Effect of formalin in early stages of the freshwater neotropical catfish, Lophiosilurus alexandri.

The objective of this study was to evaluate the acute toxicity of formalin and its level of therapeutic safety in early stages of Lophiosilurus alexandri. Experiment 1, larvae 7 days after hatching (DAH) were exposed to 43.2, 86.4, 172.8, 345.6, 691.2, 1404.0 mg/L of formalin. Experiment 2, juveniles with 22 DAH exposed to 54, 108, 216, 432, 648 mg/L. Experiment 3, 45 DAH exposed to 86.4, 172.8, 345.6, 691.2, 1036.8 mg/L. The experiments had a control without addition of formalin and all were carried out in duplicate. The LC50-12 h were: Experiment 1 = 108.86 mg/L; 2: 152.74 mg/L; 3: 244.38 mg/L of formalin. The respective safety levels were: Experiment 1 = 66.22 mg/L (1 h), 10.89 mg/L (12 h); 2 = 49.17 mg/L (2 h), 15.27 mg/L (12 h); 3 = 68.89 mg/L (2 h), 24.44 mg/L (12 h). The results showed that the developmental stage influenced the sensitivity of animals to formalin.

Production of the Kappaphycus alvarezii extract as a leaf biofertilizer: technical and economic analysis for the north coast of São Paulo-Brazil / Produção do extrato da macroalga Kappaphycus alvarezii como biofertilizante foliar: análise técnica e econômica para o litoral norte de São Paulo-Brasil

The extract of the Kappaphycus alvarezii seaweed is used as a leaf biofertilizer and several studies had proven its efficiency in several crops. This study aimed to analyze the economic viability of fresh seaweed production and the chemically characterized seaweed extract cultivated as a technical-economic alternative for coastal communities. Yields of the solid and liquid fractions were quantified, the extract was characterized in its chemical composition of macronutrients and micronutrients and it was classified according to the criteria of the Brazilian legislation of agricultural fertilizers. For the study of economic feasibility, different sales price scenarios were considered and compared with the commercialization of fresh seaweed. The average yield obtained from the processing of the Brazilian adapted strains of the K. alvarezii for the liquid fraction or fresh seaweed extract was 0.71 ± 0.0080 L kg-1 and the moisture solid production was 295 ± 0.0126 g kg-1. The production of fresh seaweed was unfeasible and for the handmade extract was economically viable for the scenario with the sale price of US$ 2.77 with a net present value of US$ 35,300.13 and an internal rate of return (IRR) of 38.99% over ten year.

O extrato da alga marinha Kappaphycus alvarezii é utilizado como biofertilizante foliar e vários estudos comprovaram sua eficiência agronômica em diversas culturas agrícolas. Este estudo teve como objetivo analisar a viabilidade econômica do cultivo de algas marinhas frescas e seu extrato quimicamente caracterizado como alternativa técnico-econômica para comunidades costeiras. Os rendimentos das frações sólida e líquida foram quantificados, o extrato foi caracterizado em sua composição química de macronutrientes e micronutrientes e foi classificado de acordo com os critérios da legislação brasileira de fertilizantes agrícolas. Para o estudo de viabilidade econômica, diferentes cenários de preços de venda foram considerados e comparados com a comercialização das algas marinhas frescas. O rendimento médio obtido pelo processamento das linhagens brasileiras de K. alvarezii para a fração líquida ou extrato de alga marinha fresca foi de 0,71 ± 0,0080 L kg-1 e o rendimento da fração sólida foi de 295 ± 0,0126 g kg-1. A produção de algas marinhas frescas foi inviável economicamente e, para o extrato artesanal, foi viável para o cenário com preço de venda de US$ 2,77, com valor presente líquido de US$ 35.300,13 e taxa interna de retorno (TIR) de 38,99% para um horizonte de dez anos.

