Can in vitro embryo production be estimated from semen variables in Senepol breed by using artificial intelligence?

Thoroughly analyzing the sperm and exploring the information obtained using artificial intelligence (AI) could be the key to improving fertility estimation. Artificial neural networks have already been applied to calculate zootechnical indices in animals and predict fertility in humans. This method of estimating the results of reproductive biotechnologies, such as in vitro embryo production (IVEP) in cattle, could be valuable for livestock production. This study was developed to model IVEP estimates in Senepol animals based on various sperm attributes, through retrospective data from 290 IVEP routines performed using 38 commercial doses of semen from Senepol bulls. All sperm samples that had undergone the same procedure during sperm selection for in vitro fertilization were evaluated using a computer-assisted sperm analysis (CASA) system to define sperm subpopulations. Sperm morphology was also analyzed in a wet preparation, and the integrity of the plasma and acrosomal membranes, mitochondrial potential, oxidative status, and chromatin resistance were evaluated using flow cytometry. A previous study identified three sperm subpopulations in such samples and the information used in tandem with other sperm quality variables to perform an AI analysis. AI analysis generated models that estimated IVEP based on the season, donor, percentage of viable oocytes, and 18 other sperm predictor variables. The accuracy of the results obtained for the three best AI models for predicting the IVEP was 90.7, 75.3, and 79.6%, respectively. Therefore, applying this AI technique would enable the estimation of high or low embryo production for individual bulls based on the sperm analysis information.

Pharmacological perturbation of peroxisome-proliferator-activated receptor gamma alters motility and mitochondrial function of bovine sperm.

BACKGROUND: Sperm transit through the female reproductive relies upon maintenance of sperm motility. Peroxisome-proliferator-activated receptor gamma (PPARγ) is a ligand-activated nuclear transcription factor with roles in glucose metabolism and reproductive processes including placental function. PPARγ roles in the mammalian postejaculatory sperm function are incompletely defined. OBJECTIVES: Determine expression, localization, and functions of PPARγ in postejaculatory bovine sperm. MATERIALS AND METHODS: Frozen-thawed bovine sperm from three to four different bulls were pooled and subjected to immunofluorescence and western blot for detection and localization of PPARγ. Functions in sperm energetics were explored through the addition of pharmacological inhibition (GW; GW9662) and activation (Ros; Rosiglitazone) in the culture medium at 0 and 24 h under non-capacitating conditions. Samples were analyzed for sperm kinematics (CASA) and mitochondrial membrane potential (MMP; JC-1 fluorophore). RESULTS: PPARγ was detected in bovine sperm and co-localized to the acrosome with re-localization to the equatorial region in acrosome-compromised sperm. The addition of Ros 50 µM for 24 h maintained superior total and progressive motility of sperm compared to vehicle control (VC-DMSO 0.01%). The PPARγ antagonist GW 1 µM was detrimental to both total and progressive motility. A challenge experiment (Ros + GW) partially rescued total and progressive motility phenotypes observed with GW incubation. GW-treated samples had a lower number of sperm with high MMP at 24 h compared to Ros or VC. The negative GW MMP phenotype was reversed with the addition of Ros + GW. Likewise, GW-treated samples had more sperm with low MMP compared to VC and Ros, and this phenotype was partially restored with Ros + GW. CONCLUSION: PPARγ is expressed in post-ejaculatory bovine sperm with regulatory roles in sperm motility and MMP. These findings implicate PPARγ as a novel regulator of postejaculatory mammalian sperm energetics through non-canonical signaling mechanisms.

Frozen-Thawed Sperm Analysis of Benign Prostatic Hyperplasia Dogs Treated With Finasteride.

