The critical role of natural history museums in advancing eDNA for biodiversity studies: a case study with Amazonian fishes.

Ichthyological surveys have traditionally been conducted using whole-specimen, capture-based sampling with varied but conventional fishing gear. Recently, environmental DNA (eDNA) metabarcoding has emerged as a complementary, and possible alternative, approach to whole-specimen methodologies. In the tropics, where much of the diversity remains undescribed, vast reaches continue unexplored, and anthropogenic activities are constant threats; there have been few eDNA attempts for ichthyological inventories. We tested the discriminatory power of eDNA using MiFish primers with existing public reference libraries and compared this with capture-based methods in two distinct ecosystems in the megadiverse Amazon basin. In our study, eDNA provided an accurate snapshot of the fishes at higher taxonomic levels and corroborated its effectiveness to detect specialized fish assemblages. Some flaws in fish metabarcoding studies are routine issues addressed in natural history museums. Thus, by expanding their archives and adopting a series of initiatives linking collection-based research, training and outreach, natural history museums can enable the effective use of eDNA to survey Earth's hotspots of biodiversity before taxa go extinct. Our project surveying poorly explored rivers and using DNA vouchered archives to build metabarcoding libraries for Neotropical fishes can serve as a model of this protocol.

A systematic review of Rhinopetitia Géry (Teleostei, Characiformes, Characidae) with descriptions of four new species and redescription of R. myersi Géry.

Rhinopetitia was known for a long time only by R. myersi, its type species, described from the Ilha do Bananal, Rio Araguaia basin, Tocantins, Brazil. Another recently described species, R. potamorhachia, was considered to differ from R. myersi by having the outer and inner rows of premaxillary teeth with seven to nine cusps, a midlateral dark stripe and a dark humeral blotch (versus outer and inner rows of premaxillary and of maxillary teeth with three to five cusps, and the absence of a dark midlateral stripe). Recently collected specimens from the Araguaia and Tocantins river basins are considered to belong to R. myersi, allowing to better characterize this species which is redescribed herein. Four new species are recognized for the genus, all from Brazil: 1) Rhinopetitia paucirastra, new species, collected from the upper Rio Tocantins drainage, state of Goiás, distinguished from all its congeners mainly by the presence of rudimentary and fewer gill rakers in external row on first gill arch; 2) R. oligolepis, new species originating from the Rio Jamanxim, Rio Tapajós drainage, state of Pará, differing from its congeners by having 4 longitudinal scale rows from dorsal-fin origin to lateral line; 3) R. melanohumeralis, new species, collected from small streams tributaries of the upper Rio Tapajós, and the upper Rio Xingu basins, states of Pará and Mato Grosso, that has the body as deep as in R. paucirastra but has well-developed and more gill rakers in the external row on first branchial arch; and 4) R. nigrofasciata, new species, collected in small tributaries of the upper portions of the Rio Tapajós and Rio Xingu basins, states of Pará and Mato Grosso, with the body narrower than in R. paucirastra and R. melanohumeralis.

A new species of Knodus Eigenmann (Characiformes: Characidae: Stevardiinae) with comments on nuptial tubercles and gill gland in characiform fishes.

Knodus nuptialis n. sp. is described from the Rio Curuá drainage, Rio Xingu basin, Brazil. It can be diagnosed from its congeners by having dentary teeth decreasing gradually in size posteriorly, outer premaxillary teeth row with five cusps, 12-15 branched anal-fin rays and a single humeral spot. The species presents notable sexual dimorphism consisting of densely concentrated nuptial tubercles on head, body, and fins, gill-gland, and bony hooks in the anal fin of mature males. It was found that these sexually dimorphic features are useful and functional in males of the new species only during the reproductive season and after this period, they become atrophied, and eventually disappear. The list of characiform species presenting breeding tubercles is updated and nine species and two genera of the Characidae, Deuterodon and Bryconacidnus, are for the first time reported to have breeding tubercles.

Redescription of Mugil setosus Gilbert 1892 with comments on the occurrence of Mugil curema Valenciennes 1836 in the Pacific Ocean (Teleostei: Perciformes: Mugilidae).

