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The Paleogene fossil record of birds in Europe.

The Paleogene (Paleocene-Oligocene) fossil record of birds in Europe is reviewed and recent and fossil taxa are placed into a phylogenetic framework, based on published cladistic analyses. The pre-Oligocene European avifauna is characterized by the complete absence of passeriform birds, which today are the most diverse and abundant avian taxon. Representatives of small non-passeriform perching birds thus probably had similar ecological niches before the Oligocene to those filled by modern passerines. The occurrence of passerines towards the Lower Oligocene appears to have had a major impact on these birds, and the surviving crown-group members of many small arboreal Eocene taxa show highly specialized feeding strategies not found or rare in passeriform birds. It is detailed that no crown-group members of modern 'families' are known from pre-Oligocene deposits of Europe, or anywhere else. The phylogenetic position of Paleogene birds thus indicates that diversification of the crown-groups of modern avian 'families' did not take place before the Oligocene, irrespective of their relative position within Neornithes (crown-group birds). The Paleogene fossil record of birds does not even support crown-group diversification of Galliformes, one of the most basal taxa of neognathous birds, before the Oligocene, and recent molecular studies that dated diversification of galliform crown-group taxa into the Middle Cretaceous are shown to be based on an incorrect interpretation of the fossil taxa used for molecular clock calibrations. Several taxa that occur in the Paleogene of Europe have a very different distribution than their closest extant relatives. The modern survivors of these Paleogene lineages are not evenly distributed over the continents, and especially the great number of taxa that are today restricted to South and Central America is noteworthy. The occurrence of stem-lineage representatives of many taxa that today have a restricted Southern Hemisphere distribution conflicts with recent hypotheses on a Cretaceous vicariant origin of these taxa, which were deduced from the geographical distribution of the basal crown-group members.

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Visual lateralisation in quails (Coturnix coturnix).

Two-week-old quails (Coturnix coturnix) were trained to discriminate food grains scattered randomly on a background of small pebbles of similar size adhering to the floor and differing from the grains in texture and hue ("pebble floor task"). Quails tested binocularly or with only their right eye in use showed less pecking to the pebbles and more pecking to the grains than quails tested with only their left eye in use. Adult quails in contrast did not show lateralisation. These findings add to previous evidence for visual lateralisation in birds in the pebble floor task suggesting that neural structures fed by the right eye, mainly located to the left hemisphere, are better suited to rapid visual categorisation of food objects. Like other galliformes species such as the domestic chick (Gallus gallus), but unlike non-galliformes species such as the pigeon, behavioural lateralisation in the pebble floor task may be associated with transitory anatomical asymmetries in the thalamofugal visual pathway.

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Daily energy budgets of avian embryos: the paradox of the plateau phase in egg metabolic rate.

The metabolic rate of precocial bird eggs reaches a plateau when about 80% of the incubation period has passed. This is unexpected, as in many species the embryo continues to grow and maintenance costs must therefore increase. To investigate this paradox, daily energy budgets were constructed for embryos of four galliform species according to two models that used empirical data on egg metabolic rate and embryo growth. In the first model, embryonic synthesis costs were estimated, with an assumed synthesis efficiency, before calculating the maintenance costs. In the second model, embryonic maintenance was calculated first, and no assumptions were made on the synthesis efficiency. The calculations show that assumptions of the synthesis efficiency had a major impact on the energy budget calculations, because embryonic growth rate was high. During the plateau phase, a galliform embryo allocated energy in favor of its maintenance costs in three ways: by decreasing growth rate, by increasing synthesis efficiency, and by depressing the formation of glycogen. Our study suggests that a reduction in growth rate plays a minor role. An increase of synthesis efficiency is more likely to explain the plateau in energy expenditure, since small increases in synthesis efficiency can lead to great savings on synthesis costs.