Benign Prostatic Hyperplasia (BPH) is a pathological condition that directly interferes with the reproductive potential of senile dogs, by leading to prostate enlargement and sperm injury, which in turn may compromise sperm freezeability. Moreover, albeit finasteride treatment reduces prostatic volume and blood supply and maintains seminal quality and testicular integrity, the effects of sperm samples submitted to cryopreservation after the finasteride treatment are still unknown. Thus, the aim of this study was to evaluate frozen-thawed semen of BPH dogs, as well as dogs subjected to BPH pharmacological treatment with finasteride. For such purpose, 20 dogs were previously selected and assigned to three experimental groups, according to BPH diagnosis and treatment with finasteride: Control (n = 9), BPH Group (n = 5) and BPH-Finasteride Group (n = 6). Semen was subjected to one-step cryopreservation protocol with tris-fructose-citric acid extender with 5% glycerol and thawed at 37°C for 30 sec. Fresh and post-thaw sperm samples were evaluated for macroscopic parameters, sperm concentration, sperm motility kinetics, sperm mitochondrial activity and potential, oxidative stress, plasmatic and acrosome membrane integrity, sperm DNA fragmentation and sperm binding test on perivitelic membrane of chicken egg yolk. Regarding fresh semen, BPH-Finasteride group had the lowest ejaculate visual aspect (opacity), higher frequency of sperm flagellar beating (BCF) and percentage of sperm with medium velocity. Control group had the highest percentage of sperm DNA integrity compared to BPH group. For the frozen-thawed semen, Control group presented the highest percentage of spermatozoa with high mitochondrial activity. However, the BPH-Finasteride group showed higher number of sperm bound to the perivitelline membrane of chicken egg yolk compared to the BPH Group. Conversely, BPH group had higher percentage of DNA damage. In conclusion, the ejaculate of BPH dogs has higher susceptibility to cryoinjury, whereas finasteride-treated dogs have increased spermatozoa functional performance, suggesting a promising use of BPH dogs as semen donors in sperm cryopreservation programs.

Chronological characterization of sperm morpho-functional damage and recovery after testicular heat stress in Nellore bulls.

Heat stress (HS) affects spermatogenesis and sperm maturation, decreasing sperm quality. Yet sperm morpho-functional changes caused by HS in Nellore bulls are not fully elucidated. This study aimed to show the chronological effects on sperm quality of HS during spermatogenesis and sperm maturation until recovery of the seminiferous epithelium in Nellore bulls. Nine Nellore bulls were distributed into control and heat stress (HS-scrotal bags/96 h) groups. The study was divided into five Periods: 1. Control (14-7 days before HS); 2. Stored sperm (0-7 days after HS); 3. Sperm maturation and late spermatogenesis (14-42 days after HS); 4. Early spermatogenesis (49-63 days after HS), and 5. Recovery (70-77 days after HS). Semen was collected once a week and evaluated for sperm motility, morphology, plasma, acrosome, and mitochondrial membranes, lipid peroxidation, and DNA fragmentation. Sperm characteristics were similar between groups in Periods 1 (control). During Period 2, HS increased detached normal head defect and decreased mitochondrial membrane potential, denoting effects on the sperm stored at the epididymis cauda. In Period 3, HS decreased sperm motility, plasma membrane integrity, and mitochondrial membrane potential and increased abnormal sperm, lipid peroxidation, and DNA fragmentation; reflecting the effects on sperm that were in the epididymis body and head and late spermatogenesis (spermiogenesis and meiosis). In Period 4, HS maintained a reduction in the mitochondrial membrane potential and an increase in abnormal sperm; injuries that could occur during early spermatogenesis (mitosis). Finally, in Period 5, the groups were similar, confirming the recovery of the seminiferous epithelium after HS. This study provides insights on the effects of HS on the complete process of sperm maturation and spermatogenesis, until recovery in sperm from Nellore bulls.

Sperm function and oxidative status: Effect on fertility in Bos taurus and Bos indicus bulls when semen is used for fixed-time artificial insemination.

Semen quality is one of the criteria used for the selection of bulls with relatively greater fertility. In addition, bull fertility depends on the integrity and function of all sperm structures. The aim of this study, therefore, was to determine associations when there was conducting of conventional and functional techniques for the evaluation of sperm samples from bulls with known fertility history as determined when semen from these bulls was used for fixed-time artificial insemination programs. The study was designed in a 2 × 2 factorial arrangement, with one factor being breed (Angus x Nellore) and the other fertility (greater x lesser). Greater fertility groups were composed of ten Angus and 11 Nellore bulls, while lesser fertility groups were composed of ten Angus and seven Nellore bulls. Sperm were analyzed, in four cryopreserved distinct batches for each animal, for morphology, kinetics, plasma and acrosomal membrane integrity, mitochondrial activity and mitochondrial membrane potential, DNA integrity and oxidative status. There was no difference in characteristics commonly used in sperm quality conventional analysis. The results from functional analysis indicated an important association between mitochondrial dysfunctions, oxidative stress, and damage to sperm structures in lesser fertility bulls. Greater fertility bulls had greater sperm quality and indicators of functional cell structures. The associations, when there were evaluations using different techniques, indicate the importance of evaluation and correlation between different sperm functions to understand effects of distinct parameters on sperm fertilization capacity.

