Rodrigo Fortes-Silva

Daily rhythms of the expression of genes from the somatotropic axis: The influence on tilapia (Oreochromis niloticus) of feeding and growth hormone administration at different times.

The aim of this research was to investigate the presence of daily rhythms in the somatotropic axis of tilapia fed at two times (mid-light, ML or mid-dark, MD) and the influence of the time of day of growth hormone (GH) administration on the response of this axis. Two different GH injection times were tested: ZT 3 (3h after lights on) and ZT 15 (3h after lights off). In both experiments, the mRNA expression levels of hypothalamic pituitary adenylate cyclase-activating polypeptide (pacap), pituitary growth hormone (gh), liver insulin-like growth factors (igf1 and igf2a), and liver and muscle growth hormone receptors (ghr1 and ghr2) and IGF receptors (igf1ra and igf2r) were evaluated by means of qPCR. Daily rhythms were observed in the liver for ghr1, ghr2 and igf2r but only in fish fed at ML, with the acrophases located in the light phase (ZT 3:30, 3:31 and 7:38 h, respectively). In the muscle, ghr1 displayed a significant rhythm in both groups and ghr2 in ML fed fish (acrophases at ZT 5:29, 7:14 and 9:23h). The time of both GH administration and feeding influenced the response to GH injection: ML fed fish injected with GH at ZT 15 h showed a significant increase in liver igf1, igf2a and ghr2; and muscle ghr2 expression. This is the first report that describes the existence of daily rhythms in the somatotropic axis of tilapia and its time-dependent responses of GH administration. Our results should be considered when investigating the elements of the somatotropic axis in tilapia and GH administration.

Synchronization to light and mealtime of daily rhythms of locomotor activity, plasma glucose and digestive enzymes in the Nile tilapia (Oreochromis niloticus)

The light-dark cycle and feeding can be the most important factors acting as synchronizers of biological rhythms. In this research we aimed to evaluate synchronization to feeding schedule of daily rhythms of locomotor activity and digestive enzymes of tilapia. For that purpose, 120 tilapias (65.0±0.6g) were distributed in 12 tanks (10 fish per tank) and divided into two groups. One group was fed once a day at 11:00h (zeitgeber time, ZT6) (ML group) and the other group was fed at 23:00h (ZT18) (MD group). The fish were anesthetized to collect samples of blood, stomach and midgut at 4-hour intervals over a period of 24h. Fish fed at ML showed a diurnal locomotor activity (74% of the total daily activity occurring during the light phase) and synchronization to the feeding schedule, as this group showed anticipation to the feeding time. Fish fed at MD showed a disruption in the pattern of locomotor activity and became less diurnal (59%). Alkaline protease activity in the midgut showed daily rhythm with the achrophase at the beginning of the dark phase in both ML and MD groups. Acid protease and amylase did not show significant daily rhythms. Plasma glucose showed a daily rhythm with the achrophase shifted by 12h in the ML and MD groups. These results revealed that the feeding time and light cycle synchronize differently the daily rhythms of behavior, digestive physiology and plasma metabolites in the Nile tilapia, which indicate the plasticity of the circadian system and its synchronizers.

Synchronization to light and mealtime of the circadian rhythms of self-feeding behavior and locomotor activity of white shrimps (Litopenaeus vannamei).

