Description of Ewiss cheese, a new ewe's milk cheese processed by Swiss cheese manufacturing techniques: microbiological, physicochemical and sensory aspects.

Typically, Swiss-type cheese is made from cow's milk. However, in the present work an attempt to expand the sheep supply chain and product offering in this field was made by developing a new type of cheese using Swiss-type cheese technology. The cheese was manufactured under industrial conditions, and fermentations were carried out using freeze-dried commercial starters that are traditionally used in the production of Swiss cheese. Two experimental "Ewiss cheese" (EC) products were produced using raw milk (RM-EC) and pasteurized milk (PM-EC), respectively. Fourteen microbial groups were investigated by plate counts from curd until ripened cheeses. According to microbiological analyses, no statistically significant differences were found between the 2 productions with respect to the group of lactic acid bacteria (LAB). The curds were mainly characterized by mesophilic LAB cocci (7.45 log10 cfu/g in RM-EC and 7.33 log10 cfu /g in PM-EC). However, at the end of the ripening period (9 mo), the cheeses exhibited a higher presence of mesophilic LAB rods. Undesired microbiological groups were found only in the curd of raw milk cheese in the range of 104-105 cfu/g, but reaching undetectable levels by plate count in the cheese at the end of ripening. RM-EC and PM-EC were characterized by 76% and 68% of dry matter, respectively. These cheeses contained 29.30% and 34.36% of protein, and 51.31% and 50.38% of fat, respectively. Textural analysis showed differences in terms of hardness, chewiness, and gumminess between the experimental cheeses and Swiss cheese sold on the market. These differences could be attributed to the higher protein content of ewe's milk. The main fatty acids in the cheeses were palmitic acid, myristic acid, oleic acid, and capric acid. Among the organic acids, RM-EC had higher concentrations of lactic acid, while PM-EC was higher in propionic acid. The ewe's cheeses emitted 46 volatile compounds, including acids, aldehydes, ketones, esters, alcohols, and other compounds. PM-EC was characterized by the main compounds of Swiss-type cheese: acetic acid, butyric acid, ethyl butyrate, ethyl caproate, propanoic acid, and tetramethylpyrazine. Sensory evaluation showed that the new dairy products were generally appreciated, and PM-EC was the most preferred by the judges. This research has enabled the development of new ewe's milk products, which could stimulate the valorization of a sector that has been long neglected and still has a large margin of improvement.

The effect of human menopausal gonadotropin and equine chorionic gonadotropin on the reproductive performance of treated ewes with short-term progesterone injections and sponge during the non-breeding season.

This study investigated the effect of human menopausal gonadotropin (hMG) on reproductive efficiency of synchronized ewes with the sponge and progesterone (P4) injection-based protocols. In study 1, anoestrous ewes (n = 120) were used. Sixty ewes were treated with sponge (S) for 12 days. The injection of eCG (SeCG group, n = 30) or hMG (ShMG, n = 30) was given at the time of sponge removal. Thirty ewes received IM injection of P4, three times every 48 h and the injection of hMG was given 24 h after the third P4 injection (3PhMG group, n = 30), and 30 ewes were used as control group. Pregnancy was diagnosed on day 50 after the release of ram. In study 2, 60 ewes were randomly divided into two equal groups. In the treated group with antibiotics (n = 30), before inserting, the sponges were impregnated with the antibiotic penicillin G sodium (5,000,000 IU) and in the control group (n = 30), there was no added antibiotics. Before inserting and after removing sponges, a vaginal cytology sample was taken with a sterile cotton swab. The number of neutrophils in each sample was counted and analysed. The rate of oestrus and total pregnancy was greater in SeCG (96.7, 93.3%), ShMG (82.8, 93.1%) and 3PhMG (67.9, 89.3%) groups compared with the control group (13.8, 41.4%) (p < .05). No significant difference was found in single, twin and total lambing and pregnancy rates after injection of eCG and hMG during the non-breeding season (p > .05). A higher percentage of control ewes had the vaginal smear with neutrophils more than 50% (96.7% vs. 76.7%; p < .05). In conclusion, a single dose of hMG can induce fertile oestrus in synchronized ewes with P4 administered by either injection or intravaginally. Purulent discharge and percentage of neutrophils were significantly reduced in the synchronized ewes by the impregnated sponges with the antibiotic penicillin.

