New occurrences of the endangered Notholebias minimus (Cyprinodontiformes: Rivulidae) in coastal plains of the State of Rio de Janeiro, Brazil: populations features and conservation

Notholebias minimus is an endangered annual killifish endemic to the coastal plains of the State of Rio de Janeiro, Brazil. This study aimed to present new occurrences in the Atlantic Forest biome, provide unprecedented population features (body and egg size, fecundity, sexual ratio, and length-weight relationship - LWR), and compare changes in land use and coverage between 1985 and 2021 in biotopes located inside and outside protected areas. Three new occurrence localities were found in shallow temporary wetlands with acidic pH (6.4 ± 0.2) and low concentrations of dissolved oxygen (2.0 ± 0.9 mg/L). Males and females total length ranged from 11.1 to 31 mm and 11 to 26 mm, respectively. Batch fecundity ranged from 18 to 40 oocytes (24.8 ± 8.8), corresponding to oocytes with sizes between 800-1,006 µm (905 ± 56). Males were significantly larger than females (W = 2193.5, p = 0.0067), but both sexes occurred in similar proportions (p = 0.472). LWR showed positive allometry (b = 3.18). Biotopes located within protected areas exhibited higher conservation. Our discoveries expand the knowledge about habitat and population features of N. minimus and reinforce the importance of establishing protected areas for the conservation of annual fish biotopes.

Notholebias minimus é um peixe anual ameaçado de extinção, endêmico das planícies costeiras do Estado do Rio de Janeiro, Brasil. Neste estudo, objetivamos apresentar novas ocorrências no bioma Mata Atlântica, fornecer características populacionais inéditas (tamanho do corpo e dos ovos, fecundidade, proporção sexual e relação peso-comprimento), e comparar mudanças no uso e cobertura do solo entre 1985 e 2021 em biótopos localizados dentro e fora de unidades de conservação. Registramos três novos locais em áreas úmidas temporárias rasas com pH ácido (6,4 ± 0,2) e baixas concentrações de oxigênio dissolvido (2,0 ± 0,9 mg/L). O comprimento total de machos e fêmeas variou de 11,1 a 31 mm e de 11 a 26 mm, respectivamente. A fecundidade do lote variou entre 18-40 oócitos (24,8 ± 8,8), correspondendo a diâmetros entre 800-1.006 µm (905 ± 56). Os machos foram significativamente maiores que as fêmeas (W = 2193,5; p = 0,0067), mas ocorreram em proporções similares (p = 0,472). A relação peso-comprimento detectou alometria positiva (b = 3,18). Biótopos localizados dentro de áreas protegidas exibiram maior preservação ambiental. Nossas descobertas ampliam o conhecimento sobre as características do habitat e da população de N. minimus e reforçam a importância do estabelecimento de áreas protegidas para a conservação dos biótopos dos peixes anuais.

Salinity and prey concentration on larviculture of killifish Hypsolebias radiseriatus (Cyprinodontiformes: Rivulidae)

The aim of this study was to investigate the tolerance of Hypsolebias radiseriatuslarvae to different salinities, and the effects of different prey concentrations and water salinities on the larviculture of this species. Salinity tolerance was tested by subjecting newly-hatched larvae to 96 hours of osmotic shock testing (experiment I) and gradual acclimatization (experiment II) of the following salinities: freshwater (control), 2, 4, 6 and 8 g of salt L-1. A third experiment (experiment III) evaluated three water salinities (S0-freshwater, S2-2 g of salt L-1and S4–4 g of salt L-1) and three initial daily prey concentrations (100, 300 and 500 artemia nauplii larva-1). In experiments I and II, survival was only influenced by the salinity of 8 g of salt L-1(p < 0.01). After 35 days, weight was only influenced by prey concentration (p < 0.05), with the highest value being with 500 artemia nauplii larva-1. The lowest survival was for 4 g of salt L-1and for 100 artemia nauplii larva-1. H. radiseriatuslarviculture can be carried out in salinity of up to 2 g of salt L-1and initial daily prey concentrations with 500 artemia nauplii larva-1.(AU)

Salinity and prey concentration on larviculture of killifish Hypsolebias radiseriatus (Cyprinodontiformes: Rivulidae)

