Trophic morphology and diet of the endangered fish Tlaloc hildebrandi (Cyprinodontiformes: Profundulidae) / Morfología trófica y dieta de Tlaloc hildebrandi (Cyprinodontiformes: Profundulidae), especie amenazada de extinción

Abstract Introduction: Tlaloc hildebrandi is a freshwater killifish, endemic to Southern Mexico and under threat of extinction; the knowledge of the trophic morphology and diet is needed by conservation managers. Objective: To analyse and describe the anatomy of the visceral skeleton, visceral musculature, digestive tract and its adjoining glands of T. hildebrandi; as well as its diet. Methods: We performed the trophic anatomy on 20 adult specimens of both sexes, through manual dissection; as well as gut content analysis in 60 individuals to describe the diet. Results: As notable characters of the visceral skeleton of T. hildebrandi we found the posterior notch of the premaxillary, the presence of the "coronoid cartilage", the tricuspid shape of the gill rakers of the first branchial arch, and the presence of the coronomeckelian bone; some outstanding characters of the visceral musculature are the origin of the retractor dorsalis muscle from the first four vertebral centra, and the division of the pharyngoclavicularis externus muscle into two sections. The notable characters of the digestive tube are the absence of stomach and pyloric caeca, and the presence of the "intestinal valve". Insects (IVI = 66.6 %) and ostracods (13 % IVI) were the dominant prey items of the Tlaloc hildebrandi diet; larvae and adults of the family Chironomidae were the most dominant insects in the diet (53 % IVI). Conclusions: The organization of the digestive system of T. hildebrandi corresponds to the general morphologic pattern of the Cyprinodontiformes; however, we register as new information for these fish, the presence of the "coronoid cartilage" and the "intestinal valve". The structures of the trophic morphology and the components of the diet, confirms us that T. hildebrandi is a carnivorous-insectivorous fish.

Resumen Introducción: Tlaloc hildebrandi es un killi de agua dulce, endémico del sur de México y bajo amenaza de extinción; el conocimiento de la morfología trófica y la dieta son necesarios para los administradores de la conservación. Objetivo: Analizar y describir la anatomía del esqueleto visceral, la musculatura visceral, el tracto digestivo y las glándulas adyacentes de T. hildebrandi; así como los componentes de su dieta. Métodos: Mediante la técnica del descarnado manual, realizamos la descripción de la anatomía trófica en 20 especímenes adultos de ambos sexos, y el análisis del contenido estomacal en 60 individuos para describir la dieta. Resultados: Como caracteres sobresalientes del esqueleto visceral de T. hildebrandi está la escotadura posterior del premaxilar, la presencia del "cartílago coronoides", la forma tricúspide de las branquiespinas del primer arco branquial y la presencia del hueso coronomeckeliano; como caracteres de la musculatura visceral sobresalen el origen del músculo retractor dorsalis de los cuatro primeros centros vertebrales, y la división del músculo pharyngoclavicularis externus en dos secciones. Los caracteres notables del tubo digestivo son la ausencia de estómago y de ciegos pilóricos y la presencia de la "válvula intestinal". Los insectos (IVI = 66.6 %) y ostrácodos (13 % IVI) fueron los componentes dominantes de la dieta de T. hildebrandi; particularmente las larvas y adultos de la familia Chironomidae fueron los insectos más abundantes en la dieta (53 % IVI). Conclusiones: La organización del sistema digestivo de T. hildebrandi corresponde al patrón morfológico general de los Cyprinodontiformes, sin embargo, se registra como nueva información para estos peces, la presencia del cartílago coronoides y la válvula intestinal. Las estructuras de la morfología trófica y los componentes de la dieta nos confirman que T. hildebrandi es un pez carnívoro-insectívoro.

Profundulus chimalapensis, una nueva especie de pez ciprinodóntido (Cyprinodontiformes: Profundulidae) del Río Coatzacoalcos, México / Profundulus chimalapensis, a new species of cyprinodontid fish (Cyrpinodontiformes: Profundulidae) from the Coatzacoalcos River, Mexico