Tracing of persistent Listeria monocytogenes contamination in ewe's milk farm.

Ewe's milk farm production is permanently associated with the risk of contamination by pathogenic bacteria, including Listeria monocytogenes. In the present study, the prevalence and diversity of L. monocytogenes strains repeatedly isolated from tank ewe's milk and the milking environment on a farm in Slovakia during a prolonged period were investigated to identify the source of potentially persistent contamination. A total of 140 samples along the milk production chain were collected during an 18-month period. From all these samples, 45 samples were found L. monocytogenes positive with 90.3% positivity of tank milk samples (28 positive samples from 31 analysed). Pulsed-field gel electrophoresis profiling resulted in strain discrimination into six profiles with one pulsotype (NS1) corresponding to MLST-ST14 being predominant. A total of 17 proportionally selected L. monocytogenes isolates, including 11 NS1/ST14 isolates, were subjected to whole genome sequencing. Resulted data were used to compare the genomes diversity and to confirm the persistent contamination when <10 allelic differences threshold in cgMLST analysis was applied. The source of persistent contamination was localized inside the milking apparatus, probably in shelters that were very difficult to clean. Despite great efforts, the ewe's milk contamination could not be eliminated during the reporting period.

Effect of dietary protein on growth performance, and serum biochemical index in late pregnant Hu ewes and their offspring.

The purpose of this study was to investigate the effects of different protein levels in late pregnancy on ewe and lamb growth performance, serum biochemical indexes. Thirty-three ewes (46.4 ± 1.38 kg initial weight) were randomly divided into 3 groups, with 11 ewes in each group. The protein levels of three diets formulated to provide components to meet 10.00 MJ/kg ME requirements diets were: 10.12%, 11.26%, 12.4%. Ewes were raised from the 90th day of pregnancy to the end of delivery, and the lambs were weaned at 60 days. Dietary protein levels had significant effects on blood urea nitrogen, glucose, ammonia nitrogen and triglyceride of ewes (p < 0.05). The height, chest depth, chest circumference, straight crown hip length and curved crown hip length of lambs decreased at first and then increased with the increase of protein. The body length, chest circumference, head width and head length of weaned lambs decreased at first and then increased with the increase of protein. The results showed that when the dietary protein level was increased to 12.4%, the amino acid, glucose and fat metabolism of ewes were affected. The body size development of lambs was better than 10.12% and 11.26% proteins.

Feed restriction around parturition does not affect colostrum immunoglobulin G concentration in dairy fat-tailed sheep but does affect performance and blood metabolites in newborn lambs.

This study aimed to investigate the effect of prepartum and postpartum feed restriction of fat-tailed dairy sheep on colostrum IgG concentration, and performance and blood metabolites of newborn fat-tailed lambs. Twenty fat-tailed dairy sheep were randomly allocated into control (Ctrl; n = 10) and feed restriction (FR; n = 10) groups. The Ctrl group received a diet that met 100% of energy requirements, both prepartum (from wk -5 to parturition) and postpartum (from parturition to wk 5). The FR group received a diet equivalent to 100, 50, 65, 80, and 100% of the energy requirements in wk -5, -4, -3, -2, and -1 relative to parturition, respectively. After parturition, the FR group received a diet equivalent to the 100, 50, 65, 80, and 100% of the energy requirements in wk 1, 2, 3, 4, and 5, respectively. At birth, lambs were assigned to their dam's experimental group. Both the Ctrl lambs (n = 10) and the FR lambs (n = 10) were allowed to suck colostrum and milk from the dams. Colostrum samples (50 mL) were collected at parturition (0 h) and then at 1, 12, 24, 36, 48, and 72 h postpartum. Blood samples were collected from all lambs before suckling colostrum (0 h) and then at 1, 12, 24, 36, 48, and 72 h after birth and weekly until the end of the experimental period (i.e., wk 5 relative to birth). The data were evaluated using the MIXED procedure of SAS (SAS Institute Inc.). The model included feed restriction, time, and the interaction feed restriction × time as fixed effects. The individual lamb was set as a repeated subject. Variables measured in colostrum and plasma were considered dependent variables, and significance was set at P < 0.05. Prepartum and postpartum feed restriction in fat-tailed dairy sheep did not affect colostrum IgG concentration. Consequently, no differences in blood IgG concentrations were observed in the lambs. In addition, the prepartum and postpartum feed restriction experienced by fat-tailed dairy sheep caused decreased body weight and milk intake in lambs from the FR group compared with the Ctrl group. Feed restriction also promoted increased concentration of blood metabolites such as triglycerides and urea in FR lambs compared with control lambs. In conclusion, prepartum and postpartum feed restriction in fat-tailed dairy sheep did not affect either colostrum IgG concentration or blood IgG concentration of the lambs. However, prepartum and postpartum feed restriction decreased lamb milk intake and, therefore, lamb body weight gain during the first 5 wk after birth.