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Phylogenetic relationships of the enigmatic hoatzin (Opisthocomus hoazin) resolved using mitochondrial and nuclear gene sequences

The hoatzin (Opisthocomus hoazin) is a bizarre, long-tailed, crested bird that inhabits the riparian lowlands of South America. Among its peculiar attributes are (1) microbial foregut fermentation to convert plant cellulose in consumed foliage into simple sugars, (2) a highly modified skeleton to accommodate its large crop, and (3) in the young of this species, wing claws at the wrist joint which are used to climb among the branches of the nest tree. Consequently, the taxonomic position of this unusual bird has perplexed systematists since its description over 200 years ago. Traditionally classified among the fowl-like birds (Galliformes), recent studies have favored its placement with the cuckoos (Cuculiformes: Cuculidae). To help resolve this systematic uncertainty, we sequenced six mitochondrial genes (cytochrome oxidase I, II, and III, ATPase 8, ATPase 6, and cytochrome b) and one nuclear gene (c-mos), totaling 5,487 base pairs. With this large data set and an appropriate range of outgroup taxa, we demonstrate that the hoatzin should not be classified among the cuckoos or Galliformes. Instead, our analyses indicate that the hoatzin is most closely related to the turacos (Musophagiformes: Musophagidae), a small family of arboreal, frugivorous birds inhabiting sub-Saharan Africa. This phylogenetic relationship is also supported by osteological behavioral, and fossil evidence.

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Neuroanatomical localization of sex steroid-concentrating cells in the Japanese quail (Coturnix japonica): autoradiography with [3H]-testosterone, [3H]-estradiol, and [3H]-dihydrotestosterone.

Steroid autoradiography was undertaken to determine the neuroanatomical loci which might be involved in the activation of steroid-sensitive behaviors in the Japanese quail (Coturnix japonica). Male and female quail were either surgically gonadectomized or photically regressed and implanted with androgen or estrogen to restore normal sexual and courtship behavior. After gonadectomy or implant removal, each quail was injected with 250 microCi of [3H]-testosterone (3H-T), [3H]-estradiol (3H-E2), or [3H]-dihydrotestosterone (3H-DHT), sacrificed, processed for autoradiography, and the telencephalon, diencephalon, mesencephalon, and rhombencephalon were examined for labelled cells. Following 3H-T or 3H-E2 injection and autoradiography, labelled cells were found in nucleus septalis lateralis (SL), nucleus preopticus medialis (POM), nucleus paraventricularis (PVN), regio lateralis hypothalami (LHy), nucleus inferior hypothalami (IH), nucleus infundibuli (IN), nucleus intercollicularis (ICo), substantia grisea centralis (GCt), nucleus taeniae (Tn), and in the reticular formation near nucleus motorius nervi trigemini (MV). In addition, following 3H-E2 autoradiography, labelled cells were found around nucleus accumbens (Ac). Following 3H-DHT autoradiography, labelled cells were found only in SL, PVN, Tn, LHy, ICo, and CGt. No labelled cells were found in Ac, POM, IH, IN, or MV even after long exposure times. These results suggest that the nuclei labelled following 3H-E2 but not 3H-DHT administration bind exclusively the aromatized metabolites of T. Since quail show a sex difference in male-typical copulatory behavior in response to E2, labelled cells were counted in POM, LHy, IH, and Tn of male and female quail following 3H-E2 injection and autoradiography. No sex differences in the number of labelled cells were found in POM, LHy, or IH. Males were found to have more labelled cells than females in Tn. These results show that sex differences in male-typical copulatory behavior are not due to sex differences in the number of cells binding estrogens in POM. The results reported here constitute the most neuroanatomically extensive report of steroid binding cells to date for a galliform brain, the first comparison in a galliform bird of the distributions of cells labelled following injection of 3H-T, 3H-E2, and 3H-DHT and the first analysis of sex differences in numbers of estrogen-binding cells in four nuclei in the avian brain.

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Serotyping of US isolates of Chlamydophila psittaci from domestic and wild birds.