Introducción: El género Profundulusse considera endémico de la región Mesoamericana, desde el centro-sur del estado de Guerrero hasta la parte central de Honduras, habita principalmente en las cuencas superiores y cabeceras de los ríos de la vertiente Pacífica, con una menor diversidad en la Atlántica. Objetivo: En este trabajo, basado en la comparación morfológica, proponemos una nueva especie, Profundulus chimalapensis sp. nov., confinada a México. Métodos: Se recolectaron ejemplares en los tributarios de la cuenca superior del Río Coatzacoalcos y se depositaron en colecciones de referencia. Se tomaron datos morfológicos (conteos, mediciones y características esqueléticas) y se compararon con especies relacionadas. Resultados: La nueva especie se diferencia de sus congéneres por tener la siguiente combinación de caracteres: 12-15 radios en la aleta dorsal, 14-17 radios anales y 13-16 pectorales; banda de color oscuro en los costados; aleta anal con un margen distal claro; margen anterior y posterior del proceso alveolar del premaxilar cóncavo; procesos epióticos cortos y anchos; base de la aleta anal grande, su longitud igual o mayor a la del pedúnculo caudal. Conclusión: La descripción de esta nueva especie de Profundulus de la vertiente atlántica del Istmo de Tehuantepec, sugiere que esta área ha sido un centro de especiación para el género.

Introduction: The genus Profundulus is considered a Mesoamerican endemic fish which spreading occurs, from the center-south of the state of Guerrero, Mexico, to the central part of Honduras. They mainly inhabit the upper basins and headwaters of the rivers of the Pacific slope, with less diversity in the Atlantic. Objective: This paper, based on morphological comparison we propose a new species, Profundulus chimalapensis sp. nov., confined to Mexico. Methods: Specimens were collected in the tributaries of the upper basin of the Coatzacoalcos River and were deposited in scientific collections. Morphological data (counts, measurements and skeletal features) were taken and compared with related species. Results: The new species differs from its congeners by having the following combination of characters: 12-15 dorsal-fin rays, 14-17 anal- and 13-16 pectoral-fin rays; dark colored stripe on the flanks; anal fin with a clear distal margin; anterior and posterior margin of the alveolar process of the premaxilla concave; pair of short and wide epiotic processes; and large anal fin base, equal to or greater than length of caudal peduncle. Conclusion: The description of this new Profundulus species from Atlantic side of the Isthmus of Tehuantepec suggests that this area has been a center of speciation for the genus.

Edad y crecimiento del pez Fundulus lima (Cyprinodontiformes: Fundulidae) en el río La Purísima, Baja California Sur, México

Introducción: El estudio de la edad y crecimiento en peces es un prerrequisto básico para generar información sobre la longevidad, tasas de crecimiento y estructura poblacional, entre otros atributos, todos ellos para la aplicación de políticas enfocadas en el manejo y conservación poblacional. Evaluamos estos atributos en la sardinilla peninsular en peligro de extinción (Fundulus lima) que habita los oasis de Baja California Sur, con la perspectiva de generar información base de este pez endémico. Objetivo: Determinar la edad, crecimiento somático y estructura de las poblaciones de F. lima en una de las cuencas hidrológicas más importantes de su ámbito de distribución. Métodos: De octubre 2002 a julio 2004 se muestreó F. lima en 4 sitios del río La Purísima por métodos de captura pasiva y activa. Los individuos fueron medidos y pesados en el campo, donde submuestras fueron seleccionadas para el análisis de la edad en escamas. Todos los individuos fueron liberados vivos a sus sitios originales de captura. Estimamos edad, relaciones longitud-peso y longitud-longitud, y estructura poblacional, basados en 802 individuos examinados por técnicas no invasivas. Resultados: El crecimiento somático de F. lima fue alométrico positivo (b > 3.0) en todas las poblaciones estudiadas, con valores de "b" variando de 3.126 (Ojo de Agua) a 3.420 (El Pilón). Tres clases de edad (0, 1 y 2 años) fueron reconocidas, de las cuales la edad 1 fue la más frecuente (49 %). El método polimodal basado en datos de frecuencia de tallas identificó tres clases de edad. Longitudes totales retrocalculadas en edades previas basadas en incrementos de crecimiento en escamas fue entre 38.61 y 68.81 mm para individuos de 1 y 2 años, respectivamente. Conclusiones: Tres clases de edad (0, 1 y 2 años) fueron determinadas para las poblaciones de F. lima en el río La Purísima, basadas en la interpretación de anillos de crecimiento anual en escamas y en la distribución modal de frecuencia de tallas observadas. Todas las poblaciones examinadas demostraron un crecimiento alométrico positivo (b > 3.0).