Plasma and milk metabolomics in lactating sheep divergent for feed efficiency.

Enhancing the ability of animals to convert feed into meat or milk by optimizing feed efficiency (FE) has become a priority in livestock research. Although untargeted metabolomics is increasingly used in this field and may improve our understanding of FE, no information in this regard is available in dairy ewes. This study was conducted to (1) discriminate sheep divergent for FE and (2) provide insights into the physiological mechanisms contributing to FE through high-throughput metabolomics. The ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) technique was applied to easily accessible animal fluids (plasma and milk) to assess whether their metabolome differs between high- and low-feed efficient lactating ewes (H-FE and L-FE groups, respectively; 8 animals/group). Blood and milk samples were collected on the last day of the 3-wk period used for FE estimation. A total of 793 features were detected in plasma and 334 in milk, with 100 and 38 of them, respectively, showing differences between H-FE and L-FE. The partial least-squares discriminant analysis separated both groups of animals regardless of the type of sample. Plasma allowed the detection of a greater number of differential features; however, results also supported the usefulness of milk, more easily accessible, to discriminate dairy sheep divergent for FE. Regarding pathway analysis, nitrogen metabolism (either anabolism or catabolism) seemed to play a central role in FE, with plasma and milk consistently indicating a great impact of AA metabolism. A potential influence of pathways related to energy/lipid metabolism on FE was also observed. The variable importance in the projection plot revealed 15 differential features in each matrix that contributed the most for the separation in H-FE and L-FE, such as l-proline and phosphatidylcholine 20:4e in plasma or l-pipecolic acid and phosphatidylethanolamine (18:2) in milk. Overall, untargeted metabolomics provided valuable information into metabolic pathways that may underlie FE in dairy ewes, with a special relevance of AA metabolism in determining this complex phenotype in the ovine. Further research is warranted to validate these findings.

Effect of glucose infusion on glucose and insulin metabolism in early- and mid-lactation ewes and goats fed diets differing in starch and highly digestible fiber concentration.

This study aimed to test possible metabolic differences between ewes and goats in response to an intravenous glucose infusion. Thirty-six animals, 18 mature Sarda dairy ewes and 18 mature Saanen goats (from 15 to 150 ± 11 d in milk [DIM], mean ± SD; body weight: 49.8 ± 6.8 kg for ewes, 60.6 ± 7.3 kg for goats) were compared simultaneously. In early lactation, both species received the same high-starch diet (HS: 20.4% starch, 35.4% neutral detergent fiber [NDF], on dry matter [DM] basis), whereas from 92 ± 11 DIM both species were randomly allocated to 2 dietary treatments: HS (20.0% starch, 36.7% NDF, on DM basis) and low-starch (LS: 7.8% starch, 48.8% NDF, on DM basis) diets. At 50 and 150 ± 11 DIM, ewes and goats were challenged with an intravenous glucose tolerance test and peripheral concentrations of glucose and insulin were determined 15 min before and 5, 10, 15, 30, 45, 90, and 180 min after glucose infusion. In early lactation, baseline plasma glucose and insulin concentrations tended to be higher in ewes than in goats (glucose: 55.8 vs. 42.9 ± 7.3 mg/dL; insulin: 0.13 vs. 0.05 ± 0.04 µg/L). After glucose infusion, glucose and insulin concentrations were higher in ewes than in goats (278.6 vs. 247.6 ± 13.1 mg/dL; 0.82 vs. 0.46 ± 0.12 µg/L). In mid-lactation, the dietary treatment (HS vs. LS) did not affect glucose and insulin metabolism. Baseline plasma glucose was numerically highest in ewes, while baseline insulin was higher in ewes than in goats (0.39 vs. 0.12 ± 0.099 µg/L). After glucose infusion, glucose concentration did not differ between ewes and goats, while insulin concentration was highest in ewes. Compared with goats, ewes showed in both periods a higher peak insulin, insulin increment, linear insulin area under the curve, insulin resistance index, and lower insulin sensitivity indices. In conclusion, despite the limitations associated with the use of intravenous glucose tolerance test to assess glucose regulation mechanisms, this study indicated large species differences in both early and mid-lactation and a more evident anabolic status in the ewes compared with the goats.

