Small ruminant lentiviruses (SRLV) are chronic progressive infections associated with persistent inflammation, with arthritis representing one of the main clinical manifestations in goats. Despite the immunomediated nature of SRLV-associated joint disease, integrated evaluations of systemic and local protein alterations remain limited. This cross-sectional observational study aimed to characterize serum and synovial fluid protein electrophoretic profiles in goats naturally infected with SRLV presenting chronic arthritis and to investigate associations with clinical joint parameters. Fifty adult lactating Saanen goats were allocated into two groups: SRLV-positive goats with chronic arthritis (n = 25) and SRLV-negative healthy controls (n = 25). Clinical evaluation included the clinical articular index (CAI), carpal circumference, and estimated carpal volume, followed by blood and synovial fluid collection. Protein fractions were determined by polyacrylamide gel electrophoresis (PAGE), quantifying albumin, α1-, α2-, β1-, β2-, and γ-globulin fractions. SRLV-positive goats showed significant increases (p < 0.05) in serum α1-, α2-, β1-, and β2-globulins, while all synovial protein fractions were significantly altered compared to controls. The most pronounced synovial alteration was observed in the γ-globulin fraction, which was approximately 3.1-fold higher in infected goats. Moderate positive correlations were observed between serum α2-globulin and synovial γ-globulin (r = 0.56; p < 0.01), as well as between synovial β1-, α1-, and γ-globulins and clinical indicators of joint disease severity (r = 0.52–0.64; p < 0.05). These findings indicate that SRLV-associated arthritis in goats is characterized by a systemic-articular inflammatory axis involving sustained acute-phase activation and marked local humoral immune activity. Serum and synovial fluid protein electrophoresis may provide a useful complementary approach for inflammatory profiling and for identifying protein fractions with potential as candidate biomarkers of articular disease activity in SRLV-infected goats.
Caseous lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is a chronic, contagious disease of small ruminants that impacts animal health and economic concerns globally. This retrospective study analyzed 10 years (May 1, 2015–May 1, 2025) of domestic sheep and goat diagnostic data from the Washington Animal Disease Diagnostic Laboratory (WADDL) to characterize CLA prevalence using synergistic hemolysin inhibition (SHI) and CLA-targeted aerobic cultures. Data from 139,238 sample submissions (120,958 individual animals; 20,492 herds) were standardized and analyzed at the sample, individual, and herd level. Herd-level prevalence exceeded individual prevalence and the number of animals tested per herd was associated with herd-level positivity for both species. The prevalence of CLA based on SHI was lower (Chi-sq p < 0.0001) in goat samples (3.7%; 4,452/120,213) than sheep samples (9.9%; 1,779/17,990). However, the prevalence of CLA based on CLA-targeted aerobic culture was similar (Chi-sq p > 0.05) in goat samples (33.3%; 269/808) and sheep samples (32.2%; 73/227). Overall, prevalence increased with animal age (Chi-sq p < 0.0001) and was found to be higher (Chi-sq p < 0.05) in females of the combined species (SHI 4.9% (2,281/46,268); aerobic culture 35.7% (251/704)) than in males (SHI 3.9% (450/11,561); aerobic culture 25.6% (61/238)). Geographic variation was observed, with SHI results indicating a lower prevalence of CLA in both goats and sheep in WA as compared to other regions in the United States and Canada (Chi-sq p < 0.0001). These findings underscore the importance of surveillance to inform epidemiological studies and herd-level control strategies aimed at reducing the burden of CLA across diverse production systems.
