Nile tilapia (Oreochromis niloticus) show high tolerance to acute ammonia exposure but lose metabolic scope during prolonged exposure at low concentration.

Ammonia is a respiratory gas that is produced during the process of protein deamination. In the unionised form (NH3), it readily crosses biological membranes and is highly toxic to fish. In the present study we examined the effects of unionized ammonia (UIA), on the resting oxygen consumption (MO2), ventilation frequency (fV), heart rate (HR) and heart rate variability (HRV) in Nile tilapia (Oreochromis niloticus). Fish were either exposed to progressively increasing UIA concentrations, up to 97 µM over a 5 h period, or to a constant UIA level of 7 µM over a 24 h period. For both treatment groups resting MO2, HR and fV were recorded as physiological variables. Relative to the control group, the fish groups exposed to the incremental UIA levels did not exhibit significant changes in their MO2, HR and fV at UIA concentrations of 4, 10, 35, or 61 µM compared to control fish. Exposure to 97 µM UIA, however, elicited abrupt and significant downregulations (p < 0.05) in all three responses, as MO2, HR and fv decreased by 25, 54 and 76 % respectively, compared to control measurements. Heart rate became increasingly irregular with increasing UIA concentrations, and heart rate variability was significantly increased at 61 and 97 µM UIA. Prolonged exposure elicited significant changes at exposure 7 µM UIA. Standard (SMR) and maximum metabolic rate (MMR) were significantly reduced, as was the corresponding fV and HR. It is evident from this study that Nile tilapia is tolerant to short term exposure to UIA up to 61 µM but experience a significant metabolic change under conditions of prolonged UIA exposures even at low concentrations.

Relatedness of hypoxia and hyperthermia tolerances in the Nile tilapia (Oreochromis niloticus) and their relationships with cardiac and gill traits.

In fish, thermal and hypoxia tolerances may be functionally related, as suggested by the oxygen- and capacity-limited thermal tolerance (OCLTT) concept, which explains performance failure at high temperatures due to limitations in oxygen delivery. In this study the interrelatedness of hyperthermia and hypoxia tolerances in the Nile tilapia (Oreochromis niloticus), and their links to cardiorespiratory traits were examined. Different groups of O. niloticus (n = 51) were subjected to hypoxia and hyperthermia challenges and the O2 tension for aquatic surface respiration (ASR pO2) and critical thermal maximum (CTmax) were assessed as measurement endpoints. Gill filament length, total filament number, ventricle mass, length and width were also measured. Tolerance to hypoxia, as evidenced by ASR pO2 thresholds of the individual fish, was highly variable and varied between 0.26 and 3.39 kPa. ASR events increased more profoundly as O2 tensions decreased below 2 kPa. The CTmax values recorded for the O. niloticus individuals ranged from 43.1 to 44.8 °C (Mean: 44.2 ± 0.4 °C). Remarkably, there was a highly significant correlation between ASR pO2 and CTmax in O. niloticus (r = -0.76, p < 0.0001) with ASR pO2 increasing linearly with decreasing CTmax. There were, however, no discernible relationships between the measured cardiorespiratory properties and hypoxia or hyperthermia tolerances. The strong relationship between hypoxia and hyperthermia tolerances in this study may be related to the ability of the cardiorespiratory system to provide oxygen to respiring tissues under thermal stress, and thus provides some support for the OCLTT concept in this species, at least at the level of the entire organism.

Did bold fish spark Lake Tanganyika's diversity?

A gene mutation tied to exploratory behavior may have jump-started the evolution of hundreds of cichlid species.

Executive functions and brain morphology of male and female dominant and subordinate cichlid fish.

BACKGROUND: Living in a social dominance hierarchy presents different benefits and challenges for dominant and subordinate males and females, which might in turn affect their cognitive needs. Despite the extensive research on social dominance in group-living species, there is still a knowledge gap regarding how social status impacts brain morphology and cognitive abilities. METHODS: Here, we tested male and female dominants and subordinates of Neolamprologus pulcher, a social cichlid fish species with size-based hierarchy. We ran three executive cognitive function tests for cognitive flexibility (reversal learning test), self-control (detour test), and working memory (object permanence test), followed by brain and brain region size measurements. RESULTS: Performance was not influenced by social status or sex. However, dominants exhibited a brain-body slope that was relatively steeper than that of subordinates. Furthermore, individual performance in reversal learning and detour tests correlated with brain morphology, with some trade-offs among major brain regions like telencephalon, cerebellum, and optic tectum. CONCLUSION: As individuals' brain growth strategies varied depending on social status without affecting executive functions, the different associated challenges might yield a potential effect on social cognition instead. Overall, the findings highlight the importance of studying the individual and not just species to understand better how the individual's ecology might shape its brain and cognition.