The aim of this study was to investigate the tolerance of Hypsolebias radiseriatuslarvae to different salinities, and the effects of different prey concentrations and water salinities on the larviculture of this species. Salinity tolerance was tested by subjecting newly-hatched larvae to 96 hours of osmotic shock testing (experiment I) and gradual acclimatization (experiment II) of the following salinities: freshwater (control), 2, 4, 6 and 8 g of salt L-1. A third experiment (experiment III) evaluated three water salinities (S0-freshwater, S2-2 g of salt L-1and S4–4 g of salt L-1) and three initial daily prey concentrations (100, 300 and 500 artemia nauplii larva-1). In experiments I and II, survival was only influenced by the salinity of 8 g of salt L-1(p < 0.01). After 35 days, weight was only influenced by prey concentration (p < 0.05), with the highest value being with 500 artemia nauplii larva-1. The lowest survival was for 4 g of salt L-1and for 100 artemia nauplii larva-1. H. radiseriatuslarviculture can be carried out in salinity of up to 2 g of salt L-1and initial daily prey concentrations with 500 artemia nauplii larva-1.

The recapture of Leptopanchax opalescens (Aplocheiloidei: Rivulidae), a critically endangered seasonal killifish: habitat and aspects of population structure

Leptopanchax opalescens (Myers, 1942) is a small seasonal killifish endemic to coastal drainages in the state of Rio de Janeiro, Brazil. Leptopanchax opalescens was recaptured in temporary open vegetation pools (22°42.35S, 43°41.59W) in the hydrographic basin of the Guandu River, state of Rio de Janeiro, Brazil, after eight years without records. Information on habitat and population parameters (Capture per unit area (CPUA), size, sex ratio and length-weight relationship (LWR)) are given to provide subsidies for the conservation of this species. Specimens were captured during the rainy season (January-April 2020) in shallow pools (mean depth = 21 ± 12 s.d. cm), with relatively acidic pH (5.2 ± 0.8) and low concentrations of dissolved oxygen (3.5 ± 1 mg/L). The estimated CPUA of the population was 0.89 fish/m2. The total length (TL) of males ranged from 21.1 to 28.8 cm, and females ranged from 17.0 to 31.2 mm. Males were significantly larger (t-test = 2.04, p 0.05) than females, which were in greater proportion (2.83Female: 1Male, p 0.001). A positive allometric coefficient (b = 3.19) was found through length-weight relationship curve, indicating that individuals of the species gain proportionally more increment in weight than in length. Leptopanchax opalescens is a critically endangered species, and the results of this study contribute to the knowledge of the distribution, population structure and conservation of the species and its natural habitats.

The recapture of Leptopanchax opalescens (Aplocheiloidei: Rivulidae), a critically endangered seasonal killifish: habitat and aspects of population structure

Leptopanchax opalescens (Myers, 1942) is a small seasonal killifish endemic to coastal drainages in the state of Rio de Janeiro, Brazil. Leptopanchax opalescens was recaptured in temporary open vegetation pools (22°42.35S, 43°41.59W) in the hydrographic basin of the Guandu River, state of Rio de Janeiro, Brazil, after eight years without records. Information on habitat and population parameters (Capture per unit area (CPUA), size, sex ratio and length-weight relationship (LWR)) are given to provide subsidies for the conservation of this species. Specimens were captured during the rainy season (January-April 2020) in shallow pools (mean depth = 21 ± 12 s.d. cm), with relatively acidic pH (5.2 ± 0.8) and low concentrations of dissolved oxygen (3.5 ± 1 mg/L). The estimated CPUA of the population was 0.89 fish/m2. The total length (TL) of males ranged from 21.1 to 28.8 cm, and females ranged from 17.0 to 31.2 mm. Males were significantly larger (t-test = 2.04, p 0.05) than females, which were in greater proportion (2.83Female: 1Male, p 0.001). A positive allometric coefficient (b = 3.19) was found through length-weight relationship curve, indicating that individuals of the species gain proportionally more increment in weight than in length. Leptopanchax opalescens is a critically endangered species, and the results of this study contribute to the knowledge of the distribution, population structure and conservation of the species and its natural habitats.(AU)

SALINIDADE E CONCENTRAÇÃO DE PRESAS NA LARVICULTTURA DO KILLIFISH ANUAL HYPSOLEBIAS RADSERIATUS( CYPRIINODONTIFORMES: RIVULIDAE).

