Impact of antibiotics on spermatozoa quality and bacterial load of chilled-stored camels (Camelus dromedarius) semen.

This experiment was conducted to assess the effect of different antibiotics in tris-fructose egg yolk-based diluent on bacterial load and sperm quality of dromedary camels during processing and cold storage. Ten semen ejaculates were collected from five male dromedary camels. Each sample was fractioned into four equally divided aliquots and diluted in one of four tris-fructose egg yolk. The first extender contained no antibiotic (NC). The second extender included streptomycin sulphate (1000 µg/ml) and benzyl penicillin (1000 IU/ml) (SP). The third extender was supplied with 250 µg/ml gentamicin sulphate (Gent). The fourth extender contained 500 µg/ml gentamicin sulphate,100 µg/ml tylosin tartrate, 300 µg/ml lincomycin hydrochloride and 600 µg/ml spectinomycin hydrochloride (GTLS). After dilution, the extended semen samples were cooled to 5 °C within 2 h and finally stored at 5 °C for 72 h. Microbial concentration, motility of spermatozoa, live spermatozoa, plasma membrane and acrosome integrity percentages were evaluated just after dilution at 35 °C, 0, 24, 48 and 72 h from the start of cooling to 5 °C. The results revealed that the diluent containing gentamicin had significantly (P<0.05) maximum motility percentage at the different examination intervals. The pattern of live spermatozoa percentage was varied between the different treatments at different examination intervals. The diluent supplied with Gent was distinguished with a significant peak percentage (P<0.05) of swelled spermatozoa among the other antibiotics supplied diluents. The number of colony-forming units isolated from the semen samples kept in diluent containing no antibiotics was significantly (P<0.05) higher than that isolated from the diluents supplemented with antibiotics. In conclusion, the semen diluents fortified with gentamicin generally keep the motility, acrosomal and plasma membrane integrity and live spermatozoa for 72-h preservation of dromedary semen.

Effect of heavy metals arsenic, cadmium, and lead on the semen variables of dromedary camels (Camelus dromedarius).

In this study, there was investigation of the effect of heavy metals on the fertility of dromedary camels. Fourteen camels at the Camel Research Center, King Faisal University, and 41 infertile dromedaries admitted to the Veterinary Teaching Hospital were used for semen evaluation during the breeding season. Seminal plasma and blood serum were collected from all males until analysis. Concentrations of three heavy metals [arsenic (As), cadmium (Cd), and lead (Pb)] were determined in the seminal plasma and serum using an atomic absorption spectrophotometer. The results indicate there are differences (P < 0.05 - P < 0.01) in pH, sperm motility, sperm concentration, and sperm abnormalities between the fertile and infertile male camels. In seminal plasma, there were marked differences (P < 0.01- P < 0.0001) between the control and infertile male camels in As, Cd, and Pb concentrations. In serum, there were differences (P < 0.01 - P < 0.001) between the fertile and infertile camels in serum As, Cd, and Pb concentrations. There was a positive correlation (P < 0.05; r = 0.77 and r = 0.94, respectively) between serum and seminal plasma concentrations of both As and Cd in the infertile dromedaries. In the control group, there was a positive correlation (P < 0.05; r = 0.70) between seminal plasma concentrations of Cd and percent sperm abnormalities. In conclusion, relatively greater seminal plasma and serum concentrations of As, Cd, and Pb are associated with lesser values for semen quality variables and infertility in dromedary camels.

Effect of dystocia on some hormonal and biochemical parameters in the one-humped camel (Camelus dromedarius).

The present study compared some of the hormonal and biochemical constituents of serum from eutocic and dystocic one-humped camels (Camelus dromedarius). Sera were harvested from eutocic (n = 9) and dystocic (n = 20) camels within the first 15 minutes after delivery. Although there were no differences in the concentrations of estradiol-17ß (E2) and prostaglandin F2α (PGF2α) between the eutocic and the dystocic animals, the level of progesterone (P4) and cortisol was significantly higher (P < 0.01) in animals that experienced dystocia than those that had a normal birth. There were no differences between the concentrations of alkaline phosphatase, aspartate aminotransferase, calcium, cholesterol, creatine kinase, creatinine, or magnesium (Mg) in eutocic and dystocic animals. The nitric oxide concentration was significantly higher (P < 0.01) in the serum from animals with dystocia than those that had normal births. By contrast, the serum concentrations of glucose, phosphorus (P), and triglycerides were significantly lower (P < 0.01) in eutocic camels compared with dystocic camels. As the delayed decline of P4 is reported to be the major hormonal difference between eutocic and dystocic camels, we propose that the insensitivity of corpus luteum to luteolytic action may be a cause of dystocia. Moreover, stress and hormonal changes may affect the metabolic traits in dystocia camels.

