Induced-ovulation in female dromedary camel involves kisspeptin neuron activation by ß nerve growth factor†.

The dromedary camel (Camelus dromedarius) is a short-day desert breeder in which female ovulation is induced by mating. Current data indicate that male-induced ovulation is triggered by its seminal plasma nerve growth factor beta (ß-NGF), but the exact mechanisms involved in the induction of ovulation are still unknown. In this study, we report that an intramuscular injection of ß-NGF in sexually active short-day-adapted female camels induces an ovulation attested by a surge of circulating LH (2-6 h after treatment) followed by an oocyte release with its cumulus oophorus (confirmed by ultrasonography 72 h after treatment) and a large and progressive increase in circulating progesterone (significant from the 2nd to the 10th days after ß-NGF injection). In addition, this ß-NGF treatment induces a broad nuclear c-FOS activation in cells located in various hypothalamic areas, notably the preoptic area, the arcuate nucleus, the dorso- and ventromedial hypothalamus, the paraventricular nucleus, and the supraoptic nucleus. A double immunostaining with neuropeptides known to be involved in the central control of reproduction indicates that ~28% kisspeptin neurons and 43% GnRH neurons in the proptic area, and ~10% RFRP-3 neurons in the dorso- and ventromedial hypothalamus are activated following ß-NGF injection. In conclusion, our study demonstrates that systemic ß-NGF induces ovulation in the female dromedary camel and indicates that this effect involves the central activation of hypothalamic neurons, notably the kisspeptin neurons.

Sleep pattern in the dromedary camel: a behavioral and polysomnography study.

STUDY OBJECTIVES: To investigate sleep patterns in the camel by combining behavioral and polysomnography (PSG) methods. METHODS: A noninvasive PSG study was conducted over four nights on four animals. Additionally, video recordings were used to monitor the sleep behaviors associated with different vigilance states. RESULTS: During the night, short periods of sporadic sleep-like behavior corresponding to a specific posture, sternal recumbency (SR) with the head lying down on the ground, were observed. The PSG results showed rapid shifts between five vigilance states, including wakefulness, drowsiness, rapid eye movement (REM) sleep, non-REM (NREM) sleep, and rumination. The camels typically slept only 1.7 hours per night, subdivided into 0.5 hours of REM sleep and 1.2 hours of NREM sleep. Camels spent most of the night being awake (2.3 hours), ruminating (2.4 hours), or drowsing (1.9 hours). Various combinations of transitions between the different vigilance states were observed, with a notable transition into REM sleep directly from drowsiness (9%) or wakefulness (4%). Behavioral postures were found to correlate with PSG vigilance states, thereby allowing a reliable prediction of the sleep stage based on SR and the head position (erected, motionless, or lying down on the ground). Notably, 100% of REM sleep occurred during the Head Lying Down-SR posture. CONCLUSIONS: The camel is a diurnal species with a polyphasic sleep pattern at night. The best correlation between PSG and ethogram data indicates that sleep duration can be predicted by the behavioral method, provided that drowsiness is considered a part of sleep.

Seasonal variations in locomotor activity rhythm and diurnal activity in the dromedary camel (Camelus dromedarius) under mesic semi-natural conditions.

The dromedary camel (Camelus dromedarius) is a large ungulate that copes well with the xeric environment of the desert. Its peculiar adaptation to heat and dehydration is well-known. However, its behavior and general activity is far from being completely understood. The present study was carried out to investigate the ecological effect of the various seasons on the locomotor activity (LA) rhythm and diurnal activity of this species. Six adult female camels were maintained under mesic semi-natural conditions of the environment during four periods of 10 days in each season: autumn, winter, spring and summer. In addition, three female camels were used to test the effect of rain on the LA rhythm during a period of 18 days during the winter. The animal's LA was recorded using the locomotion scoring method. Camels displayed a clear 24.0h LA rhythm throughout the four seasons. Activity was intense during Day-time (6-22 fold higher in comparison to night) and dropped or completely disappeared during nighttime. Mean daytime total activity was significantly higher in the summer as compared to winter. Regardless of the season, the active phase in camels coincided with the time of the photophase and thermophase. Furthermore, the daily duration of the time spent active was directly correlated to the seasonal changes of photoperiod. The diurnal activity remained unchanged over the four seasons. For each season, the start and the end of the active phase were synchronized with the onset of sunrise and sunset. At these time periods, temperature remained incredibly stable with a change ranging from 0.002 to 0.210°C; whereas, changes of light intensity were greater and faster with a change from 0.1 to 600 lux representing a variation of 3215-7192 fold in just 25-29 min. Rainfall affected the pattern of the LA rhythm with occurrence of abnormal nocturnal activity during nighttime disturbing nocturnal rest and sleep. Here we show that the dromedary camel exhibits significant seasonal changes of its activity within daylight hours. However, the diurnal pattern remains unchanged regardless of the season; whereas, abnormal nocturnal activity is observed during periods of rain. The activity onset and offset in this species seems to be primarily driven by the changes in light intensity at dusk and dawn.

