Colloid Centrifugation Selects Normal Spermatozoa from Polymorphic Bull Ejaculates: A Case Study

Semen from a Western Finncattle bull exhibiting a highly polymorphic spermiogram was processed by colloid centrifugation using Androcoll‐B, a species‐specific silane‐coated silica colloid. In the first experiment, Single Layer Centrifugation (SLC) was used to identify which density colloids were needed to separate different cell populations. Colloids of the two chosen densities were then used in a density gradient resulting in two sperm subpopulations, one containing nearly all normally sized spermatozoa and the other enriched for the macrocephalic spermatozoa. Microcephalic spermatozoa did not appear in either of the selected subpopulations. Using a combination of SLC and DGC with this species‐specific colloid, it was possible to separate the spermatozoa into different subpopulations, that is, a subpopulation containing nearly all normally sized spermatozoa, and another one enriched for the macrocephalic spermatozoa. Thus, colloid centrifugation could be used to select sufficient normal spermatozoa from a highly polymorphic ejaculate for AI, if desired.     

Endometrial Expression of IFNAR‐1 and Oxytocin Receptor (OTR) is not Improved by Prostaglandin Analogues when Compared to Progestagens in Ewes

The objective of this study was to investigate differences on the endometrial immunoexpression of type I IFN receptor subunit 1 (IFNAR1) and oxytocin receptor (OTR) during the time of maternal recognition of pregnancy in sheep, when oestrus is synchronized with either prostaglandin analogues (group PG) or conventional progestagens (group P). Plasma progesterone was measured from day 0 to 21 post‐coitus (pc) (day 0 = day of oestrus). Immunohistochemistry was performed in samples of uterine horns from pregnant sheep on days 9pc, 13pc, 15pc, 17pc and 21pc to locate IFNAR1 and OTR expression in different endometrial compartments. Mean levels of plasma progesterone were different between treatments, obtaining higher levels in the PG group than in the P group (p < 0.05). Comparing days of pregnancy, IFNAR1 protein expression was different in the luminal epithelium (LE) (p < 0.05), while OTR was different in the LE and in the superficial glandular epithelium (SG) (p < 0.05). Temporal variation on the expression of both proteins from day 9pc to 21pc has been evidenced. IFNAR1 and OTR expression did not show significant differences between treatments. However, the response observed in the endometrium was highly inconsistent when prostaglandin analogues were used. Therefore, the protocol based on prostaglandin analogues still needs to be optimized before being considered as a better alternative to progestagens for oestrous synchronization in sheep.     

Pyometra and unilateral uterine horn torsion in a sheep

A 13‐year‐old Romanov sheep presented for evaluation of vaginal discharge, depression and anorexia. Blood work demonstrated mature neutrophilia and marked hyperlactatemia. Transrectal and transabdominal ultrasound revealed echogenic fluid (pus) within the uterus. Purulent uterine fluid culture yield Escherichia coli. A diagnosis of pyometra was made. After medical treatment without clinical improvement, an exploratory laparotomy was performed, and a 360‐degree uterine left horn torsion was identified with the pyometra and multiple corpora lutea on both ovaries. Ovariohysterectomy was performed and the ewe recovered uneventfully.     

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.     

Vaccine efficacy for reducing turbinate atrophy and improving growth rate in piggeries with endemic atrophic rhinitis

Two vaccines, based on formalin-killed whole cells of toxigenic Pasteurella multocida type D and Bordetella bronchiseptica combined with a partially toxoided cell extract of P multocida, were prepared with Freund's incomplete adjuvant (vaccine 1) or by alum precipitation (vaccine 2). Each was tested for safety and efficacy in reducing the severity of nasal turbinate atrophy and improving the growth rate of pigs in three Western Australian commercial piggeries with endemic atrophic rhinitis. In safety experiments with vaccine 1, no adverse clinical effects were observed in vaccinated sows or their progeny. Piglets receiving vaccine 2 showed no injection site abnormalities, pyrexia or turbinate atrophy. In field trials, vaccine 1 significantly reduced the prevalence of moderate to severe nasal turbinate atrophy (Done score 3 to 5) when used in two piggeries (A and B). Progeny from vaccinated sows in piggery B also grew significantly faster than controls. When vaccine 2 was used in piggery A at a later date and in another piggery (C), growth rate was not improved in either piggery and the prevalence of moderate to severe turbinate atrophy was reduced only in piggery C.     

Timing of Early Foetal Loss for Single and Twin Pregnancies in Dairy Cattle

The incidence of early foetal loss is increasing under intensive management systems for dairy cattle. The aims of the present study were to determine whether there is any peak period of pregnancy loss during the early foetal period and to evaluate possible differences between single and twin pregnancies. The study population consisted of 1442 pregnant cattle from a single herd. Pregnancy was diagnosed by transrectal ultrasonography between 36 and 42 days after insemination, and then weekly until day 90 of gestation or until pregnancy loss. A total of 1310 cows (90.8%) bore single embryos and 132 (9.2%) carried twins. Pregnancy loss was registered in 139 (9.6%) cows before day 90 of pregnancy: 101 (7.7%) in single and 38 (28.8%) in twin pregnancies. The average time of pregnancy loss for all animals was 58.4 ± 12.6 days and ranged from 45 to 90 days. Seventy‐five per cent of the pregnancy losses were registered between 45 and 60 days of gestation. The average time of pregnancy loss for cows with singletons was 52.1 ± 4.1 days and ranged from 45 to 61 days and that for those with twins was 75.1 ± 12.4 days and ranged from 46 to 90 days. Seventy‐five per cent of the twin pregnancy losses were registered between 68 and 90 days of gestation. Our data show that the foetal loss in singleton pregnancies occurs earlier than in twin pregnancies. Assessment of normal development of gestation on days 60 and 90 after insemination is suggested.