This study assessed the effects of pelleted high-concentrate dietary strategies formulated with different fiber sources on intake, nutrient digestibility, growth performance, and carcass traits of finisher lambs from four genotypes. Eighty male lambs were used in a 4 × 4 factorial arrangement with four genotypes [Santa Inês (SI), ½ Dorper × ½ Santa Inês (DS), ½ Texel × ½ Santa Inês (TS), and ½ Somalis × ½ Santa Inês (SS)] and four dietary fiber sources [sugarcane bagasse (SCB), alfalfa hay (ALH), citrus pulp (CPP), and soybean hulls (SBH)]. Genotype influenced dry matter intake (DMI), with TS lambs exhibiting lower intake than the other groups, although DMI as %BW did not differ. Fiber source significantly affected nutrient utilization; lambs fed SCB and ALH diets had higher crude protein intake and digestibility than those fed CPP and SBH. Average daily gain (ADG) was highest in lambs fed the SCB diet, particularly when compared to those fed SBH. Genotype was the primary driver of performance, with TS lambs showing the best feed-to-gain ratio (3.20kg:kg) and DS lambs requiring fewer days to reach slaughter weight. Significant genotype × fiber source interactions were observed for carcass yields, viscera, offal, internal fat, neck, and rib yields. ALH+SS and ALH+TS combinations produced the lowest carcass yields (~44.9% hot; ~43.4% cold), while SCB+SS exhibited the highest offal yield (18.8% hot; 19.3% cold). Furthermore, internal fat peaked in SS lambs fed ALH and SBH (~15.9%), whereas neck (5.2–10.5%), rib (5.5–7.0%), and viscera yields varied moderately across specific treatment combinations. Genotype also affected commercial cuts, with TS lambs presenting superior shoulder and leg yields. In conclusion, pelleting ensured uniform intake across fiber sources, but sugarcane bagasse promoted slightly better growth performance. While dietary responses can vary by genotype, especially carcass yield, the fiber source (sugarcane bagasse, citrus pulp, and soybean hulls) can effectively replace conventional forages like alfalfa in pelleted diets for finisher lambs without compromising productive efficiency.
This controlled crossbreeding study evaluated the reproductive performance, milk yield, and milk physicochemical properties of F₁, F₂, and B₁ generations derived from Alpine × Hair (AH) and Saanen × Hair (SH) crosses raised under high-altitude, pasture-based conditions in the mountainous regions of Konya, Türkiye. The growth traits of offspring from native Hair, F1, F2, F3, B1 and B2 generations were also evaluated, and differences among generations were identified. Reproductive traits were evaluated in 828 does from three flocks, whereas milk production and milk composition traits were evaluated using 5,118 test-day milk records collected from 769 lactating does over two consecutive years. Crossbred genotypes demonstrated 26–58% higher lactation milk yield compared to Hair goats, with the B₁ generation showing the greatest milk production performance. While crossbreeding increased milk yield and lactation length, it was accompanied by a partial reduction in milk macronutrient content and the fat-to-protein ratio, indicating a genetically regulated balance between yield and composition. The highest twinning rates were observed in the B₁ generation, followed by the F₁ generation. The live weights of the goats were highest during the postpartum and mating periods in the AHF1 generation, and lowest in the SHF2 generation. Live weight gain at weaning was highest in the F1 generation and lowest in the Hair generation, followed by the B2 generation. Comparisons between the F₁ and F₂ generations showed that Saanen-derived crossbreds exhibited larger declines in milk yield and early growth traits than Alpine-derived crossbreds, whereas Alpine-derived crossbreds maintained more stable performance across generations. These findings highlight the importance of considering yield, milk quality and environmental adaptability together when designing crossbreeding strategies for the sustainable production of dairy goats in high-altitude, pasture-based livestock systems.
The aim was to evaluate the classifiability of Kangal Akkaraman and Kivircik breed lamb carcasses using machine learning (ML) methods based on carcass characteristics, carcass dimensions, meat quality, and fatty acid composition data. Data from a total of 155 lamb carcasses, including 60 Kangal Akkaraman and 95 Kivircik breeds, were used. Linear Discriminant Analysis (LDA), Quadratic Discriminant Analysis (QDA), K-Nearest Neighbors (KNN), Support Vector Machines (SVM), Random Forest (RF), XGBoost, and Decision Tree (DT) algorithms were applied. In Dataset-1, where carcass conformation and fatness measures, percentages of carcass joints, and fat colour parameters are predictors, all carcasses could be accurately classified using SVM algorithm with LOOCV validation. In Dataset-2, where carcass dimensions are the predictors, classification was performed with LDA, QDA, SVM, and XGBoost algorithms with an accuracy of 0.994. Classification was performed with Dataset-3, where meat quality characteristics were the predictor, with an accuracy of 0.942, and with Dataset-4, where percentages of individual fatty acids were the predictor, with an accuracy of 0.974. According to LDA results, the most important features in breed differentiation were fatness and conformation in Dataset-1; hind limb length and chest width in Dataset-2; meat yellowness and cooking loss in Dataset-3; and C22:0 and C15:1 in Dataset-4. According to the SVM model, the most important features for Datasets 1-4 were conformation, chest width, meat yellowness, and C20:5 n3, respectively. In conclusion, this study provides a robust proof-of-concept for the high-accuracy classification of the investigated breeds using ML methods. The results suggest that LDA and SVM models based on carcass traits offer significant potential as objective tools for breed characterisation.
