Molecular phylogeny of early vertebrates: monophyly of the agnathans as revealed by sequences of 35 genes

Extant vertebrates are divided into three major groups: hagfishes (Hyperotreti, myxinoids), lampreys (Hyperoartia, petromyzontids), and jawed vertebrates (Gnathostomata). The phylogenetic relationships among the groups and within the jawed vertebrates are controversial, for both morphological and molecular studies have rendered themselves to conflicting interpretations. Here, we use the sequences of 35 nuclear protein-encoding genes to provide definitive evidence for the monophyly of the Agnatha (jawless vertebrates, a group encompassing the hagfishes and lampreys). Our analyses also give a strong support for the separation of Chondrichthyes (cartilaginous fishes) before the divergence of Osteichthyes (bony fishes) from the other gnathostomes.     

Psychological and social well-being of bony fishes in zoos and aquariums

Research on the behavior of animals in zoos has been conducted for decades and observations have provided information that has improved the psychological and social well-being of animals. However, research on fishes in zoos and aquariums seems to be lacking. Here we assess the current state of research on fishes in zoos and aquariums by surveying peer-reviewed literature. Our assessment differs from previous surveys in that we examine the taxonomic classes Chondrichthyes (sharks and rays) and Osteichthyes (bony fishes) separately. Our survey finds that bony fishes have been drastically underrepresented in zoo journals, more so than chondrichthyans, revealing an urgent need for zoos and aquariums to conduct research on the behavior of the bony fishes in their care, to ensure a positive state of psychological and social well-being. To plot a course for the future, we discuss the few studies that have been conducted on the behavior of bony fishes in zoos and aquariums, and we discuss research conducted on bony fishes in food-fish aquaculture and biomedicine to identify the types of studies that could be conducted in zoos and aquariums. We conclude that data-driven analyses of fish behavior could aid in development of evidence-based practices that enhance the well-being of bony fishes in zoos and aquariums, just as they already do for terrestrial animals.     

Piscine macrophage aggregates: A review

Piscine macrophage aggregates (MAs), alternatively known as melano-macrophage centers (MMC), are focal accumulations of macrophages usually containing the pigments hemosiderin, lipofuscin and ceroid, and melanin. The structures are not confined to fish and have been observed in other poikilothermic vertebrates. The aggregations are most commonly present in the spleen, pronephros, mesonephros and liver but may be found in other organs, especially in relation to inflammation. In the lower fishes (Agnatha, Chondrichthyes) the pigmented cells tend to be solitary or in small (<30 cells/aggregation), irregularly shaped aggregations primarily in hepatic tissue. In the Osteichthyes, greater numbers of cells and aggregates are present than in the lower fishes. These aggregates are more nodular, and they occur more commonly in the spleen and kidney rather than the liver.An exception to these observations if found in the Clupeiformes and the Salmoniformes, considered lower teleosts, who have poorly organized and irregularly shaped aggregations that are smaller than the higher teleosts.Macrophage aggregates function in normal physiological processes and in the body's defense against injurious agents. Evidence indicates that these functions are multiple, complex, and not well understood. They may be classified as, (a) immune, including humoral and inflammatory responses; (b) storage, destruction, or detoxification of exogenous and endogenous substances; and (c) iron recycling. There is a growing body of information that elucidates these functions.Macrophage aggregates qualify as anatomical and cytological biomarkers since they are known to change in number, size, and pigment content in relation to fish health and environmental degradation. Their value lies in their ubiquity, availability, and ease of measurement. There have been few extensive, controlled attempts to produce MAs or to study their kinetics by chronic exposure to contaminants known to exist in polluted environments. Clearly, without such investigations the value of MAs as monitors (biomarkers) of fish health and environmental degradation remains questionable.     

Conservation of all three p53 family members and Mdm2 and Mdm4 in the cartilaginous fish

Analysis of the genome of the elephant shark (Callorhinchus milii), a member of the cartilaginous fishes (Class Chondrichthyes), reveals that it encodes all three members of the p53 gene family, p53, p63 and p73, each with clear homology to the equivalent gene in bony vertebrates (Class Osteichthyes). Thus, the gene duplication events that lead to the presence of three family members in the vertebrates dates to before the Silurian era. It also encodes Mdm2 and Mdm4 genes but does not encode the p19^Arf gene. Detailed comparison of the amino acid sequences of these proteins in the vertebrates reveals that they are evolving at highly distinctive rates, and this variation occurs not only between the three family members but extends to distinct domains in each protein.     