Cheese yield and nutrients recovery in the curd predicted by Fourier-transform spectra from individual sheep milk samples.

The objectives of this study were to explore the use of Fourier-transform infrared (FTIR) spectroscopy on individual sheep milk samples for predicting cheese-making traits, and to test the effect of the farm variability on their prediction accuracy. For each of 121 ewes from 4 farms, a laboratory model cheese was produced, and 3 actual cheese yield traits (fresh cheese, cheese solids, and cheese water) and 4 milk nutrient recovery traits (fat, protein, total solids, and energy) in the curd were measured. Calibration equations were developed using a Bayesian approach with 2 different scenarios: (1) a random cross-validation (80% calibration; 20% validation set), and (2) a leave-one-out validation (3 farms used as calibration, and the remaining one as validation set) to assess the accuracy of prediction of samples from external farms, not included in calibration set. The best performance was obtained for predicting the yield and recovery of total solids, justifying for the practical application of the method at sheep population and dairy industry levels. Performances for the remaining traits were lower, but still useful for the monitoring of the milk processing in the case of fresh curd and recovery of energy. Insufficient accuracies were found for the recovery of protein and fat, highlighting the complex nature of the relationships among the milk nutrients and their recovery in the curd. The leave-one-out validation procedure, as expected, showed lower prediction accuracies, as a result of the characteristics of the farming systems, which were different between calibration and validation sets. In this regard, the inclusion of information related to the farm could help to improve the prediction accuracy of these traits. Overall, a large contribution to the prediction of the cheese-making traits came from the areas known as "water" and "fingerprint" regions. These findings suggest that, according to the traits studied, the inclusion of water regions for the development of the prediction equation models is fundamental to maintain a high prediction accuracy. However, further studies are necessary to better understand the role of specific absorbance peaks and their contribution to the prediction of cheese-making traits, to offer reliable tools applicable along the dairy ovine chain.

Oxidative response of sheep to transcervical applications.

This study aimed to investigate the stress-related changes that may occur in sheep following non-surgical cervical line passage. A total of 20 sheep underwent transcervical transition, and their heart rate, respiratory rate, body temperature, and blood samples were collected at specific time points: 1 h prior to the procedure, immediately after the procedure, and at 1, 2, 3, 4 and 5 h post-procedure. The study found that there was a statistically significant decrease in body temperature immediately after the procedure (p < .05); however, there were no significant differences in body temperature at other time points (p > .05). Respiratory rate remained similar throughout the study, and no significant differences were observed (p > .05). On the other hand, there was a significant increase in heart rate at the 1-h mark after the procedure (p < .05), which subsequently decreased to a similar level as the control group at the 4-h mark after the procedure (p > .05). The cortisol levels increased significantly after the application (p < .05) but decreased to a level similar to the control after 1 h (p > .05). The level of nitric oxide increased significantly in the first hour (p < .05) and decreased by the 5th hour (p < .05). There was a significant increase in malondialdehyde (MDA) and total oxidant capacity (TOC) levels 2 h after the application (p < .05), but this increase ceased at the 4th and 5th hours for MDA (p > .05) and at the 3rd, 4th and 5th hours for TOC (p > .05). The total antioxidant capacity decreased after the 2nd hour of application (p < .05) but remained similar at the fourth and 5th hours (p > .05). In conclusion, based on the cortisol data, transcervical manipulations create short-term stress. Therefore, it may be necessary to use various antioxidants before such procedures to reduce the effects of oxidative stress.

Induction of ovulation using repeated doses of sulpiride, a dopamine antagonist, in ewe lambs.

