Hematological reference intervals in newborn dromedary calves in the first week after birth: Age and sex-related variations.

The establishment of hematological reference intervals (RIs) is an important tool to assess the health status of animals and to evaluate the impact of diseases at individual and population levels. Nowadays, specific RIs of hematological parameters in newborn dromedary camel calves at birth and during the first week after birth, are lacking. Therefore, RIs for the hematological variables from a complete blood cell count were established in 47 healthy newborn dromedary calves (18 females and 29 males). Blood samples were collected within 2 h after birth (d0), at 24 h (d1), at 3 (d3) and 7 days (d7) after birth, and analyzed within 24 h. The RIs were described based on the 95% confidence interval, and possible differences among mean values due to age (sampling time) and sex were investigated. Statistical analysis showed that age affected all the hematological variables except MCV, MCH, and MCHC, indicating that the adaptational process to the extrauterine life continues for several days after birth; sex affected most of the hematological variables, with higher RBC and PLT count, HGB, PCV, neutrophil population and neutrophil:lymphocyte ratio at d7 in females compared to males. These findings suggest possible sex-based differences in the physiological maturation mechanisms and deserves further investigations. To the best of the authors' knowledge, this is the first report of hematological RIs for newborn dromedary calves at birth up to 7 days of age; the RIs registered in the present study in newborns differ from those reported in adult dromedaries in literature, thus confirming the need for the adoption of separated reference ranges according to age also in the dromedary camel, as previously reported for other species.

Collection of epididymal semen in the tomcat (Felix catus) by stereomicroscope-aided retrograde flushing (SARF) improves sample quality.

In small and laboratory animals epididymal spermatozoa can be collected only by the mincing technique however, blood and cell debris contamination may adversely affect the quality of the samples. This study aimed at performing the retrograde flushing technique, for the first time, in the tomcat. In the first experiment attempts to insert a blunted needle into tomcats' ductus deferentes were made and the cannulation time and the success rate were recorded. In the second experiment, insertion of the needle into the ductus was carried out under a stereomicroscope for one testis, while the other was submitted to mincing for sperm recovery and the samples obtained from paired testes were compared. In the first study, only twenty-eight percent of the cannulation attempts were successful whereas in the second experiment, 91% of the deferentes were successfully cannulated: 80% of the ducts were cannulated in less than three minutes. The stereomicroscope-aided retrograde flushing samples showed significantly higher sperm viability, motility and velocities, as compared to those resulted from mincing. The domestic cat serves as a model for more than 36 species of wild felids; the developed stereomicroscope-aided retrograde flushing technique has the potential to become the gold standard method for harvesting spermatozoa of endangered species that have a small ductus deferens.

The Effect of Cushioned Centrifugation, with and without Enzymatic Reduction of Viscosity, on the Motility Pattern and Kinematic Parameters of Dromedary Camel Bull Spermatozoa.

In order to contribute to the development of semen processing procedures in camelids, the aims of the present study were to evaluate (i) the effect of 35% seminal plasma incubation on dromedary camel epididymal sperm motility and kinematic parameters, (ii) the effects of centrifugation, with cushion fluid and enzymatic reduction of viscosity (Papain + E64) during ejaculate processing, on the motility and kinematic parameters of dromedary camel ejaculates. The incubation with seminal plasma significantly reduced the percentage of progressively motile spermatozoa as well as the proportion of medium progressive spermatozoa whilst increasing the percentage of non-progressive spermatozoa. The centrifugation procedure improved the sperms' kinematic parameters, and the highest values were observed for samples centrifugated with cushion fluid. The samples treated with Papain + E64 showed a significant increase in both total and medium progressive spermatozoa, along with a reduction of non-progressive spermatozoa (p < 0.05). The results of this investigation show that a simple, cheap, and effective procedure, such as cushioned centrifugation, could improve the motility patterns of dromedary camel spermatozoa; in combination with enzymatic reduction of viscosity, this method leads to the best results in terms of recovery rates and sperms' kinematic parameters.

New Strategies for Conservation of Gentile di Puglia Sheep Breed, an Autochthonous Capital of Millennial Tradition in Southern Italy.

