Enhancing cryopreserved ram sperm quality at -80 °C with Spirulina platensis and Salvia verbenaca extracts.

This study was conducted in two steps to evaluate the influence of freezing methods and natural extracts on cryopreserved ram sperm quality. Initially, the research compared the effects of two freezing methods: liquid nitrogen (LN2) versus -80 °C, on post-thawed ram semen on total and progressive motilities and velocity parameters. Experiment I revealed no significant differences (P > 0.05) between the LN2 and -80 °C freezing methods, indicating similar effects on the analyzed parameters. Experiment II aimed to examine the influence of Spirulina platensis (SP) and Salvia verbenaca (SV) extracts added to egg yolk extender on cryopreserved sperm quality, utilizing the -80 °C freezing method. Various concentrations (1.25, 3.75, 6.25 and 8.75 µg*mL-1) of acetone (Ac-SP and Ac-SV) and hexanoic (Hex-SP), as well as methanolic (MeOH-SV) extracts, were added into the extender. A thorough assessment of post-thawed sperm quality parameters, encompassing motility, velocity parameters, viability, membrane integrity, abnormality and lipid peroxidation was conducted. The outcomes demonstrated that 1.25 and 3.75 g*mL-1 of Ac-SP and Hex-SP and 1.25 µg*mL-1 of AC-SV and MeOH-SV increased the post-thawed ram sperm quality. In conclusion, this study emphasizes the antioxidant properties of SP and SV extracts, highlighting their potential to protect cryopreserved sperm cells from oxidative stress at -80 °C.

Inclusion of Spirulina platensis and Salvia verbenaca extracts to boost semen quality and fertilization ability in sheep.

The aim of this study was to investigate the effect of Spirulina platensis (SP) and Salvia verbenaca (SV) extracts added to skimmed milk (SM) extender on ram sperm quality and fertility. Semen was collected using an artificial vagina, extended in SM to reach a final concentration of 0.8 × 109 spermatozoa/mL, stored at 4°C and evaluated at 0, 5 H and 24 H. The experiment has been performed in three steps. Firstly, from four extracts (methanol: MeOH, acetone: Ac, ethyl acetate: EtOAc and hexane: Hex) of SP and SV, only acetonic and hexanoic extracts of SP and acetonic and methanolic extracts of SV showed the highest in vitro antioxidant activities and were then selected for the following step. Thereafter, the effects of four concentrations (1.25, 3.75, 6.25, and 8.75 µg/mL) of each selected extract on stored sperm motility were evaluated. The output of this trial led to select the best concentrations having beneficial effects on sperm quality parameters (viability, abnormalities, membrane integrity, and lipid peroxidation) and fertility after insemination. The results showed that the same concentration (1.25 µg/mL) of both Ac-SP and Hex-SP, as well as 3.75 µg/mL of Ac-SV and 6.25 µg/mL of MeOH-SV, maintain all sperm quality parameters at 4°C during 24 H of storage. Besides, no difference was found in fertility between the selected extracts and the control. In conclusion, SP and SV extracts were shown to improve the quality of ram sperm and to maintain fertility rate after insemination as similar or competitive to many previous studies published in the field.

Effects of Origanum majorana essential oil and antibiotics on the quality of frozen thawed Beni Arouss buck semen.

This study aims to investigate the effects of Origanum majorana (OM) essential oil (EO) at different concentrations and antibiotics on post-thawed Beni Arouss buck semen quality. Semen collection was performed using artificial vagina from eight Beni Arouss bucks. Ejaculates were pooled, divided into 12 equal aliquots, washed and diluted to 400 × 106 sperm/ml (with 7% of glycerol). Skim milk-based extender was supplemented with different concentrations of OM EO (0%; 0.01%; 0.02%; 0.03%; 0.04% and 0.05%) without antibiotics, marked as (CTR-), (M1-), (M2-), (M3-), (M4-) and (M5-), and with 50 mg of streptomycin and 50,000 IU of penicillin per 100 ml, marked as (CTR+), (M1+), (M2+), (M3+), (M4+) and (M5+), respectively. Aliquots were cooled to 4°C, then frozen in 0.25 ml straws with a programmable freezer and finally stored in liquid nitrogen for 48 h. Thawing was performed at 37°C for 30 s. Motility, live sperm, sperm abnormalities, membrane integrity, lipid peroxidation and bacterial growth were evaluated after thawing. Among the tested extenders, M2+ improved all semen quality parameters. Sperm motility, live sperm and membrane integrity increased significantly, while the number of abnormal sperm and bacterial growth decreased significantly. The toxic effect of OM EO, with and without antibiotics, appeared beyond 0.03%. In conclusion, M2+ is recommended to improve the cryopreservation of Beni Arouss buck semen.

