P. A. A. Góes

Reproductive characteristics of captive greater rhea (Rhea americana) males reared in the state of São Paulo, Brazil

Rheas (Rhea americana) belongs to the ratite group. Considering the commercial significance of this birds, some techniques, such as semen collection, were standardized. In this study, 107 male rheas (3 to 4 years of age) reared in commercial farms in the state of Sao Paulo, Brazil, were used. Semen was collected during the breeding and off-breeding seasons of 2001, 2002, and 2003. Bird hierarchical behavior was observed. Birds were restrained performed using a box and a black hood. Semen was collected by digital pressure on the base of the phallus, which size was measured, and the presence or absence of spiral shape was observed. Immediately after collection, semen samples were evaluated for volume, motility, sperm concentration, and morphology. In a limited number of birds, blood samples were collected to measure testosterone levels. Among the 69 birds studied during the breeding season, 44 presented large phalluses, out of which 26 showed spiral shape. The method of semen collection was efficient. The following semen parameter results were obtained: volume (0.68 ±0.14 ml), motility (61.11±11.54%), sperm concentration (3.29±1.33 x109 sptz/ml), and number of spermatozoa per ejaculate (2.40±1.38x109 sptz/ml). Morphological abnormalities were analyzed and recorded. Testosterone levels were statistically different (p = 0.0161) between the breeding and non-breeding season (53.28±18.41 ng/ml and 5:57±3.81 ng/ml, respectively). Variations in phallus size were also found between the breeding and non-breeding seasons. Larger phalluses and higher testosterone levels were correlated with dominant behavior. The results of the present experiment confirmed that it is possible to collect semen from rheas, allowing the future use of biotechnologies such as artificial insemination.

Susceptibility of Stallion Spermatozoa to Different Oxidative Challenges: Role of Seminal Plasma

Abstract Seminal cryopreservation provides several advantages in horse breeding. However, improvement on post‐thaw sperm survival is still necessary. One of the main factors known to impair post‐thaw quality of stallion’s sperm is the oxidative damage caused by reactive oxygen species (ROS). In this context, the aim of this study was evaluated the effects of ROS on stallion sperm and the possible influence of seminal plasma (SP). Toward this aim, 13 ejaculates from adult stallions (n = 13) were divided into two aliquots which were centrifuged (600g/10 minutes), and the SP was removed and reserved. Pellets were suspended in physiological saline solution (A) or SP (B). Each of these solutions was divided into four aliquots and subjected to challenge with different ROS (superoxide anion, hydrogen peroxide, and hydroxyl radical [OH−]) and the toxic by‐product of lipid peroxidation (malondialdehyde [MDA]). Samples were then evaluated for the susceptibility to lipid peroxidation (thiobarbituric acid reactive substances) plasma membrane integrity (eosin–nigrosin), acrosome integrity (Fast Green/rose bengal), mitochondrial activity (3′3 diaminobenzidine), and DNA integrity. Samples incubated in the presence of SP were highly impaired by OH− with regard to motility, plasma membrane integrity, mitochondrial activity, and DNA integrity. On the other hand, in the absence of SP, MDA was highly deleterious, especially with regard to motility, plasma membrane integrity, and mitochondrial activity. Thus, these results indicate that SP may have an important role on the protection of stallion sperm against the damages caused by MDA, an important product of lipid peroxidation. HighlightsSeminal plasma (SP) has a protective effect against oxidative damages.Hydroxyl radical in the presence of SP is the most deleterious reactive oxygen species (ROS) to equine spermatozoa.Malondialdehyde is more deleterious than ROS to equine spermatozoa in the absence of SP.Hydroxyl radical is extremely deleterious to equine sperm DNA.Malondialdehyde impaired membrane integrity and mitochondrial activity.