Gentile di Puglia (GdP) is an autochthonous sheep breed of Southern Italy included among ovine breeds threatened by genetic erosion and extinction risk, which have been given attention by local and international institutions, thus emphasizing the need for germplasm conservation actions. In the present study, two assisted reproduction approaches, finalized for GdP conservation, were performed: (1) on-farm reproductive efficiency evaluation, expressed as pregnancy rate (PR), twin pregnancy rate (tPR), and body condition score (BCS), for three consecutive breeding cycles and (2) pre-pubertal lambs' immature cumulus-oocyte complex (COC) retrieval, vitrification, in vitro maturation (IVM), and assessment of meiotic stage and bioenergetic-oxidative status compared with those of other Italian and European commercial breeds. PR and tPR were progressively reduced over time. In all clinical examination times, BCS was significantly lower in nonpregnant ewes compared with pregnant ones. Fresh GdP pre-pubertal lamb COCs achieved meiotic maturation and showed healthy bioenergetic-oxidative status after IVM. Vitrification reduced the oocyte maturation rate in all groups. However, mature oocytes retained their cytoplasmic maturity, expressed as a mitochondria distribution pattern and activity, indicating promising developmental competence. In conclusion, clinical- and biotechnological-assisted reproduction approaches can support conservation strategies of GdP and other local sheep breeds in Southern Italy.

Blood Gas, Acid-Base and Electrolyte Analysis in Healthy Dromedary Camel Calves up to 21 Days of Life.

The importance of prompt evaluation and care of the newborn is essential for reducing neonatal mortality, which represents a major cause of loss in camelids. This study investigated the blood gases, acid-base and electrolyte profiles in healthy dromedary calves during the first 3 weeks of life, assessing possible associations with age. Twenty-one dromedary camel calves aged 1 to 21 days were sampled, and venous whole blood analyzed through a VETSTAT® analyzer. The following parameters were measured: sodium (Na+), potassium (K+), chloride (Cl-), hydrogen ion concentration (pH), partial pressure carbon dioxide (pCO2), partial pressure oxygen (pO2), total hemoglobin concentration (tHb), hemoglobin oxygen saturation (sO2), total carbon dioxide (tCO2), bicarbonate (HCO3-), base excess (BE) and anion gap (AG). Calves were divided in two groups; younger calves (1-10 d), and older calves (11-21 d). Statistical analysis showed an effect of age, with lower K+ (p < 0.001) and higher Na+ and Cl- (p < 0.05) mean concentrations in the younger calves compared to the older ones, and higher pCO2 and lower sO2 mean concentrations in the older group. These preliminary results firstly described the blood gas, acid-base and electrolyte profiles in the healthy dromedary calf during the first 3 weeks of age, suggesting an effect of age on some parameters.

Deep-horn Artificial Insemination With Frozen Thawed Semen After re-extension in Autologous Seminal Plasma May Improve Pregnancy Rates in Jennies.

In the literature, the very low pregnancy rates after artificial insemination (AI) with frozen semen in donkeys were improved in one study after re-extension of the frozen-thawed semen in autologous seminal plasma. The aims of our study were (1) to describe in vitro post-thaw parameters of donkey jackass semen after re-extension in seminal plasma (SP) or in INRA96 and (2) to compare pregnancy rates in jennies bred with frozen-thawed semen using two different AI protocols. Semen collected from two Amiata donkey stallions, known to be fertile, was frozen in INRA96 supplemented with 2% egg yolk, and 2.2% glycerol. From the same jacks, seminal plasma was filtered, and stored at -20°C. According to the in vitro analysis, the post-thaw re-extension in SP some kinematic parameters decreased more over time. Two different AI protocols were tested: inseminating deeply into the uterine horn (DHAI) with or without thawed semen re-extension in SP. Higher pregnancy rates were obtained with re-extension in SP than when AIs were performed without SP. Further studies are needed to confirm the advantage of re-extension in autologous SP of donkey frozen-thawed semen before AI in jennies.

Effects of two different management systems on hormonal, behavioral, and semen quality in male dromedary camels.

