Simulated climate change and atrazine contamination can synergistically impair zebrafish testicular function.

Elements that interfere with reproductive processes can have profound impacts on population and the equilibrium of ecosystems. Global warming represents the major environmental challenge of the 21st century, as it will affect all forms of life in the coming decades. Another coexisting concern is the persistent pollution by pesticides, particularly the herbicide Atrazine (ATZ), which is responsible for a significant number of contamination incidents in surface waters worldwide. While it is hypothesized that climate changes will significantly enhance the toxic effects of pesticides, the actual impact of these phenomena remain largely unexplored. Here, we conducted a climate-controlled room experiment to assess the interactive effects of the projected 2100 climate scenario and environmentally realistic ATZ exposures on the reproductive function of male zebrafish. The gonadosomatic index significantly decreased in fish kept in the extreme scenario. Cellular alterations across spermatogenesis phases led to synergic decreased sperm production and increased germ cell sloughing and death. ATZ exposure alone or combined with climate change effects, disrupted the transcription levels of key genes involved in steroidogenesis, hormone signaling and spermatogenesis regulation. An additive modulation with decreased 11-KT production and increased E2 levels was also evidenced, intensifying the effects of androgen/estrogen imbalance. Moreover, climate change and ATZ independently induced oxidative stress, upregulation of proapoptotic gene and DNA damage in post-meiotic germ cell, but the negative effects of ATZ were greater at extreme scenario. Ultimately, exposure to simulated climate changes severely impaired fertilization capacity, due to a drastic reduction in sperm motility and/or viability. These findings indicate that the future climate conditions have the potential to considerably enhance the toxicity of ATZ at low concentrations, leading to significant deleterious consequences for fish reproductive function and fertility. These may provide relevant information to supporting healthcare and environmental managers in decision-making related to climate changes and herbicide regulation.

Early Neuroprotective Effects of Bovine Lactoferrin Associated with Hypothermia after Neonatal Brain Hypoxia-Ischemia in Rats.

Neonatal hypoxic-ischemic (HI) encephalopathy (HIE) in term newborns is a leading cause of mortality and chronic disability. Hypothermia (HT) is the only clinically available therapeutic intervention; however, its neuroprotective effects are limited. Lactoferrin (LF) is the major whey protein in milk presenting iron-binding, anti-inflammatory and anti-apoptotic properties and has been shown to protect very immature brains against HI damage. We hypothesized that combining early oral administration of LF with whole body hypothermia could enhance neuroprotection in a HIE rat model. Pregnant Wistar rats were fed an LF-supplemented diet (1 mg/kg) or a control diet from (P6). At P7, the male and female pups had the right common carotid artery occluded followed by hypoxia (8% O2 for 60') (HI). Immediately after hypoxia, hypothermia (target temperature of 32.5-33.5 °C) was performed (5 h duration) using Criticool®. The animals were divided according to diet, injury and thermal condition. At P8 (24 h after HI), the brain neurochemical profile was assessed using magnetic resonance spectroscopy (1H-MRS) and a hyperintense T2W signal was used to measure the brain lesions. The mRNA levels of the genes related to glutamatergic excitotoxicity, energy metabolism and inflammation were assessed in the right hippocampus. The cell markers and apoptosis expression were assessed using immunofluorescence in the right hippocampus. HI decreased the energy metabolites and increased lactate. The neuronal-astrocytic coupling impairments observed in the HI groups were reversed mainly by HT. LF had an important effect on astrocyte function, decreasing the levels of the genes related to glutamatergic excitotoxicity and restoring the mRNA levels of the genes related to metabolic support. When combined, LF and HT presented a synergistic effect and prevented lactate accumulation, decreased inflammation and reduced brain damage, pointing out the benefits of combining these therapies. Overall, we showed that through distinct mechanisms lactoferrin can enhance neuroprotection induced by HT following neonatal brain hypoxia-ischemia.

Low-molecular weight sulfated marine polysaccharides: Promising molecules to prevent neurodegeneration in mucopolysaccharidosis IIIA?