The identities of chlamydial strains, which can infect a given host, are important to know for disease prognosis, disease control, and epidemiology. The microimmunofluorescence test (MIFT) was used with a panel of 14 serovar-specific monoclonal antibodies (MAbs) to serotype 150 chlamydial isolates from domestic and wild birds. The isolates were obtained from birds submitted to diagnostic laboratories or during investigation of outbreaks. The 150 US isolates included 96 from the order Psittaciformes, 14 isolates from the order Columbiformes, 2 from the order Passeriformes, 16 from the order Galliformes, 12 from the order Struthioniformes, and 3 from the order Falconiformes. A total of 93, or 97%, of the Psittaciformes isolates were of serovar A; 11, or 79%, of the Columbiformes isolates were of serovar B; 64% of the Galliformes isolates were of serovar D, and all the Struthioniformes isolates were of serovar E. The 3 Falconiformes isolates did not react with any of the MAbs to the avian and mammalian isolates and are presumed to represent a new strain. The results show that specific chlamydial strains are usually associated with certain types of birds and that some serovars may be unusually virulent for certain species of birds. The MIFT using serovar-specific MAbs provides a rapid method to serotype new isolates, making it a useful system for epidemiological studies.

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Identification of mosquito bloodmeals using polymerase chain reaction (PCR) with order-specific primers.

A polymerase chain reaction (PCR) protocol was developed to identify host bloodmeals from mosquitoes. Primers for the cytochrome b gene were designed to distinguish between mammalian and avian bloodmeals and further differentiate among four avian orders: passeriformes, falconiformes, columbiformes, and galliformes. The assay was validated by testing tissues from 18 species of passeriformes, three species of falconiformes, three species of columbiformes, and two species of galliformes. American crows were distinguished from other passeriformes by restriction enzyme digestion. Host bloodmeals from engorged mosquitoes collected in New York State were identified to avian order level. PCR was able to detect the mosquito bloodmeal for up to 3 d after feeding on a quail. Significantly, these studies use order-specific primers in a single PCR test to identify mosquito bloodmeals.

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[Distribution of lethal haemosporidoses pathogens in poultry].

Peculiarities of geographical distribution and distribution among hosts of the haemosporidian parasites, which cause lethal diseases in poultry, are specified. So far, 11 species of such parasites have been described. This represents about 5% of bird haemosporidian parasites. The haemosporidians cause the lethal epizooties in turkeys, chickens, ducks, geese, pigeons and domesticated ostriches. One species of parasites belongs to the family Haemoproteidae, six--to Plasmodiidae, and four--to Leucocytozoidae. The agents of lethal haemosporidioses have been recorded in domestic birds of the orders Galliformes, Anseriformes, Columbiformes and Struthiorniformes. The majority of these parasites (eight species of 73%) occurs in the galliform birds. Each other order obtaines only one species of parasites. Five species parasitize turkey, which is the main host of the agent of lethal haemosporidioses. The majority of highly virulent haemosporidian species have been recorded throughout a global territory, which includes the Holarctic, Ethiopian and Oriental zoogeorgaphical regions. Three species of these parasites have been found in the Neotropical region, while none species has been recorded in Australia. The majority of highly virulent haemosporidian species have quite clear outlined areas, that is important to know for the prophilactic purposes including the quarantine of bird introducing from endemic territories.

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[The characteristics of the morphofunctional organization of the lampbrush chromosomes from the oocytes of the rock dove].

Nuclei and chromosomes isolated from growing oocytes of the pigeon C. livia have been studied by light microscope techniques. The pigeon chromosomes in diplotene assume a lampbrush form with a well expressed chromomere-loop organization. The lampbrush chromosomes (LC) with most extended lateral loops were observed in the oocytes 0.5-1.5 mm in diameter. A mean loop contour length on the preparations from oocytes of such a diameter varied within 11-16 mkm. A comparison of the pigeon LC with those of representatives of the order Galliformes described earlier (domestic fowl and Japanese quail) revealed essential differences. The LC of the pigeon are longer than those of the domestic fowl or Japanese quail. This fact well compares with the differences in DNA contents in the genomes of these species (haploid DNA quantities in genomes of the pigeon and the fowls are 1.9 and 1.2 pg, respectively). A specific feature of the pigeon LC is an even distribution of lateral loops along the chromosomal length. The central parts of the bivalents have a chromomere-loop organization typical of LC, but no chromomeres were observed, and the lateral loops were absent in the prolonged terminal regions. These regions are likely to be transcriptionally inactive. A specific structures of different types form on the pigeon LC. There are no such structures on the LC of Galliformes. Taking into account relative lengths of the chromosomes, the number and distribution of chiasmata, the location of the marked structures, an attempt was made to identify the macrobivalents of the pigeon, including the sex Z/W-bivalent.

