RESUMO

Estuaries are constantly subject to continuous environmental impacts of human activities, such as fisheries, port or industry, and domestic sewage, with fish being one of the most affected aquatic animals, reflecting the impacts directly on their bodies. Thus, the aim of this study was to carry out the biomonitoring of an estuary located on the Amazonian Equatorial Coast through analysis of PAHs (Polycyclic Aromatic Hydrocarbons) in the water, in addition to trace metals, histopathological alterations and analysis of erythrocyte micronuclei in Sciades herzbergii. S. herzbergii was used as a model species, due to its estuarine-resident behavior. Gonad and gill samples were subjected to histopathological evaluations. The quantification of trace metals was performed in samples of skeletal muscles of the animals collected, where concentrations of Lead (Pb), Copper (Cu), Zinc (Zn), Cadmium (Cd), Magnesium (Mg), Iron (Fe) and Aluminum (Al) were found. Except for Cadmium (Cd), all the concentrations were above the recommended limits. The PAHs analysis revealed the presence of Naphthalene and Acenaphthene in the water samples Histopathological and genotoxic analyses revealed of lesions in 100% of the study specimens. Thus, the histological and genotoxic alterations found in 100% of S. herzbergii specimens captured in São José Bay-MA are potentially associated with PAH concentrations present in the water. These results are potentially associated with the presence of PAH and trace metals, both in water and in animal tissues, inferring a general scenario of environmental contamination which directly implies a risk to the health and survival of the local biota. This study shows the relevance of continuous biomonitoring of estuarine ecosystems, in order to guide authorities regarding sewage management and ensure the evolutionary development of estuarine species, especially fishes of importance in the local cuisine, therefore related to human food security.

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RESUMO

The aim of the current study is to evaluate gene expression patterns of LH (lhr) and estrogen (er) receptors and plasma steroid levels during testicular development in Genyatremus luteus. Males were histologically classified as immature (n=7), maturing (n=7) and mature (n=7), based on the cellular structure of their testes. Plasma 11-KT concentration recorded peak at the final maturation stage. The highest plasma 17α-OHP concentrations were observed at the immature stage; they decreased at the maturation and mature stages. On the other hand, 17ß-estradiol (E2) recorded higher concentrations at the maturation stage. Er expression has significantly increased along the maturational development of animals' testes. The mRNA observed for the LH receptor has decreased from immature to maturing stage; it presented expression peak at the mature stage. There was high association between receptor gene expression and plasma steroid levels, mainly E2. The current study was the first to feature different reproductive maturation stages in male G. luteus specimens, based on cellular, endocrine and molecular aspects. In addition, it has shown that the gene expression profile for er and lhr receptors, as well as plasma 11-KT and E2 concentrations, are directly linked to testicular maturation, although they are not necessarily associated with the gonadosomatic index.

Assuntos

RESUMO

The aim of the current study is to evaluate gene expression patterns of LH (lhr) and estrogen (er) receptors and plasma steroid levels during testicular development in Genyatremus luteus. Males were histologically classified as immature (n=7), maturing (n=7) and mature (n=7), based on the cellular structure of their testes. Plasma 11-KT concentration recorded peak at the final maturation stage. The highest plasma 17α-OHP concentrations were observed at the immature stage; they decreased at the maturation and mature stages. On the other hand, 17ß-estradiol (E2) recorded higher concentrations at the maturation stage. Er expression has significantly increased along the maturational development of animals' testes. The mRNA observed for the LH receptor has decreased from immature to maturing stage; it presented expression peak at the mature stage. There was high association between receptor gene expression and plasma steroid levels, mainly E2. The current study was the first to feature different reproductive maturation stages in male G. luteus specimens, based on cellular, endocrine and molecular aspects. In addition, it has shown that the gene expression profile for er and lhr receptors, as well as plasma 11-KT and E2 concentrations, are directly linked to testicular maturation, although they are not necessarily associated with the gonadosomatic index.

O objetivo deste estudo foi avaliar os padrões de expressão gênica dos receptores de LH (lhr) e de estrogênio (er) e dos níveis de esteróides plasmáticos durante o desenvolvimento testicular de Genyatremus luteus. Os machos foram classificados histologicamente em imaturos, em maturação e maduros, de acordo com a estrutura celular dos testículos. A concentração plasmática de 11-KT apresentou um pico na fase de maturação final (P<0.05). As maiores concentrações plasmáticas de 17α-OHP foram encontradas no estádio imaturo (P<0.05), com consequente diminuição nos estádios em maturação e maturo. O 17ß-estradiol (E2) apresentou maiores níveis de concentração no estádio em maturação (P<0.05). A expressão de er aumentou significativamente ao longo do desenvolvimento maturacional dos testículos (P<0.05). O mRNA para o receptor de LH diminuiu do estádio imaturo para o estádio em maturação (P<0.05) com consequente pico de expressão no estádio maduro. Houve alta relação entre a expressão gênica dos receptores e os níveis de esteróides plasmáticos, especialmente com E2. Em conclusão, este estudo caracterizou pela primeira vez, sob os aspectos celular, endócrino e molecular, os diferentes estádios de maturação reprodutiva em machos de G. luteus, demonstrando que o perfil da expressão gênica para os receptores er e lhr, bem como as concentrações plasmáticas de 11-KT e E2 foram diretamente relacionados à maturação testicular, apesar de não se relacionarem necessariamente com o índice gonadossomático.