Evaluating core body temperature is critical for monitoring livestock welfare during climatic extremes; however, traditional rectal measurement induces handling stress, creating a pressing need to validate non-invasive thermal windows. This study aimed to evaluate correlations among environmental indices, body surface temperatures (BSTs) and rectal temperature (RT) to identify reliable non-invasive thermal indicators. This study represents one of the first controlled Psychrometric chamber studies comparing Jamunapari and Barbari goats using infrared thermography. Twelve goats (six per breed) underwent a repeated-measures design, experiencing progressive cold (24°C to 9°C) and heat (24°C to 42°C; Temperature-Humidity Index (THI) 67–95) stress phases. Following 10-day exposures per thermal level, RT and infrared thermographic BSTs (eyeball, muzzle, ear and perineum) were recorded (n=156 total observations). Data were analyzed utilizing a Repeated Measures General Linear Model (GLM), independent t-tests (for breed comparisons) and Pearson correlations. Under heat stress, pooled data showed strong ambient temperature (AT) and THI correlations with RT (r ≥ 0.860) and BSTs. Muzzle temperature exhibited the highest correlation with AT (r = 0.923), while ear temperature strongly predicted RT (r = 0.902). Breed-specific variations revealed that Barbari goats mirrored the pooled trend, but Jamunapari goats showed peak environmental correlation at the perineum (r = 0.932), likely reflecting distinct morphological adaptations. During cold stress, physiological responses were consistent across breeds; muzzle maintained the strongest correlation with AT (pooled r = 0.948), and ear remained the best predictor of RT (pooled r = 0.825). Despite the limitation of a small sample size, these findings suggest muzzle temperature may serve as a promising indicator of external environmental load, while ear temperature appears to be the most reliable non-invasive predictor of core body temperature. Integrating these specific thermal windows has the potential to support precision livestock management.
Photoperiod influences reproductive maturation in male goats; however, strategies to enable earlier use of young males as breeders remain limited. This study evaluated whether a long-day photoperiod during autumn and winter advanced reproductive maturation and reproductive performance in young bucks. In Experiment 1, September-born bucks (8 weeks old) were assigned either to a natural photoperiod (control; n = 6) or to a long-day photoperiod (treated; n = 6). Testosterone concentrations were measured. In March, five bucks from each group were joined with seasonally anestrous females (n = 37 per group). Treated bucks exhibited an earlier increase in testosterone concentrations and higher values in February and March than control males (P < 0.05). More females kidded when exposed to treated than control bucks (76% vs. 65%; P < 0.05). In Experiment 2, a new group of photoperiod-treated young bucks (n = 6) was compared with photoperiod-treated adult bucks (5 years old; n = 6). Testosterone concentrations increased in both groups during March, although values were higher in adults (P < 0.05). Five bucks from each group were joined with seasonally anestrous females (n = 50 per group). The rates of females that ovulated, became pregnant, and kidded did not differ between females joined with young or adult males (P > 0.05). Long-day photoperiodic stimulation advanced reproductive maturation in young bucks and enabled their successful use as breeders during the seasonal anestrous period. Despite their younger age, photoperiodically stimulated males achieved reproductive outcomes comparable to those obtained with adult bucks.
Genetic evaluations of milk production traits in dairy goats often do not explicitly account for major gene variability, such as αs1 casein polymorphisms, despite their substantial impact on milk protein synthesis. This study assessed the effect of incorporating αs1 casein genotypes on the reliability of genetic evaluations in the Palmera goat breed. The dataset included 37,875 test-day records from 3168 goats, 4342 animals in the pedigree, and genotypes for 156 bucks. Traits analyzed were milk yield (MY) and protein (PC) and fat contents (FC). A gene-content multi-trait BLUP model (GCMTBLUP), treating αs1 casein as a correlated trait, was compared with a baseline animal model. Heritabilities were moderate and similar between models, ranging from 0.19 (FC) to 0.31 (PC). Incorporation of gene content increased reliability across traits, with overall gains from 0.07% (PC) to 1.82% (MY), with higher gains in males (3.05–4.27%) than in females, in genotyped animals (5.52–9.54%) compared to non-genotyped animals, and in animals with low baseline reliability (10.27–20.17%). These results indicated that including αs1 casein information may improve genetic evaluation reliability in the Palmera breed. Future work should expand the genotyped population, including females, to account for potential dominance effects at this locus.
