Activity of glycosidases (beta-N-acetyloglucosaminidase, alpha-mannosidase, and beta-galactosidase) in the uterine luminal fluid of cows after multiple ovulation.

This study investigated the activity of beta-N-acetyloglucosaminidase (beta-NAGASE), alpha-mannosidase, and beta-galactosidase in the uterine luminal fluid of cows after superovulation treatment, along with the possible associations between the activity of these 3 glycosidases and the superovulatory response. Embryos and a sample of fluid flushed from each uterine horn were collected on day 7 after artificial insemination (on estrus day 0) from 32 cows in which superovulation was induced with porcine follicle-stimulating hormone. Glycosidase activity was assayed colorimetrically. The cows were classified as to superovulatory response according to the number of corpora lutea per ovary (group 1, 1 to 4; group 2, > 4) and according to the total number of embryos per horn (T1, 0; T2, 1 to 2; T3, 3 to 4; T4, > 4) and the number of transferable embryos per horn (TR1, 0; TR2, 1 to 2; TR3, 3 to 4; TR4, > 4). The mean activity of beta-NAGASE was significantly lower (P < 0.05) in group 2 than in group 1, at 95.99 (standard error 20.43) versus 226.72 (46.77) IU/L. It was also significantly lower (P < 0.01) in group T4 compared with groups T1, T2, and T3, at 50.09 (8.21) versus 129.25 (34.60), 222.27 (62.62), and 290.26 (93.77) IU/L, respectively, as well as in group T1 compared with group T3. There was a positive relationship between beta-NAGASE activity and both the total number of embryos (P = 0.047) and the number of transferable embryos per horn (P = 0.013) when 1 to 4 corpora lutea developed per ipsilateral ovary. No difference in alpha-mannosidase or beta-galactosidase activity was detected among the groups.     

A study on ghrelin and LH secretion after short fasting and on ghrelin levels at perioestrual period in dairy cattle

In two experiments, we studied (a) the changes of LH secretion in heifers under different feeding schedules and (b) total ghrelin concentration at oestrus in cows and heifers. In experiment one, synchronized heifers were allocated in three groups (R, regularly fed controls; F, fasted; and F‐F fasted‐fed). One day after the completion of the oestrous induction protocol, group F and F‐F animals stayed without feed for 24 hr; thereafter, feed was provided to R and F‐F cattle; 2 hr later, GnRH was administered to all animals. Blood samples were collected for ghrelin, progesterone, LH and cortisol concentrations. Fasting caused increased ghrelin concentrations in groups F and F‐F, while in response to GnRH, LH surge was significantly attenuated in groups F and F‐F compared to R. In experiment 2, lactating cows and heifers were used. On day 9 of a synchronized cycle, PGF2α was administered, and blood samples were collected twice daily until the third day after oestrus and analysed for progesterone, estradiol, ghrelin, glucose and BHBA concentrations. No difference was recorded between groups in steroids and BHBA concentrations. In comparison to mid‐luteal values, ghrelin concentrations significantly increased at perioestrual period in cows, but not in heifers. This study provides evidence that starving‐induced elevated ghrelin concentrations can have suppressing effect on LH secretion, even after ghrelin's restoration to basal values and that during oestrus, ghrelin secretion is differently regulated in cows and heifers, likely being independent from oestradiol concentrations. Further research is required to identify the determining factors that drive the different regulation of ghrelin secretion in cows and heifers.     

Functional proteomic analysis of crossbred (Holstein Friesian × Sahiwal) bull spermatozoa

Male infertility is one of the prime concerns of dairy cattle production. The study was designed to find out differentially expressed proteins in categorized crossbred (Holstein Friesian × Sahiwal) bull semen to serve as potential biomarkers for male infertility. Frozen crossbred bull semen with satisfactory phenotypic records were defined as “good” and “poor” based on their fertility rates. A total of 1,547 proteins were detected in bull spermatozoa using liquid chromatography–mass spectrometer (LC‐MS/MS) analysis. Results revealed that 558 (36.1%) and 653 (42.2%) proteins were expressed to good and poor quality bull spermatozoa, respectively. A total of 336 proteins (21.7%) were reported to be unique for both good and poor quality bull semen, and among the common proteins, 224 (66.7%) and 112 (33.3%) were up‐ and downregulated in good and poor quality categorized bull semen, respectively. Gene Ontology analysis of global proteomes identified different signalling pathways, and most of them were related to cellular motility, immune systems as well as cellular metabolisms. The distinctive presence of some of the proteins may provide an insight into the molecular mechanistic role played by these proteins in crossbred bull infertility.     

The association of sub-clinical paratuberculosis with the fertility of Greek dairy ewes and goats varies with parity

Our cross-sectional study investigated the association of sub-clinical Mycobacterium avium subsp. paratuberculosis (MAP) infection with failing to produce a live offspring the season of lambing/kidding (November 2001 to January 2002) before testing (in April-May 2002), in four dairy-sheep and/or goat flocks in Greece (369 animals >or=1.5-year-old). From each selected animal 10 ml of blood and 10 g of feces from the rectum were obtained. The harvested sera were tested for antibodies to MAP with a commercial ELISA test kit; the feces were cultured on Herrold's egg-yolk medium supplemented with mycobactin J and antibiotics. An animal was considered sub-clinically infected when found either seropositive or culture positive. The true prevalence of sub-clinically infected animals, adjusted for the sensitivity and specificity of the parallel testing, was 14% (0.1-28%) and 35.9% (9.2-62.7%) in sheep and goats, respectively. The association of fertility of sheep and goats with sub-clinical paratuberculosis was investigated in random-effects logistic models. Sub-clinically infected animals (compared to uninfected) had OR for live offspring the previous year of 5.4 for parity <4, OR=0.05 for parity >6, and a non-significant OR for the middle parity category.     