The flagellar apparatus of spermatozoa in fish. Ultrastructure and evolution

Chondrichthyes possess an evolved type of spermatozoa. Their flagellar apparatus is characterized by the presence of flagellar roots which form the axis of the midpiece, and the existence of one or two lateral elements associated with the axoneme. Osteichthyes, mainly teleosteans, show a great diversity of spermatic forms. The primitive spermatozoon with a 9 + 2 pattern flagellum is common. The primitive spermatozoon has evolved along different lines. The spermatic diversity which results from this is mainly evident in the structure of the flagellar apparatus. In the animal kingdom the primitive spermatozoon with a 9 + 2 pattern flagellum, present in primitive metazoa, is retained in phyla where external fertilization is maintained. The main evolutionary tendencies--elongation, aflagellarity or biflagellarity--are generally connected with the acquisition of internal fertilization. These evolutionary tendencies are found in teleosteans. It is not possible to link aflagellarity or biflagellarity of the gamete in certain fishes to this method of fertilization. Only the elongation of the spermatozoon is connected, in certain cases, with internal fertilization, but this cannot be taken as general.     

Placoderm fishes,pharyngeal denticles,and the vertebrate dentition

The correlation of the origin of teeth with jaws in vertebrate history has recently been challenged with an alternative to the canonical view of teeth deriving from separate skin denticles. This alternative proposes that organized denticle whorls on the pharyngeal (gill) arches in the fossil jawless fish Loganellia are precursors to tooth families developing from a dental lamina along the jaw, such as those occurring in sharks, acanthodians, and bony fishes. This not only indicates that homologs of tooth families were present, but also illustrates that they possessed the relevant developmental controls, prior to the evolution of jaws. However, in the Placodermi, a phylogenetically basal group of jawed fishes, the state of pharyngeal denticles is poorly known, tooth whorls are absent, and the presence of teeth homologous to those in extant jawed fishes (Chondrichthyes + Osteichthyes) is controversial. Thus, placoderms would seem to provide little evidence for the early evolution of dentitions, or of denticle whorls, or tooth families, at the base of the clade of jawed fishes. However, organized denticles do occur at the rear of the placoderm gill chamber, but are associated with the postbranchial lamina of the anterior trunkshield, assumed to be part of the dermal cover. Significantly, these denticles have a different organization and morphology relative to the external dermal trunkshield tubercles. We propose that they represent a denticulate part of the visceral skeleton, under the influence of pharyngeal patterning controls comparable to those for pharyngeal denticles in other jawed vertebrates and Loganellia.     

Changes in the fish fauna of the Ria de Aveiro estuarine lagoon (Portugal) during the twentieth century

The fish biodiversity of the Ria de Aveiro has been analysed during the 20th century, as an indicator of the biological integrity of the ecosystem. Ninety-two distinct species and 38 families of Agnatha, Chondrichthyes and Osteichthyes occurred during the period. The family and species richness showed two levels of magnitude, from the 1910s, with high values (26–28 families and 51–52 species), and during the last two decades with more variable values (20–27 families and 38–55 species). A total of 13 species (four non-sporadic species) have disappeared recently. The lagoon system has been affected by large-scale anthropogenic influences mainly the substantial development of industries and the increasing population in the watershed, in addition to overfishing, climatic changes and dredging. In general, as shown by an analysis of the ecotrophic guilds present, the area maintained a similar structure during the whole period. No significant differences in time were recorded for any of the designated guilds. The twenty-two frequent species, which occurred in six or seven surveys, were mainly estuarine resident species or marine adventitious species, living in the substratum (benthic fish), on soft substratum and vegetated bottoms. They were feeding either on invertebrates alone or on invertebrates and fish, and producing pelagic or benthic eggs. The 19 sporadically-recorded species, recorded only once, were mostly marine adventitious, demersal fishes, living above rough bottom and vegetation, feeding strictly on invertebrates, and producing eggs benthically or deposited into vegetation. It is concluded that the Ria de Aveiro estuarine coastal lagoon has supported similar fish communities during the past century and that these comprise representatives of various ecological types.     