Mucopolysaccharidosis IIIA is a hereditary disease caused by mutations in the sulfamidase enzyme that participates in catabolism of heparan sulfate (HS), leading to HS fragment accumulation and multisystemic failure. No cure exists and death occurs around the second decade of life. Two low molecular weight highly sulfated compounds derived from marine diabolican and infernan exopolysaccharides (A5_3 and A5_4, respectively) with heparanase inhibiting properties were tested in a MPSIIIA cell line model, resulting in limited degradation of intracellular HS. Next, we observed the effects of intraperitoneal injections of the diabolican derivative A5_3 from 4 to 12 weeks of age on MPSIIIA mice. Brain metabolism and microstructure, levels of proteins and genes involved in MPSIIIA brain pathophysiology were also investigated. 1H-Magnetic Resonance Spectroscopy (MRS) indicated deficits in energetic metabolism, tissue integrity and neurotransmission at both 4 and 12 weeks in MPSIIIA mice, with partial protective effects of A5_3. Ex-vivo Diffusion Tensor Imaging (DTI) showed white matter microstructural damage in MPSIIIA, with noticeable protective effects of A5_3. Protein and gene expression assessments displayed both pro-inflammatory and pro-apoptotic profiles in MPSIIIA mice, with benefits of A5_3 counteracting neuroinflammation. Overall, derivative A5_3 was well tolerated and was shown to be efficient in preventing brain metabolism failure and inflammation, resulting in preserved brain microstructure in the context of MPSIIIA.

Andrology of shortnose guitarfish Zapteryx brevirostris (Müller & Henle, 1841) (Chondrichthyes, Trygonorrhinidae).

The andrological study of a species involves the macro- and microscopic analyses of the internal reproductive organs and the evaluation of seminal parameters and ultrastructural characteristics of the spermatozoa. As in other vertebrates, the male reproductive tract in chondrichthyans consists of testes and reproductive ducts (efferent duct, epididymis, Leydig's gland, ductus deferens and seminal vesicle). In this study the authors used three adult specimens of Zapteryx brevirostris from wild capture kept at the Ubatuba Aquarium, Brazil. Semen was collected by abdominal massage over the location of the seminal vesicle, preceded by ultrasonographic evaluation. The semen collected was diluted 1:200 and subject to quantitative and morphological analyses. Ultrastructural analysis was performed using transmission and scanning electron microscopy. Correlation was observed between successful collection and ultrasonographic image of an engorged seminal vesicle, as well as testicles with easily delimitable margins and higher echogenicity. It was possible to identify free spermatozoa with helical filiform appearance, as well as spermatozeugmata. The average sperm concentration resulted in 5 million packets per millilitre and 140 million spermatozoa per millilitre. The sperm nucleus is described as follows: cone shaped, parachromatin sheath less dense than the chromatin of the nucleus, smooth depression of the nuclear fossa, abaxial axoneme 9 + 2 and accessory axonemal columns in positions 3 and 8 and oval shaped, with flattened inner surface in cross-section. These results broaden the knowledge of the andrology of this species, contributing to ex situ breeding programmes.

Fish sperm cryopreservation using biodegradable containers: New low-cost and environment-friendly methodology.

In brief: The containers used in cell cryopreservation are essential to maintain cell integrity and viability after thawing. This paper reveals the methodology of using biodegradable containers for fish sperm cryopreservation. Cryopreserved sperm in biodegradable containers showed high fertility capability. Biodegradable capsules could be alternative containers to plastic straws for sperm cryopreservation. Abstract: Containers used to cryopreserve sperm are made with non-biodegradable plastic compounds, having a high monetary and environmental cost. Therefore, the development of biodegradable alternative containers for cell cryopreservation is necessary. Thus, this study aimed to evaluate the efficiency of hard-gelatin and hard-hydroxypropyl methylcellulose (HPMC) capsules as low-cost and biodegradable alternative containers for sperm cryopreservation. Sperm from 12South American silver catfish Rhamdia quelen were individually cryopreserved in plastic straws 0.25 mL (as control), hard-gelatin, and hard-HPMC capsules. The quality of post-thaw sperm cryopreserved in the different containers was checked by measuring spermatozoa membrane integrity, kinetic parameters, mitochondrial activity, fertilization, hatching, and normal larvae rates. The samples cryopreserved in straws showed a higher percentage of membrane integrity (68%) than those frozen in hard-gelatin (40%) and hard-HPMC capsules (40%). However, we did not observe differences between the samples stored in straws and hard capsules for the rest of the tested sperm parameters. Thus, based on the high sperm fertility capability, both capsules were efficient as cryopreservation containers for maintaining sperm functionality.

Food-related substrate preference in juveniles seastar Echinaster (Othilia) brasiliensis (Müller & Troschel,1842) in captivity.