Foraging-induced craniofacial plasticity is associated with an early, robust and dynamic transcriptional response.

Phenotypic plasticity is the ability of a single genotype to vary its phenotype in response to the environment. Plasticity of the skeletal system in response to mechanical input is widely studied, but the timing of its transcriptional regulation is not well understood. Here, we used the cichlid feeding apparatus to examine the transcriptional dynamics of skeletal plasticity over time. Using three closely related species that vary in their ability to remodel bone and a panel of 11 genes, including well-studied skeletal differentiation markers and newly characterized environmentally sensitive genes, we examined plasticity at one, two, four and eight weeks following the onset of alternate foraging challenges. We found that the plastic species exhibited environment-specific bursts in gene expression beginning at one week, followed by a sharp decline in levels, while the species with more limited plasticity exhibited consistently low levels of gene expression. This trend held across nearly all genes, suggesting that it is a hallmark of the larger plasticity regulatory network. We conclude that plasticity of the cichlid feeding apparatus is not the result of slowly accumulating gene expression difference over time, but rather is stimulated by early bursts of environment-specific gene expression followed by a return to homeostatic levels.

The genetics of niche-specific behavioral tendencies in an adaptive radiation of cichlid fishes.

Behavior is critical for animal survival and reproduction, and possibly for diversification and evolutionary radiation. However, the genetics behind adaptive variation in behavior are poorly understood. In this work, we examined a fundamental and widespread behavioral trait, exploratory behavior, in one of the largest adaptive radiations on Earth, the cichlid fishes of Lake Tanganyika. By integrating quantitative behavioral data from 57 cichlid species (702 wild-caught individuals) with high-resolution ecomorphological and genomic information, we show that exploratory behavior is linked to macrohabitat niche adaptations in Tanganyikan cichlids. Furthermore, we uncovered a correlation between the genotypes at a single-nucleotide polymorphism upstream of the AMPA glutamate-receptor regulatory gene cacng5b and variation in exploratory tendency. We validated this association using behavioral predictions with a neural network approach and CRISPR-Cas9 genome editing.

Social regulation of arginine vasopressin and oxytocin systems in a wild group-living fish.

The neuropeptides arginine vasopressin (AVP) and oxytocin (OXT) are key regulators of social behaviour across vertebrates. However, much of our understanding of how these neuropeptide systems interact with social behaviour is centred around laboratory studies which fail to capture the social and physiological challenges of living in the wild. To evaluate relationships between these neuropeptide systems and social behaviour in the wild, we studied social groups of the cichlid fish Neolamprologus pulcher in Lake Tanganyika, Africa. We first used SCUBA to observe the behaviour of focal group members and then measured transcript abundance of key components of the AVP and OXT systems across different brain regions. While AVP is often associated with male-typical behaviours, we found that dominant females had higher expression of avp and its receptor (avpr1a2) in the preoptic area of the brain compared to either dominant males or subordinates of either sex. Dominant females also generally had the highest levels of leucyl-cystinyl aminopeptidase (lnpep)-which inactivates AVP and OXT-throughout the brain, potentially indicating greater overall activity (i.e., production, release, and turnover) of the AVP system in dominant females. Expression of OXT and its receptors did not differ across social ranks. However, dominant males that visited the brood chamber more often had lower preoptic expression of OXT receptor a (oxtra) suggesting a negative relationship between OXT signalling and parental care in males of this species. Overall, these results advance our understanding of the relationships between complex social behaviours and neuroendocrine systems under natural settings.

Turbidity drives plasticity in the eyes and brains of an African cichlid.