Onset of puberty and regularity of oestral cycles in ewe lambs of four breeds under high-altitude conditions in a non-seasonal country.

Twenty-four Colombian Creole, Romney Marsh, Hampshire and Corriedale ewes raised under high-altitude conditions in a non-seasonal country were used to determine and to characterize the onset of puberty and of regularity ovarian function since the age of 3months. They underwent blood collection for determination of progesterone and monthly weight assessment. Oestrus was assessed daily by a teaser ram. In all breeds, age at onset of puberty ranged from 5.4 to 6.9 months and age at onset of regular ovarian function ranged from 7.4 to 8.6 months. Colombian Creole showed a higher body development at onset of puberty: 73.5 ± 8.3% versus 56.2 ± 7.4 in Romney Marsh, 58.8 ± 10.4 in Hampshire, and 57.3 ± 8.0 in Corriedale (p < 0.05), as well as a higher relative daily weight gain (+17%, p < 0.05). A negative correlation between daily weight gain and age at onset of puberty was established. Progesterone at onset of puberty and of regularity did not differ between breeds, but characterization of the luteal phase by the progesterone area under the curve (P4-AUC) revealed at both time points significantly lower values for Creole lambs (p < 0.05). Decreased P4-AUC paralleled a higher proportion of short oestral cycles observed prior to onset of regularity in Creoles, whereas an increased proportion of extra-long cycles were recorded in Romney Marsh (p < 0.05). These results establish first reference data for economically important breeds raised under tropical conditions. In comparison with other breeds, Colombian Creole requires a higher body development to achieve puberty and that an important proportion of short characterizes its prepubertal period cycles.

Evaluation of different hormonal treatments on oestral and ovarian responses in Moroccan Beni Arouss goats during anoestrus and breeding season.

The efficacy of eight combinations of fluorogestone acetate (FGA, 20 or 40 mg as intravaginal device during 11 days), equine chorionic gonadotropin (eCG, 300 or 500 UI injected 48 hr before FGA removal) and prostaglandin F2α (cloprostenol, 0 or 50 µg injected 48 hr before FGA removal) aiming at induction and synchronization of oestrus and ovulation was evaluated during the anoestrus season in spring and during the breeding season in autumn in adult Beni Arouss goats. Oestrous behaviour was recorded between 12 and 60 hr after FGA removal. Blood samplings allowing to assess onset of the pre-ovulatory LH surge and increase of progesterone as sign of an active corpus luteum were performed, respectively, between 20 and 60 hr and 3, 5, 8 and 15 days after FGA removal. No season-related differences (spring vs. autumn) were observed for oestrous response (95% vs. 93%), pre-ovulatory LH surge (94% vs. 84%) and luteal response after 3-8 and 11-15 days post-treatment (respectively 92% vs. 66% and 92% vs. 98%). The onset of oestrus (21 [13-53] vs. 32 [12-54] hr) and LH surge (26 [20-60] vs. 38 [22-60] hr) occurred significantly later in autumn. FGA (40 vs. 20 mg) in autumn significantly delayed the onset of oestrus (36 [16-54] vs. 23 [12-47] hr) and LH surge (44 [26-58] vs. 33 [22-60] hr). Significant treatment-related differences were recorded for onset of LH surge (earliest for 20 mg FGA, 300 IU eCG, 50 µg PGF2α ) and onset of luteal phase (latest for 40 mg FGA, 300 IU eCG, 50 µg PGF2α ). In conclusion, the hormone combinations tested appeared equally effective in terms of oestrous and ovulation rates. Season has influenced significantly the onset of oestrus and LH surge, and the high dose regimen of FGA delayed the ovarian response in autumn.

