Early prediction of oestrus for herd fertility management in cattle and buffaloes - a review.

Oestrus is defined as a period when a female animal exhibits characteristic sexual behaviour in the presence of a mature male. Oestrous manifestation in dairy animals is due to the oestrogen (E2) effect on the central nervous system (CNS). It is a critical issue to be considered on a priority basis. Inefficient oestrous detection reduces the fertility status of the herd. The primary and most reliable indicator of oestrus is standing to be mounted by a bull or another female herd mate, signalling receptivity and the pre-ovulatory state in dairy cattle. Oestrous detection is primarily a management challenge requiring skill and vigilance. To improve the efficiency of oestrous detection in dairy cattle, visual observation is one of the best methods if done three times a day; however, heat detection aids, if combined, give better results. However, techniques like using teaser bulls, tail painting, chin ball markers, ultrasound (USG) examination, hormonal analysis and examination of cervicovaginal mucus (CVM) improve oestrous detection efficiency. Moreover, the changes in production systems have reduced the expression of oestrous behaviour among cows, due to higher oestrogen (E2) metabolism. Therefore, automated systems, such as pedometers, accelerometers and acoustic sensors like infrared thermography (IRT) and image processing, have significantly enhanced reproductive performance by facilitating oestrous detection and optimizing insemination schedules. From this review, we would conclude that oestrous detection alone contributes considerably to the reproductive status of the herd; therefore, applying different methods of oestrous detection reduces the incidence of missed oestrus and improves the fertility status of the herd.

Cloprostenol effect with and without male stimulation on oestrus synchronization in Peru guinea pigs.

Cloprostenol, a synthetic derivative of prostaglandin F2α, effectively triggers functional and morphological regression of the corpus luteum (luteolysis). In rural Peru, the guinea pig (Cavia porcellus) holds significance within the local economy and serves as a valuable protein source. Enhancing reproductive efficiency is crucial to achieve uniformity in weight, age, and litter size across commercial systems. Thus, our study aimed to evaluate the effect of cloprostenol with and without male stimulation on the onset, duration, and intensity of oestrus in Peru guinea pigs. A total of 128 guinea pigs (120 females and eight males) between 8 and 12 months of age, weighing between 800 and 1200 g, were distributed in cages of 15 females per treatment. Cloprostenol sodium (0 [control], 0.20, 0.25, and 0.30 mg/animal) was IM administered to the groups with and without male stimulation. Four isolated males in individual cages, different from the one used for the treatment, were considered to detect oestrus. The oestrus intensity was assessed by observing sexual behaviour signs such as restlessness, moaning, attempts to mount, pelvic elevation, loin stretching, and vulvar inflammation. The oestrus was manifested 2 days after the administration of cloprostenol sodium. At a dose of 0.30 mg/animal and with male stimulation, the earliest oestrus was observed at 46.9 h. There was a reduction in the oestrus duration (p < .05) in guinea pigs that received the three doses of cloprostenol sodium compared to the control group. The highest percentages of frank oestrus intensity (66.7%) were strongly associated with the administered doses of cloprostenol sodium (p < .01). In conclusion, the cloprostenol sodium administration was proper for rapid oestrus induction in Peru guinea pigs.

Oestrous cycle affects emergence from anaesthesia with dexmedetomidine, but not propofol, isoflurane, or sevoflurane, in female rats.