Evaluation of Blood Glucose and Lactate Concentrations in Mule and Equine Foals.

Information about mule physiology is scarce. Glucose and lactate serve as prognostic tools in neonates; thus, real-time evaluations would be beneficial. Our main objective was to measure glucose and lactate concentrations from healthy mule and equine foals from birth to 720 hours. Glucose and lactate concentrations were evaluated with a benchtop Randox Daytona analyzer (LAB) using plasma and with an Accutrend Plus system (ACP) using whole blood. Data were analyzed using PROC MIXED (P < .05), intraclass coefficient correlation and Bland-Altman analysis. Glucose and lactate concentrations from mule and equine foals were different when evaluated with LAB, but there was no difference when evaluated with ACP. Glucose pattern of variation, evaluated with both analyzers for mule and equine foals, had an increase, with subsequent decrease, whereas lactate pattern of variation had initial higher values with subsequent decrease. Intraclass coefficient correlation for glucose was low and moderate for lactate. Results of glucose and lactate with ACP from our experimental neonates were not highly correlated with LAB. However, the ACP had the same pattern of variation for glucose and lactate, thus can still be used clinically for immediate evaluations if the technique is standardized with the specific samples that will be used.

Lipid peroxidation in bull semen influences sperm traits and oxidative potential of Percoll®-selected sperm.

Although bovine embryo in vitro production (IVP) is a common assisted reproductive technology, critical points warrant further study, including sperm traits and oxidative status of sperm for in vitro fertilization (IVF). Our aim was to evaluate whether the lipid peroxidation index of commercial bull semen is influenced by sperm traits and oxidative status of sperm populations selected using Percoll® gradient. Semen straws from 48 batches from 14 Nelore bulls were thawed individually, analyzed for motility and subjected to Percoll selection. After Percoll, the lipid peroxidation index of the extender was evaluated, whereas selected sperm were analyzed for motility, acrosome and membrane integrity, mitochondrial membrane potential, chromatin resistance and oxidative potential under IVF conditions. Batches were divided retrospectively in four groups according to lipid peroxidation index. Sperm from Group 4 with the lowest index of lipid peroxidation had, after Percoll selection, greater plasma membrane integrity (81.3%; P = 0.004), higher mitochondrial potential (81.1%; P = 0.009) and lower oxidative potential (135.3 ng thiobarbituric acid reactive substances (TBARS)/ml; P = 0.026) compared with Group 1 with highest lipid peroxidation index (74.3%, 73% and 213.1 ng TBARS/ml, respectively). Furthermore, we observed negative correlations for the lipid peroxidation index with motility, membrane integrity and mitochondrial potential, and positive correlations with oxidative potential. In conclusion, oxidative stress in semen straws, as determined using lipid peroxidation in the extender, is associated with sperm traits and their oxidative potential under IVF conditions. These results provided further insights regarding the importance of preventing oxidative stress during semen handling and cryopreservation, as this could affect sperm selected for IVF. Finally, Percoll selection did not completely remove sperm with oxidative markers.

Spermatic profile of captive giant anteaters (Myrmecophaga tridactyla): Knowing more to preserve better.

The giant anteater (Myrmecophaga tridactyla) is being threatened by natural habitat destruction and fragmentation, illegal hunting and road kills. In this context, the generation of basic information on the reproductive parameters of this species is vital, aiming to improve reproductive management via, amongst others, assisted reproductive technologies. This study aimed to describe the morphological and functional features of semen collected from captive giant anteaters. Electroejaculation was performed in 13 animals housed in zoos located in São Paulo state, Brazil. Semen samples were collected from 13 animals in 16 procedures. Samples were evaluated for volume, motility, vigor, pH, concentration, sperm morphology, and functional tests. The following mean values were obtained: volume 1.28 ± 0.27 mL; motility 28.3 ± 6.2%; vigor 2.4 ± 0.25; concentration 129.4 ± 36.1 × 106 sperm/mL; pH 7.4 ± 0.2. Total acrosome, head, midpiece, and tail sperm abnormalities were 3.2 ± 0.8%, 25.4 ± 3.6%, 20.7 ± 3.2%, and 14.7 ± 2.6%, respectively. Intact acrosome was found in 83.7 ± 3.1% and intact membrane in 81.1 ± 4.0% of all samples collected. Mitochondrial activity was 66.4 ± 6.0% (Class I), 18.7 ± 2.9% (Class II), 8.0 ± 2.0% (Class III), 3.9 ± 1.0% (Class IV), and 3.0 ± 0.9% (Class V). Sperm DNA fragmentation rate was 13.2 ± 3.7%. These results indicated that electroejaculation is a feasible method for semen collection in giant anteaters, allowing a more detailed description of the semen in this species.