This study aimed to test the hypothesis that sulpiride can increase the concentration of circulating gonadotropin that can promote puberty in pre-pubertal ewe lambs. Here, 12 1-3-year-old Merino rams and 60 7-9-month-old Merino sheep were included in the study. The sheep were randomly divided into sulpiride (n = 30) and control (n = 30) groups. The sulpiride group was subcutaneously injected with 0.6 mg/kg sulpiride twice daily (morning and evening) for 9 days. During these 9 days, blood samples were taken from the sheep before drug administration and at 4 h after every drug administration. The number of ovulating animals in the sulpiride group was significantly higher than that in the control group (90% vs. 32%). No oestrous signs were observed in either group during ram release. Further, there were no differences in the levels of mean follicle-stimulating hormone in the two groups based on treatment (p = .2), time (p = .3) or treatment-by-time interaction (p = .3). After sulpiride administration, the luteinizing hormone (LH) levels of the sulpiride group rapidly increased and remained stable for a long time, whereas physiological LH fluctuations in the control group remained unchanged. Within-group changes in terms of LH concentrations were significant for both groups (p < .001), whereas LH pulse frequency was significantly different between the sulpiride group (p = .03). Therefore, it is concluded that sulpiride can be used as a non-steroidal alternative to stimulate pre-pubertal ewe lambs and sheep during anoestrus.

The effect of progesterone injection combined with prostaglandineF2α, GnRH and hCG administrations on the pregnancy and lambing rate of lactating and non-lactating fat-tailed ewes during the breeding season.

The objective was to compare the effect of human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) on the pregnancy and lambing rate of treated ewes with a short-term progesterone (P4) injection during the breeding season. In Exp 1, non-lactating ewes (n = 158) were used and received P4 , three times every 48 h and received PGF2α along with the last dose of P4. Ewe received hCG (n = 79, 3PHCG, IM) or GnRH (n = 79, 3PGnRH, IM) 24 h after the last dose of P4 treatment. In the Exp 2, lactating ewes (n = 62) received P4 and hCG (n = 24, 3PHCG) or GnRH (n = 24, 3PGnRH) similar to Exp 1, or considered as control (n = 14) and received PGF2α 48 h before ram release. Rams were released into the studied ewe's flock 24 h after the hCG or GnRH administration, and pregnancy diagnosis was performed on day 50 after ram release. In Exp 1, all reproductive indices were not significant between groups except twin lambing rate, that was higher in the 3PHCG (11.3%) compared with the 3PGnRH (1.9%) group (p = .05). There were no significant differences in overall pregnancy and lambing rates between 3PHCG (70.9% and 67.1%) and 3PGnRH (69.6% and 68.4%) groups (p > .05). In Exp 2, all reproductive parameters were not significant between 3PHCG and 3PGnRH groups (p > .05). There was significant difference in overall oestrous rate between control (35.7%) and treatment (3PHCG, 70.9% and 3PGnRH, 79.2%) groups. To conclude, administration of GnRH can be a good alternative to hCG injection under field conditions.

Development and evaluation of an injectable slow-release progesterone formulation for estrus synchronization in ewes out of the breeding season.

This study was aimed at developing a type of slow-release progesterone micro-particles useable in a single intramuscular injection for estrus synchronization in non-breeding season ewes. A total of 66 ewes were randomly assigned into four groups: CIDR (n = 16): exposed to intravaginal CIDR for 12 days, and three experimental groups, i.e., T100 (n = 16), T150 (n = 17) and T200 (n = 17), receiving a single intramuscular injection of 100, 150 and 200 mg slow-release progesterone, respectively. Blood sampling was performed on all ewes at five different times, and the ELISA method measured progesterone levels. No significant differences were observed in progesterone levels among the groups in each sampling time. More than 90% of ewes in the CIDR, T100 and T150 groups and all those in T200 showed estrus behaviour, and the rate was not significantly different between groups. The difference in the mean interval from progesterone treatment to estrus was also insignificant. The parturition rate declined by increasing the dose of injected progesterone; although it was similar in CIDR and T100 groups, it decreased significantly in T150 and T200 . Since our injectable progesterone formulation was successful in the induction and synchronization of estrus in ewes out of the breeding season, it can be applied as an alternative to the conventional progesterone containing intravaginal devices.

Pharmacokinetics of letrozole and effects of its increasing doses on gonadotropins in ewes during the breeding season.

