Impact of Buserelin Acetate or hCG Administration on the Day of First Artificial Insemination on Subsequent Luteal Profile and Conception Rate in Murrah Buffalo (Bubalus bubalis)

This study was designed to investigate the impact of buserelin acetate (BA) or human chorionic gonadotropin (hCG) administration on the day of first artificial insemination (AI) on subsequent luteal profile (diameter of corpus luteum (CL) and plasma progesterone) and conception rate in Murrah buffalo. The present experiment was carried out at two locations in 117 buffalo that were oestrus‐synchronized using cloprostenol (500 μg) administered (i.m.) 11 days apart followed by AI during standing oestrus. Based on treatment (i.m.) at the time of AI, buffalo were randomly categorized (n = 39 in each group) into control (isotonic saline solution, 5 ml), dAI‐BA (buserelin acetate, 20 μg) and dAI‐hCG (hCG, 3000 IU) group. Out of these, 14 buffalo of each group were subjected to ovarian ultrasonography on the day of oestrus to monitor the preovulatory follicle and on days 5, 12, 16 and 21 post‐ovulation to monitor CL diameter. On the day of each sonography, jugular vein blood samples were collected for the estimation of progesterone concentrations. All the buffalo (n = 117) were confirmed for pregnancy on day 40 post‐ovulation. The conception rate was better (p < 0.05) in dAI‐BA (51.3%) and dAI‐hCG (66.7%) groups as compared to their control counterparts (30.8%). Furthermore, the buffalo of dAI‐hCG group had improved (p < 0.05) luteal profile, whereas the buffalo of dAI‐BA group failed (p > 0.05) to exhibit stimulatory impact of treatment on luteal profile when compared to control group. In brief, buserelin acetate or hCG treatment on the day of first AI leads to an increase in conception rate; however, an appreciable impact on post‐ovulation luteal profile was observed only in hCG‐treated Murrah buffalo.     

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.     

Follicle Ablation Improves the Ovarian Response and the Number of Collected Embryos in Superovulated Cows During the Early Stages of Lactation

A field study was designed to compare ovarian response and embryo yield in cows during early lactation when gonadotropin administration followed one of four treatments. In group 1A (n = 19) and 1B (n = 9), the estrouses were synchronized by two prostaglandin F2alpha (PG) injections given 11 days apart, and starting from day 9 of the synchronized cycle superovulation was conducted with eight decreasing dose of FSH. In group 1B, ablation of all follicles >3 mm was carried out on day 8. In group 2A and 2B (each n = 9), a progesterone plus oestradiol intravaginal device (PRID) was inserted for 11 days and gonadotropin administration started on day 9, while cows from group 2B had a follicle ablation on day 8. In all groups, two PG injections were given along with the sixth and the seventh dose of FSH, and the cows were twice inseminated 12 and 24 h after estrus detection. Embryos were collected on day 7. In cumulative results from aspirated and non-aspirated cows, follicular ablation significantly improved: the ovarian response (10 +/- 1.23 vs 6.69 +/- 0.60 corpora lutea per donor), the mean collected embryos (6.57 +/- 0.94 vs 2.46 +/- 0.53) and the mean transferable embryos (4.43 +/- 0.89 vs 2.18 +/- 0.47). Group 1B and 2B cows had better ovarian response than 1A (6.44 +/- 0.81, 12.25 +/- 4.11 and 9.44 +/- 0.93, for groups 1A, 1B and 2B, respectively, p < 0.05). Similarly, from groups 1B and 2B more (p < 0.05) embryos were collected in comparison with their respective group, while the mean transferable embryos from group 2B (5.22 +/- 1.13) was greater (p < 0.05) than that of group 1A (1.67 +/- 0.35), and tented to be greater than those of groups 2A (3.44 +/- 1.19, p = 0.062) and 1B (3.00 +/- 1.78, p = 0.066). The highest (p < 0.05) transferable embryo collection rate was recorded in group 2B (55.29%), followed by that of group 1B (41.33%). In summary, early in lactation, an acceptable number of transferable embryos can be collected from high producing dairy cows, when follicle ablation prior to superovulation is combined with progesterone and oestradiol administration.     

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.     

Genetic Diversity Based on AIIozyme Alleles of Chinese Cultivated Rice

Genetic diversity was analyzed with 6 632 core rice cultivars selected from 60 282 Chinese rice accessions on the basis of 12 allozyme loci, Pgil, Pgi2, Amp1, Amp2, Amp3, Amp4, Sdhl, Adhl, Estl, Est2, Est5 and Est9, by starch gel electrophoresis. Among the materials examined, 52 alleles at 12 polymorphic loci were identified, which occupied 96.3% of 54 alleles found in cultivated germplasm of O. sativa L. The number of alleles per locus ranged from 2 to 7 with an average of 4.33. The gene diversity （He） each locus varied considerably from 0.017 for Amp4 to 0.583 for Est2 with an average gene diversity （Ht） 0.271, mid Shannon-Wiener index from 0.055 to 0.946 with an average of 0.468. The degree of polymorphism （DP） was in a range from 0.9 to 46.9% with an average of 21.4%. It was found that the genetic diversity in japonica （Keng） subspecies was lower in terms of allele＇s number, Ht and S-W index, being 91.8, 66.2 and 75.7% of indica （Hsien） one, respectively. Significant genetic differentiation between indica and japonica rice has been appeared in the loci Pgil, Amp2, Pgi2, and Est2, with higher average coefficient of genetic differentiation （Gst） 0.635, 0.626, 0.322 and 0.282, respectively. Except less allele number per locus （3.33） for modern cultivars, being 76.9% of landraces, the Ht and S-W index showed in similar between the modem cultivars and the landraces detected. In terms of allozyme, the rice cultivars in the Southwest Plateau and Central China have richer genetic diversity. The present study reveals again that Chinese cultivated rice germplasm has rich genetic diversity, showed by the allozyme allele variation.     

Identification of lactoferrin and glutamate receptor‐interacting protein 1 in bovine cervical mucus: A putative marker for oestrous detection

Accurate detection of oestrus is important for artificial insemination. The aim of this study was to identify oestrous‐specific bovine cervical mucus proteins that could be used to determine the optimal time for artificial insemination. Non‐oestrous and controlled internal drug release (CIDR)‐induced oestrous‐stage mucus proteins were purified and subjected to surface‐enhanced laser desorption/ionization time‐of‐flight mass spectrometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis and MALDI‐TOF/TOF. Among differentially expressed proteins, lactoferrin (LF) and glutamate receptor‐interacting protein 1 (GRIP1) showed a twofold increase during the CIDR‐induced oestrous stage compared to the levels in non‐oestrous stage in bovine cervical mucus. The RT‐PCR, Western blotting and immunohistochemistry results showed that LF and GRIP1 expression was significantly increased during the oestrous stage in the uterus. This study demonstrated that bovine LF and GRIP1 exist during the oestrous stage, but not during the non‐oestrous stage, suggesting that cervical mucus LF and GRIP1 are useful oestrous detection markers in cattle.