There are only a few studies that describe the larval development of Echinaster or aspects on culture systems for the genus. For starfishes, the choice of suitable substrates has received special attention since it could influence the acid-base balance of the water, movement capacity and predation rate. The objective of this study was to evaluate the ideal food-related substrate for the rearing of juvenile Echinaster brasiliensis. A batch of fertilized eggs released in spontaneous spawning was collected and kept in a plankton-kreisel until metamorphosis. Data on preference of food-related substrate was recorded for 10 weeks from day 58 post-release. From release to 132 days old, arm length increased from 0.81 mm to 1.31 ± 0.03 mm. Considering the sudden increase in arm length (AL), it was estimated that feeding started around 40 days of age. Regarding food-related substrate preferences, biofilm grown on "rocks" showed a significant difference among other treatments, adding up to 50% of preference (p < .05). For sponge and biofilm from bio media, there was no statistical difference for the whole period. In this study, sponges showed to be the least preferred food-related substrate for post-settlement juveniles. Considering that Echinaster and other starfish are commonly maintained on a diet of collected or cultured sponges, difficulties in sourcing a ready supply throughout the year represent limitations to their sole use within commercial or laboratory-scale production. In this sense, the use of biofilm from biological media for the feeding of juvenile starfish is not yet reported in the literature and showed to be an easy and promising option.

Can canthaxanthin intensify the colour of the blood swordtail Xiphophorus helleri?

Colouring has a great influence on the commercialization of ornamental fish. The aim of this study was to evaluate different concentrations of canthaxanthin in the diet of the blood swordtail Xiphophorus helleri in an effort to obtain a more intense red colour. Six concentrations of canthaxanthin (0, 50, 100, 250, 400 and 600 mg kg-1 diet) were used. The experiment lasted 60 days. Fish were evaluated for increased red pigmentation through the use of photographs (performed by smartphone) and imaging applications considering the Hunter method and the CMYK and productive performance. No significant differences were observed for productive performance. The use of photographs by means of a smartphone and the use of imaging applications proved to be adequate to assess differences in colour in the species. Swordtail red pigmentation did not show significant increase regardless of canthaxanthin dosages. Varieties of ornamental fish bred for red coloration may have limits for increased colour due to the storage capacity of carotenoids by chromatophores.

Correction to "Synthesis of Lysoglycerophosphocholines from Crude Soybean Lecithins as Sustainable and Non-toxic Antifouling Agents against the Golden Mussel Limnoperna fortunei".

[This corrects the article DOI: 10.1021/acsomega.2c05645.].

Birth and growth of the shortnose guitarfish Zapteryx brevirostris (Müller & Henle, 1841) (Chondrichthyes, Rhinobatidae) in captivity.

Endemic to the south-west Atlantic Ocean, the shortnose guitarfish (Zapteryx brevirostris) is a small species, classified as endangered by the IUCN. Although reproduction in captivity has been successful for some species, a range of factors can limit the success of captive breeding programmes for elasmobranchs. In Brazil, the Ubatuba Aquarium was the first public aquarium to reproduce small-sized elasmobranchs. Since 2018, at least five parturition events have been recorded for Z. brevirostris at the institution. From a total of 13 live neonates that rearing was attempted, the mean ± standard error of weight, total length (TL) and disc width at birth were 17.47 ± 1.6 g, 13.25 ± 0.7 cm and 6.53 ± 0.2 cm, respectively. The mean weight as well as mean TL were higher for females at all births, with 26.15 g and 15.07 cm for females in comparison with 17.09 g and 13.94 cm for males. Considering the increasing risk of extinction that the species is facing, age and growth studies are fundamental for the success of conservation, improving the knowledge of Z. brevirostris life cycle so that a more efficient and sustainable management can be carried out. This study provides important data, as well as directions for captive breeding of the species.

Synthesis of Lysoglycerophosphocholines from Crude Soybean Lecithins as Sustainable and Non-toxic Antifouling Agents against the Golden Mussel Limnoperna fortunei.