The evolution from asparagine or threonine to cysteine in position 146 contributes to generation of a more efficient and stable form of muscle creatine kinase in higher vertebrates

Creatine kinase, a key enzyme in vertebrate excitable tissues that require large energy fluxes, catalyzes the reversible transfer of phosphate between adenosine triphosphate and creatine. Sequence alignment indicated that the 146th amino acid is cysteine in the muscle creatine kinase of higher vertebrates including Amphibia, Reptilia, Aves and Mammalia. In fishes, it is cysteine in Agnatha and Chondrichthyes, and asparagine or threonine in Osteichthyes, which is the ancestor of Amphibia, Reptilia, Aves and Mammalia. To explore the structural and functional role of this special residue, a series of site-directed mutants of rabbit muscle creatine kinase were constructed, including C146S, C146N, C146T, C146G, C146A, C146D and C146R. A detailed comparison was made between wild-type creatine kinase and the mutants in catalytic activity, physico-chemical properties and structural stability against thermal inactivation and guanidine hydrochloride denaturation. It was found that except for C146S, the mutants had relatively lower catalytic activity and structural stability than Wt-CK. Wt-CK and C146S were the most stable ones, followed by C146N and C146T, and then C146G and C146A, and C146D and C146R were the least stable mutants. These results suggested that the 146th residue plays a crucial role in maintaining the structural stability of creatine kinase, and that the evolution in this amino acid from asparagine or threonine to cysteine contributes to the generation of a more efficient and more stable form of creatine kinase in higher vertebrates.     

Conservation of all three p53 family members and Mdm2 and Mdm4 in the cartilaginous fish

Analysis of the genome of the elephant shark (Callorhinchus milii), a member of the cartilaginous fishes (class Chondrichthyes), reveals that it encodes all three members of the p53 gene family, p53, p63 and p73, each with clear homology to the equivalent gene in bony vertebrates (class Osteichthyes). Thus, the gene duplication events that lead to the presence of three family members in the vertebrates dates to before the Silurian era. It also encodes Mdm2 and Mdm4 genes but does not encode the p19^Arf gene. Detailed comparison of the amino acid sequences of these proteins in the vertebrates reveals that they are evolving at highly distinctive rates, and this variation occurs not only between the three family members but extends to distinct domains in each protein.Key words: p53, p63, p73, Mdm2, Mdm4, elephant shark     

The fate of intraperitoneally injected carbon particles in cyprinid fish

Summary The lymphoid organs of rosy barb (Barbus conchonius) and carp (Cyprinus carpio) were investigated for their phagocytic uptake of carbon, after its intraperitoneal injection. Carbon handling was similar in both species. It was first detected in the lymphoid organs at 30 min after injection. During the first day, carbon was phagocytized by macrophages situated in the spleen within the ellipsoids and in the red pulp. In head and trunk kidney, carbon was found in macrophages scattered throughout the haemopoietic parenchyma, and in cells lining the blood sinuses. In the spleen, macrophages replete with carbon left the ellipsoidal structures and formed aggregates with pigment-containing macrophages from day 6 onwards. In all lymphoid organs, almost all carbon was ultimately concentrated in the melano-macrophage centres.     

Distribution of interstitial retinol-binding protein (IRBP) in the vertebrates

Immunoblots of interphotoreceptor matrix preparations from 20 species belonging to six vertebrate classes were probed with antibodies against bovine interstitial retinol-binding protein (b-IRBP). Each preparation displayed an immunoreactive protein band. In the Osteichthyes, the apparent Mr of this band was 67,600 +/- 2,700 (mean +/- SD, n = 8). In two of the Osteichthyes, the band was resolved into a closely spaced doublet. Including previously published data for five mammals and one amphibian, species from the other classes (Chondrichthyes, one species; Amphibia, four species; Reptilia, one species; Aves, one species; Mammalia, nine species) had IRBPs with Mr that averaged 2.0 times that of the Osteichthyes, namely 134,200 +/- 8,600 (mean +/- SD, n = 17). Frog IRBP was very similar to mammalian IRBP in terms of its immunohistochemical distribution (determined with rabbit anti-frog IRBP antibodies), its molecular weight (sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel-filtration chromatography), retinol- and concanavalin A-binding ability, and because it was synthesized and secreted in vitro by the isolated retina but not by the pigmented layers of eye. Goldfish IRBP apparently binds exogenous (3H)-retinol but does not bind concanavalin A and has about half the Mr of frog IRBP. The occurrence of IRBP-like proteins cross-reacting with anti b-IRBP antibodies in the interphotoreceptor matrix of all six major vertebrate classes is consistent with the hypothesis that IRBP is an important element in the vertebrate visual cycle.     

