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Pharmacokinetics of progesterone in dromedary camels (Camelus dromedarim).

The effect of ovariectomy or administration of progesterone (P4) on the disposition kinetics of P4 was determined in dromedary camels. The disappearance of P4 from peripheral circulation of the camel following ovariectomy or after a single intravenous (I.V.) injection of 1 mg kg(-1) body weight followed a bioexponential curve. Both curves were parallel indicating that the disappearance of injected P4 behaved similarly to endogenous P4. The mean (+/- SD) half-life calculated from the slower component of this decline, was 26.0 +/- 2.0 min after I.V. injection of P4 and 28.0 +/- 2.1 min after ovariectomy. The apparent volume of distribution (1370 ml kg(-1)) exceeded total body water suggesting extensive tissue penetration.

Animals↗

Production of dromedary (Camelus dromedarius) embryos by IVM and IVF and co-culture with oviductal or granulosa cells.

The general objective of this work was to produce dromedary embryos from cumulus-oocyte complexes (COCs) that were matured, fertilized and co-cultured in vitro. A total of 1598 COCs were recovered from 457 ovaries; 1308 were deemed suitable for IVM and were cultured at 38.5 degrees C, 5% CO2, and >95% humidity for 36 h in TCM-199 supplemented with 10% heat-treated fetal calf serum (FCS), 10 ng/ml epidermal growth factor (EGF), 1 microg/ml FSH, and 500 microM cysteamine. Matured COCs (n = 88) were denuded, fixed, and stained to determine nuclear status; 63% (56/88) had reached metaphase II (MII) at 36 h. Overall, 1135 COCs were inseminated with ejaculated fresh semen (0.5 x 10(6)spermatozoa/ml in modified TALP-solution). Inseminated oocytes (n = 155) were examined for evidence of fertilization; 68% (106/155) were penetrated by spermatozoa, including 52% (55/106) with two pronuclei and 34% (36/106) with polyspermy. Inseminated, denuded oocytes (n = 819) were co-cultured with dromedary oviductal epithelial or granulosa cells in TCM-199 supplemented with 10% heat-treated FCS. Although the rate of first cleavage (two to eight cells) was similar for the two co-culture systems (32 versus 33%, respectively), more embryos (two-cell to blastocyst stage) were obtained from oocytes co-cultured with oviductal versus granulosa cells (61 versus 45%; P < 0.05). The proportions of fertilized oocytes developing to the early morula stage were 19% (80/417) and 12% (48/402) for oocytes co-cultured for 7 days with oviductal or granulosa cells, respectively (P > 0.05). However, development to the blastocyst stage (10% of fertilized oocytes) occurred only in oocytes co-cultured with oviductal cells. In conclusion, dromedary embryos were produced in vitro using abattoir-derived oocytes, fresh (ejaculated) semen, and oviductal cell co-culture.

Animals↗

The effect of harvesting technique on efficiency of oocyte collection and different maturation media on the nuclear maturation of oocytes in camels (Camelus dromedarius).