This research aimed to produce, on a multigram scale, a new class of non-toxic, halogen- and metal-free antifouling agents from the abundant lecithin byproducts of industrial soybean oil extraction. Three glycerophospholipid analogues were prepared by a facile methanolysis of crude soybean lecithins and a subsequent solvent-free O-alkylation: lysoglycerophosphocholines (LGPCs) and its ether derivatives O-alkyl lysoglycerophosphocholines (ALPCs). As efficient antiproliferative agents, LGPCs and ALPCs are an eco-friendly alternative to current commercial antifoulants which possess significant toxicity to aquatic life. In situ immersion tests of coated stainless-steel nets with previously incorporated automotive paint products, LGPCs and ALPCs (1-O-octadecyl-2-O-acyl-sn-glycero-3-phosphocholine, ALPC18, and 1-O-hexadecyl-2-O-acyl-sn-glycero-3-phosphocholine, ALPC16), in an aquaculture reservoir in SP-Brazil revealed significant growth inhibition against macrofouling species, especially the epibiotic golden mussel (Limnoperna fortunei), when compared with the control. These results promise a more sustainable and ecologically innocuous approach to combating the biofouling phenomenon and the deeply concerning dissemination of the golden mussel which has provoked an economic crisis in the energy and aquaculture sectors.

Early neurodevelopmental reflex impairments in a rodent model of cerebral palsy.

Cerebral palsy (CP) causes sensorimotor disabilities due to injury to the developing brain. Experimental models do not always induce the CP phenotype completely. Early neurological assessment predicts future impairments and is valuable during development. Using a rodent model characterized by brain injury caused by maternal inflammation and perinatal anoxia, and sensorimotor restriction (experimental cerebral palsy [ECP]), we describe early neurodevelopmental delays by assessing reflexes in a stage corresponding to the brain development of term infants (Postnatal Day [P] 8 in rats). Pregnant Wistar rats were injected with lipopolysaccharide (LPS; 200 µg/kg) (n = 6) or saline (n = 4) on Embryonic Days 18/19. Following delivery, 87 male and female pups were used. At P0, injured animals were exposed to anoxia for 20'. From P2 to P21, ECP rats were subjected to hindlimb movement restriction for 16 h/day. ECP group had impaired righting reflex and negative geotaxis and, interestingly, performed home bedding test better than controls. From P7, ECP animals showed decreased body weight compared with controls. Overall, data provide evidence showing that this CP model based on the association of brain damage followed by sensorimotor restriction mimics CP delays and highlights the valuable information given by early neurological assessment during the establishment of the CP phenotype.

Neuroprotective Role of Lactoferrin during Early Brain Development and Injury through Lifespan.

Early adverse fetal environments can significantly disturb central nervous system (CNS) development and subsequently alter brain maturation. Nutritional status is a major variable to be considered during development and increasing evidence links neonate and preterm infant impaired brain growth with neurological and psychiatric diseases in adulthood. Breastfeeding is one of the main components required for healthy newborn development due to the many "constitutive" elements breastmilk contains. Maternal intake of specific nutrients during lactation may alter milk composition, thus affecting newborn nutrition and, potentially, brain development. Lactoferrin (Lf) is a major protein present in colostrum and the main protein in human milk, which plays an important role in the benefits of breastfeeding during postnatal development. It has been demonstrated that Lf has antimicrobial, as well as anti-inflammatory properties, and is potentially able to reduce the incidence of sepsis and necrotizing enterocolitis (NEC), which are particularly frequent in premature births. The anti-inflammatory effects of Lf can reduce birth-related pathologies by decreasing the release of pro-inflammatory factors and inhibiting premature cervix maturation (also related to commensal microbiome abnormalities) that could contribute to disrupting brain development. Pre-clinical evidence shows that Lf protects the developing brain from neuronal injury, enhances brain connectivity and neurotrophin production, and decreases inflammation in models of perinatal inflammatory challenge, intrauterine growth restriction (IUGR) and neonatal hypoxia-ischemia (HI). In this context, Lf can provide nutritional support for brain development and cognition and prevent the origin of neuropsychiatric diseases later in life. In this narrative review, we consider the role of certain nutrients during neurodevelopment linking to the latest research on lactoferrin with respect to neonatology. We also discuss new evidence indicating that early neuroprotective pathways modulated by Lf could prevent neurodegeneration through anti-inflammatory and immunomodulatory processes.

Therapeutic hypothermia for the treatment of neonatal hypoxia-ischemia: sex-dependent modulation of reactive astrogliosis.