BACKGROUND: Although sex differences in anaesthetic sensitivity have been reported, what underlies these differences is unknown. In rodents, one source of variability in females is the oestrous cycle. Here we test the hypothesis that the oestrous cycle impacts emergence from general anaesthesia. METHODS: Time to emergence was measured after isoflurane (2 vol% for 1 h), sevoflurane (3 vol% for 20 min), dexmedetomidine (50 µg kg-1 i.v., infused over 10 min), or propofol (10 mg kg-1 i.v. bolus) during proestrus, oestrus, early dioestrus, and late dioestrus in female Sprague-Dawley rats (n=24). EEG recordings were taken during each test for power spectral analysis. Serum was analysed for 17ß-oestradiol and progesterone concentrations. The effect of oestrous cycle stage on return of righting latency was assessed using a mixed model. The association between righting latency and serum hormone concentration was tested by linear regression. Mean arterial blood pressure and arterial blood gases were assessed in a subset of rats after dexmedetomidine and compared in a mixed model. RESULTS: Oestrous cycle did not affect righting latency after isoflurane, sevoflurane, or propofol. When in the early dioestrus stage, rats emerged more rapidly from dexmedetomidine than in the proestrus (P=0.0042) or late dioestrus (P=0.0230) stage and showed reduced overall power in frontal EEG spectra 30 min after dexmedetomidine (P=0.0049). 17ß-Oestradiol and progesterone serum concentrations did not correlate with righting latency. Oestrous cycle did not affect mean arterial blood pressure or blood gases during dexmedetomidine. CONCLUSIONS: In female rats, the oestrous cycle significantly impacts emergence from dexmedetomidine-induced unconsciousness. However, 17ß-oestradiol and progesterone serum concentrations do not correlate with the observed changes.

Prevalence of Oestrus ovis in small ruminants from the eastern Iberian Peninsula. A long-term study.

Oestrus ovis is an obligate parasite that causes myiasis in domestic ruminants, being commonly found in the Mediterranean area. From 2009 to 2019 a total of 3476 heads of culling sheep and goats from the Mediterranean coast of Spain were examined for the presence of O. ovis. The total prevalence was 56.3%, significantly higher in sheep than in goats (61.2% and 43%, respectively). Differences were found in the mean annual prevalence, with the highest value being registered in 2018 (61.7%) and the lowest in 2012 (50.3%). Autumn, for sheep, and winter, for goats, were the seasons with the highest number of infested specimens. Temperature, but not rainfall, was found to be associated with prevalence (p < 0.05). Most L1 were found in the anatomic region I (septum, meatus, and ventral conchae), while L2 and L3 were mainly located in regions II (nasopharynx, ethmoid labyrinth, and dorsal conchae), and III (sinuses). The overall intensity was 12.8 larvae per head, significantly higher in sheep (13.3) than in goats (3.5). Our results confirm the high prevalence of O. ovis in sheep and goats in this geographic area over the last decade, with the trend increasing in recent years in association with higher mean temperatures.

Parity affects the relationship between the insemination-ovulation interval and the conception rate in lactating dairy cows.

The aim of this study was to investigate the relationship between parity and the optimal insemination-ovulation interval (IOI) in lactating dairy cows, as well as the relationship between the optimal timing of artificial insemination (AI) based on the onset of oestrus and parity. A total of 183 postpartum lactating Holstein cows participated in the study and underwent 522 AI procedures using unsorted frozen semen between 2011 and 2018. Transrectal ultrasound was used to detect ovulation times in 326 AI procedures, and a pedometer was used to detect oestrus onset times in 470 AI procedures. An analysis was also performed on data from 265 AI procedures in which all the times of oestrus onset, oestrus end and ovulation were detected. The results showed that ovulation occurring 6-12 h after AI was associated with the numerical highest conception rate in multiparous cows. Moreover, the conception rate was higher when ovulation occurred between 6 and 12 h after AI than when it occurred between 0 and 6 h or 18 and 24 h after AI. In contrast, the effect of IOI on the conception rate in primiparous cows was insignificant. There was no effect of AI timing based on the oestrus onset on the conception rate in either primiparous or multiparous cows. Furthermore, the interval from oestrus onset to ovulation was similar in primiparous and multiparous cows. The difference in optimal IOI between primiparous and multiparous cows may be due to the larger uterus size in multiparous cows. These findings suggest that the optimal timing for AI in multiparous cows is more restricted than in primiparous cows. Therefore, it is essential to carefully determine the timing of AI when it is performed in multiparous cows.