Efficient utilization of agro-industrial co-products such as olive pomace in ruminant nutrition can contribute to long-term food and nutrition security, optimize resource utilization, minimize food waste, and mitigate environmental impacts. This study investigated the effects of dietary inclusion of yeast-fermented olive pomace (YFOP) on feed intake, total-tract nutrient digestibility, milk production, and metabolic health of lactating Damani sheep. Thirty-six multiparous ewes (30 ± 3kg) were assigned to four dietary treatments (n=12) in a randomized complete block design: a control diet (YFOP0), and diets containing 100 (YFOP100), 150 (YFOP150), or 200 (YFOP200) g/kg of YFOP on a dry matter (DM) basis. Olive pomace, produced through a two-phase centrifugation process, was biotechnologically upgraded via a two-stage anaerobic fermentation with yeast. The animals were blocked based on initial milk yield, days in milk, and parity. The trial period included a 12-day adaptation phase, followed by a 38-day data collection period. Daily intakes of diet DM and neutral detergent fiber (NDF) increased (P < 0.001) quadratically, with the highest values recorded for YFOP150 (P < 0.05). The increasing inclusion levels of YFOP had quadratic effects on all measured digestibility parameters, with the highest (P < 0.05) apparent digestibility of DM (683g/kg), crude protein (CP; 774g/kg), and NDF (642g/kg) observed for YFOP150. The increases in diet intake and digestibility were accompanied by a 4.02% increase (P < 0.05) in milk yield in YFOP150 sheep compared with the control sheep. Similarly, significant improvements (P < 0.05) in metabolic health markers were also observed for the YFOP150 group, as reflected by higher levels of total protein and high-density lipoprotein cholesterol, and lower levels of total cholesterol and low-density lipoprotein cholesterol in ewe's serum. However, the higher inclusion level (YFOP200) decreased diet intake, digestibility, and milk yield, but increased (P < 0.05) serum alanine aminotransferase and alkaline phosphatase activities, indicating hepatic stress. These findings suggest that YFOP150 optimizes nutrient utilization, improves milk production, and metabolic health in lactating Damani sheep, whereas a higher inclusion level (YFOP200) was associated with adverse physiological responses.
Integrated crop-livestock (ICL) systems have been proposed as an alternative production strategy in semiarid regions. This study evaluates nutrient intake and digestibility, metabolic profile, microbial protein synthesis, performance, carcass traits, and commercial cuts of sheep raised in different ICL systems in semiarid regions compared to those raised on an exclusive natural rangeland. The treatments included three ICL systems implemented in the Caatinga, comprised of sheep livestock integrated with (i) bean crops, (ii) maize crops, and (iii) herb-cotton crops. The control treatment consisted of exclusive Caatinga rangeland. The experiment, conducted over two years in a thinned Caatinga site enriched with Buffel and Urochloa grasses, involved 24 non-castrated male lambs (Santa Ines × Dorper F1 crossbred) each year. The experimental design was completely randomized, with four treatments (three ICL systems and one control treatment) and six replicates (six sheep per treatment per year), with years treated as a fixed effect. ICL systems increased (P<0.05) the digestibility of nutrients, microbial protein synthesis, serum glucose and urea concentrations, weight gain, and carcass traits. Thus, under the conditions of this study, the use of natural rangelands integrated with bean or herb-cotton crops is recommended, as it favors the intake and digestibility of nutrients, increases serum glucose and urea concentrations, in addition to increasing the microbial protein synthesis, resulting, consequently, in better productive performance and superior carcass traits of Santa Ines × Dorper F1 crossbred grazing lambs.
Maedi-Visna and Caprine arthritis–encephalitis viruses are common among sheep rearing countries and have been causing economic losses because of the long preclinical incubation period and fatal onset. Two viruses are members of the small ruminant lentiviruses and infection generally happens between sheep and goat without a species barrier. In this study 15 sheep with respiratory sings and suspected of having Maedi-Visna disease was investigated for antioxidant response associated with the Sirtuin-1 (SIRT1) – Nuclear factor erythroid 2-related factor 2 (NRF2) – Heme oxygenase-1 (HO‑1) axis in conjunction with 8-hydroxy-2′-deoxyguanosine (8-OhdG) levels. Diagnosis of the disease was determined with polymerase chain reaction specific to LTR region of the virus. The diagnosis was supported with histochemical findings. SIRT1, NRF2, HO‑1, 8-OHdG expression levels were determined using immunohistochemical and immunofluorescence methods. According to results Maedi infection was found to be severely suppressing antioxidant defence mechanisms and increasing oxidative damage. The findings indicate an inverse relationship between suppression of the SIRT1/NRF2/HO-1 axis and increased 8-OHdG levels, suggesting that oxidative stress may play an important role in the pathogenesis of this disease.