Natural variation in environmental turbidity correlates with variation in the visual sensory system of many fishes, suggesting that turbidity may act as a strong selective agent on visual systems. Since many aquatic systems experience increased turbidity due to anthropogenic perturbations, it is important to understand the degree to which fish can respond to rapid shifts in their visual environment, and whether such responses can occur within the lifetime of an individual. We examined whether developmental exposure to turbidity (clear, <5 NTU; turbid, ∼9 NTU) influenced the size of morphological structures associated with vision in the African blue-lip cichlid Pseudocrenilabrus multicolor. Parental fish were collected from two sites (clear swamp, turbid river) in western Uganda. F1 broods from each population were split and reared under clear and turbid rearing treatments until maturity. We measured morphological traits associated with the visual sensory system (eye diameter, pupil diameter, axial length, brain mass, optic tectum volume) over the course of development. Age was significant in explaining variation in visual traits even when standardized for body size, suggesting an ontogenetic shift in the relative size of eyes and brains. When age groups were analyzed separately, young fish reared in turbid water grew larger eyes than fish reared in clear conditions. Population was important in the older age category, with swamp-origin fish having relatively larger eyes and optic lobes relative to river-origin fish. Plastic responses during development may be important for coping with a more variable visual environment associated with anthropogenically induced turbidity.

Innervation and osteoclast distribution in the inferior pharyngeal jaw of the cichlid Nile tilapia (Oreochromis niloticus).

In addition to an oral jaw, cichlids have a pharyngeal jaw, which is used for crushing and processing captured prey. The teeth and morphology of the pharyngeal jaw bones adapt to changes in prey in response to changes in the growing environment. This study aimed to explore the possible involvement of the peripheral nervous system in remodeling the cichlid pharyngeal jaw by examining the innervation of the inferior pharyngeal jaw in the Nile tilapia, Oreochromis niloticus. Vagal innervation was identified in the Nile tilapia inferior pharyngeal jaw. Double staining with tartrate-resistant acid phosphatase and immunostaining with the neuronal markers, protein gene product 9.5, and acetylated tubulin, revealed that osteoclasts, which play an important role in remodeling, were distributed in the vicinity of the nerves and were in apposition with the nerve terminals. This contact between peripheral nerves and osteoclasts suggests that the peripheral nervous system may play a role in remodeling the inferior pharyngeal jaw in cichlids.

Effect of water calcium, copper, and silver on branchial Na+ permeability in a characid and cichlid fish.

We tested the hypothesis that water Ca2+ is involved in control of branchial Na+ permeability in low pH tolerant convict cichlids and black neon tetras. We measured Na+ efflux in water with different Ca2+ concentrations during exposure to low pH, silver, and copper, at levels which are known to stimulate Na+ efflux. For convict cichlids at pH 7.5 exposure to 0 µmol L-1 Ca2+caused Na+ efflux to rise 2.5 times above controls at 100 µmol L-1 Ca2+. However, raising [Ca2+] to 500 µmol L-1 had no effect. Upon exposure to pH 3.5 (control [Ca2+]) Na+ efflux rose almost 5× and increasing the [Ca2+] 5-fold did not reduce the magnitude of stimulation. Exposure to 1 µmol L-1 silver and 25 µmol L-1 copper stimulated Na+ efflux 7×, and 2×, respectively. Raising [Ca2+] concentration during metal exposure halved the stimulation of Na+ efflux caused by silver, and eliminated the stimulation elicited by copper. For black neon tetras raising or lowering water [Ca2+] had no effect on Na+ efflux at pH 7.5. Exposure to pH 3.5 caused Na+ efflux to rise 2.5× but changing [Ca2+] had no effect. Exposure to 1 µmol L-1 silver, or 25 µmol L-1 copper caused Na+ efflux of tetras to rise 4-fold and 3-fold, respectively. Raising [Ca2+] during silver exposure reduced the stimulation of Na+ efflux by about 50%, but during copper exposure increased [Ca2+] had no effect on stimulation of Na+ efflux. These results suggest water Ca2+ plays a role in control of branchial Na+ permeability in cichlids, but perhaps not tetras. In addition, the silver and copper concentrations required to inhibit Na+ uptake and stimulate Na+ efflux were higher than the concentrations used on non-characids and non-cichlids, which indicates that our fish are much more tolerant of these metals.

Assessing the effect of ß-glucan diets on innate immune response of tilapia macrophages against trichlorfon exposure: an in vitro study.