Mugil setosus Gilbert 1892 was originally described by Gilbert based on specimens from Clarion Island, in the western and most remote of the Revillagigedo Islands, about 1,000 km off the western Pacific coast of Mexico. Examination of the type of material and recently collected specimens from Ecuador and Peru, resulted in the redescription provided herein. Diagnostic characters of the species were mainly: tip of the pelvic fin reaching beyond the vertical through the base of the third dorsal-fin spine, the pectoral-fin rays with ii+13-14 rays, the anterodorsal tip of second (soft) dorsal fin uniformly dark, and an external row of larger teeth, and more internally a patch of scattered smaller teeth, visible mainly in adults 150 mm SL. The expansion of geographic distribution of Mugil setosus and occurrence of Mugil curema Valenciennes 1836 in the Pacific Ocean are discussed.

Anatomical, taxonomic, and phylogenetic reappraisal of a poorly known ghost knifefish, Tembeassu marauna (Ostariophysi: Gymnotiformes), using X-ray microcomputed tomography.

A detailed osteological study of the poorly known and critical endangered ghost knifefish, Tembeassu marauna, from the rio Paraná, Brazil, was conducted using X-ray microcomputed tomography (µCT scan). A redescription of the external anatomy was performed, including the unusual presence of a rostral patch of extra teeth on the region of the upper lip anterior to the premaxilla and the prominent anterior fleshy expansions in both upper and lower lips. The newly surveyed characters were included and analyzed in light of a recent morphological data matrix for Gymnotiformes. In spite of some uncertainties that remains as to phylogenetic allocation of the genus, the most probable hypothesis is that Tembeassu is the sister group of a clade that includes Megadontognathus and Apteronotus sensu stricto. The phylogenetic analysis also supports that Tembeassu is considered a valid genus of Apteronotidae. An amended diagnosis for the genus is also provided.

Poor taxonomic sampling undermines nomenclatural stability: A reply to Roxo et al. (2019).

A recent study based on genomic data by Roxo et al. (2019) provided a phylogeny of the Loricariidae, the largest catfish family and second largest Neotropical fish family with approximately 1,000 species. The study represents a valuable and innovative contribution for understanding higher-level relationships within the family. The phylogenetic tree inferred by Roxo et al. (2019) thoroughly corroborates the monophyly and relationships of most currently accepted subfamilies of Loricariidae, based on a fair taxon sampling (nearly 14% of the species in the family) representing most genera of each but one of the subfamilies, the Lithogeninae, the sister-group of the remaining members of the family (Pereira & Reis, 2017; Reis et al., 2017). In addition to a hypothesis of relationships, Roxo et al. (2019) also proposed a series of lower-level taxonomic changes, which are deemed premature considering that the taxonomic sampling of the study targeted higher-level clades, and go against one of the pillars of biological classification: nomenclatural stability (e.g., Heterick & Majer, 2018; Beninger & Backeljau, 2019). Here we (1) discuss implications of inadequate taxonomic sampling as a basis for changes in classification of species; (2) explain why the taxonomic sampling design of Roxo et al. (2019) is inadequate for the proposed nomenclatural changes; and (3) advocate that changes to classifications must be grounded on phylogenies with dense sampling of taxa at the relevant level.

Gephyrocharax machadoi, a new species of Stevardiinae (Characiformes: Characidae) from the Rio Paraguai basin, central Brazil.

Gephyrocharax machadoi, new species, is described from tributaries of the rio Sepotuba, upper rio Paraguai basin, Mato Grosso state, Brazil. The new species is distinguished from its congeners, except Gephyrocharax major, by presenting two modified scales on the ventral caudal-fin lobe (pouch scales): a larger, sexually dimorphic scale, slightly superior and anterior in position, followed by a smaller, posterior and ventrally placed accessory scale (vs. a single modified pouch scale without ventrally placed accessory scales). Gephyrocharax machadoi differs from G. major by presenting a gap (more conspicuous in mature males than in mature females) between the second and third ventral procurrent caudal-fin rays (vs. second and third ventral procurrent caudal-fin rays near each other or fused distally); premaxilla with tricuspid teeth (vs. tetra- to pentacuspid teeth); and body depth 21.5-25.4% of SL (vs. 25.9-36.8% in SL). Spermiogenesis and spermatozoa morphology are also described.

Taxonomic review of Copella (Characiformes: Lebiasinidae) with an identification key for the species.

A taxonomic review of Copella is presented based on the analysis of the type material of all nominal species and extensive material from South American drainages. Six out of ten nominal species are recognized as valid: Copella arnoldi, C. callolepis, C. compta, C. eigenmanni, C. nattereri, and C. vilmae. Copella carsevennensis is a junior synonym of C. arnoldi, C. nigrofasciata and 'Nannostomus' stigmasemion are junior synonyms of C. callolepis, C. metae is junior synonym of C. eigenmanni, and C. meinkeni is junior synonym of C. nattereri. Species of Copella occur in the rio Amazonas and Orinoco basins, and coastal drainages of Guyana, French Guiana, Surinam, and Venezuela. An identification key is provided.

