Incidence of Unilateral Fertilizations after Low Dose Deep Intrauterine Insemination in Spontaneously Ovulating Sows under Field Conditions

A new procedure for non-surgical deep intrauterine insemination (DUI) in unrestrained sows hormonally induced to ovulate, has been reported. In comparison with standard artificial insemination (AI), with this procedure, the sperm numbers inseminated can be reduced 20-fold without reducing the reproductive performance of these hormonally treated sows. The present study evaluated, using two experiments, the reproductive performance applying 20-fold different sperm numbers per AI dose using DUI or standard AI in spontaneously ovulating sows, under field conditions. In experiment 1, AI was applied to crossbred sows at 12, 24 and 36 h after onset of spontaneous oestrus using one of the following two regimes: (i) DUI (treatment) with 0.15 x 10(9) fresh boar spermatozoa in 5 ml of Beltsville thawing solution (BTS) extender (n = 95), and (ii) standard cervical AI (control) with 2.85 x 10(9) fresh spermatozoa in 95 ml of BTS extender (n = 95). The farrowing rates of the two groups of sows were statistically similar (NS). However, a decrease (p < 0.002) in litter size and the total number of pigs born alive was observed in sows inseminated with the DUI procedure. In experiment 2, 42 post-weaned oestrus sows were inseminated following the same design described for experiment 1 during spontaneous oestrus. On day 6 after onset of oestrus, the proximal segment of the uterine horns of the sows were flushed under surgery to retrieve eventual embryos and evaluate the success of fertilization per cornua (e.g. occurrence of effective uni- vs bilateral sperm transport rendering uni- or bilateral, complete or partial fertilization). Retrieved embryos were assessed for cleavage and number of accessory spermatozoa. Although identical overall pregnancy rates were achieved in both insemination groups, the percentage of sows with partial bilateral fertilization and unilateral fertilization was markedly higher (p < 0.05) in the DUI group (35%) compared with the control (standard AI) group (5%), with a consequent lower (p < 0.001) percentage of viable early embryos after DUI. The number of accessory spermatozoa in the zona pellucida of the embryos was highly variable, but higher (p < 0.001) in control animals than in DUI-AI. No accessory spermatozoa were found in oocytes retrieved from sows depicting unilateral fertilization. In conclusion, DUI in spontaneously ovulating sows with 0.15 x 10(9) spermatozoa renders similar farrowing rates but a lower litter size compared with use of standard AI with a 20-fold higher sperm dose. The lower litter size ought to be related to a decreased distribution of spermatozoa after DUI leading to a higher incidence of partial bilateral and unilateral fertilization.     

Functional Activity of Frozen Thawed Chinchilla lanigera Spermatozoa Cryopreserved with Glycerol or Ethylene Glycol

The cryopreservation of spermatozoa constitutes a valuable tool for the captive breeding management of valuable and/or threatened species. Chinchilla lanigera is a species almost extinct in the wild, and the domestic counterpart has one of the most valuable pelts in the world. The objectives of this study were to: (i) compare the functional activity of post‐thawed chinchilla spermatozoa cryopreserved at ?196°C either with glycerol (G) or ethylene glycol (EG) as cryoprotectants (1 m final concentration) and (ii) investigate the effects of incubating the gametes for 4 h in the presence or in the absence of the cryoprotectants; evaluations were performed taking into account motility, viability, response to hypo‐osmotic shock and acrosome integrity of the cells. Parameters reflecting post‐thaw (0 h) sperm functional activity were significantly lower than those of freshly ejaculated gametes. When comparing the cryoprotectant efficiency of G vs EG, neither cryoprotectant agent offered appreciable advantages. After 4 h of incubation, in the presence or absence of the cryoprotectant agent, a rapid and significant decrease was found in all functional parameters and remained at ～ 20–30% motile, viable and viable acrosome intact cells. Viability was significantly lower when the cryoprotectant was removed from the media (possibly due to the centrifugation process). With respect to the maintenance of sperm membrane integrity, only ～ 10% of cells showed membrane resistance to hypo‐osmotic conditions after the 4 h incubation period. These results constitute new insights for cryopreservation protocols and the development of assisted reproductive techniques in this species.     