Ocular diseases can negatively affect sheep welfare and productive performance. Therefore, establishing reference ocular parameters is important for both clinical practice and ophthalmic research. This study aimed to establish reference values for tear production using the Schirmer tear test (STT-1), tear crystallization using the tear ferning test (TFT), corneal sensitivity (CS) assessed with a Cochet-Bonnet esthesiometer, intraocular pressure (IOP) measured by applanation tonometry, as well as conjunctival cytological (CYT) and histological (HIS) characteristics in adult healthy Santa Inês sheep (10 males and 10 females). Data from both eyes were analyzed. Data normality was assessed using the Shapiro-Wilk test. Student’s t-test was performed for STT-1, CS, and IOP. The Kruskal-Wallis test was applied to evaluate inter-examiner agreement for the TFT. A p-value < 0.05 was considered statistically significant. No differences were observed between the right and left eyes; therefore, the mean value of both eyes was used. Mean STT-1 was 20.6 ± 5.7 mm/min, and mean CS was 25.2 ± 6.7 mm, with no sex-related differences. Mean IOP was 16.6 ± 2.7 mmHg, with a significant difference between sexes. TFT patterns were 13.1% type I, 30.6% type II, 35.0% type III, and 21.3% type IV, according to the Rolando scale. Cytological evaluation revealed a predominance of epithelial cells (489 superficial anucleated cells, 315 superficial nucleated cells, 639 basal cells and 523 intermediate cells), followed by leukocytes (396 cells), and few goblet cells (121 cells); superficial nucleated cells were significantly more numerous in males. Histological analysis showed non-keratinized stratified squamous epithelium and connective tissue in 40 samples (100%), while goblet cells and blood vessels were observed in 19 samples (47,5%). In conclusion, reference values for tear parameters (tear production and tear crystallization), corneal sensitivity, intraocular pressure, and conjunctival cytological and histological features were established in Santa Inês sheep. These data are relevant to ophthalmic evaluation in this species, which is used both as a food-producing animal and as an experimental model for ocular research.
Accurate prediction of breeding values (BVs) is critical for effective genetic improvement in livestock. This study evaluated the predictive performance of multiple machine learning (ML) algorithms, Extreme Gradient Boosting (XGBoost), Random Forest Regression (RFR), Support Vector Regression (SVR), and Artificial Neural Networks (ANN), for predicting EBVs of weaning weight (WWT) in 8405 Murciano-Granadina kids born between 2016 and 2024. The EBVs were calculated using the conventional BLUP animal model. The dataset included 389 sires and 3459 dams, providing a well-structured pedigree framework. Models were trained and validated using a structured data split, with performance assessed via coefficient of determination (R²), root mean square error (RMSE), mean absolute error (MAE), bias, and information criteria (AIC/BIC). XGBoost consistently outperformed all other models, achieving the highest predictive accuracy (R² = 0.76), lowest prediction errors (RMSE = 0.04; MAE = 0.03), minimal bias (0.003), and the strongest correlation with BLUP-derived EBVs (r = 0.89). RFR also demonstrated robust performance (R² = 0.65; r = 0.81), whereas SVR and ANN were less accurate. Feature importance analyses revealed that growth-related traits, particularly Average Daily Gain (ADG) and Birth Weight (BWT), were the most influential predictors, with tree-based ensemble methods achieving higher parsimony and explanatory power than SVR and ANN. These findings establish gradient boosting, and specifically XGBoost, as a powerful computational paradigm for high-accuracy breeding value prediction in small ruminants. The superior performance of XGBoost demonstrates that ensemble tree-based ML models can complement or augment conventional BLUP methods, supporting early selection and breeding decisions in Murciano-Granadina goats.