Evaluation of sexed semen-based artificial insemination in Tharparkar cattle under organized farm condition.

Sexed semen facilitates additional female calf production for the expansion of a herd at a faster rate and also curtails the surplus production of unwanted male calves. A study was conducted to evaluate the performance of sexed semen in indigenous Tharparkar cows based on 114 artificial inseminations (AI) performed at natural oestrus using two protocols i.e., single AI (n = 48) and double AI (n = 66). Overall, the first service conception rate (CR) was significantly higher in double (53.0%) than single (33.3%) AI protocol. The odds ratio of conception rate in double AI was 2.26 (χ2 = 4.4, df = 1, p = .04) with respect to single AI. The time that elapsed since the detection of oestrus to insemination was also analysed. In a single AI protocol, the CR was higher (p < .05) at 16 h (54.6%) than insemination at 8 h (27.0%) following the onset of oestrus. Yet, the CR using double AI protocol did not differ (p = .73) significantly when AIs were performed either at 8 h and 24 h (51.9%) or 16 h and 24 h (57.1%) post onset of oestrus. Besides, like the single AI protocol, the parity of the animals also influenced the CR, being higher in heifers (n = 22) than those of parous (n = 92) cows (72.73 vs. 40.43%, χ2 = 7.48, df = 1, p = .006) in the present study. The odds ratio of conception in heifers was 3.93 with respect to parous cows. Overall, the birth of female calf was 91.7%. In conclusion, the present study indicates a future promise of the sexed semen for the production of more female offspring from Tharparkar cattle.

Detection of changes in the circadian rhythm of cattle in relation to disease, stress, and reproductive events.

Disease and stress can disrupt the circadian rhythm of activity in animals. Sensor technologies can automatically detect variations in daily activity, but it remains difficult to detect exactly when the circadian rhythm disruption starts. Here we report a mathematical Fourier-Based Approximation with Thresholding (FBAT) method designed to detect changes in the circadian activity rhythm of cows whatever the cause of change (typically disease, stress, oestrus). We used data from an indoor positioning system that provides the time per hour spent by each cow resting, in alleys, or eating. We calculated the hourly activity level of each cow by attributing a weight to each activity. We considered 36-h time series and used Fourier transform to model the variations in activity during the first and last 24 h of these 36-h series. We then compared the Euclidian distance between the two models against a given threshold above which we considered that rhythm had changed. We tested the method on four datasets (giving a cumulative total of ~120000 cow*days) that included disease episodes (acidosis, lameness, mastitis or other infectious diseases), reproductive events (oestrus or calving) and external stimuli that can stress animals (e.g. relocation). The method obtained over 80% recall of normal days and detected 95% of abnormal rhythms due to health or reproductive events. FBAT could be implemented in precision livestock farming system monitoring tools to alert caretakers to individual animals needing specific care. The FBAT method also has the potential to detect anomalies in humans to guide healthcare intervention or in wild animals to detect disturbances. We anticipate that chronobiological studies could apply FBAT to help relate circadian rhythm anomalies to specific events.

Morphometric changes on the vulva from proestrus to oestrus of nulliparous and multiparous HYPERPROLIFIC sows.

The aim of this study was to assess whether vulvar morphometric changes occurring in female pigs during proestrus and oestrus could be objective, accurate and predictive indicators of the onset to oestrus and thus performed artificial inseminations at the most appropriate time. For that purpose, pictures of vulvas from 60 hyperprolific females (30 gilts and 30 sows) during proestrus and oestrus were taken once a day. Vulva measurements (area, perimeter, length and width) on these pictures were performed using the image processing ImageJ software. Gilts and sows showed statistical differences (p < .01) in all vulvar morphometric measurements between proestrus and oestrus. Statistical differences in vulvar metrics were detected 24 h before the onset to oestrus, affecting all vulvar measurements in gilts, whereas only vulvar width was affected in sows. The image analysis used in this study may contribute to the development of smart technology in swine farming.

