Comparison between Cloprostenol-induced and Spontaneous Oestrus Fertility in Dairy Cows

A short calving to conception interval is of main importance to achieve high economic efficiency in dairy cow industry. In order to reduce this interval, several hormonal treatments have been put on the market, in which cloprostenol, a synthetic analogue of prostaglandin F2alpha (PGF2alpha). The aim of this study was to compare fertility of cloprostenol-induced oestrus to that of spontaneous oestrus in dairy cows. In a group of 525 cows, 280 (treated group) were administered 0.5 mg cloprostenol i.m. after transrectal corpus luteum (CL) detection, and inseminated at detected oestrus during the following week. The other 245 cows (control group) were inseminated during spontaneous oestrus. Whey progesterone concentrations were checked at treatment and at insemination in order to remove from the study cows whose P4 levels indicate a non-functional CL, or a lack of luteolysis respectively. Moreover, cows that were not inseminated due to genital problems were also excluded from this study. Conception (59% vs 54.5%) and calving rates (93.7% vs 93%) were not significantly different between the two groups.     

Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.     

Refrigerated Storage of Red Deer Epididymal Spermatozoa in the Epididymis, Diluted and with Vitamin C Supplementation

We have approached the problem of refrigerated storage of epididymal sperm samples from red deer by comparing three options: storing the genital (testicles within the scrotum), diluting the semen in extender or diluting the semen in extender supplemented with an anti-oxidant. Twenty-nine pairs of testes were collected. Spermatozoa from one of each of the pairs were immediately recovered, and diluted to 400 × 10⁶ sperm/ml in Tris-citrate-fructose with 20% egg yolk. Control group was stored as such, and Anti-oxidant group was supplemented with 0.8 m m vitamin C. The remaining epididymides and the diluted samples were stored at 5°C and spermatozoa were analysed at 0, 24, 96 and 192 h for: motility [computer-assisted semen analysis (CASA)], acrosomal integrity, sperm viability (eosine/nigrosine staining), normal tails and chromatin status [sperm chromatin structure assay (SCSA)]. In general, seminal quality decreased with storage time. Vitamin C supported progressive motility better at 24 h (median 42% vs 23% Control and 15% epididymis), reduced the incidence of tail abnormalities and protected chromatin. Storing the semen in the epididymis slowed down motility loss, but slightly increased the occurrence of tail abnormalities and viability was lower at 192 h. However, regarding chromatin status, sperm stored in the epididymis was protected similarly to those diluted in the medium supplemented with vitamin C. Although the differences between the three groups were small, there were some advantages in supplementing the extender with vitamin C. Besides, refrigerating the epididymis may be a good option when immediate processing is not available.     

Mid‐gestation pregnancy is not disrupted by a 5‐day gastrointestinal mucosal cytoprotectant oral regimen of misoprostol

Reasons for performing study: To investigate effects of a 5‐day oral misoprostol regimen recommended for use in horses as a gastrointestinal mucosal cytoprotectant during colic on mid‐gestation pregnancies. Objectives: To monitor cervical tone, ultrasonographic characteristics of the uterus, cervix and conceptus, as well as serum progesterone and oestrone sulphate concentrations, and observations of general health, behaviour and comfort of mid‐gestation mares given a 5‐day course of misoprostol or control treatment. Methods: Eleven light horse and pony mares with known breeding dates were administered 5 µg/kg bwt misoprostol orally, twice daily for 5 days. General health and pregnancy status were monitored daily during treatment via general physical examination, as well as palpation and ultrasonography per rectum of the uterus, cervix and conceptus. Jugular serum was obtained during and for 5 days following treatment for assay of progesterone and oestrone sulphate concentrations. Additionally, daily 12 h video samples of the mares were obtained to evaluate behaviour and comfort. Results: All findings, including cervical tone, ultrasonographic characteristics of the uterus, cervix and conceptus, as well as progesterone and oestrone sulphate concentrations, and observations of general health, behaviour and comfort, were similar during misoprostol and control treatment. Conclusions: Treatment of pregnant mares with a gastrointestinal mucosal cytoprotectant regimen of oral misoprostol for 5 days did not disrupt pregnancy, nor adversely affect the general health and comfort of these mares. Additional investigation of treatment at earlier and later stages of gestation, for longer‐term treatment, as well as evaluating neonates for developmental disturbances, would add further information on safety of misoprostol during gestation. Potential relevance: These results provide some assurance of safety of a 5‐day gastrointestinal mucosal cytopretectant regimen of oral misprostol in mid‐gestation pregnant mares.     