Therapeutic hypothermia (TH) is the standard treatment for neonatal hypoxia-ischemia (HI) with a time window limited up to 6 h post injury. However, influence of sexual dimorphism in the therapeutic window for TH has not yet been elucidated in animal models of HI. Therefore, the aim of this study was to investigate the most effective time window to start TH in male and female rats submitted to neonatal HI. Wistar rats (P7) were divided into the following groups: NAÏVE and SHAM (control groups), HI (submitted to HI) and TH (submitted to HI and TH; 32ºC for 5 h). TH was started at 2 h (TH-2 h group), 4 h (TH-4 h group), or 6 h (TH-6 h group) after HI. At P14, animals were subjected to behavioural tests, volume of lesion and reactive astrogliosis assessments. Male and female rats from the TH-2 h group showed reduction in the latency of behavioral tests, and decrease in volume of lesion and intensity of GFAP immunofluorescence. TH-2 h females also showed reduction of degenerative cells and morphological changes in astrocytes. Interestingly, females from the TH-6 h group showed an increase in volume of lesion and in number of degenerative hippocampal cells, associated with worse behavioral performance. Together, these results indicate that TH neuroprotection is time- and sex-dependent. Moreover, TH started later (6 h) can worsen volume of brain lesion in females. These data indicate the need to develop specific therapeutic protocols for each sex and reinforce the importance of early onset of the hypothermic treatment.

Post-thaw dilution of Rhamdia quelen sperm improves the reproductive success.

The aim was to evaluate the effect of a post-thaw dilution of Rhamdia quelen sperm in 1.1% NaCl (325 mOsm kg-1; pH 7.6; 24 °C) solution on the quality and reproductive capacity. Sperm from eight males were cryopreservation in nitrogen vapor at - 170 °C for 18 h in 0.25 mL straws in a freezing medium containing 5% fructose, 5% Powdered milk, and 10% methanol. The samples were thawed and post-thaw diluted (1:20) in NaCl solution or not (control). The higher spermatozoa velocities were observed in the post-thaw diluted samples (curvilinear (VCL) - 69 ± 11 µm s-1; average path (VAP) - 45 ± 8 µm s-1; straight-line (VSL) - 43 ± 8 µm s-1) compared to the control (VCL - 47 ± 10 µm s-1; VAP - 31 ± 6 µm s-1; VSL - 30 ± 6 µm s-1). Greater straightness (STR), progression (PROG), and beat cross frequency (BCF) were observed in the post-thaw diluted samples (STR - 96 ± 7%; PROG - 666 ± 128 µm; BCF - 42 ± 2 Hz) than in control (STR - 95 ± 5%; PROG - 463 ± 92 µm; BCF - 40 ± 2 Hz). The strongly curled tail was the only morphology change that differ between the post-thaw diluted (5 ± 2%) and control (2 ± 1%). Membrane integrity, mitochondrial activity, and normal larvae rate were not different between treatments. Fertilization and hatching were higher in the post-thaw diluted sperm (93 ± 3%; 82 ± 9%) when compared to control samples (65 ± 13%; 55 ± 17%). Were used oocytes from one female, limiting these results. The post-thaw dilution improved the sperm kinetics and reproductive parameters. Thus, this methodology can be included in the sperm cryopreservation protocol for R. quelen.

Plinia trunciflora Extract Administration Prevents HI-Induced Oxidative Stress, Inflammatory Response, Behavioral Impairments, and Tissue Damage in Rats.

The disruption of redox homeostasis and neuroinflammation are key mechanisms in the pathogenesis of brain hypoxia-ischemia (HI); medicinal plants have been studied as a therapeutic strategy, generally associated with the prevention of oxidative stress and inflammatory response. This study evaluates the neuroprotective role of the Plinia trunciflora fruit extract (PTE) in neonatal rats submitted to experimental HI. The HI insult provoked a marked increase in the lipoperoxidation levels and glutathione peroxidase (GPx) activity, accompanied by a decrease in the brain concentration of glutathione (GSH). Interestingly, PTE was able to prevent most of the HI-induced pro-oxidant effects. It was also observed that HI increased the levels of interleukin-1ß in the hippocampus, and that PTE-treatment prevented this effect. Furthermore, PTE was able to prevent neuronal loss and astrocyte reactivity induced by HI, as demonstrated by NeuN and GFAP staining, respectively. PTE also attenuated the anxiety-like behavior and prevented the spatial memory impairment caused by HI. Finally, PTE prevented neural tissue loss in the brain hemisphere, the hippocampus, cerebral cortex, and the striatum ipsilateral to the HI. Taken together our results provide good evidence that the PTE extract has the potential to be investigated as an adjunctive therapy in the treatment of brain insult caused by neonatal hypoxia-ischemia.