A comparative transcriptome and proteomics study of post-partum ovarian cycle arrest in yaks (Bos grunniens).

Post-partum ovarian cycle arrest is the main factor affecting yak reproductive efficiency. There are few reports regarding the molecular regulatory mechanism of post-partum oestrus at transcriptome and proteome levels in yaks. Our previous studies focussed on the ovaries of yaks with post-partum ovarian cycle arrest and post-partum oestrus yaks. In this study, RNA sequencing transcriptomic study was combined with quantitative proteomic analyses to identify post-partum ovarian cycle-related genes and proteins. Consequently, 1,149 genes and 24 proteins were found to be up- or downregulated during post-partum oestrus. The analysis of differentially regulated genes identified three gene or protein pairs that were synchronously upregulated and no gene or protein pairs that were synchronously downregulated, suggesting that these upregulated genes may regulate the post-partum ovarian cycle. The functional classification of these differentially expressed genes and proteins indicated their connection with the oocyte meiosis, the oestrogen signalling pathway, the progesterone-mediated oocyte maturation and the gonadotrophin-releasing hormone (GnRH) signalling pathway. In this study, a total of six genes and two proteins involved in the oocyte meiosis, the oestrogen signalling pathway, the progesterone-mediated oocyte maturation and the GnRH signalling pathway were identified. The CSNK1A1, M91_09723, M91_11326, M91_21439, M91_19073, SHC2, Atf6b, M91_03062, HSPCA and calmodulin could regulate oestrus, respectively, in the post-partum so as to control the anoestrus status.

Suppression of reproductive behaviour and gonadal function in female horses-An update.

The behaviour of mares is often detrimental to their performance resulting in frequent demand for methods to suppress gonadal function. In addition, prevention of unintended reproduction especially in feral horse populations may require methods for suppression of gonadal function. Surgical ovariectomy is a safe method but not an acceptable approach in feral mares and undesired in mares where future breeding is considered. There are different approaches for artificial prolongation of the luteal phase resulting in transient inhibition of oestrus and ovulation. Among those, treatment with natural or synthetic progestogens is considered the most common and successful method. Whereas application of intrauterine devices may result in prolongation of luteal function in non-pregnant mares, intrauterine insertion of glass balls is no longer recommended because of complications in individual mares. There are several safer alternatives that may be of interest, especially for population control in free-roaming horses. Treatment with long-acting deslorelin implants inhibited ovulation and oestrus behaviour in mares for limited and variable time intervals in a dose-dependent manner. The effect of GnRH vaccines varies considerably among individual mares, is age dependent, and oestrus-like behaviour may still occur. Contraception via immunization against native porcine or recombinant zona pellucida antigen is successful, but immunocontraception is as much a result of ovarian inactivity as an antibody-based block to sperm-oocyte binding. In conclusion, several treatments for suppression of gonadal function in mares are available, but there are advantages and disadvantages associated that have to be considered. The treatment of choice will thus differ with regard to the demands.

Effect of lowering blood cortisol level along with progesterone priming on intensifying the secondary symptoms of oestrus in Murrah buffaloes during the hot summer months.