Brucella melitensis Rev.1 is widely used to control brucellosis in sheep and goats, but the route of administration may affect both protection and post-vaccination serological interference. This study compared subcutaneous (SC) and conjunctival (VC) administration of the live attenuated B. melitensis Rev.1 vaccine in small ruminants. Twenty-five local-breed sheep and goats were assigned to subcutaneous vaccination (SC, n = 10), conjunctival vaccination (VC, n = 10) or a non-vaccinated challenged control group (n = 5). The vaccinated groups differed simultaneously in route, dose and excipient (SC: 2 × 109 CFU in 2 mL of conventional diluent; VC: 1 × 109 CFU in 40 µL of diluent containing 5% Montanide Gel-01), so the comparison reflects these three factors combined rather than route alone. Serology was monitored for 15 months by indirect ELISA and the Rose Bengal test, after which all animals were challenged conjunctivally with virulent B. melitensis 16 M and euthanised 15 days later. Subcutaneous vaccination induced a persistent antibody response that remained above baseline at month 15 (S/P% 181.4 ± 49.4), with 4/10 animals still Rose Bengal-positive. Conjunctival vaccination produced a transient response that never reached the positivity threshold (maximum 36.1%) and all VC animals were Rose Bengal-negative from month 6 onward (month 6: 10/10 versus 0/10; Holm-adjusted p = 6.5 × 10−5). After challenge, Brucella was recovered from 8/10 SC, 4/10 VC and 5/5 control animals; the difference between the vaccinated groups was not significant (p = 0.17). Bacterial burden was significantly lower in VC (median 0 versus 262.5 colonies per animal; p = 0.0052), and recovery in VC was confined to the lymph nodes, whereas SC animals also yielded Brucella from spleen and liver. All isolates were identified as strain 16 M. Neither route conferred sterilising immunity. Within the limits of a small cohort and a confounded design, the conjunctival formulation combined reduced serological persistence with more restricted dissemination a combination that may be advantageous where vaccination must remain compatible with serological surveillance.
Serum proteins play central roles in the inflammatory response and maintaining homeostasis. Serum protein electrophoresis is widely used in veterinary medicine, including in small ruminants, to detect inflammatory, infectious, and metabolic disorders by evaluating the protein electrophoretogram. Among the most frequently employed techniques in clinical laboratories are agarose gel electrophoresis (AGE), a laboratory method that separates serum proteins into 5–6 fractions using a solid matrix, and capillary zone electrophoresis (CZE), a more recent approach offering greater analytical precision and resolution. This study aimed to compare AGE and CZE for the analysis of the serum protein profile of small ruminants, establish reference intervals (RI), and assess agreement between fractions. Samples from 65 goats and 68 crossbred adult sheep, all clinically healthy, were analyzed. Fractions of albumin, α-, β-, and γ-globulins were identified and quantified. Data were evaluated using Spearman/Pearson correlations, Passing–Bablok regression, and Bland–Altman analysis. The results indicate that both methods are suitable for serum protein fractionation, showing a high degree of agreement overall. AGE demonstrated more robust quantification of γ-globulins, whereas CZE enabled further subdivision of β-globulins into β1 and β2 fractions and showed greater sensitivity in detecting immunoglobulin variations. While AGE profiles were characterized by the predominance of albumin and γ-globulins, reflecting physiological transport and immune functions, CZE provided superior analytical resolution. The established reference intervals proved consistent, confirming AGE and CZE as suitable and complementary methods. The standardization of species-specific values contributes to more accurate diagnoses and improved health monitoring of small ruminants.
This study evaluated the effect of antifreeze protein type I (AFP I) on the vitrification of immature ovine cumulus-oocyte complexes (COCs), examining metabolic, structural, and molecular parameters. Slaughterhouse-derived COCs were allocated into three groups: fresh control, vitrified without AFP I (AFP-), and vitrified with AFP I (250 ng/mL; AFP+). Mitochondrial activity did not differ significantly between the Fresh (114.79 ± 8.00 AU) and AFP + (135.43 ± 7.96 AU) groups, whereas the AFP- group showed a significantly lower value (89.78 ± 4.64 AU; P < 0.05). The AFP + group exhibited higher levels of reactive oxygen species (ROS) (58.11 ± 2.57 AU) compared to the Fresh (40.61 ± 3.11 AU) and AFP- (34.26 ± 3.58 AU) groups (P < 0.05). Glutathione (GSH) levels did not differ significantly among the Fresh (15.78 ± 2.32 AU), AFP- (19.90 ± 4.00 AU), and AFP + (26.41 ± 5.60 AU) groups. Regarding actin integrity, the Fresh group (15.02 ± 1.15 AU) had higher values than AFP- (11.22 ± 0.71 AU) and did not differ from AFP + (13.09 ± 0.69 AU). In gene expression analysis, the AFP + group presented a profile more similar to the Fresh group, whereas BMP15, MATER, and ZAR-1 were overexpressed in the AFP- group. AFP I partially preserved mitochondrial activity and cytoskeletal integrity during cryopreservation, but its use was associated with increased ROS.
