Semen-induced ovulation in the bactrian camel (Camelus bactrianus)

Bactrian camels (63 female female, 8 male male) were used in the breeding season to determine the factors that will induce ovulation. After insemination of semen samples into the vagina, the ovaries were checked for ovulation by rectal palpation. The results indicated that ovulation was induced by the seminal plasma, but not by the spermatozoa, and the incidence of ovulation after insemination was 87%. Most of the females (66%) had ovulated by 36 h after insemination and the rest by 48 h, as after natural service. The least amount of semen required to elicit ovulation was about 1.0 ml. Intramuscular injections of LH, hCG and LHRH also caused ovulation, even in females that had not ovulated in response to insemination.     

Monophyletic origin of domestic bactrian camel (Camelus bactrianus) and its evolutionary relationship with the extant wild camel (Camelus bactrianus ferus)

The evolutionary relationship between the domestic bactrian camel and the extant wild two-humped camel and the factual origin of the domestic bactrian camel remain elusive. We determined the sequence of mitochondrial cytb gene from 21 camel samples, including 18 domestic camels (three Camelus bactrianus xinjiang , three Camelus bactrianus sunite , three Camelus bactrianus alashan , three Camelus bactrianus red , three Camelus bactrianus brown and three Camelus bactrianus normal ) and three wild camels ( Camelus bactrianus ferus ). Our phylogenetic analyses revealed that the extant wild two-humped camel may not share a common ancestor with the domestic bactrian camel and they are not the same subspecies at least in their maternal origins. Molecular clock analysis based on complete mitochondrial genome sequences indicated that the sub-speciation of the two lineages had begun in the early Pleistocene, about 0.7 million years ago. According to the archaeological dating of the earliest known two-humped camel domestication (5000–6000 years ago), we could conclude that the extant wild camel is a separate lineage but not the direct progenitor of the domestic bactrian camel. Further phylogenetic analysis suggested that the bactrian camel appeared monophyletic in evolutionary origin and that the domestic bactrian camel could originate from a single wild population. The data presented here show how conservation strategies should be implemented to protect the critically endangered wild camel, as it is the last extant form of the wild tribe Camelina.     

Hormone concentrations before and after semen-induced ovulation in the bactrian camel (Camelus bactrianus)

During the follicular phase of bactrian camels, basal concentrations of LH were 2.7 +/- 1.2 ng/ml. By 4 h after insemination peak values of 6.9 +/- 1.0 ng/ml occurred. In addition, a smaller LH peak (5.4 +/- 2.5 ng/ml) appeared 1 day before regression of the follicle began in unmated camels. During the follicular phase peripheral plasma progesterone values were low (0.36 +/- 0.28 ng/ml), but values increased to reach 1.73 +/- 0.74 ng/ml at 3 days and 2.4 +/- 0.86 ng/ml at 7 days after ovulation. Plasma oestradiol-17 beta concentrations were 26.8 +/- 9.0 pg/ml during the follicular phase and 30.8 +/- 5.1 pg/ml when the follicle was maximum size. Values fell after ovulation but rose to 29.8 +/- 6.5 pg/ml 3 days later.     

Distributions of antibovine-pepsinogen-positive cells in glandular regions of forestomach in the bactrian camel, Camelus bactrianus.

The present paper describes mucous cells in the forestomach glandular region which immunoreacted to antibovine pepsinogen serum in adult and young bactrian camels, Camelus bactrianus (female, 25 years old; female, 6 months old; from Hilar Inner Mongolia, People's Republic of China, in August 1988). These immunoreactive cells in forestomach glandular regions also reacted to Alcian blue (pH 1.0) and/or periodic acid-Schiff. Cross-reactivity of antibovine pepsinogen serum against extracts from the forestomach glandular region was analyzed by Western blotting, supporting that this serum could detect pepsin and/or pepsinogen in the bactrian camel. It was suggested that mucous cells in forestomach glandular regions produce pepsinogen molecules.     

Biochemical isolation and purification of ovulation-inducing factor (OIF) in seminal plasma of llamas

Background  

The objective of the present study was to isolate and purify the protein fraction(s) of llama seminal plasma responsible for the ovulation-inducing effect of the ejaculate.     

