Phylogenetic analysis and expression differences of eye-related genes in cavefish genus Sinocyclocheilus

The adaptive evolution of visual systems has been observed in many cavefish. However, little is known about the molecular mechanisms underlying these adaptations, which include regressive changes such as eye degeneration. Here, we analyzed phylogenetic and expression patterns of 6 eye-related genes (crx, foxg1b, opn1sw2, otx2, rho and sox2) in 12 Sinocyclocheilus species from China, including 8 stygobionts and 4 stygophiles, and examined photoreceptor cell morphology of these species. Those eye-degenerated species of Sinocyclocheilus were polyphyletic and showed different degrees of photoreceptor defects in responses to cave environments. The eye loss and degeneration are the result of convergent evolution. Although S. anophthalmus grouped with the eye-normal species, it displayed not only a high degree of eye degeneration but also significant expression differences in eye-related genes compared with the eye-normal species. The gene foxg1b, which was determined to be under positive selection, might play an important role in the process of eye degeneration in S. anophthalmus based on differential expression. Eye-related gene expression and selection may have contributed to the polyphyly of the cave species. We examined gene expression and duplication in 6 eye-related genes and revealed that these genes displayed considerable diversity in relative expression in Sinocyclocheilus fishes. Otx2 and sox2 were significantly up-regulated in individual cave species, while the other 4 genes (crx, foxg1b, opn1sw2 and rho) were significantly down-regulated. These findings provide a valuable resource for elucidating molecular mechanisms associated with visual system evolution in cavefish.     

Morphological structure and peripheral innervation of the lateral line system in the Siberian sturgeon (Acipenser baerii)

Light and scanning electron microscopy (SEM) were used to study the epidermal lateral line system of the Siberian sturgeon (Acipenser baerii Brandt, 1869). This system consists of mechanoreceptive neuromasts, ampullae and the electroreceptive organ. The neuromasts are located in 5 pairs of cephalic and 1 pair of trunk canals and superficially in the middle and posterior pit lines that lie dorsomedially along the top of the skull immediately adjacent to the otic ampullae field. Both canal neuromasts and pit organ superficial neuromasts have opposite polarized hair cells that are parallel along the axis of the canal and pit line, respectively. However, they differ in both size and shape and in the density and length of the hair bundles. The ampullae are confined on the head, adjacent to the neuromast lines. The morphological structure of the ampullae in the Siberian sturgeon is similar to the ampullae in elasmobranchs and other primitive fish. Nevertheless, it has a relatively large mucus-filled ampulla, and a shorter and narrower canal leading to a small opening to the outer epidermal surface. We also present new information concerning the peripheral innervation of lateral line receptors in sturgeons. The receptors of the lateral line system are innervated by 2 pairs of cranial nerves: anterior and posterior lateral line nerves. The peripheral processes of the anterior lateral line nerve form superficial ophthalmic, buccal, otic and anteroventral rami. The peripheral processes of the posterior lateral line nerve form middle, supratemporal and lateral rami.     

Fine structure of the canal neuromasts of the lateral line system in the adult zebrafish

The mechanosensory lateral line system of fish is responsible for several functions such as balance, hearing, and orientation in water flow and is formed by neuromast receptor organs distributed on head, trunk and tail. Superficial and canal neuromasts can be distinguished for localization and morphological differences. Several information is present regarding the superficial neuromasts of zebrafish and other teleosts especially during larval and juvenile stages, while not as numerous data are so far available about the ultrastructural characteristics of the canal neuromasts in adult zebrafish. Therefore, the aim of this study was to investigate by transmission electron microscopy the ultrastructural aspects of cells present in the canal neuromasts. Besides the typical cellular aspects of the neuromast, different cellular types of hair cells were observed that could be identified as developing hair cells during the physiological turnover. The knowledge of the observed cellular types of the canal neuromasts and their origin could give a contribution to studies carried out on adult zebrafish used as model in neurological and non‐neurological damages, such as deafness and vestibular disorders.     

