Incorporation of fresh leaves of wormwood (Artemisia herba alba) and/or rosemary (Rosmarinus officinalis) in the diet of rams: Effect on testicular function, sexual behavior, and blood parameters.

This study aimed to investigate the effect of wormwood and rosemary supplementation on some reproductive traits of Barbarine rams. The experiment lasted 2 months. Twenty-four adult rams were divided into four groups (n = 6) balanced for the weight (53.3 ± 1.2 kg body weight [BW] ± SD). All rams received 1200 g of straw and 600 g of barley. Control rams (C) without aromatic medicinal plant (AMP), while experimental rams received 20 g of fresh rosemary leaves (R), 20 g of fresh wormwood leaves (A), and 10 g of fresh rosemary leaves plus 10 g of fresh wormwood leaves (RA). The results revealed that the live weight of all rams increased (p < .05) in the RA group compared to the C, A, and R groups. Scrotal circumference increased in the R rams when compared to the controls rams (p < .05). For sperm parameters we showed that the A rams had higher sperm concentrations (p < .05). But, the sperm volume decreased in the R rams (p > .05). However, when the rams received rosemary plus wormwood, their sperm volume increased (p > .05). The sperm mass motility was higher for the A, R and AR rams in comparison to the C rams (p = .05). On the other hand, biochemical analysis of the seminal fluid showed no effect of diets on calcium and total proteins concentration. But the measurement of glucose and seminal insulin showed a decrease (p < .05) in these two biochemical markers in group A rams and a decrease (p < .05) in insulin without modification of the glucose concentration in R rams. Blood glucose and insulin decreased in the animals on AMP diet compared to the other groups (p < .05) while aspartate aminotransferase (AST) increased (p < .05). Rosemary leaves (R and RA groups) increased (p < .05) plasma cortisol compared to the other groups. It can be concluded that the addition of Rosmarinus officinalis and/or Artemisia herba alba in ram diet can have a positive effect on the reproductive function by increasing the concentration and motility of sperm, plasma testosterone, and sexual behavior.

Substitution of soyabean meal by faba bean improves body weight, sperm production and testosterone concentration in mature Queue Fine de l'Ouest rams.

The current study aimed at investigating the effects of partial or total substitution of soybean meal with faba bean on reproductive parameters of Queue Fine de l'Ouest rams. Eighteen adult rams (49.8 ± 3.7 kg and 2.4 ± 1.5 years old) were divided into three homogenous groups. Rams received oat hay ad libitum and 3 types of concentrate (33 g/BW0.75) where soybean meal was the main protein source (SBM diet, n = 6), partially (50%, SBMFB diet, n = 6) or totally (100% FB diet, n = 6) substituted in nitrogen basis by local faba bean. The volume of ejaculate, sperm concentration and sperm mortality rate were measured weekly by collecting semen using an artificial vagina. Serial blood samples were collected 30 days and 120 days after the beginning of the experiment in order to assess plasma testosterone concentrations. The results showed that hay intake was affected (P < 0.05) by the nature of the nitrogen source incorporated (1032.3 ± 12.2; 1026.8 ± 56.6 and 972.8 ± 39.05 g DM/d for SBM, FB and SBMFB respectively. The average live weight of the rams increased from 49.8 ± 0.4 (week 1) to 57.3 ± 0.9 kg (week 17) without effect of the diet. A positive effect of faba bean incorporation in the concentrate was observed on ejaculate volume, concentration and spermatozoa production. All these parameters were significantly higher in SBMFB and FB groups than in SBM group (P < 0.05). The protein source did not affect the percentage of dead spermatozoa and the total abnormalities which were similar for the three diets (38.7; 35.8 and 38.1% for SBM; SBMFB and FB, respectively). The mean testosterone concentration was higher (P < 0.05) in rams fed with faba bean (1.7 ± 0.7 to 1.9 ± 0.7 ng/ml for the SBMFB and the FB groups respectively) than in rams fed SBM diet (1.06 ± 0.5 ng/ml). It was concluded that the substitution of soybean meal by faba bean improved the reproductive performance and did not affect sperm quality in Queue Fine de l'Ouest rams.