Dose-response of female llamas to ovulation-inducing factor from seminal plasma

The present study was designed to determine if the dose of purified ovulation-inducing factor (OIF) from llama seminal plasma required to provoke an ovulatory response is physiologically relevant in terms of the proportion present in a normal ejaculate and to test the hypothesis that corpus luteum (CL) form and function are affected by OIF in a dose-dependent manner. Female llamas were assigned randomly to five groups (n = 10 per group) and given a single i.m. dose of 500, 250, 125, or 60 μg of purified OIF (representative of the amount present in 1/25th to 1/200th of a normal ejaculate) or 1 ml of PBS (control). Ovulation and CL development were monitored by transrectal ultrasonography. Blood samples were taken to measure plasma progesterone concentrations and to determine changes in plasma concentrations of luteinizing hormone (LH). The high dose of OIF (500 μg) was associated with the highest incidence of ovulation (P < 0.05), the greatest maximum CL diameter (P < 0.05), and the largest day-to-day profiles of CL diameter (P < 0.05) and plasma progesterone concentrations (P < 0.01). A rise in plasma LH concentration was apparent in all llamas that ovulated and was most rapid and highest in the high-dose group (P < 0.01). The low dose of OIF (60 μg) was minimally effective for induction of ovulation and the least luteotrophic, as evidenced by the smallest maximum CL diameter and the smallest day-to-day profiles for CL diameter and plasma concentrations of progesterone and LH. Responses were intermediate for the middle-dose groups (125 and 250 μg). We conclude that OIF from llama seminal plasma has a dose-dependent effect on ovulation rate and CL form and function in llamas and that the biological effect of OIF is evident at physiologically relevant doses (i.e., as little as 1/100th of that present in an ejaculate).     

Anatomical subdivisions of the stomach of the Bactrian camel (Camelus bactrianus)

Twenty stomachs of Bactrian camels (Camelus bactrianus) were studied by gross dissection. Based on the configuration of the stomach and the structure of the mucous membrane, the stomach was divided into three ventricles that differ from the arrangement described for ox and sheep. The first and second ventricles of the proventriculus of camel form one stomach rather than two different stomachs. These ventricles of the proventriculus do not correspond to the rumen and reticulum of ox and sheep. The third ventricle appears to be the abomasum. One part of the abomasum has reticular mucosal folds that indicate it is not the reticulum. A second part of the abomasum has longitudinal mucosal folds suggesting it is not the omasum. Three glandular sac areas associated with the preventriculus and abomasum are also described.     

Local versus systemic effect of ovulation-inducing factor in the seminal plasma of alpacas

Background  

Camelids are induced (reflex) ovulators. We have recently documented the presence of an ovulation-inducing factor (OIF) in the seminal plasma of alpacas and llamas. The objective was to test the hypothesis that OIF exerts its effect via a systemic rather than a local route and that endometrial curettage will enhance the ovulatory response to intrauterine deposition of seminal plasma in alpacas.     

Synchronization of follicular wave emergence prior to superovulation in Bactrian camel (Camelus bactrianus)

This study was conducted to synchronize follicle wave emergence prior to superovulation using either GnRH or progestogen treatments, in Bactrian camels. GnRH group camels (n=5) received 20 microg of the GnRH analogue Buserelin on Days -18 and -4 of the experiment (initiation of superovulation=Day 0). Camels in the progestogen group (n=5) received two consecutive treatments of progestogens, 7 days apart, on Days -14 and -8 of the experiment. On each occasion, each female received three norgestomet implants and 200mg progesterone (i.m.) and all implants were removed 14 days after the first progestogen treatment coinciding with Day -1 of superovulation. A combination of eCG and FSH was used to induce superovulation and the growth of all subsequent follicles and CLs were monitored daily by ultrasonography. Following the first GnRH injection, mature follicles ovulated within 1-2 days, and a new follicle wave emerged after 3+/-0.77 days. At the time of the second GnRH injection, a mature follicle (15.6+/-0.97 mm) ovulated and a new follicular wave emerged between 1 and 2 days after GnRH injection. Growing follicles at the time of the first progestogen treatment became either atretic (n=1) or persistent (n=4) and a new follicle wave (n=3) emerged 3-6 days later. At the initiation of superovulation, the diameters of the largest follicle in GnRH and progestogen groups were 7.4+/-0.59 and 20.5+/-2.26 mm, respectively but after superovulation and mating there was no significant differences in the number of unovulated follicles or CLs between groups. In conclusion, two GnRH injections, 14 days apart, may be used to synchronize follicle wave emergence in Bactrian camel.     