The widespread use of pesticides in some areas where fish species such as tilapia are farmed may cause damage to the environment and affect commercial fish and therefore, human health. Water leaching with the pesticide trichlorfon, during the fumigation season in the field, can affect water quality in fish farms and consequently affect fish health. At the same time, the use of immunomodulatory compounds such as ß-glucan supplied in the diet has become widespread in fish farms as it has been shown that improves the overall immune response. The present research examines the immunomodulatory impacts observed in macrophages of Nile tilapia (Oreochromis niloticus) after being fed a diet supplemented with ß-glucan for 15 days, followed by their in vitro exposure to trichlorfon, an organophosphate pesticide, at concentrations of 100 and 500 µg mL-1 for 24 h. The results showed that ß-glucan diet improved the viability of cells exposed to trichlorfon and their antioxidant capacity. However, ß-glucan did not counteract the effects of the pesticide as for the ability to protect against bacterial infection. From the present results, it can be concluded that ß-glucan feeding exerted a protective role against oxidative damage in cells, but it was not enough to reduce the deleterious effects of trichlorfon on the microbicidal capacity of macrophages exposed to this pesticide.

The role of serotonin in modulating social competence in a cooperatively breeding fish.

Behavioural interactions between conspecifics rely on the appreciation of social cues, which is achieved through biochemical switching of pre-existing neurophysiological pathways. Serotonin is one of the major neurotransmitters in the central nervous system responsible for the modulation of physiological and behavioural traits, in particular social behaviour. The relative importance of serotonin in modulating optimal social responses to the available social information (i.e., social competence) is yet unknown. Here we investigate how serotonin and the serotonin 1 A receptor (5-HT1A) modulate social competence in a competitive context. In the cooperatively breeding cichlid Neolamprologus pulcher, we pharmacologically manipulated the serotonin availability and 5-HT1A activity to test their effects on social behaviours during an asymmetric contest between the owner of a defended territory containing a shelter and an intruder devoid of a territory. In this contest, the adequate response by the intruders, the focal individuals in our study, is to show submissive behaviour in order to avoid eviction from the vicinity of the shelter. While the serotonin enhancer Fluoxetine did not affect the frequency of submission towards territory owners, reducing serotonin by a low dosage of 4-Chloro-DL-phenylalanine (PCPA) increased submissive behaviour. Furthermore, threat displays towards territory owners were reduced at high dosages of Fluoxetine and also at the lowest dosage of PCPA. 5-HT1A activation increased threat displays by intruders, indicating that this receptor may not be involved in regulating social competence. We conclude that serotonin, but not its receptor 5-HT1A plays an important role in the regulation of social competence.

Potentials of Gynura procumbens to modulate chronic stress and immunological responses in Oreochromis niloticus.

Natural products and traditional remedies have become more popular over the years since they have less harmful side effects and are considered environmentally friendly. In this study we aimed to investigate the potential of Gynura procumbens extract (GPE), a well-known traditional medicinal plant extract, on the stress modulation of Oreochromis niloticus (Nile tilapia). Four different experimental groups: control, stress, prevention, and treatment were monitored for 12 weeks. Hydrocortisone (0.01% of body weight) was incorporated with the feed to induce the stress response in stress, prevention and treatment groups. Feed was also supplemented with 0.15% GPE of body weight for the prevention and treatment groups. Cortisol concentration was reduced significantly in the prevention (1870.52 pg/mL; p = 0.006) and treatment (2925.91 pg/mL; p = 0.002) groups than the stress group (7614.22 pg/mL). The result is substantiated by significant decrease in blood glucose level in prevention (29.5 mg/dL; p = 0.002) and treatment (31.5 mg/dL; p = 0.006) groups, compared to stress group (47.33 mg/dL) at the end of the experiment. Considering the current finding, we can conclude the GPE has potential to be used as therapeutic option for stress regulation however there is a room for further detailed study to understand the in-depth mechanism.

Combined effects of water salinity and ammonia exposure on the antioxidative status, serum biochemistry, and immunity of Nile tilapia (Oreochromis niloticus).