Production of the Kappaphycus alvarezii extract as a leaf biofertilizer: technical and economic analysis for the north coast of São Paulo-Brazil / Produção do extrato da macroalga Kappaphycus alvarezii como biofertilizante foliar: análise técnica e econômica para o litoral norte de São Paulo-Brasil

The extract of the Kappaphycus alvarezii seaweed is used as a leaf biofertilizer and several studies had proven its efficiency in several crops. This study aimed to analyze the economic viability of fresh seaweed production and the chemically characterized seaweed extract cultivated as a technical-economic alternative for coastal communities. Yields of the solid and liquid fractions were quantified, the extract was characterized in its chemical composition of macronutrients and micronutrients and it was classified according to the criteria of the Brazilian legislation of agricultural fertilizers. For the study of economic feasibility, different sales price scenarios were considered and compared with the commercialization of fresh seaweed. The average yield obtained from the processing of the Brazilian adapted strains of the K. alvarezii for the liquid fraction or fresh seaweed extract was 0.71 ± 0.0080 L kg-1 and the moisture solid production was 295 ± 0.0126 g kg-1. The production of fresh seaweed was unfeasible and for the handmade extract was economically viable for the scenario with the sale price of US$ 2.77 with a net present value of US$ 35,300.13 and an internal rate of return (IRR) of 38.99% over ten year.(AU)

O extrato da alga marinha Kappaphycus alvarezii é utilizado como biofertilizante foliar e vários estudos comprovaram sua eficiência agronômica em diversas culturas agrícolas. Este estudo teve como objetivo analisar a viabilidade econômica do cultivo de algas marinhas frescas e seu extrato quimicamente caracterizado como alternativa técnico-econômica para comunidades costeiras. Os rendimentos das frações sólida e líquida foram quantificados, o extrato foi caracterizado em sua composição química de macronutrientes e micronutrientes e foi classificado de acordo com os critérios da legislação brasileira de fertilizantes agrícolas. Para o estudo de viabilidade econômica, diferentes cenários de preços de venda foram considerados e comparados com a comercialização das algas marinhas frescas. O rendimento médio obtido pelo processamento das linhagens brasileiras de K. alvarezii para a fração líquida ou extrato de alga marinha fresca foi de 0,71 ± 0,0080 L kg-1 e o rendimento da fração sólida foi de 295 ± 0,0126 g kg-1. A produção de algas marinhas frescas foi inviável economicamente e, para o extrato artesanal, foi viável para o cenário com preço de venda de US$ 2,77, com valor presente líquido de US$ 35.300,13 e taxa interna de retorno (TIR) de 38,99% para um horizonte de dez anos.(AU)

Effect of alkalinity on food consumption of juvenile pacific white shrimp reared in clear water and biofloc system / Efeito da alcalinidade no consumo alimentar de juvenis de camarão branco do pacífico cultivados em sistema de água clara e bioflocos

This study aimed to evaluate the effect of alkalinity on food consumption and other performance parameters of juvenile Litopenaeus vannamei reared in water containing biofloc and clear water. For this purpose, shrimp of 4.06 ± 0.34 g were kept in containers of 3 L of biofloc and clear water during 3 days, with the concentrations of 50, 100 and 200 mg L-1 of alkalinity, plus a Control group, with 5 replicates each group. Food consumption was verified once a day and other performance parameters were evaluated at the end of the experiment. The food consumption and survival rates of the shrimp was not affected by the different levels of alkalinity and clear water and biofloc systems. The best results of weight gain and specific growth rate were observed in the highest concentrations of alkalinity in the biofloc system. Survival, as well as food consumption, was not affected between levels of alkalinity and in clear water and biofloc. The exposure to inappropriate alkalinity concentrations over long periods of time can adversely affect the animals, thus emphasizing the importance of maintaining adequate levels of alkalinity to the cultivated species.

Esse estudo objetivou avaliar o efeito da alcalinidade no consumo alimentar e demais parâmetros de desempenho de juvenis de Litopenaeus vannamei cultivados em água contendo bioflocos e água clara. Para tanto, durante 3 dias, camarões de 4,06 ± 0,34 g foram mantidos em recipientes de 3 L, sob as concentrações Controle, 50, 100 e 200 mg L-1 de alcalinidade, com 5 repetições cada, em bioflocos e água clara. O consumo alimentar foi verificado uma vez ao dia e os demais parâmetros de desempenho foram avaliados ao final do experimento. Nesse estudo, verifica-se que o consumo alimentar dos camarões não é afetado entre os níveis de alcalinidade e nos sistemas de água clara e bioflocos. Já o ganho em peso e a taxa de crescimento específico são afetados positivamente nas maiores concentrações de alcalinidade, no sistema de bioflocos, onde demonstram os melhores resultados. E a sobrevivência, assim como o consumo alimentar, não é afetada entre os níveis de alcalinidade e nos sistemas de água clara e bioflocos. Contudo, a possibilidade de exposição à concentrações de alcalinidade inapropriadas, durante longos períodos de tempo, pode afetar negativamente os animais, assim, ressaltando a importância da manutenção da alcalinidade em níveis adequados à espécie cultivada.