Morphologic,morphometric and contour shape variations of sagittal otoliths of Lepidorhombus spp. in the Aegean Sea

Otoliths are calcareous anatomical structures in the inner ear of fishes, and they can be used in the discrimination of fish species and stocks due to their species-specific shape. Sagittal otoliths in the Lepidorhombus whiffiagonis and Lepidorhombus boscii, two flatfish species of the same genus distributed in the Aegean Sea, were compared using morphology, morphometry and contour shape analyses. Blind and eyed side otoliths of Lepidorhombus species were evaluated separately due to their morphological features and statistical differences in morphometric measurements (p < 0.05). Four analysis groups were formed: blind side otolith of L. whiffiagonis, eyed side otolith of L. whiffiagonis, blind side otolith of L. boscii and eyed side otolith of L. boscii. Morphometric differences in otolith shape were performed by canonical discriminant analysis, and the first discriminant axis explained 97.4% (Wilks λ = 0.270) of the variance between the groups, and the second axis explained 2.3% (Wilks λ = 0.620). Classification success between Lepidorhombus species based on CDA is 100%, and the overall CDA classification score between groups is 76.5%. The wavelet functions obtained in the contour analysis showed high variability in the anterior, posterior-dorsal, antero-dorsal and ventral regions of the otoliths among these four analysis groups. Consequently, it is thought that this study will contribute to the taxonomic classification of fish with the morphological and morphometric differences in the blind and eyed side otoliths and the analyses made with the high discrimination success obtained in Lepidorhombus species.     

The Branching Pattern of the Brachiocephalic Trunk in the Donkey (Equus asinus)

Donkeys are a member of the horse family (Equidae) and share a common ancestor. However, in morphological terms, donkeys and horses are regarded as two separate subgenus. Given variations in the branching pattern of the brachiocephalic trunk (BCT) in different species of animals and the use of donkeys in anatomy courses at colleges of veterinary medicine, we conducted this study in order to investigate the branching patterns of BCT and to describe detailed morphological information regarding donkeys. Seventeen mature donkeys were examined following euthanasia by exsanguination from the common carotid artery under general anaesthesia. Thirteen donkeys were embalmed and injected with coloured latex from BCT origin for better visualization of vessels. Four other donkeys were freshly studied without the embalming procedure. In all cases, the BCT was the only branch of the aortic arch and branched into the left subclavian (LSb) artery, the right costocervical trunk, the right subclavian (RSb) artery and the bicarotid trunk. The main branching pattern of the RSb was the costocervical trunk, the vertebral artery, the internal thoracic artery and the superficial cervical artery. The deep cervical artery branched from the costocervical trunk. The major branching pattern of the LSb was the vertebral artery, the internal thoracic artery and the superficial cervical artery. In donkeys, the external thoracic artery branches from the internal thoracic artery. Results obtained from this study indicate that BCT branching in donkeys has some similarities and differences as compared to that in horses and in the Caspian miniature horse.     

Anatomical description of the origin and distribution of the lumbosacral plexus in one puma (Puma concolor)

Wild felids often suffer spinal and limb disorders; however, their nervous system anatomy is poorly studied. Herein, the lumbosacral plexus (Plexus lumbosacralis) of an adult puma and the motor and sensitive innervation of the pelvic limb is described. We found anatomical similarities to other felids, but also some differences. Branches L4-S3 form the lumbosacral plexus (Plexus lumbosacralis) in the puma. The femoral nerve (N. femoris) arises from the union of L4-L5, while in other felids, it is formed by L5-L6. Unlike in the cat, the sartorius muscle receives branches from the saphenous (N. saphenous) and femoral nerves (N. femoris), and the lateral head of the gastrocnemius and superficial digital flexor muscles are innervated by a branch of the soleus muscle.     

An integrative insight into the synsacral canal of fossil and extant Antarctic penguins