The role of light and feeding cycles in synchronizing self-feeding and locomotor activity rhythms was studied in white shrimps using a new self-feeding system activated by photocell trigger. In experiment 1, shrimps maintained under a 12:12h light/dark (LD) photoperiod were allowed to self-feed using feeders connected to a photoelectric cell, while locomotor activity was recorded with a second photocell. On day 30, animals were subjected to constant darkness (DD) for 12days to check the existence of endogenous circadian rhythms. In the experiment 2, shrimps were exposed to both a 12:12h LD photoperiod and a fixed meal schedule in the middle of the dark period (MD, 01:00h). On day 20, shrimps were exposed to DD conditions and the same fixed feeding. On day 30, they were maintained under DD and fasted for 7days. The results revealed that under LD, shrimps showed a clear nocturnal feeding pattern and locomotor activity (81.9% and 67.7% of total daily food-demands and locomotor activity, respectively, at nighttime). Both feeding and locomotor rhythms were endogenously driven and persisted under DD with an average period length (τ) close to 24h (circadian) (τ=24.18±0.13 and 23.87±0.14h for locomotor and feeding, respectively). Moreover, Shrimp showed a daily food intake under LD condition (1.1±0.2gday(-1) in the night phase vs. 0.2±0.1gday(-1) in the light phase). Our findings might be relevant for some important shrimp aquaculture aspects, such as developing suitable feeding management on shrimp farms.

Daily rhythms after vaccination on specific and non-specific responses in Nile tilapia (Oreochromis niloticus)

ABSTRACT We evaluated the daily changes in immunological and hematological factors in tilapia (Oreochromis niloticus) after an immunization period with a subsequent challenge. Experiments were divided into two phases: Phase 1 (immunization): 144 fish were distributed into two groups with 72 fish in six tanks. One group (T1) was immunized, comprising six vaccination time points (ZT schedule = ZT2 h, ZT6 h, ZT10 h, ZT14 h, ZT18 h, ZT22 h). The same schedule was applied to the other group, but with saline solution (non-vaccinated: T2). Both groups remained in the laboratory for 30 days (considered the immunization period). Phase 2 (challenge): on day 30, both vaccinated and non-vaccinated groups were challenged with Streptococcus agalactiae (2.0 × 107 CFU mL−1) following the same ZT schedule to stimulate the immune response without leading to widespread infection and mortality. On day 45, blood and head kidney samples were collected during the same ZT schedule. The variations in time of the following parameters within each group were evaluated: hematology, peroxidase activity, IgM, tnf-α3, tgf-β1, il-1β and il-12 gene expression. No significant mortality was observed for the groups or the ZT schedule (p > 0.05). Daily rhythms with diurnal acrophases were found in T2 for il12, tnf-α3 and tgf-β1 expression gene, while the acrophases of the peroxidase level, hematocrit and thrombocytes were at nighttime (p < 0.05). In contrast, most of the parameters in the vaccinated tilapia showed no daily rhythms (p > 0.05), except IgM. For all the parameters, the interaction effect between time and treatment (vaccinated and non-vaccinated groups) depended on ZT. Our results reveal that the humoral and non-specific immune system displayed a circadian rhythm based on the light-dark cycle, which could be affected by the vaccination procedure in tilapia.

Daily rhythms of swimming activity, synchronization to different feeding times and effects on anesthesia practice in an Amazon fish species (Colossoma macropomum)

ABSTRACT This study aimed to investigate the existence of day-night differences in the time for anesthesia and recovery in tambaqui exposed to the anesthetic eugenol and the influence of feeding time. Thus, we evaluated: (1) swimming activity; (2) food anticipatory activity (FAA) as a synchronizer of swimming activity and change to susceptibility to anesthetic; and (3) the effects of diurnal/nocturnal anesthesia exposure of fish feeding in the mid-light phase: 12:00 h (ML) and fish feeding in the mid-dark phase: 00:00 h (MD). Our findings revealed strictly nocturnal activity for tambaqui (94.2%), known as diurnal fish to date. Moreover, FAA was observed in tambaqui fed at MD, which showed a sustained increase in activity that began 2 h before feeding time and lasted until feeding. In contrast, no FAA was observed in fish fed at ML. Regarding anesthesia by day or night, the tambaqui treated with eugenol exhibited no difference in induction time. However, differences were observed in recovery times, with fish anesthetized at day recovering in 1–2 min and fish anesthetized at night recovering in 5–7 min. In short, our findings revealed for the first time the nocturnal behavior of tambaqui. These results indicated that recovery by day/night by eugenol in tambaqui has a strong dependence of behavioral patterns and the time of day.