Nematocharax costai Bragança, Barbosa & Mattos a junior synonym of Nematocharax venustus Weitzman, Menezes & Britski (Teleostei: Characiformes: Characidae).

Analysis of morphometric and meristic data from specimens obtained from various river basins along the distribution of Nematocharax venustus revealed intraspecific variation among geographically isolated populations. A comparative analysis of these data with data from the original description of N. costai, in addition to data obtained from the holotype and topotypes of the species, showed broad overlap and absence of morphological features supporting the recognition of two species within Nematocharax. Characters previously used to distinguish N. costai from N. venustus, such as presence of hooks or spinules on dorsal and pelvic fins, number of hooks on anal-fin rays, occurrence of a pink horizontal mark on the caudal peduncle, number of supraneurals, and coloration of the pelvic-fin filament, are not useful to distinguish the two putative species. Thus, N. costai is considered herein as a junior synonym of N. venustus. Presence of a complete lateral line and length of fin-filaments of N. venustus are briefly discussed and a current overview of the distribution of the species is given. In view of the proposed synonymy of a so recently described species and aiming to avoid problems of similar nature in the future, we also take the opportunity to reinforce herein the importance of the examination of large population samples and of intraspecific variation of secondary sexual features.

Taxonomic review of the species of Mugil (Teleostei: Perciformes: Mugilidae) from the Atlantic South Caribbean and South America, with integration of morphological, cytogenetic and molecular data.

Analysis of morphological, molecular and cytological data helped to define and more precisely characterize the species of Mugil from the Atlantic coasts of South Caribbean and South America, allowing a correction of prior misidentifications and distributional ranges. A new species from Venezuela is described and all the species from the area are redescribed. It is demonstrated that the apparent similarity in morphological traits, which contradicted the results from recent molecular studies, is the result of the misuse of traditional morphological characters, and thus both the molecular and cytological data instead are congruent with the morphological differences that are found among mullet species. The presence of Mugil hospes Jordan & Culver in the western south Atlantic is refuted based on the comparison of type material of this species with specimens from this area that also indicated a very significant morphological difference, what on the other hand justifies the recognition of these specimens as Mugil brevirostris (Ribeiro). The distribution of Mugil incilis Hancock is restricted and the similarities among the species formerly depicted in a prior dendrogram is modified following the inclusion of recently obtained molecular data for Mugil curvidens Valenciennes.

A new species of Knodus (Characiformes: Characidae: Stevardiinae) from the rio Aripuanã, rio Madeira basin, Brazil

A new species of the characid genus Knodus is described from the rio Aripuanã (rio Madeira basin). It can be distinguished from its congeners by its very low body depth, the presence of tri- to pentacuspid teeth on the outer premaxillary series, with the median cuspid larger than the lateral ones, the teeth of the inner premaxillary series pentacuspid, distinctly larger than those of the outer series, the maxillary teeth tri- to pentacuspid, with the median cusp slightly larger than the lateral ones, the four anteriormost dentary teeth pentacuspid, the smaller posterior teeth tri- to pentacuspid, a complete lateral line with 36-38 scales, 3 longitudinal scale series from pelvic fin origin to lateral line, and 11-12 circumpeduncular scales. The new species is also compared to incertae sedis species of 'Bryconamericus' from northern South America since phylogenetic studies suggest a closer relationship of those species with Knodus.(AU)

Uma nova espécie de um caracídeo do gênero Knodus é descrita do rio Aripuanã (bacia do rio Madeira). Esta espécie se distingue de suas congeneres por possuir o corpo muito estreito, a presença de dentes tri- a pentacúspides na série externa do pré-maxilar, com a cúspide mediana ligeiramente maior que as cúspides laterais, dentes da série interna pentacúspide, distintamente maiores que os da série externa, dentes do maxilar tri- a pentacúspides com a cúspode central discretamente maior que as laterais, os quatro dentes mais anteriores do dentário pentacuspidados, os dentes mais posteriores tricuspidados a pentacuspidados, a linha lateral complete com 36-38 escamas, 3 séries longitudinais de escamas entre a origem da nadadeira pélvica e a linha lateral, e 11-12 séries longitudinais de escamas circumpedunculares. A nova espécie é comparada com espécies incertae sedis de 'Bryconamericus' do norte da América do Sul uma vez que estudos filogenéticos sugerem a próxima relação daquelas espécies com Knodus.(AU)