Rearing Management and Its Impact on Caseous Lymphadenitis in Sheep.

Two surveys were conducted to assess the prevalence and risk factors of ovine caseous lymphadenitis (CL) and explore the association between its superficial and visceral forms in Sardi sheep in Settat province, Morocco. A total of 1521 sheep, including 318 lambs < 6 months, 572 young sheep aged 6-24 months, and 631 adult sheep > 24 months, were examined from 57 flocks. Superficial CL prevalence levels were as follows: 69/318 in lambs, 292/572 in young sheep, and 114/631 in adult sheep. Additionally, 2124 sheep, including 1813 young and 311 adult sheep, were inspected in slaughterhouses ante- and post-mortem. Among all infected animals, the prevalence of visceral abscesses was notably higher in adult sheep (83%, OR = 9.8, 95% CI = 5.5-17.2) compared to young sheep (35%). Data on flock size, sex, age, body condition score, rearing methods, and hygienic measures were collected. Suspected CL cases were confirmed using quadruplex-PCR. Poor barn ventilation, lack of abscess control, and younger age were identified as key risk factors for CL. Additionally, a high percentage (89%) of animals with thoracic abscesses did not display superficial lesions. While age and abscess control have been reported in previous studies, barn ventilation appears to be a new, but important, risk factor. In conclusion, the identified risk factors in Settat province are linked to breeding management practices. Implementing regular abscess control measures and improving barn ventilation are essential for CL prevention.

Carvacrol and Thymol Enhance the Quality of Beni Arouss Buck Semen Stored at 4 °C Thanks to Their Antimicrobial Properties.

This study aims to investigate the impact of carvacrol and thymol on the quality of Beni Arouss buck semen stored in skim milk at 4 °C. Ejaculates were collected from eight Beni Arouss bucks weekly for 11 weeks, pooled, and then divided into three equal parts. Samples were diluted to 400 × 106 sperm/mL in skim milk (control) and skim milk supplemented with a single dose of 200 µM carvacrol and thymol each. Evaluations of sperm motility, viability, abnormalities, membrane integrity, lipid peroxidation, and bacterial growth were conducted at 0, 6, 24, and 48 h of liquid storage at 4 °C. After 48 h of storage, the results indicate that the addition of carvacrol positively influences total and progressive motility and viability. However, it also leads to a decrease in lipid peroxidation and bacterial growth compared to the control group (p < 0.05). Thymol showed similar results to carvacrol, except for progressive motility (p > 0.05). Bacterial growth was negatively correlated with total and progressive motility and viability (p < 0.05), while no correlation between lipid peroxidation and these parameters was observed (p > 0.05). In conclusion, the addition of carvacrol and thymol to skim milk extender moderately improves the quality of Beni Arouss buck semen after 48 h storage at 4 °C due to its antimicrobial activity.

Analysis of the Genetic Diversity and Population Structure of Four Senegalese Sheep Breeds Using Medium-Density Single-Nucleotide Polymorphisms.