The lumbosacral-canal system in birds most likely operates as a sense organ involved in the control of balanced walking and perching, but our knowledge of it is superficial. Penguins constitute interesting objects for the study of this system due to their upright walking, but only the Humboldt penguin, Spheniscus humboldti, and some incomplete fossil penguin synsacra have been studied in this respect. Here, we give an integrative comparative insight into the synsacral canal of extant Emperor penguin, Aptenodytes forsteri, Adelie penguin, Pygoscelis adeliae, and Eocene giant Anthropornis and/or Palaeeudyptes Antarctic penguins, using computed tomography imaging and associated data-extraction methodologies, complemented by analytical approaches ranging from geometric morphometrics to modularity, curvature, and wavelet analyses. We document that the variability in the number of synsacro-lumbar vertebrae is evolutionarily conserved, and all studied synsacra possess osteological correlates of the lumbosacral-canal system. We also found that Eocene and extant Antarctic penguins were separable on the basis of the main direction of the shape-related (size-independent) variability within said system, and A. forsteri was unique in the entire studied set in terms of the relative cranial shift of this compound structure. Moreover, we suggest that the evolutionary processes, shaping both the terrestrial posture and gait, were responsible, in extant penguins, for the increased simplicity and stability of the synsacral canal cross-sectional periodic patterns, as well as pave the way for the lumbosacral-canal system modularity characterized by reduced atomization/complexity.     

The side wall of the braincase in cynodont therapsids,and a note on the homology of the mammalian promontorium

The side wall of the cynodont braincase is not sufficiently well understood to form a sound basis for morphological or phylogenetic inference. This paper describes in detail the trigeminal region of a series of cynodonts from lateral and mesial aspects. Genera dealt with are Thrinaxodon, Trirachodon, Diademodon and Tritylodon. A reinterpretation of aspects of the braincase of the tritylodontid Bienotherium is presented. Attention is drawn to a fundamental dichotomy between herbivorous cynodonts and the carnivorous line leading to mammals. Homology between the cynodont parasphenoid wing and mammalian promontorium is demonstrated.     

水貂奇异变形杆菌的分离鉴定及16S rRNA基因序列分析

从辽宁某貂场发病水貂中分离到1株致病菌,命名为PMSD株,通过形态学观察和生化试验等常规鉴定发现符合奇异变形杆菌特性,进一步经VITEK 2Compact 30全自动细菌鉴定及药敏分析系统鉴定该株细菌为奇异变形杆菌。药敏试验结果显示PMSD株对氨基糖苷类药物、喹诺酮类药物等敏感,而对β-内酰胺类药物和磺胺类药物等不敏感。以细菌16SrRNA基因为模板应用通用引物进行PCR扩增,得到PMSD株的16SrRNA基因序列,长约1 504bp,提交到GenBank中,登录号:KM229530。将该序列与GenBank中序列进行BLAST比对,结果发现与其匹配度最高的均是奇异变形杆菌各株系的16SrRNA序列,均高达99%以上。选取其中前20株作为参考序列,运用生物学软件构建系统发育树并进行同源性比对,结果表明,分离菌PMSD株与20个代表菌株的同源性为98.9%~99.7%,其中与BB2000株、HI4320株、B1株和FCC141株同源性最高,为99.7%。本研究为预防和控制奇异变形杆菌引起的水貂疾病奠定了一定的基础。     

Three-dimensional modelling of computed tomography images of limb bones in gazelles (Gazella subgutturosa)

The objective of the present study was to model in 3D the detailed morphometric features of the thoracic and pelvic limb bones of the gazelle (Gazella subgutturosa), whose natural habitat is only in the Sanliurfa province, by using multidetector computerized tomography (MDCT) images. Materials from 14 (7 male and 7 female) dead gazelles were collected from the Harran University Veterinary Medicine Faculty clinics during or after treatment. MDCT images of the gazelles were taken and saved as DICOM (Digital Imaging and Communications in Medicine) files. MIMICS 20.1 software was used to generate a 3D reconstruction of the data, and morphological and morphometrical evaluation was performed. In respect of morphological features, the combined metacarpus (Mc) III and metacarpus Mc IV were the main weight-bearing metacarpal bone, while Mc II and Mc V were rudimentary. On the femur, the lateral condyle was bigger than the medial condyle at the caudodistal end and the medial wing of the trochlea femoris was larger than the lateral. As with the thoracic limb, the combined metatarsus (Mt) III and metatarsus Mt IV were the main weight-bearing metacarpal bone and they displayed a synostosis structure. There was no secondary metatarsus in the gazelles’ pelvic limb. Based on digital measurements, the tibia was the longest bone of the gazelles’ skeletal system, and there was a statistically significant sexual dimorphism throughout the skeleton at the levels of p < .05, p < .01 and p < .001. As a result, the present study's findings are useful as reference data for further studies on anatomy, surgery and archaezoology, while also being useful for classifying species and for forensic sciences.     