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[Comparative histology, histochemistry and innervation of the striated muscle fibers of the peroneus longus muscle].

The morphology of the peroneus longus muscle was compared in three Galliformes and five Passeriformes in relation to partial behavioral characteristics. In the quail two fibre types are found, while the muscle of the other species is composed of three fibre types. The frequencies of these fibres are different, especially between Galliformes and Passeriformes. The peroneus longus muscle in the quail is innervated only from the phasic system. The other species show phasic and tonic innervation. There is a correlation between the muscle fibre calibre and the extent of the synaptic gutter.

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Mapping the antigenic epitope for a monoclonal antibody against lysozyme.

A monoclonal antibody (HyHEL-5), prepared to chicken lysozyme c by the method of Köhler and Milstein, identified an antigenic site (epitope) that was shared by the lysozymes of seven different species of galliform birds. The lysozymes of two galliform species, bobwhite quail and chachalaca, shared only partial antigenic identity with the epitope defined by this antibody. Duck lysozyme did not react with the antibody at all. Amino acids that determined the epitope structure were tentatively identified by comparing the amino acid sequences of these lysozymes and assuming the antigenic changes produced by evolutionary substitutions are not due to long-range conformational changes. Arg 68 was identified as a determining amino acid. Arg 68 is hydrogen-bonded to Arg 45, and together these two amino acids form a basic cluster that may be a subsite of the epitope. The antibody inhibited lysis of Micrococcus lysodeikticus by chicken lysozyme. Additionally, Biebrich Scarlet, a dye that binds to the catalytic site, inhibited antibody binding to this lysozyme, which indicates that the epitope extends into the cleft region between Arg 45 and Arg 114. The epitope was hypothesized to involve a region measuring at least 13 x 6 x 15 A including the Arg 68-Arg 45 complex that borders the enzymatic catalytic site. Four other monoclonal antibodies to lysozyme have been partially characterized; each had a distinct pattern of binding specificity for various species of bird lysozymes.

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Blood parasites of birds in Central Europe. 4. Trypanosoma, "Atoxoplasma", Microfilariae and other rare haematozoa.

The present paper, the last in the series on avian blood parasites in Central Europe, summarizes the data on parasites of the genera Trypanosoma, "Atoxoplasma", "Lankesterella", microfilariae and spirochaetes found in this region. Trypanosomes occur most frequently in Columbiformes, Corvidae, Galliformes and Strigiformes, while "Aloxoplasma" is encountered only in passerines, most frequently in Fringillidae, Passeridae and Hirundinidae; "Lankesterella" is to be found only in two species of passerines. Microfilariae occur in Corvidae and Galliformes, and spirochaetes were encountered in the blood smears from four birds only. All these organisms are very rarely found in birds due to the fact that the blood smears made from peripheral blood of birds are unsuitable for the diagnosis of these parasites.

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Ultrastructure of ostrich (Struthio camelus) spermatozoa: I. Transmission electron microscopy.