Growing Nile tilapia in brackish water showed promising results, but the possibility of ammonia exposure can interrupt health status and productivity. Herein, the study tested the combined effects of water salinity and ammonia exposure on the antioxidative status, serum biochemistry, and immunity of Nile tilapia. Fish were assigned to eight groups where fish were reared in saline water (5, 10, and 15 ppt) with continuous or intermittent (every 3 days) total ammonia (TAN) exposure (5 mg TAN/L) (2 × 4 factorial design). After 30 days, the water salinity, TAN, and their interaction were markedly (P < 0.05), affecting the growth performance (final weight, weight gain, and specific growth rate) and survival rate of Nile tilapia. The growth performance and survival rate were markedly lower in tilapia grown in 15 ppt with continuous TAN exposure than in the remaining groups. The results showed that fish exposed to higher salinity levels (10 and 15 ppt) and continuous TAN exposure had a more robust antioxidative response, as evidenced by higher superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities and lower malondialdehyde (MDA) levels in the homogenates of the gills, intestines, and livers. The gills were notably affected, with congestion of primary filaments blood vessels and degeneration or shedding of secondary filaments epithelium, especially at salinity levels of 10 and 15 ppt. Additionally, the intestines displayed hyperplasia and inflammatory cell infiltration of intestinal mucosa at 5-10 ppt salinity, degeneration and sloughing of the intestinal epithelium at 15 ppt saline water, and increased goblet cell number at salinity of 10 ppt. The study found that continuous TAN exposure had a more significant impact on the fish, especially at higher salinity levels. Water salinity, TAN, and their interaction significantly affected all measured blood bio-indicators (total, albumin, globulin, ALT, AST, creatinine, urea, glucose, and cortisol levels). The phagocytic activity and index were markedly lowered in fish reared in 15 ppt with continuous TAN exposure, while the lysozyme activity was decreased in fish grown in 5, 10, and 15 ppt with continuous TAN exposure. In conclusion, Nile tilapia showed the possibility of growth with normal health status in brackish water (5-10 ppt); however, continuous TAN exposure can impair the productivity of tilapia, especially with high salinity (15 ppt).

Cognitive flexibility in a Tanganyikan bower-building cichlid, Aulonocranus dewindti.

Cognitive flexibility, the ability to modify one's decision rules to adapt to a new situation, has been extensively studied in many species. In fish, though, data on cognitive flexibility are scarce, especially in the wild. We studied a lekking species of cichlid fish in Lake Tanganyika, Aulonocranus dewindti. Males create sand bowers as spawning sites and maintain them by removing any objects falling into it. In the first part of our experiment, we investigated the existence of spontaneous decision rules for the maintenance of the bowers. We showed that if a snail shell and a stone are placed in their bower, fish prefer to remove the shell first. In the second phase of our experiment, we took advantage of this spontaneous decision rule to investigate whether this rule was flexible. We tested five individuals in a choice against preference task, in which the fish had to modify their preference rule and remove the stone first to be allowed to then remove the shell and have a clean bower. While there was no overall trend towards flexibility in this task, there was variation at an individual level. Some individuals increased their preference for removing the shell first, deciding quickly and with little exploration of the objects. Others were more successful at choosing against preference and showed behaviours suggesting self-regulatory inhibition abilities. Bower-building cichlids could therefore be a promising model to study cognitive flexibility, and other aspects of animal cognition in the wild.

Activation patterns of dopaminergic cell populations reflect different learning scenarios in a cichlid fish, Pseudotropheus zebra.

Dopamine is present in all vertebrates and the functional roles of the subsystems are assumed to be similar. Whereas the effect of dopaminergic modulation is well investigated in different target systems, less is known about the factors that are causing the modulation of dopaminergic cells. Using the zebra mbuna, Pseudotropheus zebra, a cichlid fish from Lake Malawi as a model system, we investigated the activation of specific dopaminergic cell populations detected by double-labeling with TH and pS6 antibodies while the animals were solving different learning tasks. Specifically, we compared an intense avoidance learning situation, an instrumental learning task, and a non-learning isolated group and found strong activation of different dopaminergic cell populations. Preoptic-hypothalamic cell populations respond to the stress component in the avoidance task, and the forced movement/locomotion may be responsible for activation in the posterior tubercle. The instrumental learning task had little stress component, but the activation of the raphe superior in this group may be correlated with attention or arousal during the training sessions. At the same time, the weaker activation of the nucleus of the posterior commissure may be related to positive reward acting onto tectal circuits. Finally, we examined the co-activation patterns across all dopaminergic cell populations and recovered robust differences across experimental groups, largely driven by hypothalamic, posterior tubercle, and brain stem regions possibly encoding the valence and salience associated with stressful stimuli. Taken together, our results offer some insights into the different functions of the dopaminergic cell populations in the brain of a non-mammalian vertebrate in correlation with different behavioral conditions, extending our knowledge for a more comprehensive view of the mechanisms of dopaminergic modulation in vertebrates.