Sperm evaluation and morphological description of male genitalia of meerkats (Suricata suricatta).

In ex situ conditions, little is known about the reproductive biology of meerkats. The aim of present study was to describe the morphological aspects of male genital organs and accessory glands using macroscopic evaluation, ultrasonography, and radiography, as well as describing semen characteristics post-electroejaculation. The results indicated anatomical characteristics of meerkats are very similar to those of cats, having a prostate, accessory bulbourethral glands, and an elongated and radiopaque structure in the penis, which is indicative of there being a baculum. The testicular volume was 0.81 cm³ (± 0.10) and the relative testis weight was 1.37 cm³/kg (± 0.15). Both testicles are present in the scrotum, which has an ellipsoidal shape, homogeneous texture, hypoechoic parenchyma and are encased in a hyperechoic tunica albuginea. Electroejaculation was effectively induced in all animals for semen collection with utilization of medetomidine and ketamine. The values semen samples variables were as follows for volume - 0.125 ±â€¯0.193 mL, motility - 19.8 ±â€¯18.6 %, vigor - 1.9 ±â€¯1.0, concentration - 40.5 ±â€¯25.2 × 106 sperm/mL and morphologically normal sperm - 10.8 ±â€¯6.6 %. This is the first study in which there is a description of morphological and imaging aspects of the male reproductive tracts of meerkats, as well as the seminal characteristics after using electroejaculation for semen collection. Knowledge of anatomical and seminal characteristics is essential for implementation of assisted reproduction programs, as well as reproductive management in the species.

Seasonal effects on testes size and sustained semen quality in captive golden-headed lion tamarins, Leontopithecus chrysomelas.

The golden-headed lion tamarin (Leontopithecus chrysomelas) is an endangered primate that, according to timing of births, there seems to have seasonal reproductive patterns in captive populations inhabiting the Southern Hemisphere. The extent to which male tamarins have circannual cyclic fluctuations in testicular functions, however, remains unknown. Changes in testis size and seminal variables, therefore, were evaluated during the rainy and dry seasons of the year in seven males. There was estimation of mating and birth seasons from the breeding colony using a 27 year database (229 birth records). Births were distributed throughout the year with peaks occurring in August-October and January-March (39.7 % and 30.5 % of all births, respectively). Semen collection using penile vibrostimulation was successfully induced in more than 96 % of the attempts regardless of the season (total of 75 ejaculates). Body mass did not vary significantly between seasons, but relative testes size was larger during the dry season. Values for none of the seminal variables (total sperm count, total and progressive motility, plasma membrane and acrosome integrity, and total sperm defects), however, were different during the rainy and dry seasons. These results indicate that testicular function in golden-headed lion tamarins may not be affected by daylength changes, and that seasonal patterns of female reproduction is perhaps more relevant for the reproductive timing of the species. Furthermore, the possibility of year-round production of ejaculates containing viable sperm broadens our perspective of preserving genetic diversity within the species because there is a greater opportunity for semen collection and freezing.

Changes in fertilization medium viscosity using hyaluronic acid impact bull sperm motility and acrosome status.