Effect of alkalinity on food consumption of juvenile pacific white shrimp reared in clear water and biofloc system / Efeito da alcalinidade no consumo alimentar de juvenis de camarão branco do pacífico cultivados em sistema de água clara e bioflocos

This study aimed to evaluate the effect of alkalinity on food consumption and other performance parameters of juvenile Litopenaeus vannamei reared in water containing biofloc and clear water. For this purpose, shrimp of 4.06 ± 0.34 g were kept in containers of 3 L of biofloc and clear water during 3 days, with the concentrations of 50, 100 and 200 mg L-1 of alkalinity, plus a Control group, with 5 replicates each group. Food consumption was verified once a day and other performance parameters were evaluated at the end of the experiment. The food consumption and survival rates of the shrimp was not affected by the different levels of alkalinity and clear water and biofloc systems. The best results of weight gain and specific growth rate were observed in the highest concentrations of alkalinity in the biofloc system. Survival, as well as food consumption, was not affected between levels of alkalinity and in clear water and biofloc. The exposure to inappropriate alkalinity concentrations over long periods of time can adversely affect the animals, thus emphasizing the importance of maintaining adequate levels of alkalinity to the cultivated species.(AU)

Esse estudo objetivou avaliar o efeito da alcalinidade no consumo alimentar e demais parâmetros de desempenho de juvenis de Litopenaeus vannamei cultivados em água contendo bioflocos e água clara. Para tanto, durante 3 dias, camarões de 4,06 ± 0,34 g foram mantidos em recipientes de 3 L, sob as concentrações Controle, 50, 100 e 200 mg L-1 de alcalinidade, com 5 repetições cada, em bioflocos e água clara. O consumo alimentar foi verificado uma vez ao dia e os demais parâmetros de desempenho foram avaliados ao final do experimento. Nesse estudo, verifica-se que o consumo alimentar dos camarões não é afetado entre os níveis de alcalinidade e nos sistemas de água clara e bioflocos. Já o ganho em peso e a taxa de crescimento específico são afetados positivamente nas maiores concentrações de alcalinidade, no sistema de bioflocos, onde demonstram os melhores resultados. E a sobrevivência, assim como o consumo alimentar, não é afetada entre os níveis de alcalinidade e nos sistemas de água clara e bioflocos. Contudo, a possibilidade de exposição à concentrações de alcalinidade inapropriadas, durante longos períodos de tempo, pode afetar negativamente os animais, assim, ressaltando a importância da manutenção da alcalinidade em níveis adequados à espécie cultivada.(AU)

Biological responses of Neotropical freshwater fish Lophiosilurus alexandri exposed to ammonia and nitrite.

This study aimed to elucidate the responses of the Neotropical fish Lophiosilurus alexandri exposed to ammonia and nitrite, following a period of recovering. Acute toxicity tests lasted 96h, subchronic toxicity tests lasted eight days and the detoxification trial lasted four days. Groups of 12 juveniles were maintained in 90-L tanks and treated with increasing concentrations of ammonia and nitrite, except during the recovery test. All treatments were performed with two replicates. The median lethal concentrations (LC50) of 24, 48, 72 and 96h were estimated at 30.12; 24.35; 19.24 and 18.68mg·L-1 TA-N; 5.37; 4.57; 3.75 and 3.66mg·L-1 NH3-N and 20.37; 7.78; 7.09 and 5.86mg·L-1 NO2--N, respectively. The NO2- caused significant decrease in hematocrit and increase in the urea levels during short-term exposure, with recovery of homeostasis after the subchronic and detox period. Acute exposure to ammonia increased the enzyme profile of transaminases, glucose and urea. Urea concentration remained high in the subchronic and detox tests. Histopathologies were observed in animals exposed to ammonia in both toxicity tests. It was highlighted detachment of epithelium, hyperemia and necrosis in the gills. Exposure to NO2- caused epithelium detachment and aneurysm. Vacuolization and swelling of hepatocytes were the most common injury for both nitrogenous compounds. We concluded that the L. alexandri has moderate tolerance to ammonia and nitrite. The recovery period revealed remedial response to ammonia and nitrite exposure.