The purpose of this investigation was to develop an efficient method for harvesting oocytes from dromedary camel ovaries and to examine the effect of different maturation media on their subsequent maturation in vitro. Oocytes were collected by aspirating the follicular contents using a needle attached to a syringe (Method I, n=163 ovaries) or to a constant aspirating pressure, applied by a vacuum pump (Method II, n=117 ovaries). Individual follicles were excised from ovaries and follicles were punctured with two needles (Method III, n=117). Oocytes were matured in vitro for 40-42 h. At the end of maturation period, oocytes were denuded of cumulus cells and the proportion of oocytes in metaphase-II (MII) stage was determined. In the second experiment, oocytes collected by the dissection method were matured in Tissue Culture Medium199 (TCM), CR1 or modified Connaught Medical Research Laboratories medium-1066 (CMRL) and their nuclear maturation was evaluated after 40-42 h. The recovery rate of oocytes was higher (P<0.01) with Method III compared with Method I or II (94, 31 and 33%, respectively). A higher proportions of oocytes collected with Method I or II were either completely or partially denuded compared with Method III (31, 14% versus 1%). The proportions of viable oocytes (78, 60 and 70%, respectively) and those showing metaphase II was not different (39, 50 and 46%, respectively, P>0.05) among the three treatment groups. Oocyte maturation rate was higher (P<0.05) when TCM was used compared with CMRL or CR1 medium. There was, however, no difference in the maturation rate for oocytes cultured in CMRL or CR1 medium. It may be concluded that a higher proportion of cumulus enclosed oocytes may be recovered by follicle dissection method compared to aspiration using syringe or pump. The higher recovery rate with a comparable proportion of viable and matured oocytes resulted in the overall increase in the number of matured (MII) oocytes/ovary with follicle dissection procedure compared with aspiration procedures. For in vitro maturation of oocytes, TCM is superior to CR1 and CMRL as basic maturation medium for this species.

Animals↗

Offspring resulting from transfer of cryopreserved embryos in camel (Camelus dromedarius).

The dromedary embryos, collected at hatched blastocyst stage, survived freezing and thawing in the presence of a high concentration of ethylene glycol (7.0 mol/L) with sucrose (0.5 mol/L) and direct plunging in liquid nitrogen. The rate of survival, as judged by the morphological appearance of the embryos after thawing, was high (92%). The transfer of frozen-thawed embryos into the recipients during the breeding (n=20) and non-breeding season (n=25) resulted in two and one pregnancy, respectively. One of the two pregnant recipients, with embryos transferred during the breeding season, delivered a normal healthy male calf at term. To our knowledge, this offspring is the first camelid produced following transfer of a frozen-thawed embryo.

Animals↗

Kinetics of nuclear maturation and effect of holding ovaries at room temperature on in vitro maturation of camel (Camelus dromedarius) oocytes.

Experiments were conducted to investigate kinetics of in vitro nuclear maturation and the effect of storing ovaries at room temperature on initial chromatin configuration and in vitro maturation of dromedary camel oocytes. Cumulus oocyte complexes (COCs) were collected from slaughterhouse ovaries and matured in vitro for 4-48h. At every 4h interval (starting from 0 to 48 h), groups of oocytes were fixed, stained and evaluated for the status of nuclear chromatin. Oocytes were categorized as germinal vesicle (GV), diakinesis (DK), metaphase-I (M-I), anaphase-I (A-I), metaphase-II (M-II) stage and those with degenerated, fragmented, activated or without a visible chromatin as others. At the start of culture, 74% (66/89) oocytes were at GV stage, 13% (12/89) at DK and 12% (11/89) were classified as others. Germinal vesicle breakdown started spontaneously in culture and at 20 h of culture 97% oocytes had already completed this process. After 8 and 16 h of maturation the highest proportion of oocytes (42%, 48/114 and 41%, 51/123) were at DK and M-I stage, respectively. The proportions of oocytes reaching M-II stage at 32 (42%, 50/118), 36 (45%, 47/104), 40 (49%, 57/117), 44 (52%, 103/198) and 48 h (46%, 55/120) of culture were not different from each other (P>0.05). The proportion of oocytes categorized as others, however, increased after 40 h of culture and was higher (P<0.05) at 48 h compared with other maturation periods. There was no difference (P>0.05) in the proportion of oocytes reaching M-II stage from the ovaries collected and stored in normal saline solution (NSS) at room temperature for 12h (43%, 64/148) and those collected in warm NSS (37 degrees C) and processed immediately after arrival in laboratory (49%, 57/117). However, low number of oocytes reached M-II stage from ovaries collected in warm NSS but stored at room temperature (29%, 37/128) compared with other two groups (P<0.05). It may be concluded that dromedary oocytes require 32-44h of in vitro culture to have an optimum number of oocytes in M-II stage. However, further studies are required to find out the most appropriate maturation period, which will result in the further development of these oocytes after IVF, ICSI, parthenogenetic activation or nuclear transfer. Ovaries can be collected and stored in normal saline solution at room temperature for 12h without any appreciable effect on the nuclear maturation of the oocytes.