The female reproductive tract, in particular the composition of the uterine and oviduct fluids, is responsible, at least in part, for triggering sperm cell modifications, essential for the acquisition of fertilization ability. Hyaluronic acid (HA) is a glycosaminoglycan present in these fluids, and its role in the fertilization process and sperm functionality is still barely understood. This work was designed to (a) determine the rheological characteristics of the fertilization medium by the addition of HA and (b) determine the HA influence on sperm motility and functional status. To that end, the in vitro fertilization medium was supplemented with 4 doses of HA (6, 60, 600 and 6,000 µg/ml) and analysed for viscosity and adhesion strength characteristics. Then, thawed semen from 6 bulls were incubated in these media and assessed at 4 different moments for morphological and functional parameters (plasma and acrosomal membrane integrities, mitochondrial membrane potential, capacitation, acrosomal reaction, and motility). The rheological evaluation showed that the addition of HA was able to increase both the viscosity and the adhesion strength of the fertilization medium, especially in the 6,000 µg/ml group in which the effect was more pronounced. No influence of HA could be observed on mitochondrial potential, and acrosomal and plasma membrane integrities. However, HA supplementation, at lower doses, led to an increase in the number of reacted sperm, as well as changes in motility parameters, with increase in the number of motile, rapid and progressive spermatozoa. In conclusion, the addition of HA alters the rheological properties of the fertilization medium and leads to the improvement of the properties related to sperm motility and capacitation, without compromising other functional aspects of the cell.

Heat stress effects on bovine sperm cells: a chronological approach to early findings.

Testicular heat stress affects sperm quality and fertility. However, the chronology of these effects is not yet fully understood. This study aimed to establish the early sequential effects of heat stress in bull sperm quality. Semen and blood samples of Nellore breed bulls were collected and distributed into control and testicular heat stress (scrotal bags/96 h) groups. Semen samples were evaluated for sperm motility, abnormalities, plasma membrane integrity, acrosomal membrane integrity, mitochondrial membrane potential, sperm lipid peroxidation, seminal plasma lipid peroxidation, and DNA fragmentation. Blood plasma was also evaluated for lipid peroxidation. An increase in sperm abnormalities was observed 7 days following heat stress. After 14 days, sperm lipid peroxidation increased and mitochondrial membrane function, sperm motility, and plasma membrane integrity decreased. Heat stress effects were still observed after 21 days following heat stress. An increase in sperm DNA fragmentation was observed as a late effect after 28 days. Thus, the initial effects of heat stress (i.e., increasing sperm abnormalities and lipid peroxidation) suggest the presence of oxidative stress in the semen that alters mitochondrial function, sperm motility, plasma membrane integrity, and belatedly, DNA fragmentation. Although sperm abnormalities persisted and increased over time, sperm lipid peroxidation, in turn, increased only until 21 days after heat stress. In this regard, these findings provide a greater understanding of the chronological effects of experimentally induced heat stress on bovine sperm, providing valuable insights about spermatogenesis during the first 28 days following heat stress.

Semen cryopreservation in golden-headed lion tamarin, Leontopithecus chrysomelas.

Wild animal genetic resource banking (GRB) represents a valuable tool in conservation breeding programs, particularly in cases involving endangered species such as the golden-headed lion tamarin (Leontopithecus chrysomelas). Thus, we aimed to assess a sperm freezing protocol for golden-headed lion tamarins using two different exenders: BotuBOV® (BB) and Test Yolk Buffer® (TYB). Ejaculates were collected by penile vibrostimulation from animals housed at São Paulo Zoological Park Foundation, São Paulo, Brazil, and after immediate analysis, two aliquots were diluted in BB and TYB. Postthawing samples were evaluated for total and progressive motility, plasma membrane and acrosome integrities, mitochondrial activity, susceptibility to oxidative stress, and sperm-egg-binding. No differences between BB and TYB were found for most seminal parameters, except for acrosome integrity and susceptibility to oxidative stress (in both cases BB showed higher values). However, in spite of these differences and regardless of the extender used, postthaw sperm motility and viability with the described protocol were encouraging (on average >50% and >80%, respectively), indicating that sperm cryopreservation may be a short-term measure for the conservation of golden-headed lion tamarins.

Spermatic mitochondria: role in oxidative homeostasis, sperm function and possible tools for their assessment.

SummaryDespite sperm mitochondrial relevance to the fertilization capacity, the processes involved in the production of ATP and functional dynamics of sperm mitochondria are not fully understood. One of these processes is the paradox involved between function and formation of reactive oxygen species performed by the organelle. Therefore, this review aimed to provide data on the role of sperm mitochondria in oxidative homeostasis and functionality as well the tools to assess sperm mitochondrial function.

Induced sperm oxidative stress in dogs: Susceptibility against different reactive oxygen species and protective role of seminal plasma.