Larviculture of siamese fighting fish Betta splendens in low-salinity water / Larvicultura do betta em água levemente salinizada

The low-salinity water may improve live food utilization during larviculture, mainly when larvae are fed with salt water organisms. This study aimed to determine the median lethal concentration (LC50) of NaCl in water for larvae of Betta splendens, an important ornamental species, and to evaluate the effect of low-salinity on the larviculture during the first 15 days of exogenous feeding. In the first experiment, 400 larvae were stocked in forty 250 mL aquariums, and exposed to ten saline concentrations. In the second experiment, 360 larvae were distributed in 24 1 L aquariums, in a factorial design 2x3 comprising two increasing prey densities, starting with 50 and 100 Artemia nauplii larva-1, and three concentrations of NaCl (0, 2 and 4 g NaCl L-1). After 24, 48, 72 and 96 h of exposure, the LC50 were 11.7, 10.1, 8.2 and 7.1 g NaCl L-1, respectively. At the end of the experiment 2, larvae reared in salinity of 2 and 4 g NaCl L-1 and fed with the initial prey density of 100 nauplii larvae-1 were bigger and heavier. The use of low-saline water (2 to 4 g NaCl L-1) is a safe protocol for larviculture of Siamese fighting fish as it does not affect the survival and optimizes the use of Artemia nauplii when higher prey densities are used.

A água levemente salinizada melhora o aproveitamento do alimento vivo durante a larvicultura, principalmente quando as larvas são alimentadas com organismos de água salgada. Este estudo objetivou determinar a concentração letal (CL50) de NaCl na água para larvas de Betta splendens, uma importante espécie ornamental, e avaliar os efeitos de salinidades baixas na larvicultura durante os primeiros 15 dias de alimentação exógena. No primeiro experimento, 400 larvas foram estocadas em 40 aquários (250 mL) e expostas a dez concentrações salinas. No segundo experimento, 360 larvas foram distribuídas em 24 aquários de 1 L (15 larvas aquário-1), em esquema fatorial 2x3 com duas densidades crescentes de presas, começando com 50 e 100 náuplios de Artemia larva-1, e três concentrações salinas (0, 2 e 4 g NaCl L-1). Após 24, 48, 72 e 96 h de exposição, a CL50 foi de 11,7; 10,1; 8,2 e 7,1 g NaCl L-1, respectivamente. No final do experimento 2, as larvas mantidas nas salinidades de 2 e 4 g NaCl L-1 e alimentadas na densidade inicial de 100 náuplios larva-1 apresentaram crescimento superior. O uso da água levemente salinizada (2 a 4 g NaCl L-1) é um protocolo seguro para a larvicultura de B. splendens, não afeta a sobrevivência das larvas e otimiza o uso dos náuplios de Artemia quando densidades elevadas de presas são utilizadas.

Larviculture of siamese fighting fish Betta splendens in low-salinity water / Larvicultura do betta em água levemente salinizada

The low-salinity water may improve live food utilization during larviculture, mainly when larvae are fed with salt water organisms. This study aimed to determine the median lethal concentration (LC50) of NaCl in water for larvae of Betta splendens, an important ornamental species, and to evaluate the effect of low-salinity on the larviculture during the first 15 days of exogenous feeding. In the first experiment, 400 larvae were stocked in forty 250 mL aquariums, and exposed to ten saline concentrations. In the second experiment, 360 larvae were distributed in 24 1 L aquariums, in a factorial design 2x3 comprising two increasing prey densities, starting with 50 and 100 Artemia nauplii larva-1, and three concentrations of NaCl (0, 2 and 4 g NaCl L-1). After 24, 48, 72 and 96 h of exposure, the LC50 were 11.7, 10.1, 8.2 and 7.1 g NaCl L-1, respectively. At the end of the experiment 2, larvae reared in salinity of 2 and 4 g NaCl L-1 and fed with the initial prey density of 100 nauplii larvae-1 were bigger and heavier. The use of low-saline water (2 to 4 g NaCl L-1) is a safe protocol for larviculture of Siamese fighting fish as it does not affect the survival and optimizes the use of Artemia nauplii when higher prey densities are used.(AU)