Assuntos

RESUMO

Estuaries are constantly subject to continuous environmental impacts of human activities, such as fisheries, port or industry, and domestic sewage, with fish being one of the most affected aquatic animals, reflecting the impacts directly on their bodies. Thus, the aim of this study was to carry out the biomonitoring of an estuary located on the Amazonian Equatorial Coast through analysis of PAHs (Polycyclic Aromatic Hydrocarbons) in the water, in addition to trace metals, histopathological alterations and analysis of erythrocyte micronuclei in Sciades herzbergii. S. herzbergii was used as a model species, due to its estuarine-resident behavior. Gonad and gill samples were subjected to histopathological evaluations. The quantification of trace metals was performed in samples of skeletal muscles of the animals collected, where concentrations of Lead (Pb), Copper (Cu), Zinc (Zn), Cadmium (Cd), Magnesium (Mg), Iron (Fe) and Aluminum (Al) were found. Except for Cadmium (Cd), all the concentrations were above the recommended limits. The PAHs analysis revealed the presence of Naphthalene and Acenaphthene in the water samples Histopathological and genotoxic analyses revealed of lesions in 100% of the study specimens. Thus, the histological and genotoxic alterations found in 100% of S. herzbergii specimens captured in São José Bay-MA are potentially associated with PAH concentrations present in the water. These results are potentially associated with the presence of PAH and trace metals, both in water and in animal tissues, inferring a general scenario of environmental contamination which directly implies a risk to the health and survival of the local biota. This study shows the relevance of continuous biomonitoring of estuarine ecosystems, in order to guide authorities regarding sewage management and ensure the evolutionary development of estuarine species, especially fishes of importance in the local cuisine, therefore related to human food security.

Os estuários estão constantemente sujeitos a impactos ambientais de atividades antropogênicas, como pesca, movimentação portuária, indústria ou esgoto doméstico, onde os peixes são um dos animais aquáticos mais atingidos, sendo capazes de refletir os impactos diretamente em seu organismo. Assim, o objetivo desse estudo foi realizar o biomonitoramento de um estuário da Costa Equatorial Amazônica por meio de análises de PAHs (Hidrocarboneto Policiclico Aromáticos) na água, além de metais traços, alterações histopatológicas e análises de micronúcleos eritrocitários em Sciades herzbergii.. O S. herzbergii foi utilizado como espécie modelo, devido seu comportamento estuarino residente. Amostras de gônadas e brânquias foram submetidas a avaliações histopatológicas. A quantificação de metais traços foi realizada em amostras de músculos esqueléticos dos animais coletados, onde foram encontrados teores de Chumbo (Pb), Cobre (Cu), Zinco (Zn), Cádmio (Cd), Magnésio (Mg), Ferro (Fe) e Alumínio (AL), e foram encontradas em concentrações acima dos limites recomendados, à exceção do Cádmio (Cd). A análise dos PAHs revelou a presença de Naftaleno e Acenafteno nas amostras de água. Análises histopatológicas e genotóxicas revelaram lesões em 100% dos espécimes estudados, resultados estes, potencialmente associados à presença de PAH e de metais traços, tanto na água quanto nos tecidos animais, comprovando um cenário geral de contaminação ambiental implicando diretamente em risco à saúde e sobrevivência da biota local. Esse estudo mostra a relevância do biomonitoramento contínuo dos ecossistemas estuarinos, afim de nortear autoridades no manejo de efluentes e assegurar o desenvolvimento evolutivo de espécies estuarinas, sobretudo dos peixes de importância na culinária regional, diretamente relacionados à segurança alimentar humana.