Serum progesterone and 17 beta-estradiol concentrations during pregnancy of Bactrian camel (Camelus bactrianus)

Serum progesterone and 17 beta-estradiol during pregnancy in the Bactrian camel were measured by radioimmunoassay. Serum progesterone concentrations increased by 15 d after artificial insemination (AI) and remained elevated throughout most of gestation, the mean concentrations (3.06 +/- 0.49 to 8.51 +/- 4.80 ng/mL) were similar to those reported for many species during the same stage of pregnancy. Serum 17 beta-estradiol increased significantly from 11 m.o. of pregnancy with peak mean concentrations of 617.47 +/- 32.56 pg/mL at the 11.5 m.o.     

Luteotrophic effect of ovulation-inducing factor/nerve growth factor present in the seminal plasma of llamas

The hypothesis that ovulation-inducing factor/nerve growth factor (OIF/NGF) isolated from llama seminal plasma exerts a luteotrophic effect was tested by examining changes in circulating concentrations of LH and progesterone, and the vascular perfusion of the ovulatory follicle and developing CL. Female llamas with a growing follicle of 8 mm or greater in diameter were assigned randomly to one of three groups (n = 10 llamas per group) and given a single intramuscular dose of PBS (1 mL), GnRH (50 μg), or purified OIF/NGF (1.0 mg). Cineloops of ultrasonographic images of the ovary containing the dominant follicle were recorded in brightness and power Doppler modalities. Llamas were examined every 4 hours from the day of treatment (Day 0) until ovulation, and every other day thereafter to Day 16. Still frames were extracted from cineloops for computer-assisted analysis of the vascular area of the preovulatory follicle from treatment to ovulation and of the growing and regressing phases of subsequent CL development. Blood samples were collected for the measurement of plasma LH and progesterone concentrations. The diameter of the dominant follicle at the time of treatment did not differ among groups (P = 0.48). No ovulations were detected in the PBS group but were detected in all llamas given GnRH or OIF/NGF (0/10, 10/10, and 10/10, respectively; P < 0.0001). No difference was detected between the GnRH and OIF/NGF groups in the interval from treatment to ovulation (32.0 ± 1.9 and 30.4 ± 5.7 hours, respectively; P = 0.41) or in maximum CL diameter (13.1 ± 0.4 and 13.5 ± 0.3 mm, respectively; P = 0.44). The preovulatory follicle of llamas treated with OIF/NGF had a greater vascular area at 4 hours after treatment than that of the GnRH group (P < 0.001). Similarly, the luteal tissue of llamas treated with purified OIF/NGF had a greater vascular area than that of the GnRH group on Day 6 after treatment (P < 0.001). The preovulatory surge in plasma LH concentration began, and peaked 1 to 2 hours later in the OIF/NGF group than in the GnRH group (P < 0.05). Plasma progesterone concentration was higher on Day 6 in the OIF/NGF group than in the GnRH group (P < 0.001). Results support the hypothesis that OIF/NGF exerts a luteotrophic effect by altering the secretion pattern of LH and enhancing tissue vascularization during the periovulatory period and early stages of CL development.     

Isolation and purification of an organ-specific autoantigen from rabbit seminal plasma

A specific autoantigen of rabbit accessory glands was isolated and purified from seminal plasma using DEAE-Sephadex batch adsorption and Sephadex G-100 chromatographic fractionations. The autoantigen appeared homogeneous by polyacrylamide electrophoresis and by immunodiffusion. It has an extinction coefficient at 284 mμ (E_1cm^1%) of 6.42. It contains 87.73% of polypeptides. The carbohydrate moiety is composed of 9.21% hexose, 1.03% hexosamines, 0.33% fucose. From these chemical values the autoantigen has been identified as glycoprotein.     