The present study sought to intensify oestrus symptoms in heat-stressed pre-pubertal Murrah buffalo heifers. The first experiment aimed at lowering the blood cortisol level. Twenty pre-pubertal buffalo heifers approximately 36-40 months of age were randomly allocated to four groups of five buffaloes. Group Gly received 100 ml glycerol orally for 8 days (day 0 is the start of treatments), group Gly-E received intra-muscular injections of 500 mg tocopherol (vitamin E activity) and 150 mg sodium selenite on alternate days (four treatments) in addition to treatments received by group Gly, group Gly-E + B-Ch received intra-muscular injections of vitamin B-complex and choline daily for 8 days in addition to the treatments received by group Gly-E. The control group Co-1 did not received any treatments. Blood cortisol and glucose level was measured on day 7 post-treatment. The second experiment sought to test the effect of lowering the blood cortisol level and progesterone priming on the intensity of oestrus symptoms in heat-stressed pre-pubertal buffalo heifers. Eighty buffaloes were allocated into four groups of twenty buffaloes. P4 group received intra-vaginal progesterone CIDR for 5 days (day 0 is the day of start of treatment). Gly-E group received the same treatment as in the first experiment for 7 days (day 0-6) and P4 + Gly-E received the combination of P4 and Gly-E groups. The control group Co-2 did not receive any treatments; however, on day 6, all groups received intra-muscular injection of 2 mg estradiol benzoate. The buffaloes were graded for oestrus symptoms on the basis of mucus discharge (A-scanty and B-copious), vulvar redness (slightly pink and moist-A and deep red and moist-B), ferning (nil or poor-A and good with arborization-B) and cellularity of cervical mucus (Type A goblet cells with round nucleus and abundant cytoplasm and Type B goblet cells with cylindrical nucleus and scanty cytoplasm). The blood glucose level (mean ± SEM) of Gly group on day 7 of treatment in experiment 1 was 58.3 ± 0.4 mg/dl that was significantly greater than Co-1 (52.4 ± 0.4 mg/dl), Gly-E (53.5 ± 0.8 mg/dl) and Gly-E + B-Ch (52.7 ± 0.6 mg/dl) groups; however, the difference was found to be non-significant among Co-1, Gly-E and Gly-E + B-Ch groups. The cortisol level (mean ± SEM) on day 7 in groups Gly (11.4 ± 0.6 ng/ml), Gly-E (8.1 ± 0.5 ng/ml) and Gly-E + B-Ch (7.5 ± 0.4 ng/ml) was significantly lower than groups Co-1 (15.5 ± 0.3 ng/ml), though the difference between groups Gly-E and Gly-E + B-Ch was non-significant and the level in group Gly-E significantly lower than Gly. Thus, the combination of vitamin E and selenium along with glycerol had the best effect in the reduction of blood cortisol level in heat-stressed buffaloes. In the second experiment the blood estradiol level on day 7 in all groups was around 40 pg/ml. Cortisol reduction (group Gly-E) improved oestrus symptoms (mucus discharge and ferning) compared to control, but when compared to group P4 showed non-significant difference with respect to mucus discharge, vulvar redness and ferning of cervical mucus. However, when cortisol reduction treatments were combined with progesterone priming (group P4 + Gly-E) oestrus symptoms were improved with respect to mucus discharge and ferning thus, combination treatment had synergistic effect. This effect was also observed with cellularity of cervical mucus, where combination treatment group P4 + Gly-E produced cervical mucus with significantly lower percentage of Type B cells compared to group P4 and group Gly-E. Thus, administration glycerol and vitamin E selenium combination reduces blood cortisol level and along with progesterone priming can be effective to improve the oestrus symptoms of buffaloes during the hot summer months.

A case report: Actinobaculum suis infection associated with formation of pyogranuloma, epididymitis and azoospermia in a boar.

BACKGROUND: Actinobaculum suis is a bacterium known to cause infections of the urogenital tract of sows. Infection can occur through close contact to boars, who frequently carry the pathogen in their preputial diverticulum but do not become clinically diseased themselves. In the current case, Actinobaculum suis was isolated from pyogranuloma of inflamed epididymis in a boar with poor fertility. CASE PRESENTATION: Increased return to oestrus rate, which worsened after the purchase of a new boar, was reported in an organic farm in Switzerland. During herd examination, azoospermia of the boar was diagnosed, and slaughter, followed by examination of its urogenital tract, was carried out. Pathologically, pyogranuloma formation and epididymitis were diagnosed. Bacteriology of the pyogranulomas showed growth of Actinobaculum suis and mixed flora. After the boar was replaced, the return to oestrus rate improved tremendously. CONCLUSION: A close relative of Actinobaculum suis, namely Actinotignum schaalii, has already been associated with epididymitis in humans. Considering the present case and the parallels in human medicine, Actinobaculum suis should be included in the list of differentials of boars with poor fertility.