Letrozole is a non-steroidal, third-generation aromatase inhibitor used in humans. Although letrozole is not approved for use in animals, it is used off-label in cases of synchronization and infertility. The aim of this study was to determine the pharmacokinetics of letrozole after a single intravenous administration at three different doses in ewes during the breeding season and its effect on gonadotropins (luteinizing hormone (LH) and follicle-stimulating hormone (FSH)) at the beginning of proestrus. The study was carried out on 24 healthy Merino ewes. Ewes were randomly divided into four groups (n = 6) as control, 0.5, 1, and 2 mg/kg. Plasma concentrations of letrozole were measured using HPLC-UV and were analyzed by non-compartmental analysis. LH and FSH concentrations were measured with a commercial ELISA kit. The terminal elimination half-life (t1/2ʎz ) was significantly prolonged from 11.82 to 18.44 h in parallel with the dose increase. The dose-normalized area under the concentration-time curve (AUC) increased, and total body clearance (ClT ) decreased at the 1 and 2 mg/kg doses (0.05 L/h/kg) compared with the 0.5 mg/kg dose (0.08 L/h/kg). There were no differences in the volume of distribution at steady-state and initial (C0.083h ) plasma concentration values between dose groups. The decreased ClT , prolonged t1/2ʎz, and increased AUC at increasing doses showed the nonlinear kinetic behavior of letrozole. Letrozole significantly reduced LH concentration without affecting FSH concentration at all doses. As a result, letrozole has the potential to be used in synchronization methods and manipulation of the follicular waves due to its effect on LH secretion.

Reduction of Beta Cyclodextrin by Curd Washing in Low-Cholesterol Manchego Cheese.

Beta-cyclodextrin (ß-CD) is a cyclic oligosaccharide consisting of seven glucose units. ß-CD is increasingly used in food research to reduce cholesterol due to its affinity for non-polar molecules such as cholesterol and as a natural additive. The purpose of this study was to evaluate the effect of curd washing in ewe's milk cheese on the reduction in cholesterol by ß-CD from pasteurized ewe's milk Manchego cheese and the characteristics of its main components: milk, lipids, and flavor. An approximately 98.45% cholesterol reduction was observed in washed experimental cheeses that were treated by using ß-CD. The remaining residual ß-CD from the effect of curd washing was 0.15% in mature cheese, of the initial 1% ß-CD treatment of the milk. The chemical properties (fat, moisture, and protein) did not change as a result of the curd washing with or without ß-CD. The curd washing with or without ß-CD on the levels of the various lipid fraction (fatty acids, triglycerides, and phospholipids) were comparable in treated and untreated cheeses. The effects of curd washing and the ß-CD treatment did not significantly affect flavor components or short chain free fatty acids. The ß-CD molecules were edible and nontoxic; as a result, they could be used safely in cholesterol removal processing in cheese manufacturing, improving the reduction in residual ß-CD by curd washing by 85%. Therefore, the present study suggests that curd washing combined with ß-CD is an effective process for cholesterol removal in Manchego cheese, preserving its desirable properties.

Use of probiotic yeasts with biocontrol activity for fermentation of ewe's milk.

BACKGROUND: There are sufficient scienctific studies that support the benefit that fermented dairy products produce in those who consume them. Traditionally, cow's milk has been the most commonly used milk but there is a growing interest in the development of new dairy products, substituting cow's milk with milk from other sources, as well as in the use of microorganisms in fermentation to replace artificial preservatives or treatments that may affect the chemical and organoleptic characteristics of the product. For these reasons, the aim of the present work was to understand the behavior of five potential probiotic yeasts during the fermentation of ewe's milk and to consider their potential use as biocontrol agents. RESULTS: Saccharomyces cerevisiae 3 and Hanseniaspora osmophila 1056 provided the most promising kinetic parameters in the different salt, temperature and pH conditions tested in their technological characterization. The profiles of organic acids and volatile compounds after the fermentation period was noteworthy for contributing to the final aroma of the dairy product. Sensory analysis revealed the sour taste of all samples, and S. cerevisiae 3, Lachancea thermotolerans 1039, and H. osmophila 1056 stood out for an accentuated cheese flavor. In addition, all strains showed biocontrol activity; they reduced the mycelium of the mycotoxigenic molds. CONCLUSION: Saccharomyces cerevisiae 3 and H. osmophila 1056 could be inoculated along with bacterial starters to provide a functional fermented beverage with improved flavor. These strains also have an added value as they act as biocontrol agents. © 2022 Society of Chemical Industry.