Biochemical and hormonal analysis of follicular fluid and serum of female dromedary camels (Camelus dromedarius) with different sized ovarian follicles.

The current study aimed to compare some biochemical and hormonal constituents in follicular fluids and serum of female dromedary camels with different sized ovarian follicles. Therefore, follicular fluids from follicles sized 1.1-1.5cm (n=10), 1.6-2.1cm (n=10) and 2.2-2.5cm (n=10) and sera were harvested from 20 female camels. The concentrations of ascorbic acid, glucose, cholesterol and activities of acid phosphatase (ACP) and alkaline phosphatase (ALP) were not changed significantly neither in follicular fluids of all follicle sizes nor in sera of female camels with different sized follicles. The concentrations of estradiol-17ß (E2) in the follicular fluid of follicles sized 2.2-2.5cm were significantly lower (P<0.01) than its corresponding value in follicular fluid of other follicle sizes. The concentrations of progesterone (P4), tri-iodothyronine (T3), thyroxin (T4), cortisol and insulin-like growth factor-1 (IGF-1) remained comparable in follicular fluids of all examined different sized follicles. The concentrations of E2, P4, T3, T4, cortisol and IGF-1 were similar in the serum of camels with different sized follicles. Interestingly, mean concentrations of P4 and IGF-1 in follicular fluids were higher than their corresponding values in sera of camels with different sized follicles and the mean concentrations of glucose, cholesterol, ALP and cortisol in sera were higher than their corresponding values in follicular fluids of the examined camels. With the exception of E2, there were no significant differences in biochemical and hormonal constituents between follicular fluids from different sized follicles.

Effect of oxytocin and PGF2α on chlortetracycline absorption from the uterus of early postpartum camels (Camelus dromedarius).

Fifteen parturient camels given chlortetracycline (CTC) as intrauterine pessaries (3 g/head) were divided into the control group (n = 5), which remained untreated, oxytocin-treated group (50 IU, intramuscular; n = 5), and cloprostenol-treated group (Estrumate, 500 µg, intramuscular; n = 5). Serum samples were collected at 0, 1, 2, 4, 8, 24, 48, and 72 hours after treatment and CTC was determined. The CTC appeared in blood within 1 hour. The maximum concentration of CTC was detected in blood after 72 (543.58 ± 117.85 µg/L), 8 (520.48 ± 13.65 µg/L), and 1 hour (831.98 ± 111.01 µg/L) of administration in control, oxytocin-, and PGF2α-treated camels, respectively. There was a high significant effect of time (P < 0.001) and treatment-by-time interaction (P < 0.001) on serum CTC concentration. In the control group, there was a significant (P < 0.05) increase in CTC concentrations at 72 hours compared to the other times. In the oxytocin group, there was a significant (P < 0.05) decrease in CTC concentrations at 24, 48, and 72 hours compared to its level after 1 or 8 hours. In PGF2α, there was a significant (P < 0.001) decrease in CTC concentrations at 2, 4, 8, 24, 48, and 72 hours compared to its level after 1 hour. Treatment contrast at different time points showed a significant (P < 0.001) increase in CTC concentration after 1 hour in the PGF2α-treated group compared to oxytocin and control groups. By 72 hours, CTC concentrations were significantly (P < 0.05) lower in PGF2α and oxytocin groups than in the control group. In conclusion, serum CTC concentration in dromedary camels increases within 1 hour after intrauterine administration and remains elevated for at least 72 hours in control, oxytocin-, and PGF2α-treated animals.

Relationship between the size of the dominant follicle, vaginal electrical resistance, serum concentrations of oestradiol and progesterone and sexual receptivity during the follicular phase of the dromedary camel (Camelus dromedarius).