Entrainment of circadian rhythms of locomotor activity by ambient temperature cycles in the dromedary camel.

In the dromedary camel, a well-adapted desert mammal, daily ambient temperature (Ta)-cycles have been shown to synchronize the central circadian clock. Such entrainment has been demonstrated by examining two circadian outputs, body temperature and melatonin rhythms. Locomotor activity (LA), another circadian output not yet investigated in the camel, may provide further information on such specific entrainment. To verify if daily LA is an endogenous rhythm and whether the desert Ta-cycle can entrain it, six dromedaries were first kept under total darkness and constant-Ta. Results showed that the LA rhythm free runs with a period of 24.8-24.9 h. After having verified that the light-dark cycle synchronizes LA, camels were subjected to a Ta-cycle with warmer temperatures during subjective days and cooler temperatures during subjective nights. Results showed that the free-running LA rhythm was entrained by the Ta-cycle with a period of exactly 24.0 h, while a 12 h Ta-cycle phase advance induced an inversion of the LA rhythm and advanced the acrophase by 9 h. Similarly, activity onset and offset were significantly advanced. All together, these results demonstrate that the Ta-cycle is a strong zeitgeber, able to entrain the camel LA rhythm, hence corroborating previous results concerning the Ta non-photic synchronization of the circadian master clock.

Melatonin rhythm and other outputs of the master circadian clock in the desert goat (Capra hircus) are entrained by daily cycles of ambient temperature.

In desert areas, mammals such as camel and goat are exposed to harsh environmental conditions. The ambient temperature (Ta) cycles have been shown to entrain the circadian clock in the camel. In the present work, we assumed that, in the goat living in a desert biotope, Ta cycles would have the same synchronizing effect on the central clock. Therefore, the effects of Ta cycles on body temperature (Tb), locomotor activity (LA) and melatonin (Mel) rhythms as outputs of the master circadian clock have been studied. The study was performed on bucks kept first under constant conditions of total darkness (DD) and constant Ta, then maintained under DD conditions but exposed to Ta cycles with heat period during subjective day and cold period during subjective night. Finally, the Ta cycles were reversed with highest temperatures during the subjective night and the lowest temperatures during the subjective day. Under constant conditions, the circadian rhythms of Tb and LA were free running with an endogenous period of 25.3 and 25.0 hours, respectively. Ta cycles entrained the rhythms of Tb and LA to a period of exactly 24.0 hours; while when reversed, the Ta cycles led to an inversion of Tb and LA rhythms. Similarly, Ta cycles were also able to entrain Mel rhythm, by adjusting its secretion to the cooling phase before and after Ta cycles inversion. All together, these results show that the Ta cycles entrain the master circadian clock in the goat.

The dromedary camel displays annual variation in hypothalamic kisspeptin and Arg-Phe-amide-related peptide-3 according to sex, season, and breeding activity.

The dromedary camel (Camelus dromedarius) is a desert mammal whose cycles in reproductive activity ensure that the offspring's birth and weaning coincide with periods of abundant food resources and favorable climate conditions. In this study, we assessed whether kisspeptin (Kp) and arginine-phenylalanine (RF)-amide related peptide-3 (RFRP-3), two hypothalamic peptides known to regulate the mammalian hypothalamo-pituitary gonadal axis, may be involved in the seasonal control of camel's reproduction. Using specific antibodies and riboprobes, we found that Kp neurons are present in the preoptic area (POA), suprachiasmatic (SCN), and arcuate (ARC) nuclei, and that RFRP-3 neurons are present in the paraventricular (PVN), dorsomedial (DMH), and ventromedial (VMH) hypothalamic nuclei. Kp fibers are found in various hypothalamic areas, notably the POA, SCN, PVN, DMH, VMH, supraoptic nucleus, and the ventral and dorsal premammillary nucleus. RFRP-3 fibers are found in the POA, SCN, PVN, DMH, VMH, and ARC. POA and ARC Kp neurons and DMH RFRP-3 neurons display sexual dimorphism with more neurons in female than in male. Both neuronal populations display opposed seasonal variations with more Kp neurons and less RFRP-3 neurons during the breeding (December-January) than the nonbreeding (July-August) season. This study is the first describing Kp and RFRP-3 in the camel's brain with, during the winter period lower RFRP-3 expression and higher Kp expression possibly responsible for the HPG axis activation. Altogether, our data indicate the involvement of both Kp and RFRP-3 in the seasonal control of the dromedary camel's breeding activity.