Dose-Dependent Neuroprotective Effects of Bovine Lactoferrin Following Neonatal Hypoxia-Ischemia in the Immature Rat Brain.

Injuries to the developing brain due to hypoxia-ischemia (HI) are common causes of neurological disabilities in preterm babies. HI, with oxygen deprivation to the brain or reduced cerebral blood perfusion due to birth asphyxia, often leads to severe brain damage and sequelae. Injury mechanisms include glutamate excitotoxicity, oxidative stress, blood-brain barrier dysfunction, and exacerbated inflammation. Nutritional intervention is emerging as a therapeutic alternative to prevent and rescue brain from HI injury. Lactoferrin (Lf) is an iron-binding protein present in saliva, tears, and breast milk, which has been shown to have antioxidant, anti-inflammatory and anti-apoptotic properties when administered to mothers as a dietary supplement during pregnancy and/or lactation in preclinical studies of developmental brain injuries. However, despite Lf's promising neuroprotective effects, there is no established dose. Here, we tested three different doses of dietary maternal Lf supplementation using the postnatal day 3 HI model and evaluated the acute neurochemical damage profile using 1H Magnetic Resonance Spectroscopy (MRS) and long-term microstructure alterations using advanced diffusion imaging (DTI/NODDI) allied to protein expression and histological analysis. Pregnant Wistar rats were fed either control diet or bovine Lf supplemented chow at 0.1, 1, or 10 g/kg/body weight concentration from the last day of pregnancy (embryonic day 21-E21) to weaning. At postnatal day 3 (P3), pups from both sexes had their right common carotid artery permanently occluded and were exposed to 6% oxygen for 30 min. Sham rats had the incision but neither surgery nor hypoxia episode. At P4, MRS was performed on a 9.4 T scanner to obtain the neurochemical profile in the cortex. At P4 and P25, histological analysis and protein expression were assessed in the cortex and hippocampus. Brain volumes and ex vivo microstructural analysis using DTI/NODDI parameters were performed at P25. Acute metabolic disturbance induced in cortical tissue by HIP3 was reversed with all three doses of Lf. However, data obtained from MRS show that Lf neuroprotective effects were modulated by the dose. Through western blotting analysis, we observed that HI pups supplemented with Lf at 0.1 and 1 g/kg were able to counteract glutamatergic excitotoxicity and prevent metabolic failure. When 10 g/kg was administered, we observed reduced brain volumes, increased astrogliosis, and hypomyelination, pointing to detrimental effects of high Lf dose. In conclusion, Lf supplementation attenuates, in a dose-dependent manner, the acute and long-term cerebral injury caused by HI. Lf reached its optimal effects at a dose of 1 g/kg, which pinpoints the need to better understand effects of Lf, the pathways involved and possible harmful effects. These new data reinforce our knowledge regarding neuroprotection in developmental brain injury using Lf through lactation and provide new insights into lactoferrin's neuroprotection capacities and limitation for immature brains.

Successive seminal collections from Leiarius marmoratus during the reproductive period.

This study aim was to verify whether milt quality of male Leiarius marmoratus is maintained among successive samples collected during the same reproductive period. Ten reproductively mature males were used to evaluate four successive sperm samples collected at 10-day intervals. For these collections, seven males were injected with carp pituitary homogenate (CPH) at a dosage of 3.0 mg/kg body weight, in two applications (30% and 70%), at an interval of 10 h. The other three males were administered only saline (control). Injection with CPH or saline occurred prior to each of the four collections. Only one male from the control group released a small volume of milt (0.33 mL), and only during the first collection period. Of the seven males treated with CPH, five released milt during all four collections. Milt volume of the first sample collected (0.63 mL) did not differ from that of other samples (0.59-1.38 mL; P > 0.05). Sperm concentration was greater in the first samples collected (1.98 × 109 spermatozoa/mL) compared to the other samples (0.35 × 109 at 0.92 × 109 spermatozoa/mL; P < 0.05). Sperm motility, curvilinear velocity, straightness, and morphological normality did not differ among the consecutive samples (P > 0.05). Average path velocity, straight-line velocity, oscillation, linearity, progression, and membrane integrity decreased slightly in the samples collected subsequent to the first sample (P > 0.05). In conclusion, milt quality decreased among successive collections; however, quality of all samples from all collections was sufficient for use for fertilization of oocytes.

Early warming stress on rainbow trout juveniles impairs male reproduction but contrastingly elicits intergenerational thermotolerance.

