Repeat Ovum Pick-up and In Vitro Embryo Production from Adult Ewes with and without FSH Treatment

Ovum pick-up (OPU) was performed three times on adult ewes after synchronization with (n = 4) or without (n = 4) FSH treatment to investigate the effects of FSH treatment on the number of ovarian follicles, oocytes recovered, oocyte quality and development in vitro. FSH treatment increased the number of ovarian follicles (85 vs 162) and oocytes recovered (33 vs 91), although recovery rate was similar for ewes with and without FSH (91/162, 56.2% and 33/85, 38.8% respectively). Of the oocytes recovered, those classified as grades I and II were similar between ewes with (78/91, 85.7%) and without FSH treatment (27/33, 81.8%). The number of ovarian follicles was similar after repeated OPU for ewes treated with FSH, but for ewes not treated with FSH the number of ovarian follicles decreased with repeated OPU. The number of oocytes recovered decreased for FSH-treated ewes only, while the oocyte recovery rate and proportion of oocytes classified as grades I and II were not affected by repeated OPU. Oocyte cleavage (46/78, 58.9% and 19/24, 79.2%) and blastocyst formation (35/46, 76.1% and 12/19, 63.2% respectively) were similar for ewes with and without FSH treatment. The number of ovarian follicles varied between ewes (p < 0.05) although the number of oocytes recovered and oocyte development in vitro were similar between ewes.     

Production and processing studies on calpain-system gene markers for beef tenderness: consumer assessments of eating quality

We investigated the effects of calpain-system genetic markers on consumer beef quality ratings, including interactions of marker effects with hormonal growth promotant (HGP) use and tenderstretch hanging. Brahman cattle in New South Wales (NSW; n = 164) and Western Australia (WA; n = 141) were selected at weaning from commercial and research herds to achieve balance and divergence in calpastatin (CAST) and calpain 3 (CAPN3) gene marker status. Genotypes for μ-calpain (CAPN1-4751 and CAPN1-316) were also determined. Angus cattle (49 in NSW, 17 in WA) with favorable CAST and CAPN3 alleles, balanced for CAPN1-316 status, were also studied. Half the cattle at each site had HGP (Revalor-H, containing 200 mg trenbolone acetate and 20 mg 17β-estradiol) implants during grain finishing. One side of each carcass was suspended from the Achilles tendon (AT) and the other from the pelvis [tenderstretch (TS)]. Meat Standards Australia consumer panels scored 7-d aged striploin steaks from both AT and TS sides, and 7-d aged rump and oyster blade steaks from the AT side of each carcass. Two favorable CAST alleles increased tenderness ratings of AT-striploin, TS-striploin, rump, and oyster blade steaks by, respectively, 6.1, 4.2, 4.2, and 3.1 units, and overall liking by 4.7, 2.8, 2.9, 3.7 (all P < 0.04). Two favorable CAPN1-4751 alleles increased tenderness of AT-striploin, TS-striploin, and rump steaks by 6.5, 4.3, and 3.9 units, and overall liking by 5.6, 3.1, and 4.1 units. Two favorable CAPN3 alleles improved rump steaks by 3.7, 3.3, 3.7, and 3.5 units, for tenderness, juiciness, liking the flavor, and overall liking. There were no significant CAPN1-316 effects. The effect of HGP was greatest for the AT-striploin (reducing tenderness and overall liking by 8.2 units, P < 0.001), then TS-striploin (-5.6 for tenderness, -5.0 for overall liking, P < 0.001), and then rump (-4.4 for tenderness, -3.3 for overall liking, P < 0.007). Processing conditions differed considerably between NSW and WA. Rump steaks from NSW scored about 10 units greater than those from WA, but Angus and Brahman steaks from the same location with the same marker alleles had similar scores. In contrast, NSW Angus striploin steaks scored about 15 units greater for tenderness and overall liking (P < 0.001) than cattle with the same marker alleles at the other 3 location × breed combinations, which had generally similar scores. Therefore, calpain-system gene markers have beneficial effects on eating quality, consistent with our previous findings for objective meat quality.     