Dataset concerning haematological and biochemical parameters changes in show jumping horses subjected to exercise and plasmapheresis session.

This article presents data on the effect of plasmapheresis on clinical, haematological and biochemical parameters in horses following exercise and after a plasmapheresis session. This blood filtration technique was realised on six jumping horses (plasmapheresis group) that underwent three consecutive days of graded physical exercise. The control group (n = 6) went through the same exercise but was not subjected to the plasmapheresis session. Blood was sampled before and after each exercise, also at the beginning and the end of plasmapheresis session. The presented data was obtained by measuring clinical and haemato-biochemical parameters in both groups. The heart and respiratory rates and rectal temperature were recorded. In addition, the number of red blood and white cells, platelets also of lymphocytes, monocytes, eosinophils and granulocytes were counted. Other haematological parameters including, hemoglobin concentration, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration were determined. Concerning the biochemical parameters, the concentrations of albumin, globulin, total protein, glucose, alkaline phosphatase, aspartate aminotransferase, gamma glutamyl transferase, total bilirubin, lactate, creatinine kinase, urea, creatinine, calcium, sodium and potassium were measured. All parameters data were analyzed by a two-way repeated measures analysis of variance followed by Holm-Sidak post-hoc procedure to evaluate the effect of plasmapheresis and time. This paper contains data related to and supporting research articles currently published entitled "Plasmapheresis effect on haematological and biochemical parameters in athletic horses subjected to exercise" (Daden et al., 2019) [1].

Plasmapheresis Effect on Hematological and Biochemical Parameters in Athletic Horses Subjected to Exercise.

To evaluate the effect of plasmapheresis on clinical, hematological, and biochemical parameters after exercise, a plasmapheresis session was realized on six jumping horses (plasmapheresis group) that underwent three consecutive days of physical graded exercise. The control group (n = 6) went through the same exercise but not subjected to the plasmapheresis session. Seventeen milliliters of plasma/kg of body weight was harvested from each horse. The procedure was well tolerated by the horses. The plasmapheresis leads to a significant increase of the hemoglobin, hematocrit, red blood cell, white blood cell, and lymphocytes counts. Plasmapheresis induced a very significant decrease (P < .001) of albumin and globulin levels and of total protein, which were reestablished 24 hours later. Plasmapheresis also generated a very significant increase (P < .001) in sodium levels and a significant decline of potassium (P < .05) and calcium (P < .01) levels. Several other biochemical variables remained unchanged. Results also showed that, after a significant rise of lactate, aspartate aminotransferase, and creatine kinase levels which are subsequent to the exercise, the plasmapheresis session induced a very significant continuous decrease (P < .001) of these parameters. The present work has demonstrated that the plasmapheresis is able to modify the physiology after exercise and to affect both the hematology and the biochemistry of the blood hematobiochemical parameters in horses subjected to physical exercise.

Smartphone and a freely available application as a new tool to record locomotor activity rhythm in large mammals and humans.

Daily pattern of locomotor activity (LA), one of the most studied rhythms in humans and rodents, has not been widely investigated in large mammals. This is partly due to the high cost and breakability of used automatic devices. Since last decade, smartphones are becoming ubiquitous. Meanwhile, several applications detecting activity by using internal sensors were made available. In this study, we assumed that this device could be a cheaper and easier way to measure the LA rhythm in humans and large mammals, like camel and goat. A smartphone application (Nokia Mate Health), normally used to quantify physical activities in humans, was chosen for the study. To validate the rhythm data obtained from the smartphone, LA rhythm was simultaneously recorded using an automatic device, the Actiwatch-Mini®. Results showed that the smartphone provided a clear and significant daily rhythm of LA. The visual assessment of the superimposed LA rhythm's curves in all three species showed that the smartphone application displayed similar rhythms as those recorded by the Actiwatch-Mini. Highly significant positive correlation (p≤ 0.0001) exists between the two recording rhythms. The daily periods were both the same at 24.0 h. Acrophases were also significantly similar and occurring around mid-day: 11:40 ± 0.35 h vs 11:41 ± 0.35 h for the camel, 11:25 ± 0.19 h vs 11:37 ± 0.25 h for the goat and 13:04 ± 0.11 h vs 13:51 ± 0.28 h for humans using smartphone and Actiwatch, respectively. The related mesor and amplitude were also close between the two recording devices. Results indicate clearly that using smartphones constitutes a reliable cheap tool to study LA rhythm for chronobiology studies, especially in laboratories facing lack of funding.

The Suprachiasmatic Nucleus of the Dromedary Camel (Camelus dromedarius): Cytoarchitecture and Neurochemical Anatomy.