Major differences in the larval anatomy of the digestive and excretory systems of three Oestridae species revealed by micro-CT.

Oestrid flies (Diptera: Oestridae) do not feed during the adult stage, so they depend on an efficient assimilation and storage of nutrients during their parasitic larval stage. We describe the general morphology and provide volumetric data for the digestive and excretory organs of the three larval instars of the nasal bot fly Oestrus ovis L., using micro-computed tomography. The size of the digestive and excretory organs greatly increased across larval instars. In all instars, the two salivary glands were remarkably large and formed a 'glandular band' by coming together, but without lumina uniting, at their posterior ends. The distal region of the anterior Malpighian tubules was greatly enlarged and full of highly radio-opaque concretions. Moreover, the anatomy of O. ovis third-instar larva was compared to that of two species of, respectively, similar and different feeding habits: Cephenemyia stimulator (Clark) and Hypoderma actaeon Brauer. Whereas the general morphology and arrangement of the digestive and excretory systems of C. stimulator was similar to that of O. ovis, some differences were observed in H. actaeon: a swollen anterior region of the midgut, salivary glands shorter and not forming a 'band' and anterior Malpighian tubules narrowly uniform throughout their entire length.

Pregnancy rate per cycle is heritable and reduces with cycle in naturally mated tropically adapted beef cows.

Pregnancy in cattle is the outcome of the complex process of initiation of cycling, fertilization, maternal recognition of pregnancy and foeto-placental development. Though much is known about initiation of cycling and associated risk factors, there are virtually no data on pregnancy rate per cycle for naturally mated cattle, especially for extensively managed, tropically adapted genotypes, which this study aimed to determine. Tropical composite (Bos indicus and African Sanga crosses with Bos taurus) and Brahman cattle (n = 2,181) of known pedigree in four-year groups at four sites were mated annually for 84 days. Body condition, ovarian function, pregnancies, calving and lactation were monitored through six full reproductive cycles using 4-8 weekly ultrasound of the reproductive tract outside the calving period and daily monitoring during calving. From this, dates of commencement of cycling and conception in each year were estimated for each animal, enabling calculation of established pregnancy for consecutive 21-day periods while cycling and of pregnancies within four months of calving while lactating (P4M). Pregnancy per 21-day period (cycle) during mating for cycling animals averaged 63%, 71%, 41% and 28% in four consecutive cycles. Pregnant per cycle was 2%-11% higher in tropical composites than in Brahmans. The only other consistently significant risk to becoming pregnant was if cycling commenced later than three weeks before mating commenced. P4M averaged 62% and was lower for cows in sub-optimal body condition and in first-parity and later-calving cows. Pregnant per cycle was moderately heritable (~20%), while heritability was moderate to high (33%) for P4M. Selection for pregnant per cycle could be achieved indirectly by selection for P4M, a trait that is readily measured.

Effects of age and season on conception rate of Mediterranean Italian Dairy Buffalo (Bubalus bubalis) following oestrus synchronization and fixed-time artificial insemination.