Effects of two different management systems on male dromedary camel hormones, behaviors, and semen parameters were documented. Camels (n=6) were tested under two management systems: (i) housed in single boxes with 1-h freedom (H23); (ii) exposed to females for 17 h (from 3.30 p.m. to 8.30 a.m.) and then housed (ConExF). Blood was collected every morning; camel behavior was recorded twice a day: (i) from 7:00 to 8:00 a.m. to determine the short effects; (ii) from 2:00 to 3:00 p.m. to determine the long effects. Each camel underwent a female parade and semen collection thrice a week; sexual behavior, libido, and semen parameters were assessed. Testosterone and cortisol concentrations were higher in ConExF than H23. Compared to the H23 group, ConExF group spent more time walking, standing tripods, and looking outside their pen/box but they spent less time eating, ruminating, resting, standing, and showing stereotypical behaviors. In the morning, ConExF group spent more time walking, ruminating, and showing typical sexual behaviors compared to themselves during afternoon time and the H23 group. However, in the afternoon time, ConExF camels put more time their heads outside the box through the window and showed higher frequencies of stereotypies, probably due to a higher level of frustration. While the sexual behavioral score was higher and ejaculates showed a higher fraction of milky white and white-colored semen in ConExF than H23 group, their libido was similar. Overall, 17 h of exposure led to an increase in testosterone and cortisol levels, enhancing sexual behavior and semen color, but leading to frustration.

Seminal plasma Alters surface Glycoprofile of dromedary camel cryopreserved epididymal spermatozoa.

The high viscosity of Camelidae semen continues to present a major impediment for its application in assisted reproduction technology. The exposure of epididymal spermatozoa (ES) to seminal plasma (SP) may provide an approach to enhance the development of assisted reproductive techniques in these important domestic species. Since the sperm glycocalyx plays a key role in reproduction we aimed to evaluate whether SP exposure modifies the surface glycosylation patterns of cryopreserved dromedary ES. Epididymal sperm was collected through retrograde flushing of the cauda epididymidis that were obtained from orchidectomized mature dromedary bulls. The collected samples were then cryopreserved after dilution with a tris citrate clarified egg yolk extender, with and without the supplementation of 15% SP. Post-thaw carbohydrate surface profiles of both control and SP-treated spermatozoa were analyzed using 15 fluorescent lectins. Morpho-functional properties were also investigated via computer assisted sperm analysis. Lectin-binding analysis of the glycocalyx in control sperm revealed the presence of (1) N-glycans terminating with lactosamine (Con A, PHA-L, and RCA120), in both acrosomal and tail regions. Whilst (2) α2,3-/α2,6-linked sialic acids (MALII, SNA), and O-linked glycans terminating with a single N-acetylgalactosamine residue (Tn antigen) (HPA, SBA) along with galactoseß1,3N-acetylgalactosamine (T antigen) (PNA) were observed in the acrosomal cap. The expression of both N-acetylglucosamine (sWGA and GSA II) and terminalαgalactose (GSA I-B4) residues was also noted in the acrosomal cap region of control sperm. Compared with controls, SP treated samples displayed: 1) the appearance of bisected di-triantennary complex-type N-glycans (PHA-E), terminating with lactosamine, as well as an increase of O-glycans terminating with Tn and T antigens in both the acrosomal and tail regions; 2) an increase in glycans containing α2,6-linked sialic acid, N-acetylglucosamine, and αgalactose in the tail region. The cytoplasmic droplets of both control and seminal plasma-treated sperm bound Con A, PHA-E, PHA-L, RCA120, HPA, PNA, sWGA, GSA I-B4, and GSA II. These results indicate that SP treatment affects the glycan composition of the dromedary camel ES glycocalyx. More comprehensive studies are required in order to evaluate the fertilization capacity of SP-treated ES in order to facilitate its application in dromedary camel assisted reproduction technology.

Retrograde flushing collection and freezing of dromedary camel epididymal spermatozoa with seminal plasma.

The objectives of this study were to describe the parameters of dromedary camel epididymal spermatozoa collected by retrograde flushing (RF) technique and to evaluate the freezability of the collected sperm, diluted with and without the supplementation of seminal plasma (SP). Two experiments were conducted: in Experiment 1, ES were recovered within 6-8 h after castration; selected samples were diluted with a Tris-citrate egg-yolk glycerolated buffer and frozen. In Experiment 2, epididymides were stored for 24 h at 4 °C before RF and semen samples were frozen after dilution with a Tris-lactose egg-yolk glycerolated extender with and without 15% SP. In Experiment 1, eight semen samples were obtained from ten epididymides with a mean of 500 × 106 total spermatozoa recovered, per flushed epididymis. Mean post-thaw motility and progressive motility were 75 and 17%, respectively. In Experiment 2, 15 samples were collected, out of the 18 epididymides (mean number of collected spermatozoa: 700 × 106), and 13 of these samples were of excellent quality. Post-thaw parameters were not satisfactory but the supplementation of the freezing medium with 15% SP improved the progressive motility and kinematic parameters of the spermatozoa.