Taxonomic revision of Galeocharax (Characiformes: Characidae: Characinae)

The taxonomy of Galeocharax, a genus of freshwater fishes distributed in most South American cis-Andean river systems, except the rio Negro, rio São Francisco and rio Xingu basins and the eastern drainages of Brazil, is herein revised. A total of 1229 specimens were examined from which 680 had meristic and morphometric data taken. Osteological and morphological features were also examined through radiographs, scanning electron microscopy and in cleared and stained specimens. Three of the four species previously considered as valid are herein recognized: Galeocharax humeralis from rio Paraguay and lower rio Paraná basins; Galeocharax goeldii, from rio Madeira basin, with records of geographical distribution expanded to the río Madre de Dios, río Beni, rio Mamoré and rio Guaporé basins and Galeocharax gulo, which is widespread throughout rio Amazonas, río Orinoco, rio Tocantins, and upper rio Paraná basins. Galeocharax knerii is herein considered a junior synonym of Galeocharax gulo. A key to species of Galeocharax is presented.(AU)

A taxonomia de Galeocharax, um gênero de peixes de água doce distribuído nos principais sistemas hídricos da América do Sul cisandina, com exceção da bacia do rio Negro, do rio São Francisco, do rio Xingu e das drenagens litorâneas do leste, é aqui revisada. Foram examinados 1229 exemplares, dos quais 680 tiveram dados merísticos e morfométricos tomados. Características osteológicas e morfológicas também foram examinadas por meio de radiografias, microscopia eletrônica de varredura e exemplares diafanizados. Três das quatro espécies previamente consideradas válidas são reconhecidas: Galeocharax humeralis das bacias do rio Paraguai e baixo rio Paraná; Galeocharax goeldii da bacia do rio Madeira, com registros de distribuição geográfica ampliados para as bacias dos rios Madre de Dios, Beni, Mamoré e Guaporé, e Galeocharax gulo amplamente distribuída nas bacias dos rios Amazonas, Orinoco, Tocantins e alto rio Paraná. Galeocharax knerii foi considerada sinônimo júnior de Galeocharax gulo. Uma chave de identificação para as espécies de Galeocharax é apresentada.(AU)

Restricted-range fishes and the conservation of Brazilian freshwaters.

BACKGROUND: Freshwaters are the most threatened ecosystems on earth. Although recent assessments provide data on global priority regions for freshwater conservation, local scale priorities remain unknown. Refining the scale of global biodiversity assessments (both at terrestrial and freshwater realms) and translating these into conservation priorities on the ground remains a major challenge to biodiversity science, and depends directly on species occurrence data of high taxonomic and geographic resolution. Brazil harbors the richest freshwater ichthyofauna in the world, but knowledge on endemic areas and conservation in Brazilian rivers is still scarce. METHODOLOGY/PRINCIPAL FINDINGS: Using data on environmental threats and revised species distribution data we detect and delineate 540 small watershed areas harboring 819 restricted-range fishes in Brazil. Many of these areas are already highly threatened, as 159 (29%) watersheds have lost more than 70% of their original vegetation cover, and only 141 (26%) show significant overlap with formally protected areas or indigenous lands. We detected 220 (40%) critical watersheds overlapping hydroelectric dams or showing both poor formal protection and widespread habitat loss; these sites harbor 344 endemic fish species that may face extinction if no conservation action is in place in the near future. CONCLUSIONS/SIGNIFICANCE: We provide the first analysis of site-scale conservation priorities in the richest freshwater ecosystems of the globe. Our results corroborate the hypothesis that freshwater biodiversity has been neglected in former conservation assessments. The study provides a simple and straightforward method for detecting freshwater priority areas based on endemism and threat, and represents a starting point for integrating freshwater and terrestrial conservation in representative and biogeographically consistent site-scale conservation strategies, that may be scaled-up following naturally linked drainage systems. Proper management (e. g. forestry code enforcement, landscape planning) and conservation (e. g. formal protection) of the 540 watersheds detected herein will be decisive in avoiding species extinction in the richest aquatic ecosystems on the planet.