The objective of this project was to report the effects of season and age on conception rate amongst Italian Mediterranean Buffalo subjected to an Ovsynch/Resynch (O/R) reproductive management protocol. The study utilized nulliparous (heifers), primiparous buffalo cows (PBC) and multiparous buffalo cows (MBC). The primiparous and multiparous groups were subjected to the synchronization protocol throughout the entire year, but heifers were synchronized and inseminated only during the spring/summer seasons. The conception rate obtained following the OvSynch oestrus synchronization protocol, applied during spring, was 68.4% for heifers, 83.3% for PBC and 67.7% for MBC. The overall total conception rates following the complete O/R protocol were 84.27%, 94.4% and 79%. Conception rates achieved during summer were heifers 52%, PBC 47.2% and MBC 49%, whilst overall conception rates following the full O/R protocol were 72%, 69.8% and 58.2% respectively. In the autumn seasons, PBC conceived 58.9% and MBC 52.1% following initial Ovsynch, which improved to total overall conception rates of 87.5% and 78.7% following the full O/R protocol. Similarly, in the winter season, PBC experienced a conception rate of 47.5% following Ovsynch and 72.5% after a follow-up Resynch. MBC experienced 60.0% and 74.4% conception following Ovsynch and full O/R, respectively, during winter. Total conception rates during all seasons were quite acceptable following the O/R protocol. There was a significant decrease from spring to summer in conception rate for all parity groups, but heifers were not a severely affected as older buffalo cows. This finding agrees with that of other investigators indicating that heifer fertility is not as negatively impacted by long photoperiod and higher ambient temperature as that of older animals. The O/R protocol as utilized in this study is an effective means of reproductive management for dairy buffalo cows and is effective for improving fertility during out-of-season breeding.

Timed artificial insemination in crossbred mares: Reproductive efficiency and costs.

Timed artificial insemination (TAI) has boosted the use of conventional artificial insemination (CAI) by employing hormonal protocols to synchronize oestrus and ovulation. This study aimed to evaluate the efficiency of a hormonal protocol for TAI in mares, based on a combination of progesterone releasing intravaginal device (PRID), prostaglandin (PGF2α ) and human chorionic gonadotropin (hCG); and compare financial costs between CAI and TAI. Twenty-one mares were divided into two groups: CAI group (CAIG; n = 6 mares; 17 oestrous cycles) and TAI group (TAIG; n = 15 mares; 15 oestrous cycles). The CAIG was subjected to CAI, involving follicular dynamics and uterine oedema monitoring with ultrasound examinations (US), and administration of hCG (1,600 IU) when the dominant follicle (DF) diameter's ≥35 mm + uterine oedema + cervix opening. The AI was performed with fresh semen (500 × 106 cells), and embryo was recovered on day 8 (D8) after ovulation. In TAI, mares received 1.9 g PRID on D0. On D10, PRID was removed and 6.71 mg dinoprost tromethamine was administered. Ovulation was induced on D14 (1,600 IU of hCG) regardless of the DF diameter's, and AI was performed with fresh semen (500 × 106 cells). On D30 after AI, pregnancy was confirmed by US. The pregnancy rate was 80.0% in TAIG and 82.3% in CAIG (p > .05). The TAI protocol resulted in 65% reduction in professional transport costs, and 40% reduction in material costs. The TAI was as efficient as CAI, provided reduction in costs and handlings, and is recommended in mares.

Development and characterization of 15 novel polymorphic microsatellite loci for two important bot flies (Diptera, Oestridae) by next-generation sequencing.

Cephenemyia stimulator and Oestrus ovis are two important parasitic bot flies (Oestridae) species causing myiasis, with a potential negative impact on the welfare of the host. Using next-generation sequencing approach and bioinformatics tools, a large panel of possible microsatellites loci was obtained in both species. Primer pairs were designed for 15 selected microsatellite loci in C. stimulator and other 15 loci in O. ovis for PCR amplification. Loci amplification and analysis were performed in four populations of each species. The results demonstrated that all selected loci were polymorphic, with the number of alleles ranging from 2 to 6 per locus in C. stimulator and 3 to 13 per locus in O. ovis. This is the first time to describe these microsatellite loci for C. stimulator and O. ovis. These two sets of microsatellite markers could be further used for biogeographic and population genetics studies.

Screening for reproductive biomarkers in Bactrian camel via iTRAQ analysis of proteomes.