A água levemente salinizada melhora o aproveitamento do alimento vivo durante a larvicultura, principalmente quando as larvas são alimentadas com organismos de água salgada. Este estudo objetivou determinar a concentração letal (CL50) de NaCl na água para larvas de Betta splendens, uma importante espécie ornamental, e avaliar os efeitos de salinidades baixas na larvicultura durante os primeiros 15 dias de alimentação exógena. No primeiro experimento, 400 larvas foram estocadas em 40 aquários (250 mL) e expostas a dez concentrações salinas. No segundo experimento, 360 larvas foram distribuídas em 24 aquários de 1 L (15 larvas aquário-1), em esquema fatorial 2x3 com duas densidades crescentes de presas, começando com 50 e 100 náuplios de Artemia larva-1, e três concentrações salinas (0, 2 e 4 g NaCl L-1). Após 24, 48, 72 e 96 h de exposição, a CL50 foi de 11,7; 10,1; 8,2 e 7,1 g NaCl L-1, respectivamente. No final do experimento 2, as larvas mantidas nas salinidades de 2 e 4 g NaCl L-1 e alimentadas na densidade inicial de 100 náuplios larva-1 apresentaram crescimento superior. O uso da água levemente salinizada (2 a 4 g NaCl L-1) é um protocolo seguro para a larvicultura de B. splendens, não afeta a sobrevivência das larvas e otimiza o uso dos náuplios de Artemia quando densidades elevadas de presas são utilizadas.(AU)

Capture, adaptation and artificial control of reproduction of Lophiosilurus alexandri: A carnivorous freshwater species.

The present study describes the capture adaptation and reproduction of wild Lophiosilurus alexandri broodstock in laboratory conditions. There were two periods when capturing was performed in natural habitats. The animals were placed in four tanks of 5m(3) with water temperatures at 28°C with two tanks having sand bottoms. Thirty days after the temperature increased (during the winter) the first spawning occurred naturally, but only in tanks with sand on the bottom. During the breeding season, there were 24 spawning bouts with egg mass collections occurring as a result of the spawning bouts that occurred in the tanks. The hatching rates for eggs varied from 0% to 95%. The spawning bouts were mainly at night and on weekends. In the second reproductive period, the animals were sexed by cannulation and distributed in four tanks with all animals being maintained in tanks with sand on the bottom at 28°C. During this phase, there were 36 spawning bouts. Findings in the present study contribute to the understanding of the reproductive biology of this endangered species during captivity.

Genotoxicity and histological alterations in grey mullet Mugil liza exposed to petroleum water-soluble fraction (PWSF).

Petroleum hydrocarbons are considered one of the main organic chemicals found in water bodies. In the present study, the median lethal concentration (LC50) was estimated for mullet Mugil liza after acute exposure to petroleum water-soluble fraction (PWSF). Furthermore, histopathological studies and micronuclei frequency were also performed in order to observe deleterious effects of medium-term exposition to PWSF. Mullets (25 ± 2.3 g) were exposed to chronic concentrations (1.7, 3.5 and 7 % of PWSF), plus the control group, for 14 and 7 days of clearance time. Throughout the experimental period (1, 4, 14 and 21 days), blood samples were collected for analysis of micronucleus (MN) and liver and gills for histopathological study. For these procedures, seven fish were sampled per concentration tested. The LC50-96 h was estimated at 37.5 % of the PWSF. The time required for MN induction was 96 h of exposure. The time of clearance was sufficient to achieve a MN frequency similar to that of the control group. Histopathological studies showed severe changes in the gill and liver tissues. The most relevant histopathology in the gills was telangiectasia. Hepatic histopathology such as cholestasis, dilated sinusoids and inflammatory infiltrates were commonly described. The MN test and histological study effectively detected damages caused by medium-term exposition to the PWSF, and despite the toxicity, a few days without exposure can minimize PWSF genotoxicity in juveniles of M. liza.

Growth and histopathological effects of chronic exposition of marine pejerrey Odontesthes argentinensis larvae to petroleum water-soluble fraction (WSF).