基于16S rRNA基因序列分析法比较苏尼特双峰驼和阿拉善双峰驼自然发酵酸驼乳的微生物多样性

【目的】传统发酵乳制品是一类未经任何处理自然发酵而成的,其微生态环境未遭破坏,从而乳酸菌的生物学特性和基因多样性得到了很好的保留,具有开发和利用价值。自然发酵酸驼乳常用来治疗多种疾病且效果良好,与其中丰富的乳酸菌资源有着密不可分的联系。然而,目前有关自然发酵酸驼乳微生物菌群及多样性相关研究甚少。因此进一步挖掘内蒙古地区双峰驼自然发酵酸驼乳微生物群落结构和多样性是至关重要的。【方法】本研究采用IlluminaMiseq测序技术,测定了苏尼特和阿拉善双峰驼的自然发酵酸驼乳中微生物16S rRNA V3–V4区序列,并对群落结构和多样性进行了比较分析。【结果】多样性分析表明,苏尼特双峰驼酸驼乳中微生物群落丰富度和种群差异性比阿拉善双峰驼酸驼乳大,细菌多样性也高。在门水平上,苏尼特和阿拉善双峰驼酸驼乳中的菌群均以厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)为主。在属水平上,苏尼特双峰驼酸驼乳主要以乳杆菌属(Lactobacillus)和乳球菌属(Lactococcus)为优势菌群,阿拉善双峰驼酸驼乳以乳杆菌属(Lactobacillus)和醋酸杆菌属(Acetobacter)为优势菌属。此外,肠杆菌属(Enterobacter)、拉乌尔菌属(Raoultella)和明串珠菌属(Leuconostoc)等的含有食源性致病菌和环境污染菌的菌属被检出。综上所述,不同地区不同品种酸驼乳的乳酸菌种类及优势菌群有较大差异,存在显著的地理差异。【结论】通过本研究,不仅对苏尼特和阿拉善双峰驼自然发酵酸驼乳乳酸菌的组成和种类有了明确的认知,为评估发酵酸驼乳微生物群落对消费者身体健康的影响提供了数据基础的同时为今后筛选优势菌群和挖掘新型益生菌奠定基础。     

Immunohistochemical study of the distribution of endocrine cells in the gastrointestinal tract of the camel (Camelus bactrianus)

The regional distribution and relative frequency of endocrine cells in the gastrointestinal tract of the camel, Camelus bactrianus, were investigated using immunohistochemical methods. Ten types of immunoreactive (IR) endocrine cells were identified in this study. Among these cell types, only serotonin- and somatostatin-IR cells were detected in almost all regions of the gastrointestinal tract. Most of the cell types showed peak density in the pyloric gland region. The others showed restricted distribution: gastrin, cholecystokinin (CCK), motilin, bovine pancreatic polypeptide (BPP), and (gastric) substance P in the stomach; gastrin, CCK, BPP, gastric inhibitory polypeptide (GIP), glucagon, peptide tyrosine tyrosine (PYY) and substance P in the small intestine; and CCK, motilin, BPP, and PYY in the large intestine. Fundamentally the distribution pattern of endocrine cells in the gastrointestinal tract of the camel is similar to that of cattle. The distribution and frequency of endocrine cells in the glandular sac region are the same as those of the cardiac gland.     

Vascular supply of the testis of the camel (Camelus dromedarius).

The origin, pattern of distribution and the length of the convoluted part of the testicular artery were examined by blunt dissection and by the preparation of arterial casts with vinylite. The testicular artery originates from the abdominal aorta just cranial to its bifurcation and forms compact coils and reveals a tortuous course in the region of the pampiniform plexus and along the attached border. The artery divides at the caudal extremity of the testis into 2--4 branches. The epididymal artery, which is a branch of the testicular artery, was seen to anastomose with the deferential artery, but no anastomosis occurs between these arteries and the testicular artery. The convoluted part of the testicular artery was 20 cm in length, but when unravelled totalled 240 cm. This is not affected by seasonal variations. The possible significance of these results was discussed and compared with similar findings of other mammals.     

Isolation of basic acrosin inhibitor from bull seminal plasma (BUSI II).

An acrosin inhibitor was isolated from bull seminal plasma by gel filtration on Sephadex G-50 fine and ion-exchange chromatography on CM-Sephadex. The inhibitor is a basic polypeptide (pl greater than or equal to 10.5) of molecular weight 6 200 (calculated from amino acid composition). Its N-terminal amino group is blocked. The inhibitor is not strictly specific in its effect since it also inhibits trypsin and to a lesser degree chymotrypsin, in addition to bull and boar acrosin.     

Comparison of the effect of ovulation-inducing factor (OIF) in the seminal plasma of llamas, alpacas, and bulls

We have recently reported the presence of an ovulation-inducing factor (OIF) in the seminal plasma of llamas and alpacas-species characterized as induced ovulators. The study was designed to test the hypothesis that the seminal plasma of bulls will induce ovulation in llamas, and to compare the ovulation-inducing effect of seminal plasma of conspecific versus hetero-specific males. The seminal plasma of alpacas, a closely related induced ovulator (Lama pacos), and cattle, a distantly related ruminant species (Bos taurus) considered to be spontaneous ovulators, were compared with that of the llama (Lama glama). Ovulation and maximum corpus luteum diameter were compared by ultrasonography among female llamas (n=19 per group) treated intramuscularly with 2 mL of phosphate buffered saline (PBS, negative control) and those treated with 2 mL of seminal plasma of bulls, alpacas, or llamas (conspecific control). The diameter of the preovulatory follicle did not differ among groups at the time of treatment. Bull seminal plasma induced ovulations in 26% (5/19) of llamas compared to 0% (0/19) in PBS group (P<0.001). The proportion of females that ovulated was lower (P<0.01) in bull seminal plasma group compared to the groups treated with alpaca or llama seminal plasma (100%). A corpus luteum was detected on Day 8 (Day 0=treatment) in all llamas in which ovulation was detected earlier (Day 2) by ultrasonography. The diameter of the CL did not differ among groups. Results document the presence of an ovulation-inducing factor in the seminal plasma of B. taurus. The interspecies effects of seminal plasma on ovulation and luteal development provide rationale for the hypothesis that OIF is conserved among both spontaneous and induced ovulating species.     