Acute mancozeb-fungicide exposure induces neuro-ethology disruption, health disorders, and immune-oxidative dysfunction in Nile tilapia (Oreochromis niloticus).

An acute exposure study of mancozeb (MAZ) fungicide was applied on Oreochromis niloticus for 96-h duration. Three hundred fish (20.50 ± 1.60 g) were assigned into six groups (50 fish/ group; 10 fish/replicate) and exposed to different six concentrations (0, 4, 8, 12, 16, and 20 mg L-1) of MAZ for 96-h. The Probit analysis program was used to compute the 96-h lethal concentration 50 (96-h LC50) of MAZ. During the exposure duration, the fish's behavior, clinical symptoms, and mortalities were recorded daily. After the exposure period was ended, the hematological, biochemical, immunological, and oxidant/antioxidant parameters were evaluated. The results of this study recorded the 96-h LC50 of MAZ for O. niloticus to be 11.49 mg L-1. Acute MAZ exposure badly affected the fish's behavior in the form of increased the breath gasping and swimming activity with aggressive mode. The exposed fish showed excessive body hemorrhages and fin rot. The survival rate of the exposed fish to MAZ was 100, 80, 66, 50, 38, and 30% in 0, 4, 8, 12, 16, and 20 mg L-1 MAZ, respectively. The hematological indices (red blood cell count, hemoglobin, packed cell volume%, and white blood cell count) were significantly decreased by increasing the MAZ exposure concentration (8-20 mg L-1). The acetylcholine esterase activity and immune indices (lysozyme, nitric oxide, immunoglobulin M, complement 3) were decreased by MAZ exposure (4-20 mg L-1). Acute MAZ exposure induced hepato-renal dysfunction and elevated stress-related parameter (cortisol) by increasing the MAZ concentration. A significant reduction in the antioxidant parameters (total antioxidant activity, catalase, and superoxide dismutase) with increasing the lipid peroxidation marker (malondialdehyde) was noticed by acute MAZ exposure (4 -20 mg L-1) in O. niloticus. Based on these outcomes, the MAZ exposure induced toxicity to the fish evident in changes in fish behavior, neurological activity, hepato-renal functioning, and immune-antioxidant responses which suggest physiological disruption.

In-feed emamectin benzoate abuse affects the biological responses and erythrocytes of Nile tilapia Oreochromis niloticus.

In aquaculture, drugs are often abused to accomplish disease control without considering the negative effects on fish health. This study aimed at elucidating the pernicious effects of in-feed antiparasitic drug emamectin benzoate (EB) abuse on the haemato-biochemistry and erythro-morphometry of healthy Nile tilapia Oreochromis niloticus. The fish were fed EB at 50 µg (1×) and 150 µg/kg biomass/d (3×) for 14 d as against the recommended 7 d and periodically assessed the blood parameters. A significant dose- and time-dependent reduction in feed intake, survival, total erythrocytes (TEC), monocytes (MC), hemoglobin (Hb), hematocrit (Ht) and mean corpuscular Hb concentration were noted. The total leukocytes (TLC), thrombocytes (TC), lymphocytes (LC) and neutrophils (NC) markedly augmented. The EB-dosing altered the fish physiology by enhancing the glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and creatinine and reducing the calcium, chloride and acetylcholinesterase (AChE) levels dose-dependently. The fish recovered within 4 weeks in the 1× group post-dosing but persevered in the overdosed group. The erythro-cellular and nuclear dimensions were reduced with the increase in dose and normalized after the cessation of dosing, except for nuclear volume. The erythro-morphological alterations were more prominent in the overdosed group. The results implied the pernicious effect of oral EB medication on the biological responses of fish if abused.

Using Azadirachta indica protein hydrolysate as a plant protein in Nile tilapia (Oreochromis niloticus) diet: Effects on the growth, economic efficiency, antioxidant-immune response and resistance to Streptococcus agalactiae.