Detection of Toxoplasma gondii infection in semen of rams used for natural mating in commercial sheep farms in Tunisia.

Toxoplasmosis, a parasitic infection, disrupts sexual functions resulting in decline in reproductive and economic performance in sheep. Lacking discernible symptoms, toxoplasmosis is difficult to detect and diagnose in infected animals. Here, to estimate the prevalence of natural infection by Toxoplasma gondii, we used PCR to amplify and detect T. gondii DNA in semen from 92 rams of three breeds from four regions in Tunisia and seroprevalence was determined using a commercial ELISA kit. From the PCR amplifications, 51.09 ±â€¯10.21% of the rams were tested positive for T. gondii with an overall seroprevalence of 39.13 ±â€¯9.97%. Risk factors including ram location and number of accomplished mating seasons significantly (p < .05) affected the sero- and molecular prevalence of T. gondii in semen but, there was a fair concordance between sero- and molecular prevalence (Kappa = 0.33). Sequences of T. gondii from five positive samples were 100% identical (same haplotype). Comparison of these sequences with those archived at the GenBank showed a sequence similarity range between 95 and 100%. The haplotype defining the five Tunisian sequences was similar to the one observed in chicken, cats, European pole cat and humans from Brazil, St Kitts and Nevis, Great Britain and Tunisia, respectively. This indicates its wide geographic distribution and non-species specificity. Our findings suggest a high prevalence of toxoplasmosis in Tunisian matting rams; further studies concerning its venereal transmission capacity are needed prior to recommending a systematic screening of T. gondii DNA in rams' semen used for both natural breeding and artificial insemination.

Neospora caninum Natural Infection in Tunisian Rams: Serological Study and Molecular Identification of Infection in Semen.

PURPOSE: The aim of this study was to estimate the seroprevalence and molecular prevalence and perform a molecular identification of Neospora caninum in semen of Tunisian rams. METHODS: A total of 92 blood samples were collected from four farms located in four Tunisian governorates (Jendouba, Kairouan, Zaghouan and Ben Arous) and samples were screened with a commercial ELISA kit for N. caninum antibodies. For the same rams, semen samples were collected and tested for the presence of N. caninum ITS1 gene using PCR. Five amplicons were randomly selected for sequencing. A phylogenetic tree was constructed to compare the partial sequences of the ITS1 gene with sequences deposited in GenBank. RESULTS: The seroprevalence of N. caninum infection was 25% (23/92) and PCR revealed that the molecular infection prevalence in semen was 11.95% (11/92). Kappa test showed an average agreement between seroprevalence and parasite prevalence in semen (κ = 0.44). The highest molecular prevalence was for rams that accomplished more than two mating seasons (21.0 ± 12.1%) compared to those performed less than two mating seasons and yearling individuals (4.0 ± 5.5%) (P = 0.01). There were no differences in N. caninum molecular prevalence according to either breed or locality. Comparison of the partial sequences of the ITS1 gene revealed 99-100% similarity with those deposited in GenBank. CONCLUSION: To the best of our knowledge, this is the first detection and molecular identification of N. caninum in semen from rams in North Africa. Our findings indicate that N. caninum infection rate was high in rams.

The high prolificacy of D'man sheep is associated with the segregation of the FecLL mutation in the B4GALNT2 gene.

Mutations in the FecL locus are associated with large variation in ovulation rate and litter size in the French Lacaune sheep breed. It has been shown that the B4GALNT2 gene within the FecL locus is most likely responsible for the high fecundity in the French breed. In this study, we have highlighted the segregation of the FecLL mutation within the B4GALNT2 gene in North African sheep breeds and notably in the highly prolific D'man breed. Genotyping of a sample of 183 Tunisian D'man individuals revealed a high frequency (0.65) of the prolific allele FecLL which was attributed to the adoption of a decades-old breeding strategy based on the selection of ewe lambs born from large litter size. Homozygous LL ewes showed a significantly increased litter size compared to heterozygous and non-carrier ewes (FecLL /FecLL  = 2.47 ± 0.09 vs. FecLL /FecL+  = 2.23 ± 0.09, p < 0.05 and FecL+ /FecL+  = 1.93 ± 0.18, p < 0.01). The presence of the FecLL polymorphism in both D'man and Lacaune breeds argues for an ancestral origin of this mutation and brings an answer to the old question of the genetic determinism of the extreme prolificacy of the D'man ewes. The results of this study can help to establish planned genotype-based mating allowing both higher profit for the breeders and an optimal management of the FecLL mutation in D'man sheep populations.