The water-soluble fraction (WSF) of petroleum contains a mixture of polycyclic aromatic hydrocarbons, volatile hydrocarbons, phenols, and heterocyclic compounds, considered deleterious to aquatic biota. Marine "pejerrey" Odontesthes argentinensis (Teleostei: Atherinopsidae) has a great commercial importance in local fisheries and a high potential for aquaculture. The aim of this study was to evaluate the histopathological effects in "pejerrey" larvae exposed to different concentrations of petroleum WSF. The chronic toxicity test was conducted with newly hatched larvae exposed for 21 days to sublethal concentrations of WSF (2.5, 5, 10, and 20 % of WSF), plus one control. Survival and growth were significantly lower in the highest concentration. Several histopathological changes were found in the gills (e.g., hyperplasia, aneurisms, edema, and necrosis), kidney (e.g., nuclear alterations, decrease in the hematopoietic cells), and liver (e.g., hypertrophy, karyorrhexis, and karyopyknosis). An index of branchial lesion was proposed to standardize gill lesions to different pollutants.

Deleterious effects of water-soluble fraction of petroleum, diesel and gasoline on marine pejerrey Odontesthes argentinensis larvae.

Accidental discharges and oil spills are frequent around the world. Petroleum-derived hydrocarbons are considered one of the main pollutants of aquatic ecosystem. The importance of petroleum and refined fuels is notorious because today's society depends on them. Researches related to the toxic water-soluble fraction (WSF) of petroleum and derivatives to aquatic biota are scarce. For this reason, deleterious effects of WSF of Brazilian petroleum, automotive diesel and unleaded gasoline to marine pejerrey Odontesthes argentinensis larvae were studied employing toxicity tests and histopathological examination. Each WSF was generated in a laboratory by mixing four parts of seawater with one part of pollutant by approximately 22 h. Larvae were exposed during 96 h to different concentrations of WSF of petroleum, diesel, and gasoline, plus a control. After 96 h of exposure to the different WSFs, three larvae were sampled for histopathological studies. The median lethal concentration after 96 h (LC50) of exposure for WSF of petroleum was equal to 70.68%, it was significantly higher (P<0.05) than the values for WSF of diesel and gasoline, which were 13.46% and 5.48%, respectively. The histological examination of pejerrey larvae exposed to WSF of petroleum, diesel and gasoline after 96 h revealed a variety of lesions in the larvae. The gills, pseudobranchs and esophagus presented epithelial hyperplasia, and the liver presented dilatation of hepatic sinusoids, hepatocitomegaly, bi-nucleated and nuclear degeneration of hepatocytes, such as pyknotic nuclei. The acute toxicity of diesel and gasoline is at least fivefold higher than Brazilian petroleum. However, all toxicants induced histopathological abnormalities in pejerrey larvae. The results are of importance since much attention has been paid to large visible surfaces of petroleum spills instead of potential toxic effects of dissolved aromatic hydrocarbons, which are more available to marine biota.

Long-term ammonia toxicity to the pink-shrimp Farfantepenaeus paulensis.

Juvenile pink-shrimp Farfantepenaeus paulensis were exposed (75 days) to NH(3) (0.016-0.287 mg L(-1)) under static condition with water renewal every 24h. Experiments were performed at 20 degrees C, at a water salinity of 15 ppt, and at pH 7.8. Endpoints analyzed were survival, growth and predation rates. After 75 days of exposure, survival was >or=90% in all concentrations tested. However, growth (carapace length and wet body mass) was reduced after exposure to NH(3) concentrations as low as 0.033 mg L(-1), while the relative growth (dry body mass and ash content) was reduced after exposure to the highest NH(3) concentration (0.287 mg L(-1)). Predatory activity was inhibited after exposure to 0.144 or 0.287 mg L(-1) NH(3). Post-larvae exposed (75 days) to 0.301 mg L(-1) NH(3) under the same experimental conditions also showed a reduced growth (wet body mass) and relative growth (dry body mass). In addition, they showed decreased body lipids content and increased body glycogen and glucose contents. However, no changes in body protein, chitin and uric acid contents were observed. Also, NH(3) did not affect post-larvae feeding response. Altogether, findings suggest that F. paulensis reduces its food intake to limit the internal accumulation of nitrogenous waste products when exposed for long time to high levels of ambient ammonia. As a consequence, shrimp show a marked change in energy metabolism, characterized by a decreased content of body lipids paralleled by an increased content of body carbohydrates, resulting in a significant reduction in growth.