Animals↗

Incidence of spontaneous ovulation and development of the corpus luteum in non-mated dromedary camels (Camelus dromedarius).

The occurrence of spontaneous ovulation in dromedaries was examined in two separate studies including 20 non-lactating, barren and 12 lactating dromedaries, respectively. Lactating camels were milked twice a day with an automatic bucket milking machine. Ovarian activity was monitored by repeated ultrasonography. Blood samples for progesterone were collected daily or two to three times a week. To compare CL development after spontaneous and induced ovulations, ovulation was induced by a GnRH analogue in eight lactating dromedaries. Spontaneous ovulation was observed in one non-lactating camel (1 of 20 camels, 5%; 1 of 70 follicular waves, 1.4%), whereas, spontaneous ovulation was detected more frequently in lactating dromedaries (5 of 12 camels, 41.7%; 13 of 91 follicular waves, 14.3%). In one lactating camel, spontaneous ovulation occurred repeatedly for nine times. There was a significant effect of type of ovulation (spontaneous versus induced, P < 0.05) and day (P < 0.001) on serum progesterone concentration. Mean serum progesterone levels and total progesterone production (AUC) were higher after induced ovulation. Luteal diameter and serum progesterone concentration were positively correlated (r = 0.71, P < 0.001), but there was a significant difference between morphological and functional development of the CL. In dromedaries, morphological development starts earlier, morphological regression starts later and last longer than functional development and regression of the CL. Compared to induced ovulation, functional development of the CL after spontaneous ovulation might be altered but the morphological development is not affected.

Animals↗

Effect of season and gonadotropins on the superovulatory response in camel (Camelus dromedarius).

The purpose of the present investigation was to study the extent to which season and the gonadotropin preparation interferes with the superovulatory response in the dromedary. Adult camels were treated for superovulation during the breeding (November to April) and non-breeding season (May to October). Animals were synchronized by daily i.m. injections of progesterone (125 mg/animal/day, Jurox, UK) for 10 to 14 days. Superovulation was induced by 400mg pFSH alone (Follitropin V, Vetrepharm, Canada) administered in eight descending doses at 12h intervals or a combination of PMSG (2000IU, Folligon, Intervet, The Netherlands), injected with last injection of progesterone and 400mg pFSH in eight descending doses. The follicular development was daily assessed by ultrasonography of the ovaries. The donors were classified as per their response to the superovulatory treatment into very good (>10 follicles), good (5-10 follicle), poor (2-4 follicles) or no response (1 or no follicle) on each ovary. Ovulation was induced by injecting 3000 IU hCG (Chorulon, Intervet) at the time of first mating. The donors were mated twice at an interval of 12h when all or most of the follicles reached to a size of about 1.0-1.7 cm. Camels were flushed non-surgically on Day 6 or 7 after the ovulation. The proportion of camels showing very good response during the breeding as well as non-breeding season was higher (P<0.05) when a combination of pFSH and eCG was used compared with pFSH only. There was no difference (P>0.05) in the proportion of donors flushed successfully (embryos recovered) when treated either with a combination of pFSH and eCG or pFSH alone during the breeding and non-breeding season. The rate of recovery of ova/embryos and proportion of transferable embryos was higher (P<0.05) when donors were treated with pFSH+eCG compared with pFSH only during the breeding as well as non-breeding season. The results may indicate that ova/embryo recovery rate of the dromedary is influenced by the gonadotropin preparation but is not appreciably affected by the season.

Animals↗

The first dromedary (Camelus dromedarius) offspring obtained from in vitro matured, in vitro fertilized and in vitro cultured abattoir-derived oocytes.