Heterogeneity of macrophage populations in human lymphoid tissue and peripheral blood

Human lymphoid tissue and peripheral blood leukocytes were stained with six monoclonal antibodies directed against monocyte/macrophage populations. The staining pattern described by each of these monoclonal reagents was compared with the distribution of morphologically distinguishable tissue macrophages. The results show that there exists considerable heterogeneity of tissue macrophages based on the expression of surface and/or cytoplasmic antigens; furthermore, the distribution of cells bearing particular antigenic determinants is associated with distinct regions in normal lymphoid tissue. Double staining methods demonstrated that these antibodies bind to different, as well as to identical, macrophage populations. OKM-1 antibody binds predominantly to sinus histiocytes and tingible body macrophages. The Leu M-1 reagent stains interdigitating reticulum cells, while the KiM-4 antibody labels follicular dendritic cells. Leu M-3 antibody identifies cells predominantly in the germinal center, and histiocytes lining the sinuses. Both CM-1 and BRL-M.1 appear to stain tissue macrophages distributed throughout the lymphoid tissue.     

First Shark from the Late Devonian (Frasnian) Gogo Formation,Western Australia Sheds New Light on the Development of Tessellated Calcified Cartilage

BackgroundLiving gnathostomes (jawed vertebrates) comprise two divisions, Chondrichthyes (cartilaginous fishes, including euchondrichthyans with prismatic calcified cartilage, and extinct stem chondrichthyans) and Osteichthyes (bony fishes including tetrapods). Most of the early chondrichthyan (‘shark’) record is based upon isolated teeth, spines, and scales, with the oldest articulated sharks that exhibit major diagnostic characters of the group—prismatic calcified cartilage and pelvic claspers in males—being from the latest Devonian, c. 360 Mya. This paucity of information about early chondrichthyan anatomy is mainly due to their lack of endoskeletal bone and consequent low preservation potential.Conclusions/SignificanceThe Meckel’s cartilages show a jaw articulation surface dominated by an expansive cotylus, and a small mandibular knob, an unusual condition for chondrichthyans. The scapulocoracoid of the new specimen shows evidence of two pectoral fin basal articulation facets, differing from the standard condition for early gnathostomes which have either one or three articulations. The tooth structure is intermediate between the ‘primitive’ ctenacanthiform and symmoriiform condition, and more derived forms with a euselachian-type base. Of special interest is the highly distinctive type of calcified cartilage forming the endoskeleton, comprising multiple layers of nonprismatic subpolygonal tesserae separated by a cellular matrix, interpreted as a transitional step toward the tessellated prismatic calcified cartilage that is recognized as the main diagnostic character of the chondrichthyans.     

鲤鱼和鲫鱼脾脏显微结构和亚显微结构的研究

鲤鱼和鲫鱼的脾脏中都没有小梁,其红髓和白髓是混合的。脾脏中都有椭圆体,其末端向脾髓开放。脾髓的纤维网眼中缺乏血管壁,椭圆体毛细血管中的血液可直接流入脾髓网状结缔组织的腔隙中。腹腔注射墨汁的鲫鱼,2小时后即可在椭圆体的巨噬细胞中看到碳粒的积累,可见脾脏的过滤作用主要在椭圆体中进行。在注射墨汁的鲫鱼脾脏中,黑色素巨噬细胞中心比未经注射鱼的脾脏明显,且数量增多,它和哺乳动物淋巴结和脾脏中的生发中心结构和功能都不相同。       

The in situ distribution of glycoprotein-bound 4-O-Acetylated sialic acids in vertebrates