Thirteen dromedaries were used to study the relationship between the size of the dominant follicle, vaginal electrical resistance (VER), sexual receptivity, and serum concentrations of oestradiol-17ß (E2) and progesterone (P4) throughout the follicular phase. On a daily basis, the animals experienced teasing with a vasectomised camel, trans-rectal ultrasound examination of the ovaries, and measurement of VER and blood collection for serum E2 and P4. Results revealed no significant differences between the mean VER in the animals that had a follicle of 5-10mm (group I, n=11), 11-15mm (group II, n=12) and 16-20mm (group III, n=13). The VER did not correlate with the follicular size. The E2 concentrations in the animals in groups II (60.14pg/ml) and III (66.52pg/ml) were significantly (P<0.05) higher than those of animals in group I (48.31pg/ml). E2 was positively correlated (r=0.50; P<0.05) with the overall size of the follicles. The P4 concentration was significantly (P<0.05) lower in the animals of group II than in those in groups I and III. Serum P4 concentrations were inversely correlated (r=-0.40; P<0.05) with the overall size of the follicles. Complete sexual receptivity was reported in 63.64%, 16.67% and 76.92% of the animals in groups I, II and III, respectively. In group III, significant (P<0.05) complete sexual receptivity was confirmed. However, 23.08% of the animals were incompletely receptive. In conclusion, impedometric characteristics of the vaginal mucosa were not a reliable method for predicting the dominant follicular size during the follicular phase in dromedary camels.

Seminal plasma and serum fertility biomarkers in dromedary camels (Camelus dromedarius).

Eight healthy fertile (control) and 11 infertile male dromedaries were used to investigate whether specific seminal plasma and serum fertility biomarkers could be related to their in vivo fertility. Eight fertility biomarkers and testosterone were determined in both seminal plasma and serum of all studied camels during the rutting season using commercial kits. Results revealed a significant (P < 0.01) difference in semen parameters between the control and infertile camels. There was a significant (P < 0.01) difference between the control and infertile dromedaries in seminal plasma glutathione peroxidase (GPx) activity (15.04 ± 1.14 vs. 4.55 ± 0.96 nmol/min/mL, respectively) and both phospholipase A2 (sPLA2; 50.66 ± 6.28 vs. 23.56 ± 4.29 pg/mL, respectively) and testosterone concentrations (732.14 ± 57.12 vs. 396.36 ± 79.34 pg/mL, respectively). A significant (P < 0.05) difference was found between the control and infertile camels in seminal plasma concentrations of osteopontin, cysteine-rich secretory protein 3 (CRISP3), and prostaglandin D synthase (lipocalcin-type). There was a significant (P < 0.01) difference between the fertile and infertile camels in serum GPx activity 67.81 ± 12.41 vs. 21.31 ± 4.63 nmol/min/mL, respectively) and both testosterone (599.57 ± 110.90 vs. 176.09 ± 24.05 pg/mL, respectively) and clusterin concentrations (137.29 ± 14.15 vs. 253.00 ± 17.14 ng/mL, respectively). A significant (P < 0.05) difference existed between the control and infertile male dromedaries in serum concentrations of sPLA2, CRISP3, malonialdehyde, and insulinlike growth factor 1. In conclusion, CRISP3, sPLA2, GPx, and testosterone are fertility-associated biomarkers in both seminal plasma and serum of dromedary camels. Seminal plasma osteopontin is positively correlated and prostaglandin D synthase (lipocalcin-type) is negatively correlated with camels' fertility. Serum malonialdehyde, insulinlike growth factor 1, and clusterin are negatively correlated with fertility of male dromedary camels.

Evaluation of the microbial quality of fresh ejaculates of camel (Camelus dromedarius) semen.