A taxonomic review of the species of charax scopoli, 1777 (teleostei: characidae: characinae) with description of a new species from the rio negro bearing superficial neuromasts on body scales, amazon basin, Brazil

The species of Charax were reviewed by Lucena (1987) who a little later (1989) added three new species to the genus. Examination of recently collected specimens from museum collections documented the more extensive geographic distribution for most species and revealed the existence of a new species in which superficial neuromasts were discovered and are herein described. These skin structures were also detected in C. metae Eigenmann, 1922. Anal- and pelvic-fin hooks previously observed only on the anal-fin of one species of the genus are described herein in some other species. Charax unimaculatus Lucena is considered a junior synonym of C. michaeli Lucena. All the species are redescribed and the new species described.

As espécies de Charax foram revistas por Lucena (1987), que pouco depois (1989) acrescentou três espécies novas ao gênero. O exame de exemplares recentemente coletados e depositados em coleções de vários museus possibilitaram a extensão da distribuição geográfica da maioria das espécies e revelou a existência de uma espécie nova, na qual foram descobertos neuromastos superficiais, caracterizados neste trabalho. Estas estruturas da pele também foram detectadas em Charax metae Eigenmann, 1922. Ganchos das nadadeiras anal e pélvica, anteriormente observados apenas na nadadeira anal de uma das espécies do gênero, são agora descritos em algumas outras espécies. Charax unimaculatus Lucena é considerada sinônimo júnior de C. michaeli Lucena Todas as espécies são redescritas, juntamente com a descrição da espécie nova.

Two new species and a review of the inseminating freshwater fish genus Monotocheirodon (Characiformes: Characidae) from Peru and Bolivia

Two new species of inseminating freshwater fishes of the genus Monotocheirodon, family Characidae, are described from Peru. Males and females of both new species have an external, visually obvious urogenital papilla that was not detected in the females in previous studies, with this longer in males, which use it as an inseminating organ. A third inseminating species from Bolivia, Monotocheirodon pearsoni, unstudied in any detail since its original description in 1942, is redescribed. This latter species lacks an inseminating. Monotocheirodon is redescribed, its phylogenetic relationships are briefly discussed and it is suggested that it is possibly related to the stevardiin genera Ceratobranchia, Ohonocheirodus, and Odontostoechus.

New species of miniature fish from Marajo Island, Para, Brazil, with comments on its relationships (Characiformes: Characidae)

A new miniature species of the family Characidae from Marajó Island, Pará State, Brazil is described and assigned to the genus Tyttobrycon. The new species can be distinguished from its congeners primarily by having multicuspid teeth on jaws, and additionally by having the combination of 5-7 premaxillary teeth, dorsal fin only with scattered melanophores and the presence of an adipose fin.

Uma espécie nova miniatura da família Characidae da Ilha do Marajó, Pará, Brasil, é descrita e atribuída ao gênero Tyttobrycon. A espécie nova pode ser diferenciada dos congêneres principalmente por ter dentes multicuspidados nas maxilas e adicionalmente por ter a combinação de 5-7 dentes no pré maxilar, nadadeira dorsal apenas com pequenos cromatóforos escuros dispersos e presença de nadadeira adiposa.

A new genus and two new species of Stevardiinae (Characiformes: Characidae) with a hypothesis on their relationships based on morphological and histological data

Lepidocharax, new genus, and Lepidocharax diamantina and L. burnsi new species from eastern Brazil are described herein. Lepidocharax is considered a monophyletic genus of the Stevardiinae and can be distinguished from the other members of this subfamily except Planaltina, Pseudocorynopoma, and Xenurobrycon by having the dorsal-fin origin vertically aligned with the anal-fin origin, vs. dorsal fin origin anterior or posterior to anal-fin origin. Additionally the new genus can be distinguished from those three genera by not having the scales extending over the ventral caudal-fin lobe modified to form the dorsal border of the pheromone pouch organ or to represent a pouch scale in sexually mature males. In this paper, we describe these two recently discovered species and the ultrastructure of their spermatozoa.

O trabalhocontém a descrição de um gênero novo, Lepidocharax, e duas espécies novas, Lepidocharax diamantina e L. burnsi do leste do Brasil. Lepidocharax é considerado um gênero monofilético de Stevardiinae e pode ser diferenciado dos outros membros desta subfamília, exceto Planaltina, Pseudocorynopoma e Xenurobrycon principalmente porque as duas espécies nele incluídas têm a origem da nadadeira dorsal verticalmente alinhada com a origem da nadadeira anal, vs. origem da nadadeira dorsal anterior ou posterior a origem da nadadeira anal. Adicionalmente, o novo gênero pode ser distinguido dos mesmos três gêneros por não possuir escamas modificadas localizadas no lobo ventral da nadadeira caudal, formando a borda dorsal do órgão produtor de feromônio ou uma escama modificada do mesmo órgão em machos sexualmente maduros. A ultraestrutura dos espermatozóides das duas espécies recentemente descobertas também é descrita.