Bactrian camel is an ancient and precious species of livestock; that is, unique resources exist in the desert and have important economic and scientific value. In recent years, the number of Bactrian camels has declined sharply. Due to its long reproductive cycle and seasonal oestrus, the mechanism of oestrus is unknown. To identify candidate biomarkers of reproduction, we performed a comprehensive proteomic analysis of serum from Bactrian camel in oestrus and non-oestrus, using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with tandem mass spectrometry. We identified 359 proteins, of which 32 were differentially expressed: 11 were up-regulated and 21 were down-regulated in samples from camels in oestrus. We validated the differential expression of a subset of these proteins using qPCR and Western blot. Gene ontology annotation identified that the differentially expressed proteins function in cellular processes, metabolic processes and immune system processes. Notably, five of the differentially expressed proteins, PCGF5, histone H1.2, RBP4, FOLR1 and ANTXR2, are involved in reproductive regulatory processes in other animals. KEGG enrichment analysis demonstrated significant enrichment in several cardiac-related pathways, such as 'dilated cardiomyopathy', 'hypertrophic cardiomyopathy', 'cardiac muscle contraction' and 'adrenergic signalling in cardiomyopathy'. Our results suggest that candidate biomarker (PCGF5, histone H1.2, RBP4, FOLR1 and ANTXR2) discovery can aid in understanding reproduction in Bactrian camels. We conclude that the profiling of serum proteomes, followed by the measurement of selected proteins using more targeted methods, offers a promising approach for studying mechanisms of oestrus.

Comparative efficiency of oestrus synchronization in sheep with progesterone/eCG and progesterone/GnRH during breeding and non-breeding season.

The present study compared the occurrence of oestrus behaviour and ovulation in response to the insertion of CIDR devices plus a classical treatment with equine chorionic gonadotrophin (eCG; single dose at CIDR removal) or alternative treatments with gonadotrophin-releasing hormone (GnRH, either in a single dose at 56 hr after CIDR removal, or in one dose at CIDR insertion and another dose at 56 hr after CIDR removal). The appearance of oestrus behaviour during reproductive season ranged between 84% and 95% and all females showing oestrus signs had subsequent ovulations. The response, during seasonal anoestrus, was similar in the group treated with eCG, but less than half of the females in the groups treated with GnRH showed oestrus signs in response to the treatment, although more than 80% of them showed resumption of ovulatory activity after the treatment. In conclusion, protocols based on GnRH administration offer similar yields to eCG-based protocols during the reproductive season but occurrence of oestrus in response to GnRH-based treatments is highly compromised during seasonal anoestrus.

Higher doses of Melatonin affect ovarian and middle uterine arteries vascular blood flow and induce oestrus earlier in acyclic ewes.

The present study aimed to evaluate the effect of a single intramuscular injection of melatonin on response of oestrus, rate of conception, ovarian and uterine blood supply during the non-breeding season. Melatonin powder was dissolved in dimethyl sulfoxide (DMSO) and olive oil and injected in two dose of 25 mg (MEL-1; n = 10) and 50 mg (MEL-2; n = 10) in May. Ten ewes were served as the control (CON). Results showed that oestrous response was higher in the MEL-2 group than in other groups (p < .01). The time to onset of oestrus in both the MEL-1 and MEL-2 groups was significantly shorter, compared with the CON group (p < .01). Pregnancy rates reached 80% and 90% at 45 days of gestation. Doppler indices of the right and left ovarian artery were lower in the melatonin groups than in normal groups (p < .01). Both Doppler indices in uterine arteries were not different among two treatment groups (p > .05), but there was a significant difference when compared with the CON group (p < .01), no difference found between two treatment group in the Doppler velocities of middle uterine arteries (p > .05). In conclusion, the definite changes in the studied parameters showed two novel findings; (a) the increase in melatonin dose results in increasing the blood flow expressed by increased Doppler velocities in fatty tailed ewes, (b) as well as, melatonin can initiate the breeding season regardless of the season earlier by 1-2 months in ewes.