A series of five factorial arranged experiments were conducted to investigate the effect of different management during semen collection on the microbial quality (bacterial load, type of microbes and frequency of isolation) of dromedaries' semen. Microbial analysis of seventy-nine fresh ejaculates from twenty-two camels showed the presence of nine variant colonies. The most prevalent organisms in the dromedary semen were species of Staphylococcus, Bacillus and Streptococcus. One yeast species was represented among the isolates. The middle aged camels (9-13 years) had significantly (P>0.05) higher mean bacterial loads than young (4-8 years) and old aged (14-18 years) animals. The mean bacterial populations of ejaculates collected by an electro-ejaculator were significantly (P>0.05) higher than those collected by an artificial vagina. Candida spp. was identified in 53.8% of the samples collected by an electro-ejaculator and was not detected in ejaculates collected by an artificial vagina. The mean semen bacterial load detected during the breeding season was significantly (P<0.01) higher than that collected during the non-breeding season. No fungi were isolated from semen samples collected in the non-breeding season. The difference between the mean semen bacterial loads in the first and the second ejaculate was highly significant (P<0.01). The preputial wash significantly (P<0.01) reduced the bacterial load. This study revealed that the microbial contamination of dromedaries' semen is found in different intensities during different management procedures of semen collection.

Evaluation of the breeding soundness of male camels (Camelus dromedarius) via clinical examination, semen analysis, ultrasonography and testicular biopsy: a summary of 80 clinical cases.

Male camel infertility is a heterogeneous disorder. A variety of factors may adversely affect sperm production and function and impair fertility. This study was designed to evaluate the sensitivity and specificity of ultrasonography and testicular biopsy in the evaluation of the breeding soundness of male dromedaries compared with results obtained by clinical examination and semen analysis. Eighty-four male dromedary camels (5-15 years old) were used in this study during the rutting season (November-May). Four sexually mature male camels were used as controls. These animals were apparently healthy and had histories of normal fertility. Eighty infertile male camels were subjected to an algorithmic approach based on information collected during careful examinations of the camels' breeding histories, clinical examinations, testicular evaluations, testicular ultrasonographies, the results of the semen analyses and testicular biopsies to diagnose the camels' infertilities. The differences in the semen parameters between the control and infertile male camels were highly significant (p < 0.01). Regarding the diagnoses of male camel infertility, the results of testicular ultrasonographies and biopsies were compared with those from the semen analyses, and the accuracies of these tests were 92.5% and 90%, respectively. Additionally, the results of the testicular ultrasonographies were matched with those of the testicular biopsies of the infertile animals, and this comparison resulted in 85% accuracy. Testicular biopsy is a promising method that, along with a carefully performed history, clinical examination, an appropriate testicular ultrasonography procedure and semen analysis, can afford veterinarians the opportunity for more precise diagnosis and treatment of many dromedary infertility disorders.

Impact of seminal plasma superoxide dismutase and glutathione peroxidase on cryopreserved buffalo spermatozoa.

Fifty semen samples were collected from sixteen buffalo-bulls (4-10 years old) and evaluated before cryopreservation. The activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) as well as the levels of glutathione (GSH) and malondialdehyde (MDA) were assayed in the seminal plasma before freezing. Aspartate aminotransferase (AST) activity and cholesterol content were assayed in seminal plasma before freezing and after thawing. Results revealed the presence of SOD and GPx activities (0.07 ± 0.01 U/ml and 14.59 ± 0.50 nmol/min/ml, respectively) in buffaloes' seminal plasma. SOD activity was positively correlated with both of GSH level and GST activity in seminal plasma, and showed an inverse relationship with both cholesterol efflux and post-thaw abnormal tails of buffalo spermatozoa. A positive correlation was found between GPx activity in seminal plasma and abnormal tails and an inverse relationship with both post-thaw viability indices and increased motility in response to PTx. GST activity showed a positive correlation with the increased motility after addition of PTx and negative correlations with both of cholesterol level and AST activity. MDA levels were negatively correlated with motility after addition of PTx and positive correlations with both post-thaw abnormal acrosomes and tails. Buffalo seminal plasma contains high activities of SOD, GPx and GST enzymes and GSH levels that have an influence on the functional competence of cryopreserved spermatozoa.

Comparison of some biochemical and hormonal constituents of oversized follicles and preovulatory follicles in camels (Camelus dromedarius).