Prevalence of dental and mandibular disorders in culled sheep in Spain

At the slaughterhouse, 1465 culled sheep heads were studied in order to identify disorders of the mandibular cheek teeth. Of these, 227 (15.5%) had evidence of osteomyelitis. The lesional profile showed that the mandible was affected in a similar proportion on both sides (46.7% on the right side vs 50.7% on the left side), mainly in the middle region (55.3%) and with most of the lesions closed without fistulisation (89.4%). In addition, swelling was palpable, with an increase in thickness in the area of the affected body (2.65 ± 0.065 cm vs 1.74 ± 0.030 cm). In 78.6% of the animals, the regional lymph nodes were enlarged. Concerning the mandibular cheek teeth, more than half of the animals lacked at least one tooth (57.0%), with the first premolar being the most frequently missing tooth (34.8%) and the third molar the least (8.2%). The impaction of food around the teeth was very common with the posterior molars being more frequently affected. In the knowledge of the authors, this is the first study that analyses the prevalence of mandibular molar and premolar disorders in sheep, and these are revealed as an important condition affecting culled sheep.     

Immunohistochemical Expression of Cyclooxygenase‐2 (COX‐2) in Feline Endometrial Adenocarcinoma and in Normal and Hyperplastic Endometria

Cyclooxygenase‐2 (COX‐2) is overexpressed in several human and animal neoplasms, including the human endometrial carcinoma. It has been suggested as a prognostic marker and a potential therapeutic target. This study aimed to (i) clarify histological aspects of feline endometrial adenocarcinomas (FEA) of the papillary serous type and (ii) characterize COX‐2 immunohistochemical expression in normal, hyperplastic and neoplastic endometrium in this species. Archived paraffin‐embedded tissue samples of 33 FEA, eight cystic endometrial hyperplasias (CEHs) and 21 samples of normal, healthy endometrium in the follicular (FS; n = 10) and luteal (LS; n = 11) stages were evaluated. Histological evaluation of haematoxylin and eosin‐stained sections of the FEA revealed a papillary proliferation of neoplastic cells of serous type, accompanied by clear and multinucleated cells. Other architectural arrangements mainly included solid and tubular growth. Randomly distributed areas of necrosis within the tumours were commonly observed. Invasion of the myometrium, of the serosa and of the vascular and/or lymphatic vessels was not constant features. The mean number of mitoses was higher in FEA compared to non‐neoplastic endometrium. COX‐2 scores were lower in FEA (p = 0.003) and CEH (p = 0.05) when compared to normal epithelium (NE). The loss of the membrane apical reinforcement in epithelial cells was observed in FEA samples, which was accompanied by the dislocation of COX‐2 labelling into the cytoplasm and the perinuclear area; in contrast, in epithelial cells in the healthy and hyperplastic endometria, the immunoreaction showed the characteristic pattern of apical membrane reinforcement, suggestive of the membrane polarization. COX‐2 epithelial scores were higher in the FS than in the LS. No differences were found in stromal COX‐2 expression between normal, CEH and FEA groups, but it was higher in the LS than in the FS. In summary, loss of COX‐2 compartmentalization in neoplastic epithelial cells might be one of the molecular events underlying endometrial carcinogenesis.     

Unusual ovarian activity in a mare preceding the development of an ovarian granulosa cell tumour

An 8-year-old mare, with a foal at foot, was inseminated on foal heat with frozen semen, with the resultant pregnancy lost between days 34 and 41. The right ovary developed a large anovulatory follicle that was non-responsive to multiple doses of ovulating agents. The follicle eventually appeared to luteinise, although plasma progesterone concentrations did not reflect this. Another follicle developed, responded to GnRH and resulted in a pregnancy from frozen semen that went to term with a healthy foal. When the mare was examined after foaling, the structure on the right ovary appeared to be a granulosa cell tumour; the left ovary was smaller than normal and non-functional. Surgical removal of the right ovary before increasing photoperiod resulted in a return to function of the left ovary and a pregnancy to frozen semen on the second cycle following removal. Figures showing concentrations of inhibin, progesterone, androstenedione, oestradiol and testosterone are presented for this entire period. Unusual ovarian activity in the mare might be a prelude to the development of a granulosa cell tumour.     

Cattle temperament: persistence of assessments and associations with productivity, efficiency, carcass and meat quality traits