Post-thawing effects of three cryopreservation diluents on Rusa deer (Rusa timorensis) spermatozoa.

The aim of this study was to evaluate home-made and commercial extenders for the cryopreservation of Rusa deer semen. After collection by electroejaculation, six ejaculates were diluted and frozen in TES-based, Tris-based and Triladyl® extenders. Subjective motility, viability, morphology, acrosome integrity and membrane functionality were assessed post-thawing and after 1-hr incubation at 37°C (Thermal stress test). Total and progressive motility, and kinematic parameters were also assessed through CASA system. Post-thawing sperm progressive motility (PM), velocity according to the straight path (VSL) and linearity (LIN) showed significant differences, and higher values were detected for spermatozoa diluted with Triladyl® and TES (p < 0.05) as compared with Tris (PM of Triladyl® 14.7% vs. 3.2% TES and 2.5% Tris; VSL 56 for Triladyl® , 59.2 for TES and 41.7 for Tris; LIN 45.6 for Triladyl® , 52 for TES and 36.5 for Tris). Triladyl® and TES extender led to better post-thawing sperm parameters, but these preliminary results need to be verified through artificial insemination trials.

THE ACUTE-PHASE AND HEMOSTATIC RESPONSE IN DROMEDARY CAMELS ( CAMELUS DROMEDARIUS).

Acute-phase reactants indicate inflammation and are increasingly used in veterinary medicine to indicate and to monitor progression of disease. Hemostasis and inflammation have interconnected pathophysiologic pathways and influence each other on different levels. This study established observed normal ranges for acute-phase reactants and for coagulation and thromboelastographic (TEG) parameters in 49 dromedary camels ( Camelus dromedarius) and assessed the response to chronic and acute inflammation. Chronically infected animals suffering from lymph abscessation due to Corynebacterium spp. had significantly higher concentrations of the acute-phase reactants haptoglobin ( P < 0.005) and fibrinogen ( P < 0.013) and an increased clot strength characterized by an increase of the TEG parameters MA ( P < 0.039), representing the maximum amplitude of the clot strengths, and G, the global clot strength ( P < 0.022), compared to healthy animals. When the acute-phase and hemostatic responses of 10 males receiving a gonadotropin-releasing hormone vaccine and of 9 males that were surgically castrated over 7 days were studied, haptoglobin proved to be a minor positive acute-phase protein, with moderate levels in healthy animals. It increased significantly after both vaccination and castration and remained elevated 7 days postinsult. The negative reactant iron significantly decreased over the 7-day period after castration, whereas a similar decrease following vaccination lasted less than 3 days. Fibrinogen reacted as a positive, minor reactant, with a significant increase and a peak on days 3-5, with higher values seen after castration. Prothrombin time showed a slight shortening at days 5-7, and the TEG parameters MA and G showed significantly increased values, similar to fibrinogen. The acute-phase protein serum amyloid A showed poor repeatability, suggesting that the assay was not reliable.

Physical and kinematic properties of cryopreserved camel sperm after elimination of semen viscosity by different techniques.

This investigation aimed to determine the influence of using different techniques for liquefaction of semen on post-thaw physical and dynamic characteristics of camel spermatozoa. A total of 144 ejaculates were collected from 3 adult camels, Camelus dromedarius, twice-weekly over 3 consecutive breeding seasons. A raw aliquot of each ejaculate was evaluated for physical and morphological properties, whereas the remaining portion was diluted (1:3) with glycerolated Tris lactose egg yolk extender, and was further subjected to one of the following liquefaction treatments: control (untreated), 5µl/ml α-amylase, 0.1mg/ml papain, 5u/ml bromelain, or 40-kHz nominal ultrasound frequency. The post-thaw objective assessment of cryopreserved spermatozoa, in all groups, was performed by a computer-assisted sperm analysis (CASA) system. The results revealed that all liquefaction treatments improved (P<0.05) post-thaw motility, viability and sperm motion criteria. However, an adverse effect (P<0.05) was observed in acrosome integrity, sperm cell membrane integrity and percent of normal sperm in all enzymatically-treated specimens compared to both control and ultrasound-treated semen. These results elucidate the efficiency of utilizing ultrasound technology for viscosity elimination of camel semen. In addition, developing enzymatic semen liquefaction techniques is imperious to benefit from when applying assisted reproductive technologies, particularly AI and IVF, in camels.