A larvicultura é considerada a fase crítica na produção de peixes. Alguns killifishes são anuais e são os únicos vertebrados onde os embriões podem, em certo momento, dependendo das condições ambientais, optarem por caminhos distintos de desenvolvimento, podendo este ser direto ou por diapausas. Para a espécie Hypsolebias radiseriatus ainda não existe nenhum dado de produção de juvenis. O objetivo deste trabalho foi avaliar a tolerância a diferentes gradientes de salinidade e realizar a larvicultura de H. radiseriatus em diferentes concentrações de presa e salinidade da água. Para os testes de tolerância, foram utilizadas larvas recém-eclodidas, onde foi realizado o choque osmótico (experimento I) e aclimatação gradual (experimento II) às seguintes salinidades: água doce, 2, 4, 6 e 8 g de sal/L. As observações foram realizadas até 96 horas. No experimento III, larvas recém eclodidas foram estocadas na densidade de 4 larvas L-1 e submetidas a três salinidades da água a saber: S0 - água doce, S2 - 2 g de sal L-1 e S4 - 4 g de sal L-1, e três concentrações diárias iniciais de presas: 100, 300 e 500 náuplios de artêmia larva-1 em esquema fatorial 3 × 3 com três repetições cada. A larvicultura teve duração de 35 dias. No experimento I e II a sobrevivência sofreu influência apenas na salinidade de 8 g de sal L-1 (P<0,01). Para o peso houve redução a partir de 6 g de sal L-1 (experimento I) e 8 g de sal L-1 (experimento II). No experimento III, a amônia total foi maior para a maior concentração de presas testada independente da salinidade da água. Após 15 dias, a sobrevivência foi menor a 4 g de sal L-1 e verificou-se efeitos tanto da salinidade, como da concentração de presas e da interação entre estes fatores no desempenho das larvas com menores valores para S4P100. Após 35 dias o comprimento não apresentou efeito da salinidade, concentração de presas e da interação entre os fatores (P>0,05). O peso foi influenciado somente pela concentração de presas (P<0,05) com maiores valores para P500. A sobrevivência foi influenciada pela salinidade (P<0,01) e pela concentração de presas (P<0.05), sem efeito da interação (P>0,05). A pior sobrevivência foi para S4 e para P100. Logo, a larvicultura de H. radiseriatus pode ser realizada em salinidade de até 2 g de sal L-1, com a maior concentração de presas testada neste experimento.

he aim of this study was to investigate the tolerance of Hypsolebias radiseriatus larvae to different salinities and the effects of different prey concentrations and water salinities on their larviculture. Salinity tolerance was tested by subjecting newly-hatched larvae to 96 hours of osmotic shock testing (experiment I) and gradual acclimatization (experiment II) with 3 replicates each of the following salinities: freshwater (control), 2, 4, 6 and 8 g of salt L-1. A third experiment (experiment III) subjected newly-hatched larvae, at a density of 4 larvae L-1, to three water salinities (S0 - freshwater, S2 - 2 g of salt L-1 and S4 4 g of salt L-1) and three initial daily prey concentrations (100, 300 and 500 artemia nauplii larva-1) in a 3×3 factorial scheme with three replicates each for 35 days. In experiments I and II, survival was found to be influenced by only the salinity of 8 g of salt L-1 (P <0.01). Weight was found to decrease at 6 g of salt L-1 (experiment I) and 8 g of salt L-1 (experiment II). In experiment III, total ammonia was higher for the highest prey concentration tested independent of water salinity. After 15 days, survival was lowest at 4 g of salt L-1 and the effects of salinity, prey concentration and their interaction decreased the performance of larvae for 4 g of salt L-1 and for 100 artemia nauplii larva-1. After 35 days, total length did not exhibit an effect of salinity, prey concentration or their interaction (P >0.05), while weight was influenced only by prey concentration (P <0.05), with the highest value being with 500 artemia nauplii larva-1. Survival was influenced by salinity (P <0.01) and prey concentrations (P <0.05), but without an effect from the interaction of the two (P >0.05). The lowest survival was for 4 g of salt L-1 and for 100 artemia nauplii larva-1. Therefore, H. radiseriatus larviculture can be carried out in salinity of up to 2 g of salt L-1 and initial daily prey concentrations with 500 artemia nauplii larva-1.