FecX Bar a Novel BMP15 mutation responsible for prolificacy and female sterility in Tunisian Barbarine Sheep.

BACKGROUND: Naturally occurring mutations in growth and differentiation factor 9 (GDF9) or bone morphogenetic protein 15 (BMP15) genes are associated with increased ovulation rate (OR) and litter size (LS) but also sterility. Observing the Tunisian Barbarine ewes of the "W" flock selected for improved prolificacy, we found prolific and infertile ewes with streaky ovaries. Blood genomic DNA was extracted from a subset of low-ovulating, prolific and infertile ewes of the "W" flock, and the entire coding sequences of GDF9 and BMP15 were sequenced. RESULTS: We evidenced a novel polymorphism in the exon 1 of the BMP15 gene associated with increased prolificacy and sterility. This novel mutation called FecX Bar is a composite polymorphism associating a single nucleotide substitution (c.301G > T), a 3 bp deletion (c.302_304delCTA) and a C insertion (c.310insC) in the ovine BMP15 cDNA leading to a frame shift at protein position 101. Calculated in the "W" flock, the FecX Bar allele increased OR by 0.7 ova and LS by 0.3 lambs (p = 0.08). As for already identified mutations, homozygous females carrying FecX Bar exhibited streaky ovaries with a blockade at the primary stage of folliculogenesis as shown by histochemistry. CONCLUSIONS: Our investigation demonstrates a new mutation in the BMP15 gene providing a valuable genetic tool to control fecundity in Tunisian Barbarine, usable for diffusion program into conventional flocks looking for prolificacy improvement.

Effect of the infection with the nematode Haemonchus contortus (Strongylida: Trichostrongylidae) on the haematological, biochemical, clinical and reproductive traits in rams.

This study aimed to investigate the effect of Haemonchus contortus infection on rams' haematological, biochemical and clinical parameters and reproductive performances. A total number of 12 Barbarine rams (control and infected) were included in the experiment. The infected group received 30 000 H. contortus third-stage larvae orally. Each ram's ejaculate was immediately evaluated for volume, sperm cell concentration and mortality rate. At the end of the experiment (day 82 post-infection), which lasted 89 days, serial blood samples were collected in order to assess plasma testosterone and luteinising hormone (LH) concentrations. There was an effect of time, infection and their interaction on haematological parameters (p < 0.001). In infected rams, haematocrit, red blood cell count and haemoglobin started to decrease from 21 days post-infection. There was an effect of time and infection for albumin. For total protein, only infection had a statistically significant effect. For glucose, only time had a statistically significant effect. Concentrations were significantly lower in infected rams compared to control animals. A significant effect of infection and time on sperm concentrations and sperm mortality was observed. The effect of infection appears in time for sperm concentrations at days 69 and 76 post-infection. Sperm mortality rate was significantly higher in infected animals at day 46 post-infection when compared to control group (p < 0.05). Finally, plasma testosterone traits (average concentration, cumulated levels during the sampling period and pulse frequency) were depressed in infected rams when compared to control counterparts; none of these endocrine traits were affected for plasma LH.

Melatonin administration enhances the reproductive capacity of young rams under a southern Mediterranean environment.