Dromedary offspring have never been produced fully in vitro. We have previously demonstrated that embryos obtained by culture in semi-defined medium (mKSOMaa) have better in vitro development ability than those cultured with oviductal epithelial cells. The aim of the present experiment was to study the pregnancy rate after embryo transfer of in vitro-produced (IVP) dromedary embryos cultured in semi-defined modified medium (mKSOMaa). IVM/IVF procedures were conducted on six hundred and sixty four (664) cumulus oocytes complexes (COCs) aspirated from ovaries collected at a local slaughterhouse and cultured in vitro (38.5 degrees C; 5% CO2, and maximum humidity >95%). Maturation was completed by incubation in TCM-199 medium supplemented with 10% heat-treated Fetal Calf Serum (FCS), 10 ng/mL EGF, 1 microg/mL FSH, 1 microg/mL E2 and 500 microM cysteamine for 30 h. In vitro fertilization was performed using fresh semen (0.5 x 10(6) spermatozoa/mL in modified TALP-solution). Fertilized oocytes were cultured in mKSOMaa, under 38.5 degrees C, 5% CO2 and 90% N2 with maximum humidity (>95%). All IVC steps were done in seven replicates. The cleavage rate (two cells to blastocyst stage) was 64% (425/664) and the percentage of oocytes reaching the blastocyst stage was 23% (155/664). The hatching rate of blastocyst obtained after culture was 46% (71/155). Good quality hatched blastocysts (n = 66) were transferred individually to synchronized recipients. Pregnancy rates, determined by ultrasonography at 15, 60 and 90 days after embryo transfer (ET), were 38%, 32% and 27%, respectively. Out of 18 pregnant females 5 aborted between the fifth and seventh month of pregnancy and 13 females (20%) remained pregnant. After 385 days of pregnancy, the first healthy and normal male-dromedary offspring produced fully in vitro was born at a birth weight of 38 kg. More dromedary calves (n = 4) were born later on. The remaining pregnant females (n = 8) are due to calf within the next months. In conclusion, this is the first reported offspring in camelids obtained by transfer of embryos produced by IVM, IVF and IVC using abattoir-derived oocytes, fresh semen and culture in a semi-defined medium.

Animals↗

Comparison of pregnancy rates in dromedary camels (Camelus dromedarius) after deep intra-uterine versus cervical insemination.

The ovarian follicular wave patterns of sixty adult female camels were monitored by serial trans-rectal ultrasound examinations and when the dominant follicle reached 1.3-1.8 cm in diameter they received a single intravenous injection of 20 microg buserelin, to induce ovulation, and were inseminated with a known number of spermatozoa 24 h later. Ejaculates were collected from the male camels and diluted 1:1 in Green Buffer with 20% egg yolk (v:v) added. Sperm concentration and motility were assessed and a dose of 40, 80 or 150 x 10(6) motile spermatozoa were deposited either just through the cervix into the uterine body or at the tip of the horn ipsilateral with the ovary containing the dominant follicle. Insemination of 150, 80 and 40 x 10(6) spermatozoa into the uterine body resulted in conception rates of 53, 7 and 0%, respectively, whereas insemination at the tip of the uterine horn resulted in conception rates of 43, 40 and 7%, respectively.

Animals↗

Comparison of the effect of Sporobolus virginicus and Rhodes (Chloris gayana) hay diets on the absorption pattern of phenylbutazone in the camel (Camelus dromedarius).

The effect of feeding Sporobolus and Rhodes hay on phenylbutazone (4 g) relative absorption was examined in six camels using a two-period, two-sequence, two-treatment crossover design. Serum concentration of the drug was measured by high performance liquid chromatography. The measured values (means+/-SD) for Rhodes and Sporobolus hay, respectively, were Cmax 35.59+/-22.36 and 36.55+/-18.99 microg/mL, Tmax 26+/-2.53 and 26.3+/-1.97 h and AUC0-72 h 1552+/-872.6 and 1621+/-903.6 microg h/mL. Broad plateau concentrations of phenylbutazone in serum were observed between 12 and 36 h. There was no significant difference in any parameter between the two feeding regimens. Multiple peaks in serum concentration-time curve were observed, regardless of the type of grass available to and the animals prior to drug administration. It was concluded that the phasic absorption of phenylbutazone was a particular feature of hay feeding in camels, and the Sporobolus hay can be fed to camels without any effect on the rate and extent of phenylbutazone absorption compared to Rhodes grass hay.