A feeding trial for 90 days was conducted on Nile tilapia (Oreochromis niloticus) (average weight: 25.50 ± 0.05 g) to evaluate the effect of dietary inclusion of Azadirachta indica seed protein hydrolysate (AIPH). The evaluation included the impact on the growth metrics, economic efficiency, antioxidant potential, hemato-biochemical indices, immune response, and histological architectures. A total of 250 fish were randomly distributed in five treatments (n = 50) and received diets included with five levels of AIPH (%): 0 (control diet, AIPH0), 2 (AIPH2), 4 (AIPH4), 6 (AIPH6) or 8 (AIPH8), where AIPH partially replace fish meal by 0, 8.7%, 17.4%, 26.1%, and 34.8%, respectively. After the feeding trial, a pathogenic bacterium (Streptococcus agalactiae, 1.5 × 108 CFU/mL) was intraperitoneally injected into the fish and the survival rate was recorded. The results elucidated that AIPH-included diets significantly (p < 0.05) enhanced the growth indices (final body weight, total feed intake, total body weight gain, and specific growth rate) and intestinal morpho-metrics (villous width, length, muscular coat thickness, and goblet cells count) in comparison to the control diet, with the AIPH8 diet recording the highest values. Dietary AIPH inclusion significantly improved (p < 0.05) the economic efficacy indicated by reduced feed cost/kg gain and increased performance index. The fish fed on the AIPH diets had noticeably significantly higher (p < 0.05) protein profile variables (total proteins and globulin) and antioxidant capabilities (superoxide dismutase and total antioxidant capacity) than the AIPH0 group. The dietary inclusion of AIPH significantly (p < 0.05) boosted the haematological parameters (haemoglobin, packed cell volume %, and counts of red blood cells and white blood cells) and immune indices (serum bactericidal activity %, antiprotease activity, and immunoglobulin M level) in a concentration-dependent manner. The blood glucose and malondialdehyde levels were significantly (p < 0.05) lowered by dietary AIPH (2%-8%). The albumin level and hepatorenal functioning parameters (aspartate aminotransferase, alanine aminotransferase, and creatinine) were not significantly (p > 0.05) altered by AIPH diets. Additionally, AIPH diets did not adversely alter the histology of the hepatic, renal or splenic tissues with moderately activated melano-macrophage centres. The mortality rate among S. agalactiae-infected fish declined as dietary AIPH levels rose, where the highest survival rate (86.67%) was found in the AIPH8 group (p < 0.05). Based on the broken line regression model, our study suggests using dietary AIPH at the optimal level of 6%. Overall, dietary AIPH inclusion enhanced the growth rate, economic efficiency, health status, and resistance of Nile tilapia to the S. agalactiae challenge. These beneficial impacts can help the aquaculture sector to be more sustainable.

Deleterious Effects of Polypropylene Microplastic Ingestion in Nile Tilapia (Oreochromis niloticus).

The effect of daily ingestion of polypropylene microplastic on the health of tilapia, Oreochromis niloticus, was evaluated. 60 fish (± 200 g) were placed in 6 aquariums (n = 10, 100 L each), constituting the following treatments: Control (without the addition of polymer), fed with 100 and 500 µg of polypropylene/kg of body weight (b.w.), respectively. After 30 days of feeding, the animals were submitted to blood collection for hemogram and biochemical study and later euthanized for gut microbiological analysis, somatic index of liver, spleen, heart, kidney, stomach, and intestine. In the serum biochemical study, an increase in cholesterol and serum Aspartate Aminotransferase (AST) activity levels was observed in animals treated with 500 µg of polypropylene. Tilapia-fed polypropylene in the diet showed an increase in thrombocyte and total leukocyte counts, marked by a significant increase in the number of circulating lymphocytes. The results of the somatic study revealed a significant increase in the stomach, liver, and heart of tilapia fed with the polymer. Increase in the number of Gram-negative microorganisms and decrease in mesophilic aerobic microorganisms were observed in the gut of fish exposed to the polymer, including a dose-response effect was observed for these analyses. Therefore, tilapias fed daily with diets containing polypropylene for 30 consecutive days showed deleterious effects, resulting in systemic inflammatory disturbs by altering liver functions, leukocyte profile, and organ morphometry, as well as changes in the intestinal microbiota. Such results demonstrate the impairment of fish health, highlighting the need for further studies that evaluate the impact of microplastics on aquatic organisms.