Análise espermática de perdizes (Rhynchotus rufescens) criadas em cativeiro e suplementadas com selênio

Due to the commercial importance of the red-winged tinamou (Rhynchotus rufescens), for the past few years, the employment of reproductive biotechnologies has been attempted. Thirty animals were randomly assigned into two groups: control group (no selenium) and treatment group (supplemented with 0,2 a 0,8 mg selenium/ 1000 kg ration). Animals were allocated at the FCAV - UNESP/Jaboticabal (2007-2008). Semen collections were performed by digital manipulation and divided in pools of at least 150 µL. After the immediate evaluation of motility, vigour, concentration and morphology, an aliquot of 20 µL was diluted in 300 µL of physiologic solution in order to test acrosome and membrane integrities, which were performed by counting 200 cells for each test. Cells were evaluated as follows: 1) Intact acrosome: lilac acrosome; Non-intact acrosome: pink acrosome; 2) Live cells: non stained; Dead: stained. Data was statistically analysed using the SAS System for Windows. No differences were found between treatment and control groups for volume, motility, vigour, mean number of spermatozoa per animal, concentration, Intact acrosome, Intact membrane. The difference found on midpiece sperm defect (Se = 1,33 ± 0,53 and control = 3,78 ± 0,69, p = 0.0107) may be due to the damages caused by the selenium deficiency to the architecture of the midpiece, which compromises sperm mobility and fertilization capacity.

Extender Supplementation with Antioxidants Selected after the Evaluation of Sperm Susceptibility to Oxidative Challenges in Goats

ABSTRACT This study aimed to detect the most deleterious ROS for goat sperm and then supplemented the extender with a proper antioxidant. For this, 12 adult goats (aged 1–7) were used. Fresh samples were submitted to challenge with different ROS (superoxide anion, hydrogen peroxide, and hydroxyl radical) and malondialdehyde (MDA—toxic product of lipid peroxidation). After experiment 1, sperms were cryopreserved in extenders supplemented to glutathione peroxidase (Control: 0 UI/mL; GPx1: 1 UI/mL; GPx5: 5 UI/mL, and GPx10: 10 UI/mL) and catalase (Control: 0 UI/mL; CAT60: 60 UI/mL; CAT120: 120 UI/mL, and CAT240: 240 UI/mL). Each sample was evaluated by motility, plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, assay of the sperm chromatin structure, mitochondrial activity (3,3-diaminobenzidine), and measurement of lipid peroxidation (thiobarbituric acid reactive substances [TBARS]). It was possible to observe a mitochondrial dysfunction (DAB—Class IV) and low membrane integrity after hydrogen peroxide action. However, the high rates of TBARS were observed on hydroxyl radical. CAT240 presents the lower percentage of plasma membrane integrity. It was possible to attest that hydrogen peroxide and hydroxyl radical are the more harmful for goat sperm. Antioxidant therapy must be improving perhaps using combination between antioxidants.

Effect of Vitamin E and Polyunsaturated Fatty Acids on Cryopreserved Sperm Quality in Bos taurus Bulls Under Testicular Heat Stress

ABSTRACT Taurine bulls are highly susceptible to heat stress, leading to increased oxidative stress (OS) and impaired sperm viability. Polyunsaturated fatty acids (PUFAs) supplementation can be an alternative to improve semen quality, which also results in more sperm susceptibility to lipid peroxidation. Moreover, this deleterious effect can be exacerbated in animals affected by heat stress. Vitamin E is a key antioxidant that counteracts lipid peroxidation of sperm membrane caused by OS. Thus, combining PUFAs with vitamin E may improve sperm quality. In this context, this study aimed to evaluate the effect of interaction between PUFAs and vitamin E on sperm quality in Bos taurus bulls under testicular heat stress. Sixteen taurine bulls under testicular heat stress were randomly assigned in four groups: Control, Vitamin E, PUFA, and PUFA + Vitamin E. All groups lasted for 60 days. Samples were cryopreserved/thawed and analyzed for motility variables (CASA), membrane and acrosome integrity, mitochondrial activity, susceptibility to oxidative stress, DNA integrity, and sperm-binding capacity. Results showed that vitamin E had a beneficial effect on some sperm characteristics, whereas PUFA supplementation had an adverse effect when the two treatments were evaluated separately. Finally, the association between PUFAs and vitamin E did not improve sperm quality.