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Two experiments were conducted to assess a hormonal strategy developed to reduce animal handling for timed artificial insemination (TAI) with sex-sorted semen. Four-hundred ninety-one (491) suckled beef cows received a progesterone (P4) intravaginal device and 2 mg intramuscular (im) injection of estradiol benzoate (EB) on a randomly chosen day of the estrus cycle (Day 0) in Experiment 1. Cows were treated with 500 µg of sodic cloprostenol (PGF2α) and with 300 IU of eCG at P4 device removal (Day 8); these cows were also randomly assigned to receive 1 mg of estradiol cypionate (EC) administered at P4 device removal (treatment EC-0h) or 1 mg of EB 24 h after P4 device removal (treatment EB-24h). Both treatments were timed inseminated (TAI) with sex-sorted semen 60 h after P4 device removal. Cows treated with EC-0h presented higher pregnancy rate per AI (P/AI) [45.0% (113/251)] than the ones treated with EB-24h [35.4% (85/240); P = 0.03)]. A subset of cows (n = 26) were subjected to ultrasound examination every 12 h after P4 device removal for 96 h in the row in order to determine the time of ovulation. Similar interval between device removal and ovulation was recorded for EB-24h = 70.0 ±â€¯2.9 h vs. EC-0h = 66.0 ±â€¯2.8 h (P = 0.52). Five-hundred ninety-one (591) cows were subjected to the same synchronization protocols and treatments (EC-0h or EB-24h). In addition, they were randomly assigned to a 2 × 2 factorial arrangement aiming at determining the effects of treatment with estradiol (EC-0h vs. EB-24h) and of semen type (Sex-sorted vs. Non-sex-sorted semen). All animals were timed inseminated 60 h after P4 device removal. There was no interaction (P = 0.07) between the ovulation inducer and semen type. The EC protocol led to greater P/AI than EB (P = 0.03). Greater (P = 0.01) P/AI was achieved through treatments with non-sex-sorted semen rather than with sex-sorted semen [sex-sorted (EB-24h = 49.0%; EC-0h = 51.0%) vs. non-sex-sorted semen (EB-24h = 52.4%; EC-0h = 68.2%)]. Therefore, EC administered at P4 device removal resulted in greater P/AI. Furthermore, the EC-0h protocol allowed reducing suckled beef cow handing for timed artificial insemination with sex-sorted semen.

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The embryo production technologies are used to enhance genetic progress through female and male lineages. Advances in the control of ovarian follicular wave emergence, superstimulation and ovulation with self-appointed treatments have facilitated donor and recipient management. However, these procedures can be influenced by several factors related to the animals and their management. Therefore, researchers continue to investigate the ideal reproductive environments and treatments to maintain the viability of the techniques and field applicability.(AU)

Assuntos

RESUMO

The embryo production technologies are used to enhance genetic progress through female and male lineages. Advances in the control of ovarian follicular wave emergence, superstimulation and ovulation with self-appointed treatments have facilitated donor and recipient management. However, these procedures can be influenced by several factors related to the animals and their management. Therefore, researchers continue to investigate the ideal reproductive environments and treatments to maintain the viability of the techniques and field applicability.

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RESUMO

Two experiments were designed to evaluate the effect of different insemination times (12 and 24h or 18 and 30h) and different types of semen (sex-sorted or non-sorted sperm) on embryo production in Nelore (Bos indicus) and Holstein (Bos taurus) superstimulated donors. In the first experiment, hormonal superstimulation of ovarian follicular development in Nelore donors (n=71) was performed in randomly allocated animals to one of the three treatment groups, and they were inseminated at 12 and 24h after an ovulatory stimulus with pLH treatment was applied, either with sex-sorted (4.2×10(6) sperm/insemination; S12/24; n=17) or non-sorted sperm (20×10(6) sperm/insemination; NS12/24; n=18), or they were inseminated at 18 and 30h using sex-sorted sperm (4.2×10(6) sperm/insemination; S18/30; n=19). A greater number of transferable embryos were found when sex-sorted sperm was used to inseminate the animals at 18 and 30h (4.5±3.0) compared to insemination at 12 and 24h (2.4±1.8; P<0.001). However, a greater embryo production (6.8±2.6) was obtained with non-sorted sperm. In the second experiment, the same insemination times and semen types were used in lactating high-production Holstein cows (n=12). A crossover design was employed in this trial. A lesser embryo production (P=0.007) was found in Holstein donors that were inseminated using sex-sorted sperm at 12 and 24h (4.6±3.0) compared to non-sorted sperm (8.7±2.8). However, intermediate results were obtained when the inseminations with sex-sorted sperm were performed at 18 and 30h (6.4±3.1). These results supported the current hypothesis that it is possible to improve embryo production using sex-sorted sperm in B. indicus and B. taurus superstimulated donors when the inseminations are performed near the same time as time-synchronized ovulations. However, the embryo production for timed artificial insemination (TAI) with sex-sorted sperm was still less than the production with non-sorted sperm.