Channel Catfish Response to Ultraviolet-B Radiation

Abstract

Fingerling channel catfish Ictalurus punctatus exposed to simulated ultraviolet-B radiation at an average daily dose of 2.9 J/cm² were quite sensitive to the radiation. After a 24-h exposure, thinning of the most dorsal epidermis frequently was accompanied by edema. Compared with epidermis of unexposed fish, mucous cells in exposed fish were less superficial and club cells were less numerous both dorsally and high on the lateral surface of the body. Sunburn cells with pyknotic nuclei were evident in the epidermis of exposed fish. Among fish exposed for 48 h, focal necrosis and sloughing of the outer epidermal layer were widespread. A methanol-extractable skin substance that is associated with resistance to sunburn in other fish species was not detected in channel catfish.     

Successful treatment of Sarcoptes scabiei in a 33‐year‐old pony with underlying pituitary pars intermedia dysfunction

A pony presented with a severe, chronic dermatitis around the ears, neck and trunk of the body, hind quarters and ventral abdomen. The pony was intensely pruritic as was an in‐contact pony and both owners. A superficial skin scraping demonstrated multiple living mites, identified as Sarcoptes scabiei. Equine pituitary pars intermedia dysfunction (PPID) was diagnosed as an underlying, potentially immunosuppressive disorder. The pony was treated for Sacroptes scabiei as well as PPID and is doing well 2 months after initial presentation.     

Survivability of Infectious Hematopoietic Necrosis Virus in Fertilized Eggs of Masu and Chum Salmon

Abstract

The possibility of vertical transmission of infectious hematopoietic necrosis virus (IHNV) was studied with the eggs of masu (cherry) salmon Oncorhynchus masou and chum salmon O. keta. The surfaces of eggs and sperm were contaminated with IHNV (10^3.8-10^4.8 50% tissue culture infective dose [TCID50]/egg) and then the eggs were fertilized. Eggs just after fertilization and embryonated eggs also were infected by injection with IHNV (10^3.8 TCID50/egg) directly into the yolk. During incubation, eggs were held in running water at 10°C. Mortality of the eggs or hatched progeny was determined and isolation of IHNV on the surface or inside of the eggs was determined during the incubation period. No mortality occurred and no virus was detected in fertile eggs from contaminated gametes. For injected eggs, IHNV was not detected on the surface of masu and chum salmon eggs after 1 d of incubation. Infectivity of IHNV inside the eggs decreased gradually and could not be detected after 1 month of incubation. This rate of IHNV reduction in the fertilized egg was similar to that found in a mixture of IHNV and homogenized yolk contents. Several individual yolk components also showed anti-IHNV activity. When eyed eggs were injected with IHNV, the embryos of both masu and chum salmon became infected, and the concentration of virus increased rapidly and reached more than 10^6.5 TCID50/fish. The cumulative mortality of eggs injected at the eyed stage for both masu and chum salmon was 90%. The susceptibilities of hatched-out larvae of masu and chum salmon to IHNV were different; cumulative mortality was more than 90% in masu salmon and 20–30% in chum salmon artificially infected with the virus. We concluded that vertical transmission of IHNV is doubtful because the virus is apparently unable to survive in eggs before the eyed stage.     

家养观赏地图鱼肺炎克雷伯氏菌、维氏气单胞菌与黏质沙雷氏菌的分离鉴定及耐药性分析

本试验旨在通过细菌分离鉴定,明确家养观赏地图鱼的死因,筛选敏感药物。采用常规方法分离纯化细菌后,进行细菌形态学观察,并通过小白鼠致病性试验、细菌主要生化鉴定、16SrDNA序列测定分析、药敏试验及耐药基因检测等方法对分离的细菌进行鉴定及耐药分析。分离出3株革兰氏阴性短杆菌,根据细菌形态特征及理化特性,结合16SrDNA序列测定与系统发育分析结果,判定其分别为肺炎克雷伯氏菌(Klebsiella pneumoniae)、维氏气单胞菌(Aeromonas veronii)和黏质沙雷氏菌(Serratia marcescens),其中肺炎克雷伯氏菌具有较强致病性。3株细菌均对洛美沙星、氧氟沙星、阿米卡星、卡那霉素敏感,对阿莫西林、氟苯尼考、克林霉素、甲硝唑等具有较强耐药性。结果表明,肺炎克雷伯氏菌、维氏气单胞菌、黏质沙雷氏菌的混合感染是家养观赏地图鱼的死亡原因。     