This study tested the effect of melatonin treatment, initiated in late February on reproductive traits of young rams. A total of 14 young Barbarine rams were used. Seven animals were treated with three melatonin subcutaneous implants (Melatonin) on 28 February while the remaining rams remained untreated (Control). After 60 days of melatonin administration, scrotal circumference reached average values of 32.1 ± 1.54 and 29.5 ± 1.0 cm for Melatonin and Control animals, respectively (P < 0.05). Semen characteristics did not differ between groups; melatonin treatment tended (P = 0.091) to increase sperm concentration 60 days after implantation when means reached 5.87 ± 0.703 and 4.61 ± 0.654 × 10(9) spermatozoa/mL for Melatonin and Control rams, respectively. Melatonin treatment significantly affected total activity time, number of lateral approaches and mount attempts in comparison to controls. During a 6-h sampling period, mean plasma testosterone concentrations increased as a result of melatonin treatment (P < 0.001) and testosterone pulse frequency averaged 3.45 ± 2.24 and 1.25 ± 1.0 (P = 0.086) for Melatonin and Control rams. Data clearly suggest that abrupt treatment of young rams with melatonin implants in winter is sufficient to improve reproductive traits.

Merits of the fat-tailed Barbarine sheep raised in different production systems in Tunisia: digestive, productive and reproductive characteristics.

Barbarine sheep is the dominant breed in Tunisia. This fat-tailed breed present in all production systems is characterised by metabolic and digestive adaptation to the contrasting environment conditions prevailing in the country (heat stress, water deprivation, salinity etc.). The fat tail (1.5 to 7 kg) is an energy reservoir that is used in periods of feed scarcity. Moreover, the rumen of this breed harbours a microflora enabling it to valorize low-quality roughages and native range vegetation. Barbarine sheep could make benefit from a wide range of local feed resources like fodder shrubs (e.g. cactus and oldman saltbushes), tannin-containing shrubs (e.g. Acacia cyanophylla) and agro-industrial by-products (e.g. olive cake, bran etc. conserved in the form of feed blocks or pellets). It has a very shallow anoestrous, the intensity of which is dependant upon nutrition conditions rather than day length as temperate breeds. Productive and reproductive performances of late pregnant-early suckling, pre-mating ewes and also rams of this breed are not affected when some alternative feed resources replace common feedstuffs which are expensive and cannot afford regularly to smallholders. In brief, the merits of the Barbarine sheep in the production systems and other main adaptive mechanisms of this breed are discussed in this paper. The prospective of Barbarine sheep husbandry in the system dynamics, climate change and the evolution of the socioeconomic context are also envisaged.

Male-induced short oestrous and ovarian cycles in sheep and goats: a working hypothesis.

The existence of short ovulatory cycles (5-day duration) after the first male-induced ovulations in anovulatory ewes and goats, associated or not with the appearance of oestrous behaviour, is the origin of the two-peak abnormal distribution of parturitions after the "male effect". We propose here a working hypothesis to explain the presence of these short cycles. The male-effect is efficient during anoestrus, when follicles contain granulosa cells of lower quality than during the breeding season. They generate corpora lutea (CL) with a lower proportion of large luteal cells compared to small cells, which secrete less progesterone, compared to what is observed in the breeding season cycle. This is probably not sufficient to block prostaglandin synthesis in the endometrial cells of the uterus at the time when the responsiveness to prostaglandins of the new-formed CL is initiated and, in parallel, to centrally reduce LH pulsatility. This LH pulsatility stimulates a new wave of follicles secreting oestradiol which, in turn, stimulates prostaglandin synthesis and provokes luteolysis and new ovulation(s). The occurrence of a new follicular wave on days 3-4 of the first male-induced cycle and the initiation of the responsiveness to prostaglandins of the CL from day 3 of the oestrous cycle are probably the key elements which ensure such regularity in the duration of the short cycles. Exogenous progesterone injection suppresses short cycles, probably not by delaying ovulation time, but rather by blocking prostaglandin synthesis, thus impairing luteolysis. The existence, or not, of oestrous behaviour associated to these ovulatory events mainly varies with species: ewes, compared to does, require a more intense endogenous progesterone priming; only ovulations preceded by normal cycles are associated with oestrous behaviour. Thus, the precise and delicate mechanism underlying the existence of short ovulatory and oestrous cycles induced by the male effect appears to be dependent on the various levels of the hypothalamo-pituitary-ovario-uterine axis.