The exposure of adult fish to warm or high temperatures is known to impair reproduction, yet the long-term reproductive impacts for treatments at early life are not well clarified. This study aimed to evaluate the effects of warm temperature (WT) during juvenile stage on gonad maturation, gamete quality, and offspring thermotolerance in rainbow trout. While the comparison of basic reproductive parameters in WT females did not reveal any kind of impairment, many WT males showed an atrophied, undeveloped gonad, or a smaller testis with lower milt volume; sperm quality parameters in WT males and deformity rates in the respective progeny were also highly affected. However, despite of such negative effects, many of the remaining progeny presented better rates of survival and growth when exposed to the same conditions as those of parental fish (WT), suggesting that thermal stress in parr stage males elicited intergenerational thermotolerance after a single generation. The present results support that prolonged warming stress during early life stages can adversely affect key reproductive aspects, but contrastingly increase offspring performance at upper thermal ranges. These findings have implications on the capacity of fish to adapt and to cope with global warming.

Early environmental enrichment rescues memory impairments provoked by mild neonatal hypoxia-ischemia in adolescent mice.

Hypoxia-ischemia (HI) is a consequence of a lack of oxygen and glucose support to the developing brain, which causes several neurodevelopmental impairments. Environmental enrichment (EE) is considered an option to recover the alterations observed in rodents exposed to HI. The aim of this study was to investigate the impact of early EE on memory, hippocampal volume and brain-derived neurotrophic factor (Bbnf) and glucocorticoid receptor (Nr3c1) gene expression of mice exposed to HI. At P10, pups underwent right carotid artery permanent occlusion followed by 35 min of 8% O2 hypoxic environment. Starting at P11, animals were reared in EE or in standard cage (HI-SC or SHAM-SC) conditions until behavioral testing (P45). SHAM pups did not undergo carotid ligation and hypoxic exposure. Memory performance was assessed in the Y-maze, Novel object recognition, and Barnes maze. Animals were then sacrificed for analysis of hippocampal volume and Bdnf and Nr3c1 gene expression. We observed that animals exposed to HI performed worse in all three tests compared to SHAM animals. Furthermore, HI animals exposed to EE did not differ from SHAM animals in all tasks. Moreover, HI decreased hippocampal volume, while animals reared in early EE were not different compared to SHAM animals. Animals exposed to HI also showed upregulated hippocampal Bdnf expression compared to SHAM animals. We conclude that early EE from P11 to P45 proved to be effective in recovering memory impairments and hippocampal volume loss elicited by HI. Nevertheless, Bdnf expression was not associated with the improvements in memory performance observed in animals exposed to EE after a hypoxic-ischemic event.

Pre- and early postnatal enriched environmental experiences prevent neonatal hypoxia-ischemia late neurodegeneration via metabolic and neuroplastic mechanisms.

Prenatal and early postnatal periods are important for brain development and neural function. Neonatal insults such as hypoxia-ischemia (HI) causes prolonged neural and metabolic dysregulation, affecting central nervous system maturation. There is evidence that brain hypometabolism could increase the risk of adult-onset neurodegenerative diseases. However, the impact of non-pharmacologic strategies to attenuate HI-induced brain glucose dysfunction is still underexplored. This study investigated the long-term effects of early environmental enrichment in metabolic, cell, and functional responses after neonatal HI. Thereby, male Wistar rats were divided according to surgical procedure, sham, and HI (performed at postnatal day 3), and the allocation to standard (SC) or enriched condition (EC) during gestation and lactation periods. In-vivo cerebral metabolism was assessed by means of [18 F]-FDG micro-positron emission tomography, and cognitive, biochemical, and histological analyses were performed in adulthood. Our findings reveal that HI causes a reduction in glucose metabolism and glucose transporter levels as well as hyposynchronicity in metabolic brain networks. However, EC during prenatal or early postnatal period attenuated these metabolic disturbances. A positive correlation was observed between [18 F]-FDG values and volume ratios in adulthood, indicating that preserved tissue by EC is metabolically active. EC promotes better cognitive scores, as well as down-regulation of amyloid precursor protein in the parietal cortex and hippocampus of HI animals. Furthermore, growth-associated protein 43 was up-regulated in the cortex of EC animals. Altogether, results presented support that EC during gestation and lactation period can reduce HI-induced impairments that may contribute to functional decline and progressive late neurodegeneration.