Introduction: The study of age and growth in fishes is a basic prerequiste to generate information on longevity, growth rates, population structure, among other attributes, all of them for the application of policies focused on the population management and conservation. We evaluated these attributes in the endangered Baja California killifish (Fundulus lima) that inhabits the oases of Baja California Sur, Mexico, with the perspective of generating base information for the conservation of this endemic fish. Objective: To determine the age, somatic growth and population structure of F. lima in one of the most important hydrological basins of its distribution range. Methods: From October 2002 to July 2004 we sampled for F. lima in 4 sites of La Purisima river by using passive and active collection methods. Individuals were measured and weighed in the field, where subsamples were selected for analysis of age based on scales. All of the individuals were released alive to its original sites of capture. We assessed the age, length-weight and length-length relationships and population structure on the basis of 802 individuals examined by non-invasive techniques. Results: The somatic growth of F. lima was of positive allometric type (b > 3.0) in all the populations studied, with values of "b" ranging from 3.126 (Ojo de Agua) to 3.420 (El Pilón). Three age class (0, 1 and 2 years old) were recognized, of which the age 1 year was the most frequent (49 %). The polimodal method based on length-frequency data also identified three age classes. Back-calculated total lengths at previous ages based in increments of growth in scales were 38.61 and 68.81 mm for individuals of 1 and 2 years, respectively. Conclusions: Three age classes (0, 1 and 2 years old) were determined for the populations of F. lima in La Purisima river, based on the interpretation of annual growth rings in scales and in the modal distribution of observed length frequencies. All the populations examined showed a positive allometric growth (b > 3.0).

Review of the family Rivulidae (Cyprinodontiformes, Aplocheiloidei) and a molecular and morphological phylogeny of the annual fish genus Austrolebias Costa 1998

The family Rivulidae is the fourth most diverse clade of Neotropical fishes. Together with some genera of the related African family Nothobranchiidae, many rivulids exhibit a characteristic annual life cycle, with diapausing eggs and delayed embryonic development, which allows them to survive in the challenging seasonal ponds that they inhabit. Rivulidae also includes two species known as the only the self-fertilizing vertebrates and some species with internal fertilization. The first goal of this article is to review the systematics of the family considering phylogenetic relationships and synapomorphies of subfamilial clades, thus unifying information that is dispersed throughout the literature. From this revision, it is clear that phylogenetic relationships within Rivulidae are poorly resolved, especially in one of the large clades that compose it, the subfamily Rivulinae, where conflicting hypotheses of relationships of non-annual and annual genera are evident. The second goal of this work is to present an updated phylogenetic hypothesis (based on mitochondrial, nuclear, and morphological information) for one of the most speciose genus of Rivulidae, Austrolebias. Our results confirm the monophyly of the genus and of some subgeneric clades already diagnosed, but propose new relationships among them and their species composition, particularly in the subgenus Acrolebias.

a familia Rivulidae es el cuarto clado más diverso dentro de los peces Neotropicales. Junto con algunos géneros de la familia Nothobranchiidae, muchos rivulidos presentan un característico ciclo de vida anual, con huevos resistentes a la desecación y embriones con diapausas que les permiten sobrevivir en los ambientes estacionales donde habitan. Los Rivulidae presentan también dos especies consideradas como los únicos vertebrados hermafroditas suficientes y algunas especies con inseminación interna. El primer objetivo de este artículo es actualizar la sistemática de la familia considerando las relaciones filogenéticas y las sinapomorfías de los clados que la componen, reuniendo información que se encuentra dispersa en la literatura. De esta revisión surge que las relaciones filogenéticas dentro de Rivulidae están todavía sin resolver, especialmente en uno de los grandes clados que la componen, la subfamilia Rivulinae, donde relaciones conflictivas entre géneros anuales y no anuales son evidentes. El segundo objetivo de este trabajo es presentar una hipótesis filogenética, basada en datos morfológicos, mitocondriales y nucleares, de uno de los géneros más diversos de la familia, el género Austrolebias. Nuestros resultados confirman la monofilia del género y de algunos clados subgenéricos previamente definidos, y propone nuevas relaciones entre ellos, particularmente de las especies del subgénero Acrolebias(AU)

Metaplasia in swim bladder epithelium of the endangered annual fish Austrolebias nigrofasciatus (Cyprinodontiformes: Rivulidae) results in inadequate swimming and delayed growth

Laboratory breeding of annual fish is often impaired when fish fail to inflate the swim bladder after hatching. These fish, which are known as belly-sliders, cannot swim in the water column, moving mainly on the bottom of water. Therefore, we wanted to understand the factors underlying failed inflation of swim bladder in the annual fish Austrolebias nigrofasciatus and determine the implications of this condition on the growth of juveniles. To identify how the initial filling of the gaseous vesicle is compromised, and to determine the implications of this condition on juvenile growth, we analysed the histology of the swim bladder of both normal swimming fish and belly-sliders during their initial growth phase (first month of life). In addition to slower growth rate compared with fish swimming normally, we found that belly-sliders have metaplasia characterized by epithelium peeling of the swim bladder and the presence of haemorrhage. Based on our findings, it can be concluded that metaplasia of the swim bladder is recurrent in annual fish bred in the laboratory, which explains the large number of unviable larvae. However, its causes remain unknown.(AU)