Oxidative stress (OS) is characterized by an unbalance between increased levels of reactive oxygen species (ROS) and/or impaired antioxidant protection. In this context, the composition of seminal plasma (SP) plays a key role in protecting sperm against OS. However, reproductive biotechnologies applied to dogs recommend the removal of SP. Thus, antioxidant therapy may be an important alternative when applying biotechniques such as semen cryopreservation in this specie. However, in order to be efficient, the choice of the ideal antioxidant in each condition is essential since each ROS is preferably neutralized by different antioxidant systems. Therefore, this study aims to evaluate the susceptibility of canine spermatozoa to different oxidative challenges (superoxide anion [O2-], hydrogen peroxide [H2O2], hydroxyl radical [OH-] and malondialdehyde [MDA]) in the present or absence of SP. We used ejaculates of eight dogs and submitted to induce oxidative challenges (with or without SP). After incubations, samples were evaluated for the susceptibility to lipid peroxidation, motility, mitochondrial activity and function, DNA integrity, plasma membrane and acrosome integrity. Sperm with SP had mitochondrial function preserved against ROS. However, in the absence of SP, H2O2 reduced mitochondrial membrane potential. In addition, regardless on SP, H2O2 was deleterious to sperm kinetics and plasma/acrosomal membranes. Incubation with OH- reduced mitochondrial activity and increased DNA fragmentation also independent on the absence of presence of SP. Furthermore, samples with SP were more resistant to lipid peroxidation (i.e., decreased concentration of TBARS). In conclusion, H2O2 and OH- appears to be the most deleterious ROS to dog sperm and SP protects the spermatozoa against mitochondrial injuries and lipid peroxidation.

Recovery of normal testicular temperature after scrotal heat stress in rams assessed by infrared thermography and its effects on seminal characteristics and testosterone blood serum concentration.

Reestablishment of testicular normal temperature after testicular heat stress is unknown and its effect varies widely. The aim of this study was to investigate the impact of scrotal insulation (IN) on testicular temperature and its relation to semen quality and testosterone blood serum concentration. For this, 33 rams were used; 17 submitted to IN for 72 hours (using bags involving the testes) and 16 not submitted to IN (control group). The experiment was performed between August and December 2013 in Pirassununga, Brazil (21°56″13″ South/47°28'24″ West). Seminal characteristics, testosterone blood serum concentration, rectal temperature (RT), respiratory frequency, scrotal superficies mean temperature (SSMT), and eye area mean temperature (EAMT) were analyzed 7 days before IN and 21, 35, 49, 63, and 90 days afterward. Scrotal superficies mean temperature and EAMT were measured by thermography camera FLIR T620. Testosterone was evaluated by radioimmunoassay. Analysis of variance was used to determine the main effects of treatment, time, and treatment-by-time interaction using PROC MIXED of SAS software adding command REPEAT. Pearson correlation test was used to verify correlation between SSMT, EAMT, RT, and respiratory frequency. Significant difference was considered when P ≤ 0.05. At the end of IN, SSMT was higher (P < 0.05) in insulated group (32.26 ± 0.19(o)C) than in control group (30.58 ± 0.18(o)C), and the difference between rectal and testicular (deduced from SSMT) temperatures was 1.12 °C; in the other times of the evaluation this difference was between 2.91 and 4.25 °C in IN group. Scrotal superficies mean temperature was reestablished 24 hours after IN. Rectal temperature and EAMT presented correlation (r = 0.59; P < 0.0001). There was time-by-treatment interaction for total sperm (P = 0.0038) and progressive motility (P = 0.01), abnormal spermatozoa (P < 0.0001), membranes integrity (P < 0.0001), induced thiobarbituric acid reactive substances (TBARSs; P = 0.05), and DNA integrity (P = 0.0004). These semen characteristics were negatively affected 21 days after IN, and excluding induced TBARSs and abnormalities, recovered 35 days afterward; induced TBARSs just were affected after 49 days of IN; sperm abnormalities just recovered after 63 days. Testosterone blood serum concentration was lesser in insulated rams (P = 0.03). Thus, the difference of 1.12 °C between RT and testicular temperature impacts semen quality and testosterone blood serum concentration. Moreover, this study shows that rams can recover testes temperature efficiently toward IN and that infrared thermography is an efficient tool to identify differences on SSMT.