The investigation of the effects of vitamin A, vitamin E, and ß-carotene plus vitamin E on some fertility parameters in ewes.

This study was aimed at investigating the effects of vitamin A (VITA), vitamin E (VITE), and combined ß-carotene plus vitamin E (ßCAR+VITE) injections on some fertility parameters in ewes. Estrus synchronization was performed by treating the ewes with intravaginal FGA sponges impregnated with 30 mg of fluorogestone acetate. On the days of the insertion and withdrawal of the intravaginal sponges, groups VITA, VITE, and ßCAR+VITE were administered with 500 000 IU of vitamin A, 50 mg of vitamin E, and a combination of ß-carotene plus vitamin E, respectively. The ewes in the control group (C) were maintained for control purposes. Statistically significant differences were determined between groups VITA and ßCAR+VITE, groups VITE and ßCAR+VITE, and groups C and ßCAR+VITE, as well as groups VITE and C, groups VITA and C for the multiple birth rates. While significant differences were determined between groups VITA and C, groups VITE and C, and groups ßCAR+VITE and C for the lambing rates, it was ascertained that the ratio of newborn lambs to delivered ewes (litter size) significantly differed between groups VITA and ßCAR+VITE, groups VITA and C, groups VITE and ßCAR+VITE, groups VITE and C, and groups ßCAR+VITE and C. The highest MDA level and lowest GSH level were determined on day 20 after mating in the control group. In conclusion, it is suggested that both multiple birth rates and litter size can be increased by the combined administration of ß-carotene and vitamin E.

Biochemical and molecular characterization of sialylated cervical mucins in sheep†.

Sialic acid occupies terminal positions on O-glycans of cervical mucins, where they contribute to the increased viscosity of mucin thereby regulating sperm transport. This study characterized the sialylated cervical mucins from follicular phase mucus of six European ewe breeds with known differences in pregnancy rates following cervical artificial insemination (AI) using frozen-thawed semen at both synchronized and natural estrus cycles. These were Suffolk (low fertility) and Belclare (medium fertility) in Ireland, Ile de France and Romanov (both with medium fertility) in France, and Norwegian White Sheep (NWS) and Fur (both with high fertility) in Norway. Expression of mucin and sialic acid related genes was quantified using RNA-sequencing in cervical tissue from Suffolk, Belclare, Fur, and NWS only. Cervical tissue was also assessed for the percentage of cervical epithelial populated by mucin secreting goblet cells in the same four ewe breeds. Biochemical analysis showed that there was an effect of ewe breed on sialic acid species, which was represented by Suffolk having higher levels of Neu5,9Ac2 compared with NWS (P < 0.05). Suffolk ewes had a lower percentage of goblet cells than Fur and NWS (P < 0.05). Gene expression analysis identified higher expression of MUC5AC, MUC5B, ST6GAL1, and ST6GAL2 and lower expression of ST3GAL3, ST3GAL4, and SIGLEC10 in Suffolk compared with high fertility ewe breeds (P < 0.05). Our results indicate that specific alterations in sialylated mucin composition may be related to impaired cervical sperm transport.

Effect of PGF2α and GnRH administration on reproductive performance in Ghezel ewes.

The main aim of the current study was to evaluate the effect of GnRH administration on day five after mating as well as PGF2α injection at the time of CIDR removal on the reproductive performance of Ghezel ewes. Estrus synchronization was performed using an intravaginal application of CIDR for 14 days and injection of 500 IU of PMSG at the time of CIDR removal. A total of 114 healthy fat-tailed ewes were randomly allotted into three groups as follow: control group (n = 35), did not receive any additional treatment; PG group (n = 44), each ewe received a dose of PGF2α at the time of CIDR removal; and PG+GnRH group (n = 35), the ewes received a dose of PGF2α at the time of CIDR removal and a single dose of GnRH, five days post-mating (post-conceptional day (PCD)- 5). Body condition score (BCS) of total ewes was determined at the time of CIDR insert. Blood samples were collected on PCD-19 for determining the serum progesterone levels. All the ewes were examined by transrectal ultrasonography 30-35 days after mating for pregnancy diagnosis. The serum values for P4 concentration were in control, PG and PG+GnRH groups 6.34 ± 1.17, 9.19 ± 2.55 and 10.57 ± 2.0 ng/mL respectively. The PG+GnRH treatment significantly increased the litter size compared to the control group (P = 0.04), but there were no significant differences in another reproductive indices between experimental groups. The multiple birth, twin, fecundity rates and litter size (P = 0.05, p = 0.03 and p = 0.003 respectively) were significantly higher in ewes with BCS > 2 compared to ewes with BCS ≤ 2. It is concluded that GnRH on PCD-5 treatment in addition to PGF2α injection at the time of CIDR removal could improve reproductive performance of Qezel ewes during non-breeding season.

