrastructural studies were performed on lateral musculature from individual female weakfish (Cynoscion striatus Cuvier). Based upon SDH, PhR, Sudan and myosin-ATPase determinations, different kinds of red, pink and white fibres were discerned at the anterior, medium and posterior regions. Levels of glycolytic activity increased from red to white fibres, being intermediate in the pink ones. Contrarily, oxidative activity decreased from red to pink and white fibres. Histochemical stain showed a complex distribution of fibre types in each layer. A considerable variation in size and distribution was also found among fibres.     

Ultrastructure features of the surface of the gills and the lower pharyngeal jaw of the Tilapia Zilli,redbelly tilapia (Coptodon Zillii,Gervais, 1848)

The present investigation was designed to describe the surface ultrastructure of the gill system of tilapia Zilli. The gill system is formed from four gill arches and each gill arch carries a row of gill filaments on its convex border and two rows of the gill rakers on its concave border. The quadrilateral interbranchial septum has elevated part at the level of the third gill arch. By SEM observations, the gill arch was divided into three regions: rostral, middle and caudal region. The caudal region contained two characteristic structures: oval leaf‐like structure and rounded‐shaped structure. Each oval leaf‐like structure carried two lateral rows of the triangular pointed spines separated by a median groove. All surfaces of gill arches, rakers and filaments were covered with a mosaic of the polygonal pavement cells, in addition to the opening of chloride cells and mucous cells. The gill arch and gill raker had only one appearance of taste buds named type I. Meanwhile, the filaments contained two types of different appearance of the taste buds named: type I and type II. Type I was the main common and similar to that present in gill arch and raker and characterized by its blunt end, while type II had hair‐like structures that projected from the volcano‐shaped depression. The gill rakers were formed from central axis surrounded by two lateral lobulated regions which carry pointed spines, taste buds and the opening of chloride cells. The surface of triangular lower pharyngeal jaw carries numerous teeth‐like papillae which originated from the socket‐like depression.     

Hindlimb bone maturation during postnatal life in the Greater Rhea (Rhea americana,Aves, Palaeognathae): Implications for palaeobiological and zooarchaeological interpretations

The objective of this study was to study the morphological pattern of bone maturation of the hindlimb bones of Rhea americana and find out how it can affect bone morphology after a taphonomic process. Juvenile specimens (n = 10) ranging from one month old to eight months old were studied. For comparison, bones from adults and juveniles from museum specimens (n = 4 and n = 6, respectively) were studied. In fresh bones, ossification centres were identified in the proximal and distal epiphyses of the tibiotarsi and in the proximal epiphysis of the tarsometatarsi, whereas the distal region of the femora and tarsometatarsi showed abundance of cartilage. The development and extension of the ossification centres of the tibiotarsi were different. In the proximal epiphysis, the centre presented less development with respect to the distal epiphysis. In the dry tibiotarsi, the proximal centre was absent and the distal one was well preserved. Both the fresh and dry juvenile tarsometatarsi presented unfused metatarsals and bulkier and wider aspect than the adult ones. The dry femora presented a noticeable excavation between condyles, whereas the dry tarsometatarsi showed the absence of the proximal epiphysis. The femora, tibiotarsi and tarsometatarsi possessed different traits of immaturity, which differentially affect the morphology of the preserved bones during a taphonomic process.     

Structural variation in the nasal bone region of European moose (Alces alces L.)

The ontogeny of typical (normal) nasal bone region of the European moose (Alces alces L.) and the 3 variants of the pattern, was studied. The variants, named as "extra bone type", "punctured type" and "open type" referring the morphology of the internasal suture, were originally observed in Finnish male and female moose skulls in 1971. All of the variants were later found in hunting trophy exhibitions presenting male moose trophies from Sweden, Norway, Baltic Republics of USSR and Poland. The frequencies of the variants showed regional differences. By using histological, radiological and OTC bone labelling methods, all of the nasal bone types were observed in this study in embryonal (N = 36), newborn (N = 21), juvenile (N = 38) and adult (N = 12) moose. Two twin embryos showed different nasal bone structure. The variation is considered to be of congenital origin.     

Histological and histochemical study of the proventriculus (Ventriculus glandularis) of common starling (Sturnus vulgaris)

The histological and histochemical structures of the proventriculus of starling (Sturnus vulgaris) were examined using haematoxylin and eosin and special staining, that is periodic acid schiff (PAS), Masson's trichrome, Alcian blue, Orcein and Reticulin. All three cranial, middle and caudal parts of the proventriculus were also studied. The study results showed that the wall of the proventriculus consisted of mucosal, submucosal, muscular and serosal tunics. The mucosal tunic presented folds and sulci on its luminal surface. In the first third of the proventriculus, the tunica mucosa characterized by presence of folds lined by stratified squamous epithelium and presence of simple tubular glands in the lamina propria. In the middle and caudal thirds of the proventriculus, the surface was covered by a columnar epithelium, and the branched tubular glands were extended through the lamina propria. From the base of the branched tubular glands, the deep proventricular glands were observed that were compound tubuloalveolar lobules. The surface epithelium of the tunica mucosa and the cells lining the proventricular glands showed a positive reaction to PAS and Alcian blue stainings. In addition, the epithelial cells of the tubular and branched tubular glands showed Masson's trichrome-positive reaction. The submucosal tunic was thin in the proventricular wall. The tunica muscularis was formed by a thin inner layer of longitudinal smooth muscle fibres and a thick outer layer of circular fibres. The serosa consisted of loose connective tissue, rich in blood vessels and covered by mesothelium.     

Wound healing during hibernation by black bears (Ursus americanus) in the wild: elicitation of reduced scar formation

Even mildly hypothermic body or limb temperatures can retard healing processes in mammals. Despite this, we observed that hibernating American black bears (Ursus americanus Pallas, 1780) elicit profound abilities in mounting inflammatory responses to infection and/or foreign bodies. In addition, they resolve injuries during hibernation while maintaining mildly hypothermic states (30-35 °C) and without eating, drinking, urinating or defecating. We describe experimental studies on free-ranging bears that document their abilities to completely resolve cutaneous cuts and punctures incurred during or prior to hibernation. We induced small, full-thickness cutaneous wounds (biopsies or incisions) during early denning, and re-biopsied sites 2-3 months later (near the end of denning). Routine histological methods were used to characterize these skin samples. All biopsied sites with respect to secondary intention (open circular biopsies) and primary intention (sutured sites) healed, with evidence of initial eschar (scab) formation, completeness of healed epidermis and dermal layers, dyskeratosis (inclusion cysts), and abilities to produce hair follicles. These healing abilities of hibernating black bears are a clear survival advantage to animals injured before or during denning. Bears are known to have elevated levels of hibernation induction trigger (delta-opioid receptor agonist) and ursodeoxycholic acid (major bile acid within plasma, mostly conjugated with taurine) during hibernation, which may relate to these wound-healing abilities. Further research as to the underlying mechanisms of wound healing during hibernation could have applications in human medicine. Unique approaches may be found to improve healing for malnourished, hypothermic, diabetic and elderly patients or to reduce scarring associated with burns and traumatic injuries.