In mammals, biological rhythms are driven by a master circadian clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus. Recently, we have demonstrated that in the camel, the daily cycle of environmental temperature is able to entrain the master clock. This raises several questions about the structure and function of the SCN in this species. The current work is the first neuroanatomical investigation of the camel SCN. We carried out a cartography and cytoarchitectural study of the nucleus and then studied its cell types and chemical neuroanatomy. Relevant neuropeptides involved in the circadian system were investigated, including arginine-vasopressin (AVP), vasoactive intestinal polypeptide (VIP), met-enkephalin (Met-Enk), neuropeptide Y (NPY), as well as oxytocin (OT). The neurotransmitter serotonin (5-HT) and the enzymes tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC) were also studied. The camel SCN is a large and elongated nucleus, extending rostrocaudally for 9.55 ± 0.10 mm. Based on histological and immunofluorescence findings, we subdivided the camel SCN into rostral/preoptic (rSCN), middle/main body (mSCN) and caudal/retrochiasmatic (cSCN) divisions. Among mammals, the rSCN is unusual and appears as an assembly of neurons that protrudes from the main mass of the hypothalamus. The mSCN exhibits the triangular shape described in rodents, while the cSCN is located in the retrochiasmatic area. As expected, VIP-immunoreactive (ir) neurons were observed in the ventral part of mSCN. AVP-ir neurons were located in the rSCN and mSCN. Results also showed the presence of OT-ir and TH-ir neurons which seem to be a peculiarity of the camel SCN. OT-ir neurons were either scattered or gathered in one isolated cluster, while TH-ir neurons constituted two defined populations, dorsal parvicellular and ventral magnocellular neurons, respectively. TH colocalized with VIP in some rSCN neurons. Moreover, a high density of Met-Enk-ir, 5-HT-ir and NPY-ir fibers were observed within the SCN. Both the cytoarchitecture and the distribution of neuropeptides are unusual in the camel SCN as compared to other mammals. The presence of OT and TH in the camel SCN suggests their role in the modulation of circadian rhythms and the adaptation to photic and non-photic cues under desert conditions.

Daily regulation of body temperature rhythm in the camel (Camelus dromedarius) exposed to experimental desert conditions.

In the present work, we have studied daily rhythmicity of body temperature (Tb) in Arabian camels challenged with daily heat, combined or not with dehydration. We confirm that Arabian camels use heterothermy to reduce heat gain coupled with evaporative heat loss during the day. Here, we also demonstrate that this mechanism is more complex than previously reported, because it is characterized by a daily alternation (probably of circadian origin) of two periods of poikilothermy and homeothermy. We also show that dehydration induced a decrease in food intake plays a role in this process. Together, these findings highlight that adaptive heterothermy in the Arabian camel varies across the diurnal light-dark cycle and is modulated by timing of daily heat and degrees of water restriction and associated reduction of food intake. The changed phase relationship between the light-dark cycle and the Tb rhythm observed during the dehydration process points to a possible mechanism of internal desynchronization during the process of adaptation to desert environment. During these experimental conditions mimicking the desert environment, it will be possible in the future to determine if induced high-amplitude ambient temperature (Ta) rhythms are able to compete with the zeitgeber effect of the light-dark cycle.

Entrainment of the circadian clock by daily ambient temperature cycles in the camel (Camelus dromedarius).

In mammals the light-dark (LD) cycle is known to be the major cue to synchronize the circadian clock. In arid and desert areas, the camel (Camelus dromedarius) is exposed to extreme environmental conditions. Since wide oscillations of ambient temperature (Ta) are a major factor in this environment, we wondered whether cyclic Ta fluctuations might contribute to synchronization of circadian rhythms. The rhythm of body temperature (Tb) was selected as output of the circadian clock. After having verified that Tb is synchronized by the LD and free runs in continuous darkness (DD), we submitted the animals to daily cycles of Ta in LL and in DD. In both cases, the Tb rhythm was entrained to the cycle of Ta. On a 12-h phase shift of the Ta cycle, the mean phase shift of the Tb cycle ranged from a few hours in LD (1 h by cosinor, 4 h from curve peaks) to 7-8 h in LL and 12 h in DD. These results may reflect either true synchronization of the central clock by Ta daily cycles or possibly a passive effect of Ta on Tb. To resolve the ambiguity, melatonin rhythmicity was used as another output of the clock. In DD melatonin rhythms were also entrained by the Ta cycle, proving that the daily Ta cycle is able to entrain the circadian clock of the camel similar to photoperiod. By contrast, in the presence of a LD cycle the rhythm of melatonin was modified by the Ta cycle in only 2 (or 3) of 7 camels: in these specific conditions a systematic effect of Ta on the clock could not be evidenced. In conclusion, depending on the experimental conditions (DD vs. LD), the daily Ta cycle can either act as a zeitgeber or not.