Ingested versus inhaled limonene in sheep: A pilot study to explore potential different transfer to the mammary gland and effects on milk and Caciotta cheese aroma.

Concentration is a key determinant in the overall positive impact of terpenes on milk and cheese aroma; additionally, route of intake may affect the achievable concentrations of dietary terpenes in milk and cheese. In this study, we explored the possibility that the amount of the monoterpene limonene transferred to sheep milk and its corresponding cheese could differ depending on the route of intake and that the aroma profile of these products could also differ. To this aim, 12 lactating dairy ewes were repeatedly exposed to limonene by the oral or respiratory route during a 48-h test period, according to a 3 × 3 Latin square experimental design. Limonene content was measured in individual and bulk milk samples, in 1-d-old and 15-d-old Caciotta cheese obtained from that milk, in the related whey and curd, and in the air inhaled by the ewes in the respiratory treatment group (to obtain an estimate of the dose actually supplied by this route). Bulk milk and fresh (1-d-old) cheese underwent sensory analysis by ortho-olfactory evaluation. Both intake routes demonstrated transfer of limonene to milk, but the respiratory route transferred limonene with greater efficiency than the oral route. Moreover, according to the protocol used in this study, a short period of respiratory exposure induced a slightly higher limonene content in milk compared with oral exposure. As to the fate of limonene during cheesemaking, an important part of it was lost into the whey, perhaps through volatilization. The differences between milk and cheese tended to dissipate in curd and fresh cheese and disappeared completely after 15 d of ripening. Finally, it was possible to distinguish between the 2 routes of limonene intake using sensory analysis, even though no direct relationship was identified between the different aroma profiles of milks and cheeses from the oral and respiratory groups and their respective limonene contents. Overall, our results expand current knowledge on the biological pathways of terpene transfer from feed to sheep milk and cheese, as well as on the role played by terpenes in the formation of aroma in these products. Our observations may contribute to future development of strategies for external control and better standardization of the presence of odor compounds in milk and cheese from dairy ruminants.

Chronic low BPS exposure through diet impairs in vitro embryo production parameters according to metabolic status in the ewe.

Bisphenol A (BPA), an endocrine disruptor, has been replaced by structural analogues including bisphenol S (BPS). BPA and BPS exhibited similar effects regarding reproductive functions. Moreover, metabolic status and lipid metabolism are related to female fertility and could worsen BPS effects. The objective was to determine BPS in vivo effects on folliculogenesis and embryo production after chronic exposure through diet, and the influence of metabolic status in adult ewes. Sixty primiparous 2.5 year-old ewes, undergoing a restricted or well fed diet, were exposed to BPS (0, 4 or 50 µg/kg/day) for at least three months. After hormonal oestrus synchronisation and ovarian stimulation, ewes were subjected to ovum pick-up (OPU) procedures to collect immature oocytes, that underwent in vitro maturation, fertilisation and embryo production. Body weight, body condition score and plasma glucose were higher in well-fed compared to restricted ewes, while plasma NEFA was lower during the 4-5 months after the beginning of the diets. Plasma progesterone levels increased on day 5 before OPU session in well-fed compared to restricted ewes. No effect of BPS dose was observed on follicle population, plasma AMH levels and embryo production numbers and rates. However, a significant diet x BPS dose interaction was reported for cleaved embryos, > 4-cell embryos, blastocyst and early blastocyst numbers, and plasma triiodothyronine levels. Our study showed that a contrasted diet did not affect follicle population nor embryo production in adult ewes but could affect the quality and progesterone secretion of the corpus luteum. Chronic low BPS exposure had no effect on follicular population and oocyte competence. Nevertheless, the significant diet x dose interactions observed on embryo production suggest that BPS effect is modulated by metabolic status. Further studies are required to assess the risk of BPS exposure for public reproductive health.