Daily rhythms of behavioral and hormonal patterns in male dromedary camels housed in boxes.

BACKGROUND: Daily rhythmicity has been observed for a number of hormonal and behavioral variables in mammals. It can be entrained by several external factors, such as light-dark cycle and scheduled feeding. In dromedary camels, daily rhythmicity has been documented only for melatonin secretion and body temperature. In this study, the daily rhythmicity of behavioral repertoire, cortisol and testosterone levels was investigated in captive male camels. METHODS: Six clinically healthy male dromedary camels (Camelus dromedarius) were used. The animals were housed in single boxes for 24 h daily and fed twice a day. Over a period of 48 h, behavioral observations were made and blood samples taken every two hours. The data were analyzed using diurnality index, conisor analysis and PROC mixed procedure. RESULTS: The diurnality index for rumination and lying down was close to 0 (respectively, 0.09 and 0.19), while the indices for stereotypy, standing, feeding and walking were close to 1 (respectively, 0.74, 0.84, 0.92 and 0.85). Cosinor analysis revealed daily rhythmicity for all behaviors and for cortisol levels (acrophase at 12:57) but not for testosterone. Rumination and lying down (inactive behaviors) reached a peak during the scotophase, whereas feeding, walking and stereotypy (active behaviors) reached a peak during the photophase around midday. Cortisol level and expression of stereotypies peaked before and after food distribution and were negatively correlated (r =  - 0.287, P = 0.005). Testosterone levels and expression of sexual behaviors were stimulated by the visual and olfactory contacts with the females and were positively correlated (r = 0.164, P = 0.040). Testosterone was also negatively correlated with cortisol (r =  - 0.297; P = 0.003). DISCUSSION: These preliminary results provided new knowledge about the daily rhythm of behaviors in camels housed in boxes, suggesting that camels exhibit diurnal behavior pattern in the maintenance conditions outlined in the study. Daily rhythmicity seemed to be entrained not only by the light-dark cycle but also by scheduled feeding. The rise in stereotypy after food distribution could be due to the persistence of feeding motivation and frustration after the ingestion of food. Therefore, feeding practices should be improved to satisfy the foraging and feeding motivation of these camels. Behavioral and hormonal daily patterns in camels should be taken in consideration to adapt the management system, giving the animals more freedom during the light period and a diet richer in fiber, so as to improve reproductive performance, health and welfare.

Effect of α-Amylase, Papain, and Spermfluid treatments on viscosity and semen parameters of dromedary camel ejaculates.

Ejaculates from five clinically healthy dromedary camels (Camelus dromedarius) were used to evaluate the effects of different enzymatic treatments (Amylase, Papain, Spermfluid) on liquefaction and seminal parameters. After collection, ejaculates were divided into 5 aliquots: (1) kept undiluted (control); or diluted 1:1 with: (2) Tris-Citrate-Fructose (TCF), (3) TCF containing Amylase, (4) TCF containing Papain or (5) Spermfluid containing Bromelain. At 120 min after dilution, each aliquot was evaluated, at 20-min intervals, for viscosity, motility, viability and agglutination. Only the aliquots diluted with TCF containing Papain underwent complete liquefaction. Sperm motility decreased significantly during the observation times, except for the samples diluted with Spermfluid (P=0.005). Diluted samples showed different levels of agglutination, with the lowest being observed in the control and the highest in the Papain-treated samples. The viscosity of dromedary camel ejaculates could be effectively reduced by using the proteolytic enzyme Papain.

Effects of a GnRH administration on testosterone profile, libido and semen parameters of dromedary camel bulls.