A criação de peixes anuais é frequentemente comprometida pela ocorrência de peixes que não inflam a vesícula gasosa após a eclosão. Estes peixes não têm capacidade de se manter e nadar na coluna d'água e se locomovem no fundo da água, por isso são chamados belly-sliders. Portanto, objetivamos entender os fatores subjacentes à inflação falhada da bexiga natatória no peixe anual Austrolebias nigrofasciatus e determinar as implicações dessa condição no crescimento de juvenis. Para identificar de que forma o enchimento inicial da vesícula gasosa é comprometido, e determinar as implicações desta condição no crescimento dos juvenis, analisamos histologicamente a vesícula gasosa de peixes com natação normal e de belly-sliders durante o crescimento inicial (primeiro mês de vida). Verificamos que os belly-sliders apresentam metaplasia com descamação do epitélio da vesícula gasosa e presença de hemorragia, além de apresentar crescimento mais lento em relação aos peixes com natação normal. Concluímos que a incidência desta patologia é um problema recorrente e de grande relevância para a manutenção de peixes anuais em laboratório, sendo responsável por uma grande quantidade de juvenis inviáveis, no entanto suas causas permanecem desconhecidas.(AU)

First record of a male of Kryptolebias hermaphroditus Costa, 2011 (Cyprinodontiformes: Cynolebiidae)

During an ichthyological survey in September 2015 at the Ceará-Mirim River estuary, Rio Grande do Norte State, northeastern Brazil, we collected a male of Kryptolebias hermaphroditus , a cynolebiid species that had been previously described as containing exclusively self-fertilizing hermaphrodites. This is the first record of a male in this species, over 140 years after the discovery of the mangrove rivulid species from Brazil. Our discovery reinforces the need for more studies in K. hermaphroditus , as well as the potential of this species as a model for evolutionary studies due to its unique mating system.(AU)

Durante uma amostragem ictiológica em setembro de 2015 no estuário do rio Ceará-Mirim, Estado do Rio Grande do Norte, Nordeste do Brasil, nós coletamos um macho de Kryptolebias hermaphroditus , uma espécie de rivulídeo que foi descrita como contendo apenas hermafroditas auto-fertilizantes. Este é o primeiro registro de um macho dessa espécie, mais de 140 anos depois da descoberta das espécies de rivulídeos de manguezais do Brasil. Nossa descoberta reforça a necessidade de mais estudos em K . hermaphroditus , assim como, o potencial desta espécie como um modelo para estudos evolutivos devido ao seu sistema reprodutivo único.(AU)

Geographical distribution and evaluation of mosquito larvivorous potential of Aphanius dispar (Rüppell), a native fish of Gujarat, India.

Background & objectives: Certain indigenous fish can play potential role in vector control. The study recorded distribution of Aphanius dispar (Rüppell) in its native habitats in Gujarat, India and evaluated its larval propensity for Indian mosquito vectors. Methods: Fishes were surveyed in various districts of Gujarat and samples were collected from coastal habitats and were identified to species. Physicochemical properties of water samples were analysed in the laboratory. Five laboratory acclimatized adult fish (2.2–3 cm) released in chlorine-free water in glass jars were provided with 500 larvae daily. Five replicates were run. Daily consumption of I to IV instars of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus was recorded separately for three days. Anopheles stephensi larval control using the fish in industrial tanks (9–180 m2) was evaluated. Results & conclusion: The fish was found in large numbers in estuarine waters, rivulets, backwaters and small dams in the districts of Kutch, Jamnagar, Patan and Anand in Gujarat. Mean daily consumption of larvae in laboratory was as follows: An. stephensi 128 ± 0.2 to 204 ± 6; Cx. quinquefasciatus 24 ± 4 to 58 ± 10; Ae. aegypti 43 ± 5 to 68 ± 2. In water tanks, A. dispar reduced 93% larval counts by Day 7 and 98% by Day 21 (p <0.01), showing high larval propensity. A consolidated information on bionomics, tolerance and physicochemical properties of water taken from natural habitats of A. dispar indicate its high larvivorous potential. Large-scale evaluation is warranted to evaluate its potential in vector-borne disease control.