The current study was carried out to compare some biochemical and hormonal constituents in follicular fluids from oversized follicles, preovulatory follicles, and serum in camels (Camelus dromedarius). Follicular fluids from oversized follicles (N = 10), preovulatory follicles (N = 10), and sera were harvested from 20 dromedaries. The follicular fluids and sera were subjected to biochemical and hormonal analysis. The results indicated no significant differences in the concentrations of ascorbic acid, glucose, cholesterol, acid phosphatase, and alkaline phosphatase between follicular fluid from oversized follicles and preovulatory follicles. In addition, there were no significant variations in the level of ascorbic acid, glucose, cholesterol, and acid phosphatase in the serum of animals with oversized follicles and those with preovulatory follicles. Serum alkaline phosphatase was significantly greater (P < 0.05) in camels with oversized follicles. The concentrations of estradiol-17ß (E2) and insulin-like growth factor-1 (IGF-1) in the follicular fluid of oversized follicles were significantly lower (P < 0.01) than that from preovulatory follicles. There were no differences in the concentrations of progesterone, tri-iodothyronine, and thyroxin between follicular fluid from oversized follicles and that of preovulatory follicles. The concentrations of E2, progesterone, tri-iodothyronine, thyroxin, cortisol, and IGF-1 were not different in the serum of camels with oversized follicles and camels with preovulatory follicles. The current study revealed that the significant differences of biochemical and hormonal constituents between follicular fluids from oversized follicles and preovulatory follicles were restricted on E2 and IGF-1. Relaying on the aforementioned outcome we can suggest that oversized follicle phenomenon is a form of follicular atresia of anovulatory follicles.

Immunization against GnRH in the male camel (Camelus dromedarius): effects on sexual behavior, testicular volume, semen characteristics and serum testosterone concentrations.

The effect of immunization against gonadotrophin releasing hormone (GnRH) on sexual behavior, total scrotal size, semen characteristics and serum concentrations of testosterone, was evaluated for 24 wks in sexually mature camels (Camelus dromedarius). Eight bull camels were randomly divided into a treatment and control group. Four male camels were immunized using 2 mg GnRH - tandem-dimer conjugated to ovalbumin, (Pepscan Systems, the Netherlands) administered subcutaneously, 4 wks apart. Control male camels received the same amount of saline solution. Significant decline in serum testosterone level was observed in three immunized camels out of four, whereas one camel showed no effect. The testosterone levels reached to <1.0 ng/mL serum by week 4 after booster injection and remained suppressed through the course of the study. The total testicular volume was not affected until the end of the experiment. In treated animals, the sexual behavior negatively affected after the booster injection. Anti-GnRH vaccine had a seriously detrimental effect on the acrosin amidase activity and normal acrosome percentages in treated male camels. It is concluded that the vaccine was effective in reducing serum testosterone levels and libido, and it had a serious harmful effect on the acrosin amidase activity and percentages of spermatozoa with normal acrosome. The immunogen did not affect the total testicular volume.

Some biochemical characteristics and preservation of epididymal camel spermatozoa (Camelus dromedarius).

Testicles were isolated from thirty five apparently healthy dromedary camels (Camelus dromedarius), aged between 5 to 18 years, in a local slaughterhouse during the rutting season. Epididymal fluid was collected from one epididymis for determination of twelve biochemical and antioxidant parameters using ELISA commercial kits. Spermatozoa were harvested from each region of the other epididymis (head, body and tail) and stored in SHOTOR®, Green buffer® + 20% egg yolk and INRA-96® extenders at 5 and 30 °C. Results revealed that, in the epididymal fluid, concentrations of testosterone, glucose, albumin, total protein, cholesterol, fatty acids, iron, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were 5.19 ± 1.69 ng/mL, 3.10 ± 0.41 mmol/L, 6.26 ± 1.26 g/dL, 0.50 ± 0.07 mg/dL, 1.74 ± 0.09 mmol/L, 6.62 ± 0.81 nmol/ul, 926.20 ± 100.18 ug/dL, 51.17 ± 7.74 mIU/ml, and 143.16 ± 18.67 mIU/ml, respectively. The antioxidants activities of catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD) in the epididymal fluid were 121.55 ± 6.57 nmol/min/ml, 59.35 ± 10.98 nmol/min/ml and 0.18 ± 0.03 U/ml, respectively. Epididymal sperm motility and concentration were higher (P < 0.05) in the body and tail than the head. The viability indices of total and forward sperm motility, at 5 and 30 °C, obtained from the tail region were superior (P < 0.05) in both SHOTOR® and INRA-96® extenders than Green buffer extender. It may be concluded that INRA-96® extender is the best for storing dromedary epididymal spermatozoa at 5 and 30 °C.