GnRH treatment has been suggested to increase testosterone levels temporarily and to stimulate libido in stallions, but its use has not fully ascertained in dromedary camels. The aim of this work was to study the effects of administering 100 µg of GnRH on testosterone profile, libido and semen parameters in dromedary camels. The same bulls were used as self-controls and experimental group. Blood samples were collected every 20 min (T0-T12) for 4h, and semen collections were performed over a 2-hour period after T12. GnRH was administered immediately after T0. In GnRH-treated bulls, testosterone levels showed an upward trend, peaking after 140 min, and then slowly decreasing. GnRH administration also led to a decrease in mating time and an increase in spermatozoa concentration. Overall, it seems that administration of 100 µg GnRH might increase testosterone levels temporarily and enhance camel reproduction performance.

Morphological and glycan features of the camel oviduct epithelium.

This study describes regional differences in the oviduct of the one-humped camel (Camelus dromedarius) during the growth phase (GP) and the mature phase (MP) of the follicular wave by means of morphometry, scanning electron microscopy (SEM) and glycohistochemistry investigations. Epithelium height significantly increased in the ampulla and decreased in the isthmus passing from the GP to the MP. Under SEM, non-ciliated cells displayed apical blebs (secretory) or short microvilli. Cilia glycocalyx expressed glycans terminating with sialic acid linked α2,6 to Gal/GalNAc (SNA affinity) throughout the oviducts of GP and MP and sialic acid linked α2,3 to Galß1,3GalNAc (MAL II and KOH-sialidase (K-s)-PNA staining) throughout the MP oviducts. Non-ciliated cells displayed lectin-binding sites from the supra-nuclear cytoplasm to the luminal surface. Ampulla non-ciliated cells showed O-linked (mucin-type) sialoglycans (MAL II and K-s-PNA) during GP and MP and N-linked sialoglycans (SNA) during the MP. Isthmus non-ciliated cells expressed SNA reactivity in GP and MP, also K-s-PNA binders in MP, and MAL II and PNA affinity (Galß1,3GalNAc) during GP. Galß1,3GalNAc was sialilated in the non-ciliated cells of GP UTJ. Luminal surface lacked of Galß1,3GalNAc in GP and MP, whereas it expressed α2,6- and α2,3-linked sialic acids. In GP intraluminal substance reacted with SNA, MAL II, K-s-PNA in ampulla and only with MAL II in the isthmus and UTJ. These results demonstrate that the morphology and the glycan pattern of the camel oviductal epithelium vary during the follicular wave and that could relate to the region-specific functions.

Could dromedary camels develop stereotypy? The first description of stereotypical behaviour in housed male dromedary camels and how it is affected by different management systems.

Dromedary camel husbandry has recently been evolving towards a semi-intensive system, due to the changes in use of the animal and the settlement of nomadic populations. Captivity could restrict its social activities, limiting the expression of various behavioural needs and causing the manifestation of stereotypy. The aims of this trial were, firstly, to identify and describe some stereotypical behaviours in captive male dromedary camels used for artificial insemination and, secondly, to study the effects on them of the following husbandry management systems: i) housing in single boxes for 24 hours (H24), ii) housing in single boxes for 23 hours with one hour free in the paddock (H23), and iii) housing in single boxes for 22 hours 30 min with 1 h of paddock time and 30 min exposure to a female camel herd (ExF). Every day, the camels were filmed in their single box in the morning for 30 minutes to record their behavioural activities and a focal animal sampling ethogram was filled in. In this study, male camels showed both oral and locomotor stereotypy most frequently when the bulls were reared in H24. Overall, this preliminary study is a starting point in the identification of stereotypies in male camels, reporting the positive effects of spending one hour outdoor and of social interaction with females.

Effect of different management systems on rutting behavior and behavioral repertoire of housed Maghrebi male camels (Camelus dromedarius).