Animals↗

Anti-erythrocyte natural antibody activity in the unconventional 'heavy chain' immunoglobulins of Indian desert camel (Camelus dromedarius).

Members of the family Camelidae contain unconventional 'heavy chain' antibodies (HCAbs) that are devoid of light chains (LCs) in their structure and occur under physiological conditions. The spectrum of antigenic specificities in HCAbs and hence their biological significance is not known at present. Recent studies have however indicated that they contribute significantly towards the immunoglobulin (Ig) receptor diversity. The present study was planned to investigate the natural antibody (NAb) activity in camel HCAbs as further indication of the wide spectrum of their antigenic specificities. Detection of NAbs in the sera and isolated HCAbs of Indian Thar desert camels was undertaken against erythrocyte antigens (E-Ags) from eleven animal species including nine mammals, chicken and frog by using direct haemagglutination (HA) and indirect Coombs' test. HCAbs were found to behave as 'incomplete antibodies' and agglutinated erythrocytes of different animal species in indirect Coombs' test using rabbit anti-camel IgG3 (HCAbs) antiserum. Variations were noticed in the Coombs' titres against erythrocytes from different species. HCAbs also reacted against E-Ags in immunoblots. These findings provide further evidence that camelid HCAbs are produced against diverse antigens (Ags) under natural conditions, thereby contributing to camelid Ig receptor diversity.

Animals↗

Clinical and subclinical colloid goitre in adult camels (Camelus dromedarius) at Kordofan region of Sudan.

Colloid goitre was diagnosed in adult camels in the Kordofan region of the Sudan. The disease is characterized by gross enlargement of the thyroid, histopathological follicular changes, reproductive disorders, low concentrations of circulating thyroid hormones (T3, T4) and normocytic normochromic anaemia. The possible cause of the condition is discussed and iodine supplementation is recommended.

Animals↗

Natural Dermatophilus congolensis infection in camels (Camelus dromedarius) from Kenya.

Natural Dermatophilus congolensis infection is found in many species of livestock and wild animals. It is, however, rarely described in camels and there are no details of bacterial isolation. In an investigation of both arid and semi-arid areas in Kenya, an outbreak of dermatophilosis was observed in camel calves being reared on a commercial farm in a semi-arid area. Histopathology and bacterial isolation were used to diagnose the diseases. The potential impact of the disease in camels is discussed.

Actinomycetales Infections↗

Experimental copper poisoning in the camel (Camelus dromedarius).

Copper sulphate was administered by the oral or intravenous route to five dromedary camels. Two camels (1 and 2) receiving copper sulphate at 200 mg per kg per day by drench died within 8 days and camel 3, receiving 100 mg per kg per day by the same route, was slaughtered on day 172. Intravenous injection of 2 mg per kg per day caused the death of camel 4 on day 95 and camel 5, treated similarly, was slaughtered on day 138. Anorexia, dullness, diarrhoea, dehydration and recumbency in camels 1 and 2 were probably clinical signs of copper toxicity. Camels 3, 4 and 5 lost weight. Jaundice was not a prominent clinical sign. The main lesions in camels 1 and 2 were fatty change and necrosis of the liver cells, dilatation and necrosis of kidney tubules, catarrhal abomasitis, enteritis and congestion of the blood vessels of the heart. In camels 3, 4 and 5 the hepatic lesions were mild, with leucocytic infiltration and gastrointestinal and heart lesions were either mild (camel 3) or absent (camels 4 and 5). Cytoplasmic copper granules in hepatic cells were generalized in distribution but more concentrated in the centrilobular zone. In the kidney these granules were confined to the cells of the proximal convoluted tubules. Copper accumulated in the liver and kidneys of all the camels and zinc accumulated in the liver and kidneys of those receiving copper sulphate intravenously. Macrocytic hypochromic anaemia developed in camels 3, 4 and 5 and haemoconcentration in camels 1 and 2. The concentration of serum copper, zinc and iron increased in animals 1, 2 and 4, and unbound iron binding capacity decreased in four camels. There was a rise in the activity of gamma GT, GOT, LDH and CPK in the serum of all the animals. Serum ALP activity, however, increased in camels 1 and 2 and decreased in camels 3, 4 and 5.