Expression of Occludin in Testis and Epididymis of Wild Rabbits, Lepus sinensis coreanus

Tight junctions (TJs) in inter-Sertoli junctional areas and epididymal epithelia build up the blood–testis barrier (BTB) and the blood–epididymal barrier (BEB), respectively. In this study, the expression of occludin, an integral member of the TJs, was examined in testis and different regions of epididymis of Lepus sinensis coreanus , an Korean wild rabbit species. In testis, intense occludin immunoreactivity was found in the basally located inter-Sertoli junctional area together with diffused immunoreactivity of occludin in the cytoplasm of Sertoli cells. It can be suggested that occludin is one of the robust elements of BTB in seminiferous tubules of rabbit testis. In proximal and distal caput epididymis, occludin immunoreactivity was found in the lateral as well as apical contacts of epithelial cells. In corpus epididymis, intense occludin immunoreactivity was found in the basolateral as well as apical contacts of epithelial cells together with cytoplasmic signal. In cauda epididymis, occludin immunoreactivity in luminal epithelia was relatively strong but largely found in the cytoplasm. This suggests that intriguing regulatory mechanisms differentially recruit occludin to the TJ in the different regions of epididymal epithelia. The differences in the subcellular localization as well as expression levels of occludin among the epididymal segments may reflect differential paracellular permeability of epithelia along the epididymal tubules and be correlated with sperm maturation in rabbit. In Western blot, a major form of occludin was MW 62 kDa together with small fragments of MW 34–39 kDa in testis and epididymis, suggesting the peptide cleavage of occludin. This is the first report on the molecular nature of TJs in a wild rabbit testis and epididymis.     

Morphology of alpaca (Vicugna pacos) embryos in the first third of pregnancy

The breeding of South American camelids is the main economic activity of the high Andean region of South America and it, is potentially, the most profitable resource in of the Puna environmental conditions of the Puna. The duration of the gestation in alpaca is 339.7 ± 12 days. The objective of the present work was to macroscopically and microscopically describe the ontogenic development of the splanchnic cavities of the alpaca and to determine the gestational time in which the post‐cranial ossification centers are observed in the embryos/fetuses of this species, from day 21 to 107 of gestation. The documentation of normal ontogenic development, which is vacant for this period, is of the utmost importance to understand the consequences of the alterations at the different gestational times, as well as for the estimation of the gestational age in the case of abortions. Forty‐seven alpaca specimens of both sexes, at different times of their gestational development, collected during slaughter at local slaughterhouses of the Department of Huancavelica, Peru, were evaluated. Specimens were assigned to seven groups according to their morphological characteristics. The embryogenesis in the alpaca was characterized by a series of changes comparable to those occurring in other mammals with similar gestational periods. Despite these similarities, species differences were found in some organs as stomach, which are observed too in adult individuals.     

Influence of Post‐Mortem Sperm Recovery Method and Extender on Unstored and Refrigerated Rooster Sperm Variables

Many post‐mortem sperm collection techniques have been described for mammalian species, but their use in birds is scarce. This paper compares the efficacy of two post‐mortem sperm retrieval techniques ‐ the flushing and float‐out methods ‐ in the collection of rooster sperm, in conjunction with the use of two extenders, i.e., L&R‐84 medium and Lake 7.1 medium. To determine whether the protective effects of these extenders against refrigeration are different for post‐mortem and ejaculated sperm, pooled ejaculated samples (procured via the massage technique) were also diluted in the above extenders. Post‐mortem and ejaculated sperm variables were assessed immediately at room temperature (0 h), and after refrigeration at 5°C for 24 and 48 h. The flushing method retrieved more sperm than the float‐out method (596.5 ± 75.4 million sperm vs 341.0 ± 87.6 million sperm; p < 0.05); indeed, the number retrieved by the former method was similar to that obtained by massage‐induced ejaculation (630.3 ± 78.2 million sperm). For sperm collected by all methods, the L&R‐84 medium provided an advantage in terms of sperm motility variables at 0 h. In the refrigerated sperm samples, however, the Lake 7.1 medium was associated with higher percentages of viable sperm, and had a greater protective effect (p < 0.05) with respect to most motility variables. In conclusion, the flushing method is recommended for collecting sperm from dead birds. If this sperm needs to be refrigerated at 5°C until analysis, Lake 7.1 medium is recommended as an extender.