Meningoencephalomyelitis in a foal due to Salmonella agona infection

CASE HISTORY: A neonatal Thoroughbred foal was presented with rib fractures and left forelimb lameness secondary to dystocia.

CLINICAL FINDINGS: The foal developed a head tilt, seizures and watery diarrhoea during hospitalisation and died at 7 days of age. Histological examination of the brain and spinal cord revealed a suppurative meningoencephalomyelitis with vasculitis, and numerous intralesional, gram-negative bacilli. Similar microscopic lesions were noted in the lungs, renal medullary interstitium, and umbilicus. Bacilli in the brain, spinal cord and umbilicus were identified immunohistochemically as Salmonella group B. Salmonella agona was isolated in pure culture from the brain, lung, liver, kidney, and intestine.

CONCLUSION: This is the first report of meningoencephalomyelitis and septicaemia due to Salmonella infection in an equine neonate.     

Effect of Overnight Staining on the Quality of Flow Cytometric Sorted Stallion Sperm: Comparison with Tradtitional Protocols

Flow cytometry is considered the only reliable method for the separation of X and Y chromosome bearing spermatozoa in equines. The MoFlo SX DP sorter is highly efficient, allowing the production of foals of the desired sex. However, to achieve acceptable pregnancy rates the currently used protocol requires working with fresh semen obtained close to, or at, the sorting facility. An alternative protocol was tested during two consecutive breeding seasons. Fresh stallion semen was cooled for 20 h, during which staining with Hoechst 33342 took place. On the following day, this sample was flow sorted and compared with spermatozoa from the same ejaculate that had been sexed on the previous day. All sperm parameters evaluated remained unchanged when fresh sorted and refrigerated sorted semen were compared. Pre‐sorting storage at 5°C did not alter sperm velocities nor kinetics, viability or membrane permeability, production of reactive oxygen species, mitochondrial membrane potential or DNA fragmentation index of the sorted sample. The findings open for the possibility of using semen from stallions housed far from the sorting facilities. Processed and stained sperm could be shipped refrigerated on the previous day, sorted and inseminated on the next day.     

Biogeochemical evidence for dinoflagellate ancestors in the early cambrian

Dinoflagellates are single-celled organisms that reflect the ecological conditions in modern oceans and lakes. Ultrastructural data and molecular phylogeny suggest that they originated in the Neoproterozoic, yet dinoflagellate ancestors are classified only to the Middle Triassic ( approximately 240 million years ago). Examination of dinoflagellate-specific biological markers (dinosteranes and 4alpha-methyl-24-ethylcholestane) in concentrated microfossils with known morphology identified ancient dinoflagellate ancestors from the Early Cambrian ( approximately 520 million years ago).     

Modulation of diversity by grazing and mowing in native tallgrass prairie

Species diversity has declined in ecosystems worldwide as a result of habitat fragmentation, eutrophication, and land-use change. If such decline is to be halted ecological mechanisms that restore or maintain biodiversity are needed. Two long-term field experiments were performed in native grassland to assess the effects of fire, nitrogen addition, and grazing or mowing on plant species diversity. In one experiment, richness declined on burned and fertilized treatments, whereas mowing maintained diversity under these conditions. In the second experiment, loss of species diversity due to frequent burning was reversed by bison, a keystone herbivore in North American grasslands. Thus, mowing or the reestablishment of grazing in anthropogenically stressed grasslands enhanced biodiversity.     

Neptune's eccentricity and the nature of the kuiper belt

The small eccentricity of Neptune may be a direct consequence of apsidal wave interaction with the trans-Neptune population of debris called the Kuiper belt. The Kuiper belt is subject to resonant perturbations from Neptune, so that the transport of angular momentum by density waves can result in orbital evolution of Neptune as well as changes in the structure of the Kuiper belt. In particular, for a belt eroded out to the vicinity of Neptune's 2:1 resonance at about 48 astronomical units, Neptune's eccentricity can damp to its current value over the age of the solar system if the belt contains slightly more than an earth mass of material out to about 75 astronomical units.