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The objectives of this study were to determine the interval from ovulation to deviation and the diameter of the dominant (DF) and largest subordinate (SF) follicles at deviation in buffalo (Bubalus bubalis) heifers. Two methods of evaluation (observed vs. calculated) were used. FSH and LH profiles encompassing follicle deviation (Experiment 1), and the follicular diameter when the DF acquired ovulatory capacity (Experiment 2) were also determined. The time of deviation and the diameter of the DF and the largest SF at deviation did not differ between observed and calculated methods. Overall, follicle deviation occurred 2.6 ± 0.2d (mean ± SEM) after ovulation, and the diameters of the DF and SF at deviation were 7.2 ± 0.2 and 6.4 ± 0.2mm, respectively. No changes in plasma levels of FSH or LH were observed (P=0.32 and P=0.96, respectively). Experiment 2 was conducted in two phases according to the diameter of the DF during the first wave of follicular development at the time of LH challenge (25mg of pLH). In the first phase, follicles ranging from 5.0 to 6.0mm (n=7), 6.1 to 7.0mm (n=11), or 7.1 to 8.0mm (n=9) were used, and in the second phase, follicles ranging from 7.0 to 8.4mm (n=10), 8.5 to 10.0mm (n=10), or 10.1 to 12.0mm (n=9) of diameter were used. After the pLH treatment, the DF was monitored by ultrasonography every 12h for 48h. No ovulations occurred in heifers in the first phase. However, in the second phase, an effect of follicular diameter was observed on ovulation rate [7.0-8.4mm (0.0%, 0/10), 8.5-10.0mm (50.0%, 5/10), and 10.0-12.0mm (55.6%, 5/9)]. In summary, follicle deviation occurred 2.6d after ovulation in buffalo (B. bubalis) heifers, when the diameters of the DF and SF were 7.2 and 6.4mm, respectively. No significant changes in plasma concentrations of FSH or LH were detected. Finally, the acquisition of ovulatory capacity occurred when the DF reached 8.5mm in diameter.

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A total of 177 Nelore heifers were examined by ultrasonography to determine the presence or absence of a corpus luteum (CL) and received a 3mg norgestomet ear implant plus 2mg of estradiol benzoate i.m. On Day 8, implants were removed and 150 microg of d-cloprostenol i.m. was administered. At the time of norgestomet implant removal, heifers with or without CL at the time of initiating treatment were assigned equally and by replicate to be treated with 0IU (n=87) or 400IU (n=90) eCG i.m. All heifers received 1mg of EB i.m. on Day 9 and were submitted to fixed-time artificial insemination (FTAI) 30-34h later. The addition of eCG increased the diameter of the largest follicle (LF) at FTAI (10.6+/-0.2mm vs. 9.5+/-0.2mm; P=0.003; mean+/-SEM), the final growth rate of the LF (1.14+/-0.1mm/day vs. 0.64+/-0.1mm/day; P=0.0009), ovulation rate [94.4% (85/90) vs. 73.6% (64/87); P=0.0006], the diameter of the CL at Day 15 (15.5+/-0.3mm vs. 13.8+/-0.3mm; P=0.0002), serum concentrations of progesterone 5 days after FTAI (6.6+/-1.0 ng/ml vs. 3.6+/-0.7ng/ml; P=0.0009), and pregnancy per AI [P/AI; 50.0% (45/90) vs. 36.8% (32/87); P=0.04]. The absence of a CL at the beginning of the treatment negatively influenced the P/AI [30.2% (16/53) vs. 49.2% (61/124); P=0.01]. Therefore, the presence of a CL (and/or onset of puberty) must be considered in setting up FTAI programs in heifers. In addition, eCG may be an important tool for the enhancement of follicular growth, ovulation, size and function of the subsequent CL, and pregnancy rates in progestin-based FTAI protocols in Bos indicus heifers.

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The objectives of Experiment 1 were to determine the interval from ovulation to deviation, and diameter of the dominant follicle (DF) and largest subordinate follicle (SF) at deviation in Nelore (Bos indicus) heifers by two methods (observed and calculated). Heifers (n = 12) were examined ultrasonographically every 12 h from ovulation (Day 0) to Day 5. The time of deviation and diameter of the DF and largest SF at deviation did not differ (P>0.05) between observed and calculated methods. Overall, deviation occurred 2.5+/-0.2 d (mean +/- S.E.M.) after ovulation, and diameters for DF and largest SF at deviation were 6.2+/-0.2 and 5.9 +/- 0.2 mm, respectively. Experiment 2 was designed to determine the size at which the DF acquires ovulatory capacity in B. indicus heifers. Twenty-nine heifers were monitored every 24 h by ultrasonography, from ovulation until the DF reached diameters of 7.0-8.4 mm (n=9), 8.5-10.0 mm (n=10), or >10.0 mm (n=10). At that time, heifers were treated with 25 mg of pLH and monitored by ultrasonography every 12 h for 48 h. Ovulation occurred in 3 of 9, 8 of 10, and 9 of 10 heifers, respectively (P<0.05). In summary, there was no significant difference between observed and calculated methods of determining the beginning of follicle deviation. Deviation occurred 2.5 d after ovulation when the DF reached 6.2 mm, and ovulatory capacity was acquired by DF as small as 7.0 mm.

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