Camel management has been changing in recent years from an extensive to a semi-intensive or intensive system, particularly for breeding bulls and dairy dromedary camels. Captivity may affect animal welfare, and low libido is the major complaint for housed breeding bulls. Since welfare status could also affect reproductive performance, the aim of this study was to evaluate different management practices on behavior, particularly on sexual behavior, and to identify some behavioral needs of male dromedary camels reared for semen collection. The effects of the following management systems on their behavior were compared: (i) traditional: housing in a single stall for 24 h (H24), (ii) housing in a single stall for 23 h with 1 h free in the paddock (H23), and (iii) housing in a single stall for 22 h and 30 min with 1 h paddock time and 30 min exposure to a female camel herd (ExF). During the trial, blood cortisol concentrations were assessed and camels were filmed daily for 30 min in the mornings and during a female passage in the evenings. Videos were analyzed in order to fill out a focal sampling ethogram and to score sexual behavior. As a result, there were no differences between the H24 and H23 systems, whereas ExF had a significant positive impact on their sexual behavior score and behavioral repertoire, further reducing cortisol levels. Overall, it seems that male dromedary camel welfare status improves when their behavioral needs for social interaction and movement are satisfied.

Confocal fluorescence assessment of bioenergy/redox status of dromedary camel (Camelus dromedarius) oocytes before and after in vitro maturation.

BACKGROUND: Reproductive biotechnologies in dromedary camel (Camelus dromedarius) are less developed than in other livestock species. The in vitro maturation (IVM) technology is a fundamental step for in vitro embryo production (IVP), and its optimization could represent a way to increase the success rate of IVP. The aim of the present study was to investigate the bioenergy/oxidative status of dromedary camel oocytes before and after IVM by confocal microscopy 3D imaging. METHODS: Oocytes were retrieved by slicing ovaries collected at local slaughterhouses. Recovered oocytes were examined before and after IVM culture for nuclear chromatin configuration and bioenergy/oxidative status, expressed as mitochondria (mt) distribution and activity, intracellular Reactive Oxygen Species (ROS) levels and distribution and mt/ROS colocalization. RESULTS: The mean recovery rate was 6 oocytes/ovary. After IVM, 61% of oocytes resumed meiosis and 36% reached the Metaphase II stage (MII). Oocyte bioenergy/redox confocal characterization revealed changes upon meiosis progression. Immature oocytes at the germinal vesicle (GV) stage were characterised by prevailing homogeneous mt distribution in small aggregates while MI and MII oocytes showed significantly higher rates of pericortical mt distribution organized in tubular networks (P<0.05). Increased mt activity in MI (P<0.001) and MII (P<0.01) oocytes compared to GV stage oocytes was also observed. At any meiotic stage, homogeneous distribution of intracellular ROS was observed. Intracellular ROS levels also increased in MI (P<0.01) and MII (P<0.05) oocytes compared to GV stage oocytes. The mt/ROS colocalization signal increased in MI oocytes (P<0.05). CONCLUSIONS: This study provides indications that qualitative and quantitative indicators of bioenergy and oxidative status in dromedary camel oocytes are modified in relation with oocyte meiotic stage. These data may increase the knowledge of camel oocyte physiology, in order to enhance the efficiency of IVP procedures.

Evaluation of sexual behavior of housed male camels (Camelus dromedarius) through female parades: correlation with climatic parameters.

Camels are seasonal breeders, and their sexual behavior is influenced by environmental conditions, but the relationship between climatic factors and sexual behavior has been poorly described in the available literature. Nowadays, the male camel living habit is shifting towards captivity; thus, this study was carried out to evaluate the sexual behavior of housed male dromedary camel through female's parades and to correlate it with climatic parameters. Four housed sires, reared for semen collection, and one dam were used and the trial lasted 8 weeks, considering the first week as control. Six days per week and during evenings, the female was brought near each males' boxes, while two observers filled a behavioral sampling ethogram and scored the male sexual behavior. After this parade, blood samples were taken from the female to evaluate the estradiol concentration. In addition, the following meteorological parameters were recorded, everyday, at 9:00 a.m. and 19:00 p.m.: pressure, wind, temperature, humidity, and H-index. The correlation between sexual behavioral score and female estradiol concentration and climate parameters was analyzed. All the behavioral parameters showed a significant upward trend; female estradiol concentration varied during the period and picked at week 5. Male sexual behavior was negatively correlated with morning H-index, wind, and temperature, and positively correlated with pressure and evening humidity, whereas it was not correlated with estrogen. In conclusion, female parade was a successful method to evaluate and stimulate the occurrence of housed male dromedary camel sexual activity that resulted to be negatively affected by hot temperature, warm wind, and lack of rain.