Relationships between temperament and a range of performance, carcass, and meat quality traits in young cattle were studied in 2 experiments conducted in New South Wales (NSW) and Western Australia (WA), Australia. In both experiments, growth rates of cattle were assessed during backgrounding on pasture and grain finishing in a feedlot. Carcass and objective meat quality characteristics were measured after slaughter. Feed intake and efficiency during grain finishing were also determined in NSW. Brahman (n = 82 steers and 82 heifers) and Angus (n = 25 steers and 24 heifers) cattle were used in the NSW experiment. In NSW, temperament was assessed by measuring flight speed [FS, m/s on exit from the chute (crush)] on 14 occasions, and by assessing agitation score during confinement in the crush (CS; 1 = calm to 5 = highly agitated) on 17 occasions over the course of the experiment. Brahman (n = 173) and Angus (n = 20) steers were used in the WA experiment. In WA, temperament was assessed by measuring FS on 2 occasions during backgrounding and on 2 occasions during grain feeding. At both sites, a hormonal growth promotant (Revalor-H, Virbac, Milperra, New South Wales, Australia) was applied to one-half of the cattle at feedlot entry, and the Brahman cattle were polymorphic for 2 calpain-system markers for beef tenderness. Temperament was not related (most P > 0.05) to tenderness gene marker status in Brahman cattle and was not (all P > 0.26) modified by the growth promotant treatment in either breed. The Brahman cattle had greater individual variation in, and greater correlations within and between, repeated assessments of FS and CS than did the Angus cattle. Correlations for repeated measures of FS were greater than for repeated assessments of CS, and the strength of correlations for both declined over time. Average FS or CS for each experiment and location (NSW or WA × backgrounding or finishing) were more highly correlated than individual measurements, indicating that the average values were a more reliable assessment of cattle temperament than any single measure. In Brahman cattle, increased average FS and CS were associated with significant (P < 0.05) reductions in backgrounding and feedlot growth rates, feed intake and time spent eating, carcass weight, and objective measures of meat quality. In Angus cattle, the associations between temperament and growth rates, feed intake, and carcass traits were weaker than in Brahmans, although the strength of relationships with meat quality were similar.     

Selected Ovarian Ultrasonographic Characteristics During Vernal Transition are Useful to Estimate Time of First Ovulation of the Year

It is important to get mares pregnant as early as possible after vernal transition and thus, identification signs of impending 1st ovulation of the year are warranted. To identify clinical indicators of an approaching first ovulation of the year, mares were teased with a stallion for oestrous detection starting January 3 and subjected to ultrasonographic examination. Day of first appearance of uterus oedema, follicular wall invagination, intrafollicular echogenicity, double contour of the follicle wall, increase in granulosa thickness, follicular wall hyperechogenicity and appearance of pear‐shaped follicles was registered, as well as follicle diameter and number. Seventy per cent of the mares had anovulatory oestrous periods of 4.6 ± 3.6 days, with an interoestroual interval of 12.5 ± 12.2 days. Number of anovulatory oestruses per mare was 2.4 ± 2.3. Uterine oedema occurred in 77% of the mares, 32.4 ± 25.6 days before ovulation. Invagination of the follicular wall appeared in 44.4% of the animals, 24.5 ± 18.4 days before ovulation. Intrafollicular echogenicity was seen in all mares and double contour of the follicle was seen in 77% of the animals. Both last two characteristics appeared 1–72 days before ovulation. Increased thickness of the granulosa occurred in 66% of the mares, 1–19 days before ovulation. Pear‐shaped follicles and follicular wall hyperechogenicity were detected 3 or less days before the first ovulation, in 44.4% and 55.5% of mares, respectively. Mean number of follicles >15 mm decreased at least 16 days before ovulation. We concluded that no isolated characteristic was a reliable indicator. However, increase in granulosa thickness, formation of a pear‐shaped follicle and follicular wall hyperechogenicity, associated with the reduction of the number of follicles >15 mm in diameter to <3, resulted in the first ovulation of the year in 44–67% of the transitional mares, 1–19 days after the characteristics appeared.     

Expression of Genes Associated with Apoptosis in the Porcine Corpus Luteum During the Oestrous Cycle

The corpus luteum (CL) of the pig lacks luteolytic sensitivity (LS) to prostaglandin (PG) F‐2α until after day 12 of the oestrous cycle, but the mechanisms underlying this phenomenon are poorly understood. As luteolysis involves apoptosis, we hypothesized that critical apoptotic proteins may be deficient in CLs that lack LS. The specific aim of these studies was to examine mRNA expression and protein levels of apoptosis genes/proteins (BAX/Bax, BCLX/Bcl‐x, CASP3/Caspase‐3, CASP8/Caspase‐8, NFΚB1/NFκB, TP53/p53) in porcine CLs collected at different stages of the oestrous cycle. CLs were collected surgically, mRNA and protein extracted, and expression/levels analyzed by semi‐quantitative (SQ) PCR and Western blots, respectively. At the mRNA expression level, only BAX (maximal on day 4) and TP53 (maximal on day 7) showed significant variations during the oestrous cycle. At the protein level, only Bcl‐x and Caspase‐3 showed significant changes during the cycle; Bcl‐x decreased on day 13 and Caspase‐3 increased on day 13. It is concluded that apoptosis‐associated proteins (i.e. Bcl‐x and Caspase 3) may play a critical role in luteolytic sensitivity in the pig.