The origin and relationships of the tinamous (Order Tinamiformes), ratites (Order Struthioniformes, Rheiformes, Casuariiformes, Apterygiformes) and birds of the order Galliformes and Anseriformes is the subject of much debate and it has been suggested that the ultrastructural analysis of a wide variety of avian sperm may provide information relevant to this problem. This paper describes the fine structure of ostrich sperm and compares the results with published information for other non-passerine birds. Ostrich sperm display a short, conical acrosome which covers the tapered tip of the long, cylindrical nucleus. A nuclear invagination housing an acrosomal rod extends deep within the karyoplasm. A centriolar complex is situated beneath the head and consists of a short proximal centriole and a long (3.0 microns) distal centriole which extends the complete length of the midpiece. The central cavity of the distal centriole contains a pair of microtubules embedded in a rod of electron-dense material. The midpiece is surrounded by a mitochondrial sheath. Concentrations of fine granular material are present between the mitochondria. The principal-piece of the tail is demarcated from the midpiece by a distinct annulus and characterized by a ribbed fibrous sheath enclosing a typical axoneme. Rudimentary coarse fibres are observed between the fibrous sheath and the doublet microtubules of the axoneme in the proximal region of the principal-piece. The end-piece contains a disorganized collection of axonemal microtubules. Ostrich sperm differ in a number of respects from that of other non-passerine birds (the absence of a typical perforatorium; the presence of a ribbed fibrous sheath; a deep nuclear invagination; the structure and length of the distal centriole) but show a close similarity to sperm of the rhea and crested tinamou, both representatives of primitive avian families. These observations add further support to the theory that the ratites and tinamous constitute a monophyletic group. The evidence presented also reinforces the hypothesis that the ratites were the first group to branch off from the main avian stem, to be followed by the Galliformes & Anseriformes. Although it was impossible to determine whether the sperm of the tinamou are more "primitive" than those of the ostrich or rhea, it is clear that ostrich and rhea sperm are closely allied and distinct from tinamou sperm.

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Cryptic species within the Tetratrichomonas gallinarum species complex revealed by molecular polymorphism.

Tetratrichomonas gallinarum is a widespread intestinal parasite of galliform and anseriform birds. The pathogenicity of this species is controversial, presenting an unsettled problem as yet. We analysed the polymorphism and genetic relationship among 29 isolates of T. gallinarum obtained from eight bird species and five T. gallinarum-like isolates from the oral cavity and lower respiratory tract of human patients. Two methods were used for the analyses: RAPD and sequencing of 16S rRNA, 5.8S rRNA, ITS1 and ITS2 genes, both producing consistent and well-supported results. The isolates were divided into five groups, A-E, with eleven subgroups. The distance between groups E, D and the cluster A-B-C considerably exceeded usual intraspecific polymorphism seen in trichomonads. Moreover, the largest subgroup, A2 (containing 18 isolates), was divided into three branches according to the host specificity. All isolates from humans were placed into avian subgroups A2 and B2. We conclude that our isolates represent, at least, three morphospecies or rather complexes of several cryptic species. Since certain species of the T. gallinarum complex can differ in their biological characteristics and some of them can infect humans, the problem of T. gallinarum pathogenicity should be re-examined with regard to specific genetic groups and zoonotic potential of some of these lineages should be considered.

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A new genus and four new species of quill mites (Acari: Prostigmata: Syringophiudae) from phasianid birds (Galuformes: Phasianidae).

A new genus and four new species of the quill mites are described. Galliphilopsis gen. n. (G. francolinus sp. n., type species) differs from the closely related Dissonus Skoracki, 1999 by loss of leg setae dGII, the slightly divergent tips of epimeres I and the stylophore constricted posteriorly. Three new species of the genus Galliphilopsis gen. n. and one new species of the genus Mironovia Chirov and Kravtsova, 1995 are described from galliform birds (Phasianidae) collected at the Museum of Natural History, Wroclaw University (Poland): G. lophurus sp. n. from Lophura leucomelanos, G. bochkovi sp. n. from Tragopan sp. and Alectoris barbara, G. francolinus sp. n. from Francolinus levalliantoides, and Mironovia rouloul sp. n. from Rollulus rouloul.

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West Nile virus-associated mortality events in domestic Chukar partridges (Alectoris chukar) and domestic Impeyan pheasants (Lophophorus impeyanus).