Systematics of the neotropical fish subfamily Glandulocaudinae (Teleostei: Characiformes: Characidae)

The systematics of the Glandulocaudinae is reviewed in detail and justification for the recognition of the group as a subfamily is discussed. The subfamily Glandulocaudinae consists of three genera: Lophiobrycon with one species plesiomorphic in some anatomical features but some others exclusively derived relative to the species in the other genera; Glandulocauda with two species intermediate in phylogenetic derivation; and Mimagoniates with seven species (one new), all more phylogenetically derived concerning their pheromone producing caudal-fin organs and with other anatomical characters presumably more derived than in the species of the other genera. Glandulocauda melanogenys Eigenmann, 1911, is considered a junior synonym of Hyphessobrycon melanopleurus Ellis, 1911. A replacement name, Glandulocauda caerulea Menezes & Weitzman, is proposed for G. melanopleura Eigenmann, 1911. Gland cells found in the caudal-fin organs of all species are histologically indistinguishable from club cells and probably secrete a pheromone during courtship. The club cells are associated with somewhat modified to highly derived caudal scales forming a pheromone pumping organ in the more derived genera and species. This subfamily is distributed in freshwaters of eastern and southern Brazil, Paraguay, and northeastern Uruguay.(AU)

A sistemática de Glandulocaudinae é revista e a justificativa para o reconhecimento do grupo como subfamília discutida. A subfamília Glandulocaudinae consiste de três gêneros: Lophiobrycon, com uma espécie plesiomórfica com relação a alguns caracteres anatômicos, mas outros derivados e exclusivos em relação às espécies dos outros dois gêneros; Glandulocauda, com duas espécies intermediárias quanto à condição dos caracteres derivados; e Mimagoniates, com sete espécies (uma nova), todas filogeneticamente mais avançadas quanto às características dos órgãos da nadadeira caudal produtores de feromônio e outras características anatômicas presumivelmente mais derivadas do que nas espécies dos outros gêneros. Glandulocauda melanogenys Eigenmann, 1911, é considerado sinônimo junior de Hyphessobrycon melanopleurus Ellis, 1911. O nome Glandulocauda caerulea Menezes & Weitzman, é proposto em substiutição para G. melanopleura Eigenmann, 1911. Células glandulares encontradas nos órgãos da caudal são histologicamente indistinguíveis de "células club" e provavelmente secretam algum tipo de feromônio durante a corte. As "células club" são associadas a escamas da caudal pouco ou inteiramente modificadas e fazendo parte dos órgãos bombeadores de feromônio nas espécies e gêneros mais derivados. Esta subfamília distribui-se em ambientes de água doce do leste e sul do Brasil, no Paraguai e nordeste do Uruguai.(AU)

Internal fertilization, testis and sperm morphology in glandulocaudinae fishes (Teleostei: Characidae: Glandulocaudinae).

In this report, the gonads of 32 glandulocaudine species, representing 18 genera, are compared with 11 outgroup characiform species. Through the presence of spermatozoa within the ovarian cavity, internal fertilization of the female is confirmed for the 16 genera for which mature ovaries were available. No outgroup ovary studied contains spermatozoa. All mature glandulocaudine testes have a large portion of the posterior testis, which is devoid of developing germ cells and spermatocysts (aspermatogenic), devoted to sperm storage, with the degree of partitioning in that region varying greatly within the group. All outgroup species examined have spermatozoa with spherical nuclei. With the exception of the species of the genus Planaltina, which also have spherical nuclei, all glandulocaudines have elongated nuclei, which vary among the species from 3.6 µm to 31.6 µm in length. Distinct sperm packets (spermatozeugmata) are formed in five genera by two different methods. In the genera Xenurobrycon, Tyttocharax, and Scopaeocharax, all of the tribe Xenurobryconini, the spermatozeugmata are formed within the spermatocysts and released fully formed. In all genera of the tribe Glandulocaudini, which includes Glandulocauda and Mimagoniates, loose spermatozoa are released which cluster into spermatozeugmata within the posterior storage areas. These morphological specializations are discussed within a phylogenetic framework as adaptations for internal fertilization and are hypothesized to be independently derived. © 1995 Wiley-Liss, Inc.