Sialic acids are located at the terminal branches of the cell glycocalyx and secreted glycan molecules. O-Acetylation is an important modification of the sialic acids, however very few studies have demonstrated the in situ distribution of the O-Acetylated sialic acids. Here the distribution of glycoprotein bound 4-O-Acetylated sialic acids (4-O-Ac sias) in vertebrates was determined using a novel virus histochemistry assay. The 4-O-Ac sias were found in the circulatory system, i.e. on the surface of endothelial cells and RBCs, of several vertebrate species, though most frequently in the cartilaginous fish (class Chondrichthyes) and the bony fish (class Osteichthyes). The O-Acetylated sialic acid was detected in 64 % of the examined fish species. Even though the sialic acid was found less commonly in higher vertebrates, it was found at the same location in the positive species. The general significance of this endothelial labelling pattern distribution is discussed. The seemingly conserved local position through the evolution of the vertebrates, suggests an evolutionary advantage of this sialic acid modification.     

Coevolution of the Monogenoidea (Platyhelminthes) based on a revised hypothesis of parasite phylogeny

A revised hypothesis for the phylogeny of the Subclass Polyonchoinea (Monogenoidea) was contructed employing phylogenetic systematics. The Acanthocotylidae (formerly of the Order Capsalidea) is transferred to the Order Gyrodactylidea based on this analysis. The new phylogeny is used to determine coevolutionary relationships of the familial taxa of Monogenoidea with their hosts. The coevolutionary analysis suggests that the Monogenoidea apparently underwent sympatric speciation or dispersal while parasitic on ancestral Guathostomata, resulting in two primary clades: the Polyonchoinea and the Oligonchoinea + Polystomatoinea. The two parasite clades apparently cospeciated independently with divergence of the Chondrichthyes and Osteichthyes. In the Polyonchoinea, the clade associated with Chondrichthyes experienced primary extiaction within the Holocephala, but coevolved into the Loimoidae and Monocotylidae in the Galeomorphii and Squalea (Elasmobranchii), respectively. Within the Osteichthyes, polyonchoineans experienced primary extinction with the divergence of Sarcopterygii, Polypteriformes and Acipenseriformes. They demonstrate primary dispersal from the Neopterygii into the Squalea (as Amphibdellatinea), Actinistia (as Neodactylodiscinea) and Urodela (as Lagarocotylidea). Secondary dispersals of polyonchoineans occurred in the Gyrodactylidae to the Polypteriformes, Urodela and Anura; in the Acanthocotylidae to the Myxinoidea and Squalea; in the Capsalidae to the Acipenseriformes and Elasmobranchii; and in the Monocotylidae to the Helocephala. The Oligonchoinea and Polystomatoinea developed upon divergence of the Chondrichthyes and Osteichthyes. Oligonchoineans cospeciated within the Chondrichthyes, with the Chimaericolidea developing within the Helocephala and the ancestor of the Diclybothriidea + Mazocraeidea within the Elasmobranchii. Two cases of primary dispersal occurred within this clade: the Diclybothriidae to the Acipenseriformes and the ancestor of mazocracidean families to the Neopterygii (both Osteichthyes). Secondary dispersal within the Oligonchoinea includes host switching of the common ancestor of Callorhynchocotyle (Hexabothriidae) to the Holocephala. Polystomatoineans coevolved within the Osteichthyes, but experienced primary extinctions in the Actinopterygii, Actinistia, Dipnoi and Amniota. Coevolution of the Sphyranuridae and Polystomatidae occurred with divergence of the Urodela and Anura, respectively. Secondary dispersal of polystomatids to the Urodela, Dipnoi and Amniota is suggested. A preliminary phylogenetic analysis of the Polystomatoinea suggests that primary extinction with secondary dispersal of polystomatids to the Dipnoi may not be necessary to explain extant parasite distributions, since Concinnocotyla (Concinnocotylinae) appears to represent the sister taxon of the remaining Polystomatidae + Sphyranuridae.     

