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.     

Salmonella Enteritidis: the egg and I

SUMMARY: The world-wide clinical incidence of Salmonella Enteritidis has increased markedly. The increase is associated with the enhanced ability of the bacterium to systemically colonise layer chickens. Subsequent contamination and consumption of intact shell eggs from colonised layer hens, either directly or in foods containing raw or lightly cooked eggs, causes human disease. Despite investigation, no change in the biology of the bacterium has been correlated with increased colonisation in chickens. To date, no method of control at the production level has proven effective; consumer education is the best means of minimising the public health risk.     

Fertilisation and weed control effects on yield and weeds in durum wheat grown under rain-fed conditions in a Mediterranean climate

Summary A field study was undertaken to examine the effect of fertiliser type and weed control system on grain yield, yield components and weed population in durum wheat ( Triticum turgidum. var. durum) grown in a 4-year rotation [bare fallow–barley ( Hordeum vulgare )–vetch ( Vicia sativa )–wheat]. Fertilisation treatments were: no fertilisation, organic fertilisation with 2500 kg ha⁻¹ of compost (sheep manure and cereal straw) and chemical fertilisation with 100–60–60 kg ha⁻¹ of NPK. Weed-control treatments were: no control, herbicide, harrowing with long-flex spring tines and strip sowing with inter-row hoeing. Chemical fertilisation increased grain yield with respect to the other treatments, amongst which no significant differences were noted. Weed-control systems afforded no improvement in yield compared with controls; indeed, yields obtained using inter-row hoeing were lower. The weed population consisted of a large variety of species, of which the most important were Convolvulus arvensis and Polygonum aviculare . Application of weed-control systems reduced weed density. Herbicide was the most effective, reducing weed biomass by 80%, whilst harrowing and hoeing reduced weed biomass by 40% and 52% respectively. Inter-row hoeing does not appear to be a viable alternative to herbicides, when used as the sole weed control method in a non-diverse cropping system in Mediterranean climates. The lack of response to compost suggests a need for further long-term research.     

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.     

Clinical and Pathological Findings in Testis, Epididymis, Deferens Duct and Prostate following Vasectomy in a Dog

We report the case of a bilateral and multilocular spermatocele and sperm granuloma in a dog that was vasectomized 5 years before. Clinical examination revealed scrotal dermatitis and benign prostatic hyperplasia. Orchiectomy was performed, and gross and histological examination showed testicular degeneration associated with epididymal sperm granuloma. In relation to this case, the literature about long-term effects of vasectomy in dogs has been reviewed. On the basis of these results, a preventive sonogram and physical assessment in prostate and other reproductive structures before vasectomy is recommended.     

Effects of Breed on Follicular Dynamics and Oestradiol Secretion during the Follicular Phase in Sheep

Current study evaluates effects from breed and management background on follicular dynamics and endocrine output during the follicular phase of sheep oestrous cycle. Follicular phases were synchronized with three cloprostenol doses, 10 days apart, in three groups of 10 females of different non-prolific Spanish breeds (Manchega, Rubia del Molar and Negra de Colmenar). Development of all follicles reaching antral diameters >or=2 mm was assessed by daily transrectal ultrasonographies, whereas follicular function was evaluated by measurement of plasma oestradiol concentrations. All the ovulatory follicles were present at induced luteolysis in Manchega sheep, while a 93.7% were detected in Rubia del Molar and Negra de Colmenar ewes. The mean size of these ovulatory follicles was similar between breeds at 0 h, but their growth rates were higher in Manchega ewes, reaching a larger size at oestrous detection than in Negra de Colmenar and Rubia del Molar sheep (p < 0.05). Conversely, the oestradiol levels increased with time in Rubia del Molar and Negra de Colmenar (p < 0.05); whilst remained stable in Manchega females. However, the patterns of follicular turnover were similar between breeds. These results indicate that, though differences in follicular size and size distribution, patterns of follicular turnover in sheep are affected neither by the breed nor by the background of management and selection.