West Nile virus (WNV) infection was diagnosed in captive juvenile chukars (Alectoris chukar), and captive juvenile Impeyan pheasants (Lophophorus impeyanus) on the basis of necropsy, histopathology, polymerase chain reaction, and immunohistochemistry. The chukars were kept in a game bird farm that experienced two outbreaks with approximately 25% mortality in hundreds of chukars between September and October 2002 and during the same months in 2003. The submitted pheasants were part of a group of 15 juvenile Impeyan pheasants that all died within approximately 2 wk at the end of August 2002. The macroscopic lesions in the pheasants were dominated by mucosal hemorrhage at the proventricular to ventricular junction and cecal ulcers, whereas the gross lesions in the chukar partridges were nonspecific. The predominant microscopic lesion in the chukar partridges was myocardial necrosis, whereas fibrinous and necrotizing splenitis was prominent in the pheasants. Viral antigen was usually widespread in animals of both species. Spontaneously occurring WNV infection should be considered a differential diagnosis in cases of mortality among select species of galliform birds.

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Morphology of the lingual apparatus of the domestic chicken, Gallus gallus, with special attention to the structure of the fasciae.

A detailed redescription of the mechanically interacting structural elements of the lingual apparatus of the domestic chicken, Gallus gallus, revealed the functional and constructional role of organized connective tissue (i.e., ligaments and fasciae) as structural elements that ensure the proper biomechanical interactions among the various structures within the lingual apparatus (e.g., cartilaginous and bony skeletal elements, muscles, salivary glands, epithelial structures). Fasciae, together with extrinsic muscles, also connect the lingual apparatus to the other components of the feeding apparatus, such as the skull, jaw apparatus, and larynx. For example, the hyoid apparatus is attached to the skull by a sheath-like fascia (F. vaginalis), the internal structure of which is described here for the first time. Thus, the hyoid suspension in birds differs fundamentally from that in mammals. This study is the first to examine all biomechanically functioning structural elements that are part of the galliform lingual apparatus in a systematic and comprehensive manner. It also provides a set of novel characters that may be useful for future comparative studies in evolutionary and functional morphology.

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Spermatozoal ultrastructure in three species of parrots (aves, Psittaciformes) and its phylogenetic implications.

BACKGROUND: DNA-DNA hybridization studies suggest that Psittaciformes are highly, but not the most, derived nonpasserines. Multilocus protein electrophoresis indicates that cockatoos (Cacatuinae) form a monophyletic lineage distant from the other Australo-Papuan psittacids (Psittacinae). METHODS: Transmission electron microscope procedures are applied to the spermatozoa of three parrots, in the Cacatuninae and Psittacinae, to investigate these relationships. RESULTS: Psittaciform sperm have the following characteristics: (1) conical acrosome vesicle; rodlike perforatorium; cylindrical, highly condensed nucleus; proximal and distal centriole embedded in dense material; elongate periaxonemal mitochondrial midpiece, (2) nine dense peripheral axonemal fibers (coarse fibers), (3) no fibrous sheath around the axoneme, (4) mitochondria with linear cristae, lacking intra- (or inter-) mitochondrial dense bodies, (5) restriction of the endonuclear perforatorial canal to the anterior region of the nucleus, (6) a short distal centriole, and (7) nucleus abutting on but not penetrating the acrosome. CONCLUSIONS: (1) These features are tetrapod symplesiomorphies, (2) is an amniote synapomorphy; the fibers differ from those of reptiles in being uniform in size, (3) loss of the fibrous sheath is an apomorphy known elsewhere only in columbiforms, (4) are apomorphies relative to basal aminiotes (Chelonia, Sphenodon, and Crocodilia), (5) is an apomorphic condition shared with other nonpasserines (galliforms and the white-naped crane) and crocodilians, (6) the latter taxa differ from parrots in a plesiomorphic elongation of the distal centriole, and (7) is a unique apomorphy of parrot sperm relative to other nonpasserines and reptiles. The short midpiece of N. hollandicus distinguishes this cacatuine from the two psittacines.

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