Evolution of jaw depression mechanics in aquatic vertebrates: insights from Ghondrichthyes

The widely accepted phylogenctic position of Chondrichthyes as the sister group to all other living gnathostomes makes biomechanical analyses of this group of special significance for estimates of skull function in early jawed vertebrates. We review key findings of recent experimental research on the feeding mechanisms of living elasmobranchs with respect to our understanding of jaw depression mechanisms in gnathostome vertebrates. We introduce the possibility that the ancestral jaw depression mechanism in gnathostomes was mediated by the coracomandibularis muscle and that for hyoid depression by the coracohyoideus muscle, as in modern Chondrichthyes and possibly placoderms. This mechanism of jaw depression appears to have been replaced by the sternohyoideus (homologous to the coracohyoideus) coupling in Osteichthycs following the split of this lineage from Chondrichthyes. Concurrent with the replacement of the branchiomandibularis (homologous to the coracomandibularis) coupling by the sternohyoideus coupling as the dominant mechanism of jaw depression in Osteichthyes was the fusion and shift in attachment of the intcrhyoideus and intermandibularis muscles (producing the protractor hyoideus muscle, mistakenly refereed to as the geniohyoideus), which resulted in a more diversified role of the sternohyoideus coupling in Osteichthyes. The coracohyoideus coupling appears to have been already present in vertebrates where it functioned in hyoid depression, as in modern Chondrichthyes, before it acquired the additional role of jaw depression in Osteichthyes.     

The Osteichtyes,from the Paleozoic to the extant time,through histology and palaeohistology of bony tissues

The aim of this short review is to emphasize the richness of the comparative histological studies on both fossil and extant Osteichthyes. Some selected examples in both Sarcopterygii (excluding tetrapods) and Actinopterygii show how it is possible to improve our knowledge on bone biology of extinct species but also to obtain new data on their palaeobiology or on their paleobiogeography. After a brief survey of the organization of bony tissues in osteichthyes, we review some examples of skeletal peculiarities in the following extinct and extant taxa: the histological structure of polypterid scales that suggests a hypothesis on the possible age and the biogeographical history of this basal actinopterygian taxon; the ossified lung of the fossil coelacanthids, with a discussion on its potential function; the histological organization of the sarcopterygian derived elasmoid scales (of Eusthenopteron sp., Latimeria sp. and Neoceratodus sp.). These comparative palaeohistological and histological data provide the basis of a general discussion of the evolutionary trends of bony tissues and their derivatives in Osteichthyes.     

IL-17A and Serum Amyloid A Are Elevated in a Cigarette Smoke Cessation Model Associated with the Persistence of Pigmented Macrophages,Neutrophils and Activated NK Cells

While global success in cessation advocacy has seen smoking rates fall in many developed countries, persistent lung inflammation in ex-smokers is an increasingly important clinical problem whose mechanistic basis remains poorly understood. In this study, candidate effector mechanisms were assessed in mice exposed to cigarette smoke (CS) for 4 months following cessation from long term CS exposure. BALF neutrophils, CD4⁺ and CD8⁺ T cells and lung innate NK cells remained significantly elevated following smoking cessation. Analysis of neutrophil mobilization markers showed a transition from acute mediators (MIP-2α, KC and G-CSF) to sustained drivers of neutrophil and macrophage recruitment and activation (IL-17A and Serum Amyoid A (SAA)). Follicle-like lymphoid aggregates formed with CS exposure and persisted with cessation, where they were in close anatomical proximity to pigmented macrophages, whose number actually increased 3-fold following CS cessation. This was associated with the elastolytic protease, MMP-12 (macrophage metallo-elastase) which remained significantly elevated post-cessation. Both GM-CSF and CSF-1 were significantly increased in the CS cessation group relative to the control group. In conclusion, we show that smoking cessation mediates a transition to accumulation of pigmented macrophages, which may contribute to the expanded macrophage population observed in COPD. These macrophages together with IL-17A, SAA and innate NK cells are identified here as candidate persistence determinants and, we suggest, may represent specific targets for therapies directed towards the amelioration of chronic airway inflammation.     

A bibliography and categorization of bony fishes exhibiting parental care

Parental care occurs in a diversity of fishes, but predominantly among freshwater groups. Among the approximately 422 families of bony fishes (Osteichthyes), 89 are presently known to exhibit parental care. Grouping these families into eight categories, based on the sex of the care-giver(s), reveals male parental care is as common or more common than female parental care. Although unusual among vertebrates, parental care by males alone is very common among bony fishes. Lists of families, the forms of parental care exhibited, the modes of fertilization, the environments in which reproduction occurs, and the sources of documentation are presented. An extensive bibliography and index are provided.