In Senegal, sheep breeds have adapted to their environment and play a key socio-economic role. This study aimed to explore the genetic diversity and structure of four Senegalese sheep breeds (Peul-peul, Djallonke, Touabire, and Ladoum) and their relationships with global sheep breeds. To that end, forty-seven sheep were genotyped using the OvineSNP50 BeadChip, and these genotypic data were analysed with those of 73 sheep breeds representative of worldwide ovine diversity (2729 animals). The average observed heterozygosity (Ho) ranged from 0.293 in Djallonke sheep to 0.339 in Touabire sheep. The estimated Fis values were low, ranging from 0.019 for Ladoum to 0.034 for Peul-peul sheep. The estimated Fst values were low (0.003-0.044) among the trypanosusceptible breeds (Peul-peul, Touabire, and Ladoum) but high between the previous breeds and the trypanotolerant Djallonke breed (0.075-0.116), indicating better genetic conservation of the Djallonke sheep. A principal component analysis revealed clustering of the Senegalese sheep breeds according to their geographic distribution. However, owing to genetic improvement practices, the introgression of Touabire sheep blood seems to have reshaped the genetic landscape of the trypanosusceptible sheep breeds in Senegal. The Senegalese sheep breeds showed lower genetic diversity than their presumed ancestral sheep breeds of the Middle East. They also presented some relatedness with Caribbean sheep breeds, which reveals their contribution to the global genetic diversity and to the development of Caribbean sheep breeds.

Control of cystic echinococcosis in the Middle Atlas, Morocco: Field evaluation of the EG95 vaccine in sheep and cesticide treatment in dogs.

BACKGROUND: Cystic echinococcosis (CE) is an important cause of human morbidity and mortality worldwide, particularly in Morocco and other North African countries. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the potential of three strategies to reduce Echinococcus granulosus transmission: (1) 4-monthly treatment of dogs with praziquantel, (2) vaccination of sheep with the EG95 vaccine and (3) a combination of both measures. These measures were implemented during four consecutive years in different areas of the Middle Atlas Mountains in Morocco. The outcome of the interventions was assessed through hydatid cyst (viable and non-viable) counts in liver and lungs using necropsy or in vivo ultrasound examination of the liver. A total of 402 lambs were recruited for annual vaccination with the EG95 anti-E. granulosus vaccine and 395 similar lambs were selected as non-vaccinated controls. At approximately four years of age the relative risk (estimated as odds ratio) for vaccinated sheep to have viable hydatid cysts compared with non-vaccinated controls was 3% (9.37% of the vaccinated sheep were found infected while 72.82% of the controls were infected; p = 0.002). The number of viable cysts in vaccinated animals was reduced by approximately 97% (mean counts were 0.28 and 9.18 respectively; p<0.001). An average of 595 owned dogs received 4-monthly treatment during the 44 months trial, corresponding to 91% of the owned dog population. Approximately, 5% of them were examined for E. granulosus adult worms by arecoline purge or eggs in feces (confirmed by PCR). The proportion of infected dogs significantly decreased after treatment (12% versus 35%; p<0.001). Post-treatment incidence of re-infestation corresponded to a monthly risk of 4% (95% CI: 3-6%). Treatment of owned dogs on a 4-monthly basis did not reduce the level of transmission of E. granulosus to sheep, nor did it enhance the level of control generated by vaccination of sheep with EG95, possibly because of unowned dogs and wild canids were not treated. CONCLUSIONS/SIGNIFICANCE: These data suggest that vaccination of sheep with EG95 has the potential to reduce the level of CE in Morocco and in other parts of the world with similar transmission dynamics. Under the epidemiological circumstances existing in the trial area, 4-monthly treatment of owned dogs with praziquantel was insufficient to have a major impact of E. granulosus transmission to sheep.

Influence of Season and Liquid Storage at 16 °C on Beni Arouss Bucks' Semen Quality.

The study aimed at determining the effect of storage and season on fresh semen of Beni Arouss goats. Ejaculates were collected at monthly intervals from seven mature bucks and were extended at a final concentration of 800 × 106 spermatozoa. ml-1 and stored at 16 °C for 24 h. Semen motility, viability and normal morphology were assessed at 0, 4, 8 and 24 h after collection. Motility and normal morphology parameters were recorded using computer-assisted sperm analysis (CASA) and viability was analyzed using eosin-nigrosin staining. As expected, motility, viability and normal morphology parameters showed a significant reduction within 24 h of storage and during all seasons (p < 0.05). However, semen collected in summer maintained a better quality after 24 h of storage at 16 °C than semen collected during the other periods (p < 0.05). In conclusion, the storage ability of Beni Arouss bucks' semen stored at 16 °C was significantly higher during the summer.