Effectiveness of a tris-based extender (SHOTOR diluent) for the preservation of Bactrian camel (Camelus bactrianus) semen

The development of a suitable semen extender is required to extend artificial breeding programs and to preserve the genetic potential of Bactrian camel. Experiments were conducted to provide the optimal osmolality and pH of tris-based extender and to compare that with available extenders for short-term preservation of Bactrian camel semen at 4 degrees C during 24 h. In experiments I and II, the effects of varying osmolalities (270, 300, 330, 360, and 390 mOsm/kg) and pHs (5.5, 6, 6.9, 7.5, 7.9, and 8.9) of tris-based extender on sperm viability were investigated. In experiment III, the efficiency of tris-based extender (SHOTOR diluent) in preserving Bactrian camel semen was compared with lactose (10%), sucrose (10%) and Green buffer. Viability parameters including progressive forward motility (PFM), plasma membrane integrity and the percentage of live spermatozoa were assessed. The data were analyzed using general linear model procedure. In the majority of assessments using tris-based extender, the viability of spermatozoa was superior at the osmolality of 330 mOsm/kg and pH of 6.9. PFM was significantly greater at the time of semen dilution in tris-based (65.5%) and Green buffer (60.5%) compared to that of lactose (31%) and sucrose (28%) extenders (P<0.05), and remained elevated throughout the experiment. There was no significant difference in other viability parameters among 4 extenders (P>0.05). In conclusion, the utilization of a tris-based extender, having the osmolality of 330 mOsm/kg and pH of 6.9, favors the short-term preservation of the Bactrian camel spermatozoa under chilled condition.     

Heterogeneous nuclear-transferred-embryos reconstructed with camel (Camelus bactrianus) skin fibroblasts and enucleated ovine oocytes and their development H-M

This study reconstructed heterogeneous embryos using camel skin fibroblast cells as donor karyoplasts and ovine oocytes as recipient cytoplasts for investigating the developmental potential of the reconstructed embryos. Serum-starved adult camel skin fibroblast cells were used as donor somatic cells. Ovine oocytes matured in vitro were employed as recipient cytoplasts. The fusion of fibroblast cells into recipient cytoplasm was induced by electrofusion. The fused oocytes were activated by 5mM/ml inomycin with 2mM/ml 6-dimethylaminopurine (6-DMAP). The activated reconstructed embryos were co-cultured with ovine cumulus cells in synthetic oviduct fluid supplemented with amino acid (SOFaa) and 10% fetal calf serum (FCS) for 168h. A total of 300 enucleated ovine oocytes were available for xenonuclear embryo reconstruction. The results showed that 71% of the nuclear transfer couplets were successfully fused, 55% of the fused oocytes cleaved within 48h after activation, 82% of the cleaved oocytes developed to 2-16-cell embryo stages and 18% of the cleaved nuclear transfer zygotes developed to the morula stage. This study demonstrated that the xenonuclear transfer camel embryos can undergo the first embryonic division and subsequent development to morula stage in vitro.     

The two-humped camel (Camelus bactrianus): new light on its distribution, management and medical treatment in the past

Eastern and north-eastern Iran and the adjacent Kopet Dagh foothills in south-western Turkmenistan have been postulated to be the region where the domestication of the two-humped camel took place. However, none of the evidence so far brought forward for in situ domestication in this area holds up to closer serutiny. Moreover, the postpleistocene distribution of the wild camel ( Csmelus ferus ) did not include Central Asia. Consequently, we claim that the region where the two-humped camel was domesticated must lie further to the east.
Among the peoples with a long-standing tradition in camel husbandry are the Chinese, as is illustrated by the antiquity of the texts on camel diseases and their treatment. In this contribution, the oldest camel treatise handed down in literature is presented and discussed.