Alkaline Phosphatase↗

The isolation and characterization of camel (Camelus dromedarius) immunoglobulin classes and subclasses.

Immunoglobulins G and M were prepared from camel sera by ammonium sulphate precipitation, gel filtration and fast protein liquid chromatography (ion exchange). Heavy and light chains were identified by polyacrylamide gel electrophoresis and Western blotting. IgG subclasses were isolated by DEAE ion exchange chromatography and shown to have different electrophoretic motilities. No molecules corresponding to IgA were detected. Rabbit anti-serum against camel IgG was generated and its specificity shown by Western blotting.

Animals↗

Immunoglobulins of camel (Camelus dromedarius) colostrum.

Immunoglobulins G, M and A were identified in dromedary camel colostra by acid precipitation, gel filtration and fast-protein liquid chromatography (ion exchange). Heavy and light chains were identified by polyacrylamide gel electrophoresis and Western blotting. IgG subclasses (IgG1, IgG2 and IgG3) were isolated by DEAE ion-exchange chromatography and shown to have different electrophoretic mobilities. Cross-reactivity of camel IgA with IgA of other species was determined by enzyme-linked immunosorbent assay (ELISA). Most of the immunoglobulin content was IgG, but a molecule identifiable as IgA was detected and purified. It would appear that in the camel, as in cattle, IgG is the major secretory immunoglobulin of colostrum.

Animals↗

Mastitis in Camelus dromedarius and the somatic cell content of camels' milk.

Seven hundred and sixty-three camels from 400 herds of local Sudanese camel breeds were investigated for the prevalence of mastitis, identification of its bacterial causes and determination of the leucocyte contents of camel's milk. One hundred and forty-nine (19.5 per cent) of the 763 camels examined were diagnosed as mastitis cases based on clinical signs. One hundred and fifty-nine (47.3 per cent) of the 336 randomly selected milk samples were reactive in a rapid mastitis test and 16 of the 153 tested samples contained Brucella abortus agglutinating antibodies. Streptococcus, Staphylococcus, Micrococcus and Aerobacter species and Escherichia coli were found to be the main causes of mastitis (in descending order). The leucocyte contents of the 757 milk samples ranged from < 5 x 10(5) to > 7.5 x 10(6) leucocyte ml-1 and 42.8 per cent of the samples contained < 5 x 10(5) cells ml-1. Neither significant correlation between the leucocyte content of milk and isolated bacterial species nor significant variation in leucocyte contents during different stages of lactation were detected.

Animals↗

Biochemical analysis of serum and cerebrospinal fluid in clinically normal adult camels (Camelus dromedarius).

The concentrations of total protein, glucose, cholesterol, triglyceride, urea nitrogen, creatinine, sodium, potassium, chloride, calcium, inorganic phosphorus, copper, magnesium, and iron and the activities of aspartate aminotransferase (AST), alanine aminotransferase, (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LD), creatine kinase (CK) and amylase were determined in cerebrospinal fluid (CSF) and serum from 21 clinically healthy adult camels. The concentrations of sodium, potassium and chloride in CSF were similar to those of serum; whereas the values for all other constituents were significantly higher (P<0.05) in serum than in CSF.

Analysis of Variance↗