Morphological and molecular characterization of Glossidium pedatum Looss, 1899 and Orientocreadium batrachoides Tubangui, 1931 from sharptooth catfish,Clarias gariepinus (Burchell, 1822)

The superfamily Plagiorchioidea has a considerable number of genera and species with great uncertainty of their phylogenetic position. The objectives of the current study were to specifically describe the morphology and determine for the first time the phylogenetic position of Glossidium pedatum and Orientocreadium batrachoides. Examination of G. pedatum using conventional light and scanning microscopy techniques revealed undescribed features related to the digestive system, the presence of papillae-like lateral lappets, and terminal lobes covered with backwardly directed spines. Orientocreadium batrachoides revealed a pharynx that is four lobed anteriorly. Both the large (28S) and small (18S) subunits of the ribosomal genome, analysed through Bayesian inference and maximum likelihood, were used to genetically characterise these species. Phylogenies indicate that G. pedatum does not fit well into any known family within the Plagiorchioidea, as currently indicated in various systematic structures based on morphology. However, G. pedatum was closely related to Haematoloechidae according to analyses of 18S and 28S rDNA. Based on 28S rDNA, O. batrachoides formed a well-supported clade with Orientocreadium pseudobagri within the family Orientocreadiidae. In turn, Orientocreadiidae is closely related to Leptophallidae. The current study provides essential information that could be helpful to assign the family for Glossidium in future studies. Characterisation of life-cycle stages may be necessary to fully elucidate the systematic position of G. pedatum.     

PATHOLOGIC BASIS FOR RIM ENHANCEMENT OBSERVED IN COMPUTED TOMOGRAPHIC IMAGES OF FELINE NASOPHARYNGEAL POLYPS

In postcontrast computed tomographic (CT) images, feline nasopharyngeal polyps typically demonstrate enhancement of the peripheral rim. Computed tomographic images and histologic specimens of a case series of 22 cats with surgically removed nasopharyngeal polyps were reviewed retrospectively in an attempt to elucidate the origin of rim enhancement. Polyps were present in the tympanic cavity in 15 (68%) cats (three with extension into the nasopharynx), only in the nasopharynx in four (18%) cats, and only in the external ear canal in the remaining three (14%) cats. All polyps had variable degrees of epithelial injury. Hemorrhage and inflammatory infiltration were significantly more marked in the superficial stroma whereas edema was significantly more marked in the core stroma. In noncontrast CT images (n = 22), the tympanic bulla was thickened in all 15 cats with a polyp in the tympanic cavity and enlarged in eight (53%) of these cats. In postcontrast CT images (n = 15), an outer zone of relatively increased attenuation compatible with a rim was observed in 11 (73%) polyps. The magnitude and extent of rim enhancement in CT images was positively correlated with the histologic grade of inflammation in the superficial stroma and negatively correlated with the grade of edema in the superficial stroma. It appears that inflammation is the major determinant of contrast medium accumulation in feline nasopharyngeal polyps, and the tendency for inflammation to affect predominantly the superficial layers explains the frequent observation of a rim in postcontrast CT images.     

Anatomic Study of The Collateral Branches of The Abdominal Aorta of Primate Species Saguinus niger

Saguinus niger is a primate of the family Callitrichidae, with a geographical distribution limited to northern Brazil, Guyanas, Suriname and Venezuela, where it inhabits rainforests along the coast. Three adult females from the Paragominas Bauxite Mine – PA were used, donated to the Laboratory of Animal Morphology Research (LAMR) of the Federal Rural University of Amazonia. These animals had died of natural causes. The arterial system was filled with latex, and the animals were fixed in 10% formalin. Afterwards, the abdominal aorta and its collateral branches were dissected. The arrangement of the abdominal aorta branches in this species showed a similar arrangement as domestic animals but differing in the formation of the coeliac trunk. The coeliac trunk originates from the abdominal aorta, and from this trunk, we found the hepatic, left gastric and splenic arteries, where the origin of these vessels was variable. These arteries originate from the coliac trunk or a common trunk originating from the coeliac trunk, a characteristic not described in other species. The other branches of the abdominal aorta from S. niger did not show great differences in origin when compared to other species.     