Assessing the usefulness of B‐mode and colour Doppler sonography,and measurements of circulating progesterone concentrations for determining ovarian responses in superovulated ewes

The main goal of this study was to assess the usefulness of two imaging modalities, namely the B‐mode and colour Doppler sonography, and serum progesterone (P₄) concentrations for determining the ovarian response in superovulated ewes. Twenty‐four sexually mature Santa Inês ewes underwent the superovulatory treatment consisting of eight injections of porcine FSH (total dose of 200 or 133 or 100 mg; n  =  8 ewes/total dose) given at 12‐hr intervals and initiated 48 hr before CIDR ^® (Pfizer Inc., Auckland, New Zealand) removal. Six days after natural mating, the ovaries of all donor ewes were visualized and examined with transrectal ultrasonography and then with videolaparoscopy to identify and enumerate corpora lutea (CL ) and luteinized unovulated follicles (LUF s). Jugular blood samples were collected just prior to ovarian examinations. The total number of CL (r  =  .78 and 0.83, p  <  .0001) and LUF s (r  =  .74 and 0.90, p  <  .0001) enumerated using the B‐mode and colour Doppler ultrasonographic technique, respectively, were correlated with that ascertained by videolaparoscopy. Circulating concentrations of P₄ were related directly to the number of healthy CL (r  =  .73, p  =  .0002) and inversely to the number of prematurely regressing CL (r  = ?.46, p  =  .03), but the accuracy of predicting the number of short‐lived CL with serum P₄ concentrations was very poor. The present results indicate that ultrasonographic imaging and serum P₄ measurements on the day of embryo recovery are useful indicators of total/normal CL numbers and both ultrasonographic techniques can be used to quantify LUF s in superovulated ewes.     

Woodland vegetation and its implications for archaeological survey using LiDAR

Archaeological surveys in woodland have always been problematicand many woodlands contain an unrecorded archaeological resource.For other types of rural landscape, aerial photographs are oftenused to map archaeological features but woodland cover has alwaysimpeded such disclosure. Remote sensing methods are rapidlyevolving and are used both within forestry and archaeologicaldisciplines for a range of applications. This paper considersthe exciting application of the remote sensing technique ofairborne Light Detection and Ranging (LiDAR) to reveal archaeologicalevidence previously hidden below a woodland canopy. Our researchshows how different types of woodland canopy and understoreyvegetation greatly influence the effectiveness of the LiDARto perform these surveys. A simple, visual vegetation mappingassessment is tested and its ability to predict the potentialof the LiDAR considered. This work highlights the importanceof vegetation awareness when considering both a new LiDAR surveyfor a woodland, and when interpreting the data. Simple estimatesof LiDAR penetration of the woodland canopy and understoreyvegetation can be used to predict the effectiveness of a LiDARsurvey in disclosing archaeological evidence and aid the interpretationof results.     

Azole fungicides – understanding resistance mechanisms in agricultural fungal pathogens

Plant fungal pathogens can have devastating effects on a wide range of crops, including cereals and fruit (such as wheat and grapes), causing losses in crop yield, which are costly to the agricultural economy and threaten food security. Azole antifungals are the treatment of choice; however, resistance has arisen against these compounds, which could lead to devastating consequences. Therefore, it is important to understand how these fungicides are used and how the resistance arises in order to tackle the problem fully. Here, we give an overview of the problem and discuss the mechanisms that mediate azole resistance in agriculture (point mutations in the CYP51 amino acid sequence, overexpression of the CYP51 enzyme and overexpression of genes encoding efflux pump proteins). © 2015 Society of Chemical Industry     

An Inter‐Subspecies Cloned Buffalo (Bubalus bubalis) Obtained by Transferring of Cryopreserved Embryos via Somatic Cell Nuclear Transfer

The aim of this study was to explore the feasibility of cryopreservation of inter‐subspecies cloned embryos in buffalo. In our experiment, river buffalo ear fibroblast nucleus was fused into swamp buffalo oocyte cytoplasm. The blastocyst formation rate for nuclear transfer of freshly thawed cells was not different from those of growing cells, confluent or serum‐starved cells. A total of 122 cloned blastocysts derived from cryopreserved fibroblasts were cryopreserved and thawed, 37 were survived, the cryosurvival rate was 30.3%. The survived blastocysts were transferred into 15 recipient buffalos. Five of the recipients established pregnancy, but four of them aborted on day 53, 59, 145 and 179 of gestation respectively. One cross‐bred buffalo (Murrah × Swamp buffalo (2n = 49) received three embryos delivered a 40.5 kg female calf by natural delivery on day 320 of gestation. Up to now (13‐month old), the cloned calf has been growing well with no abnormity observed. These results demonstrated that cryopreservation of inter‐subspecies cloned embryos is feasible to produce buffalo offspring.