Feeding ecology of Rivulus luelingi (Aplocheiloidei: Rivulidae) in a Coastal Atlantic Rainforest stream, southern Brazil

Feeding habits of the killifish Rivulus luelingi collected in a black water stream of the Coastal Atlantic Rainforest in southern Brazil were investigated. Eight samplings were made between April 2003 and January 2004. The diet, assessed through a similarity matrix with the estimated contribution values of food items, included microcrustaceans, aquatic immature insects (larvae and pupae), aquatic adult insects, terrestrial insects, insect fragments, spiders, and plant fragments. Differences in the diet according to temporal variations (months) were registered, but changes related with size classes evaluated and high/low precipitation period were not observed. The species presented an insectivorous feeding habit, and its diet in the studied stream was composed of autochthonous (mainly aquatic immature insects) and allochthonous (mainly insect fragments) material.

Neste estudo foram investigados os hábitos alimentares do peixe anual Rivulus luelingi em um riacho de água escura da Floresta Atlântica Costeira do Sul do Brasil. Oito amostragens foram realizadas entre abril de 2003 e janeiro de 2004. A dieta, avaliada através de uma matriz de similaridade com os valores de contribuição estimados para os itens alimentares, inclui microcrustáceos, insetos imaturos aquáticos, insetos aquáticos e terrestres, fragmentos de insetos, aranhas e fragmentos de plantas. Diferenças relacionadas ao período amostral (meses) foram registradas, mas mudanças na dieta em função das classes de tamanho avaliadas e o período de alta/baixa precipitação não foram observadas. A espécie apresentou hábito alimentar insetívoro, e sua dieta no riacho estudado foi composta por itens autóctones (principalmente insetos imaturos aquáticos) e alóctones (principalmente fragmentos de insetos).

Reproductive behavior in the annual fish Austrolebias reicherti Loureiro & García 2004 (Cyprinodontiformes: Rivulidae)

Annual fishes inhabit temporary ponds that dry out seasonally and the adaptations to survive this extreme condition include high metabolic rates and an elaborate courtship behavior which ends in the deposition of drought-resistant eggs, capable of going through diapause stages in the substrate. The pronounced sexual dimorphism that these fishes show suggests that sexual selection could play a key role in the differentiation, speciation and evolution of this diverse group of fishes. However, there is scarce information about their reproductive isolation mechanisms, including detailed analysis of courtship signals. Herein, we analyzed, qualitatively and quantitatively, the reproductive behavior of Austrolebias reicherti. The behavioral units recognized in this study correspond with the previous analysis of other species of annual fishes. The most frequent unit in males was the lateral display where specific morphological and coloration patterns are displayed to the female. The female's high relative frequency and time of quietness suggest that this unit may have an evaluation role during courtship. In addition to visual displays during courtship, males perform vibrations of the dorsal and anal fins as well as body undulations; these indicate that mechanical signals may be important for attracting females. Our results support the hypothesis of multimodal signals. The conservation of behavioral patterns in courtship displays within Austrolebias suggests that species-level recognition and the barrier to hybridization may not occur at this stage, unless quantitative differences can be identified.

Os peixes anuais habitam charcos temporários que secam sazonalmente, e as adaptações para sobreviver a estas condições extremas incluem altas taxas metabólicas e um elaborado comportamento de cortejo, que culmina com a deposição de ovos resistentes à dessecação dentro do substrato, os quais são capazes de atravessar estádios de diapausa. O pronunciado dimorfismo sexual apresentado por estes peixes indica que a seleção sexual poderia estar exercendo um papel chave na diferenciação, especiação e evolução deste grupo diverso de peixes. Entretanto, informações acerca dos mecanismos de isolamento reprodutivo, incluindo uma análise detalhada dos sinais de cortejo, são escassas. Neste artigo é analisado o comportamento reprodutivo de Austrolebias reicherti de maneira qualitativa e quantitativa. As unidades comportamentais utilizadas neste estudo correspondem àquelas incluídas em análises anteriores de outras espécies de peixes anuais. A unidade comportamental mais freqüentemente observada em machos foi a "exibição lateral", onde características espécie-específicas da morfologia e colorido são exibidas para a fêmea. A alta proporção de ocorrência (em freqüência e tempo) do comportamento de "quietude" nas fêmeas indica que o mesmo poderia desempenhar um papel avaliativo durante o cortejo. Outros sinais além dos visuais estão envolvidos no cortejo dos peixes anuais; os machos vibram as nadadeiras dorsal e anal e ondulam o corpo, o que indica que os sinais mecânicos poderiam ter influência na atração das fêmeas. Estes resultados conferem suporte à hipótese de sinais multimodais, recentemente proposta. Os movimentos de cortejo altamente conservados dentro de Austrolebias indicam que o reconhecimento específico e a barreira à hibridização poderiam não ocorrer nesta etapa, a menos que se encontrem diferenças quantitativas nestes sinais comportamentais.