Anatomical Structure of the Brachial Plexus in the Merlin (Falco columbarius)

This study aimed to document the detailed features of the morphological structure and the innervation areas of the brachial plexus in Merlin (Falco columbarius). The skin and muscles of five adult male Merlins were dissected under the stereo microscope. The Merlin had two plexus trunks. The accessory brachial plexus consisted of ventral rami C10 and C11. C11 was divided into two branches: the cranial and caudal. The brachial plexus was composed of a rather complex network involving the ventral rami of C11‐C13, T1 and T2. In addition, a thin branch from the last two cervical sympathetic nerves participated in the plexus formation. C12, C13 and T1 had rather thick trunk. C12, C13 and T1 were also involved in the formation of the brachial plexus emerging after 1 cm from the foramen inter‐vertebrale as three trunk roots.     

Rate of Disappearance of Gas Bubble Trauma Signs in Juvenile Salmonids

Abstract

To assess the rate of disappearance of gas bubble trauma (GBT) signs in juvenile salmonids, we exposed spring chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss to water containing high levels of dissolved gas supersaturation (DGS) for a time period sufficient to induce signs of GBT, reduced the DGS to minimal levels, and then sampled fish through time to document changes in severity of GBT. Because of the tendency of GBT signs to dissipate at different rates, we conducted trials focusing on emboli (bubbles) in the gill filaments and lateral line and separate trials that focused on bubbles in the external surfaces (fins, eyes, and opercula). Bubbles in gill filaments dissipated almost completely within 2 h after transfer of fish to water of nearly normal DGS (104%), whereas bubbles in the lateral line dissipated to negligible levels within 5 h. Bubbles on external surfaces were more persistent through time than they were in gill filaments and the lateral line. Although typically dissipating to low levels within 48 h, external bubbles sometimes remained for 4 d. Assuming a direct relation exists between easily observable signs and direct mortality, our results suggest that fish can recover quickly from the potentially lethal effects of DGS once they move from water with high DGS to water of almost normal gas saturation. These results should be of fundamental importance to fishery managers interpreting the results of monitoring for the severity and prevalence of GBT in juvenile salmonids in the Columbia River system and perhaps elsewhere.     

Influence of track surface on the equine superficial digital flexor tendon loading in two horses at high speed trot

Reasons for performing study: Although track surfaces are a risk factor of tendon injuries, their effects on tendon loading at high speed are unknown. Using a noninvasive ultrasonic technique, it is now possible to evaluate the forces in the superficial digital flexor tendon (SDFT) in exercise conditions. Objectives: To compare the effects of an all‐weather waxed track (W) vs. a crushed sand track (S), on the SDFT loading in the trotter horse at high speed. Methods: Two trotter horses were equipped with the ultrasonic device (1 MHz ultrasonic probe, fixed on the palmar metacarpal area of the right forelimb). For each trial, data acquisition was made at 400 Hz and 10 consecutive strides were analysed. In each session, the 2 track surfaces were tested in a straight line. The speed was imposed at 10 m/s and recorded. The right forelimb was also equipped with a dynamometric horseshoe and skin markers. The horse was filmed with a high‐speed camera (600 Hz); all recordings were synchronised. Statistical differences were tested using the GLM procedure (SAS; P<0.05). Results: Maximal tendon force was significantly lower on W compared with S. In addition to maximal force peaks around mid‐stance, earlier peaks were observed, more pronounced on S than on W, at about 13%(horse 2) and 30% (both horses) of the stance phase. Comparison with kinematic data revealed that these early peaks were accompanied by plateaux in the fetlock angle‐time chart. For high tendon forces, the tendon maximal loading rate was significantly lower on W than on S. Conclusions and potential clinical relevance: The all‐weather waxed track appears to induce a lesser and more gradual SDFT loading than crushed sand. The SDFT loading pattern at high speed trot suggests proximal interphalangeal joint movements during limb loading.