The influence of temperature on the embryonic development of the annual fish Cynopoecilus melanotaenia (Cyprinodontiformes, Rivulidae)

The present study aims to provide data about the time required for Cynopoecilus melanotaenia kept at different temperatures to complete embryonic development. This information can be valuable for optimizing laboratory culture and facilitating future use of this species as a test organism in toxicity tests. Temperature effects on hatching rate are presented as well as information related to embryonic development stages. Eggs were observed daily, from start to finish of embryonic development. Thirteen developmental stages were described. Eggs were kept at two constant temperatures (20°C and 25°C) and at a variable ambient temperature (16-25°C - mean = 21°C, sd = 1.95), to determine developmental rate (velocity) at each temperature. A shorter incubation period was necessary to complete development at 25° ± 1°C. However, all embryos kept at this temperature hatched with morphological defects, which prevented their survival. No significant difference in developmental time period (p = 0.05) was observed at the 20°C and 16°-25°C (mean = 21°C, sd = 1.95) temperatures.

O presente trabalho visa estabelecer informações referentes ao tempo requerido para embriões de Cynopoecilus melanotaenia, mantidos em diferentes temperaturas, completarem seu desenvolvimento embrionário. Esses dados são importantes na otimização de cultivos dessa espécie em laboratório e podem facilitar seu futuro uso como organismos-teste em testes de toxicidade. O efeito da temperatura na taxa de eclosão e informações sobre os estágios de desenvolvimento embrionário são apresentados. Os ovos foram observados diariamente, do início até o final do desenvolvimento embrionário. Foram descritos 13 estágios de desenvolvimento. Os ovos foram mantidos a duas temperaturas constantes (20°C e 25°C) e a uma temperatura variável (16-25°C - média = 21°C, desvio = 1,95), a fim de determinar a velocidade de desenvolvimento para cada temperatura. Foi necessário menor período de incubação para completar o desenvolvimento a 25° ± 1°C. Entretanto, todos os embriões mantidos a essa temperatura eclodiram, apresentando malformações morfológicas que inviabilizavam sua sobrevivência. Não foi observada diferença estatisticamente significativa (p = 0,05) no tempo necessário para completar o desenvolvimento embrionário nos embriões mantidos nas temperaturas de 20°C e 16°-25°C (média = 21°C, desvio = 1,95).

The influence of temperature on the embryonic development of the annual fish Cynopoecilus melanotaenia (Cyprinodontiformes, Rivulidae)

The present study aims to provide data about the time required for Cynopoecilus melanotaenia kept at different temperatures to complete embryonic development. This information can be valuable for optimizing laboratory culture and facilitating future use of this species as a test organism in toxicity tests. Temperature effects on hatching rate are presented as well as information related to embryonic development stages. Eggs were observed daily, from start to finish of embryonic development. Thirteen developmental stages were described. Eggs were kept at two constant temperatures (20°C and 25°C) and at a variable ambient temperature (16-25°C - mean = 21°C, sd = 1.95), to determine developmental rate (velocity) at each temperature. A shorter incubation period was necessary to complete development at 25° ± 1°C. However, all embryos kept at this temperature hatched with morphological defects, which prevented their survival. No significant difference in developmental time period (p = 0.05) was observed at the 20°C and 16°-25°C (mean = 21°C, sd = 1.95) temperatures.

O presente trabalho visa estabelecer informações referentes ao tempo requerido para embriões de Cynopoecilus melanotaenia, mantidos em diferentes temperaturas, completarem seu desenvolvimento embrionário. Esses dados são importantes na otimização de cultivos dessa espécie em laboratório e podem facilitar seu futuro uso como organismos-teste em testes de toxicidade. O efeito da temperatura na taxa de eclosão e informações sobre os estágios de desenvolvimento embrionário são apresentados. Os ovos foram observados diariamente, do início até o final do desenvolvimento embrionário. Foram descritos 13 estágios de desenvolvimento. Os ovos foram mantidos a duas temperaturas constantes (20°C e 25°C) e a uma temperatura variável (16-25°C - média = 21°C, desvio = 1,95), a fim de determinar a velocidade de desenvolvimento para cada temperatura. Foi necessário menor período de incubação para completar o desenvolvimento a 25° ± 1°C. Entretanto, todos os embriões mantidos a essa temperatura eclodiram, apresentando malformações morfológicas que inviabilizavam sua sobrevivência. Não foi observada diferença estatisticamente significativa (p = 0,05) no tempo necessário para completar o desenvolvimento embrionário nos embriões mantidos nas temperaturas de 20°C e 16°-25°C (média = 21°C, desvio = 1,95).