Embryo transfer,a useful technique to be applied in small community farms?

The objective of this study was to determine if the technique of embryo transfer in cattle can be commercially feasible in a region situated in the humid tropics of Mexico. Twenty-six Bos taurus and twenty-six Bos indicus cows were estrous synchronized and superovulated to obtain a total of 80 embryos of both sub-species. Embryos were classified using stereoscopic microscopy based on established criteria. Nine dual-purpose farms situated in the tropics of Mexico were chosen to provide ten recipient cows each to transfer one embryo per cow. The females were transferred using a fixed-time protocol after verifying the presence of a corpus luteum on the seventh day after the end of hormonal treatment. Pregnancy diagnosis was carried out 28 days after embryo transfer by ultrasonography. Estimation of the cost was determined by calculating the expenses for preparation of the donor and embryo recovery, which were US 633 and US633 and US 589 for B. taurus and B. indicus, respectively. The cost of each embryo was determined considering the number of transferable embryos recovered, which was 3.8 on mean. The cost of each conception was calculated taking into account the percentage of pregnant animals (27% on mean), and the cost for preparing donor and recipient cows, for transferring embryo. The overall cost per gestation was US 1,447. Considering a 50:50 ratio of male to female born, the cost for a replacement heifer calf was US1,447. Considering a 50:50 ratio of male to female born, the cost for a replacement heifer calf was US 2,894, which surpassed by far the commercial cost of a crossbred ready-to-bred heifer normally used as replacement (approximately US $900).     

Seroprevalence and bacteremia [corrected] of Anaplasma phagocytophilum in cats from Bavaria and Lower Saxony (Germany)

Anaplasma (A.) phagocytophilum is a tick-transmitted obligate intracellular bacterium and has been identified in a wide range of mammalian species, causing febrile disease in some. Few reports show that it can also cause granulocytic anaplasmosis in cats. As data on the occurrence of A. phagocytophilum in cats from Germany is limited, a total of 326 serum and 306 EDTA-blood samples from cats from Germany were screened by direct (Giemsa-stained blood/buffy coat smears, real-time PCR) and indirect (IFAT) methods. Of 274 Giemsa-stained blood smears which could be evaluated none was positive for morulae, but one blood sample (< or =0.1%; 1/306) was positive for A. < or = phagocytophilum-DNA in PCR. Antibodies (cutoff > or = 1:64) were detected in 53 out of 326 samples (16.2%). Altogether, the results show a high seroprevalence rate of anti-A. phagocytophilum antibodies in cats in Germany while the low detection rate of this bacterial agent by direct methods is similar to those of other studies on A. phagocytophilum infections in cats.     

Isoflavonoid composition of a callus culture of the relict tree Maackia amurensis Rupr. et Maxim

Isoflavonoids, an interesting and restricted group of secondary metabolites of legumes, exhibit estrogenic, antiangiogenic, and anticancer activities and are now popular as dietary supplements. Plant cell cultures that possess an increased ability to synthesize these metabolites were examined. During the investigation, cell cultures of the Far Eastern relict tree Maackia amurensis (Leguminosae) were established. A selection of seed-derived cell aggregates yielded the callus line designated A-18. This culture produces 20 isoflavonoids, namely, the isoflavones genistein, daidzein, formononetin, calycosin, derrone, and pseudobaptigenin and their glycosylated conjugates genistin, 6'-O-malonylgenistin, ononin, 6'-O-malonylononin, daidzin, 3'-methoxydaidzin, 4'-O-beta-D-glucopyranosyldaidzin, 4'-O-beta-D-glucopyranosylgenistin, and 7-O-beta-D-glucopyranosylcalycosin; the pterocarpans maackiain and medicarpin and their glycosylated conjugates 6'-O-malonyl-3-O-beta-D-glucopyranosylmaackiain and 6'-O-malonyl-3-O-beta-D-glucopyranosylmedicarpin; and the new pterocarpan glucoside 6'-O-malonyl-3-O-beta-D-glucopyranosyl-6,6a-dehydromaackiain. These isoflavonoids, possessing a hepatoprotective activity, were stably produced by the A-18 calli for prolonged periods of observation.     

Diversity among isolates ofVerticillium lecanii as expressed by DNA polymorphism and virulence towardsBemisia tabaci

Thirty-six isolates ofVerticillium lecanii andVerticillium sp. were taken from different hosts (both insects and rusts) and geographical locations. The isolates were analyzed for genomic variability, as expressed by random amplified polymorphic DNA (RAPD), in relation to virulence onBemisia tabaci. Virulence on larvae ofB. tabaci within these isolates ranged from 0% to 83%. RAPD analysis was performed employing two different arbitrary decamer primers and the calculated similarity coefficients were subjected to cluster analysis using the unweighted average linkage (UPGMA) algorithm. The dendrograms obtained with each of the two primers were identical. Eight cluster groups and three unclustered isolates were obtained by selecting a similarity level of 80%. The amplification pattern of DNA obtained by RAPD for the various isolates suggested thatV. lecanii is a highly diverse species. No correlation could be established between virulence and either RAPD polymorphism of the fungal isolates or the insect host from which they were isolated. Generally, no correlation could be established between the clustering ofV lecanii strain and geographical location although a limited number of strains obtained from Russia and Georgia were assembled in the same cluster and those from Kazakhstan were clonal.     

Evidence of Damage in Cryopreserved and Fresh Bovine Embryos Using the Tunel Technique

The objective of the present study was to evaluate the quality of bovine embryos cryopreserved in different years in Chiapas, Mexico. The embryos were obtained from a government institution (FIMEGEN) dedicated to promoting embryo transfer among dual-purpose cattle farmers. Forty-three embryos frozen in 1988, 1989, 2000 and 2002 were analysed with the Tunel technique to detect programmed cell death (apoptosis). Eleven fresh embryos were used as controls. Analysis of variance was used in embryos stored in the different years with averages tested using Tukey's test. Student's t-test was employed to compare fresh and frozen cells. Embryos with shorter storage time presented a lower number (p < 0.001) of Tunel-positive cells compared with embryos stored for longer time. On the contrary, when comparing the number of apoptotic cells between frozen and fresh embryos a higher number of positive cells (p < 0.05) were found in the former. The present results suggest that the cryopreservation per se caused damage that compromises the viability of the embryo. Another explanation for the lower pregnancy rate found in the tropics could be irreversible damage caused by poor storage technique in these large operations.     

Cytotoxic Drimane-Type Sesquiterpenes from Co-Culture of the Marine-Derived Fungi Aspergillus carneus KMM 4638 and Beauveria felina (=Isaria felina) KMM 4639

Chemical investigation of a coculture of the marine-derived fungi Beauveria felina KMM 4639 and Aspergillus carneus KMM 4638 led to the identification of three new drimane-type sesquiterpenes, asperflavinoids B, D and E (2, 4, 5), and nine previously reported related compounds. The structures of these compounds were established using spectroscopic methods and by comparison with known analogues. We also investigated the cytotoxic activity of the isolated compounds against several cancer and normal cell lines. Asperflavinoid C (3) and ustusolate E (9) exerted a significant effect on human breast cancer MCF-7 cell viability, with IC₅₀ values of 10 µM, and induced in caspase-dependent apoptosis and arrest of the MCF-7 cell cycle in the G2/M phase in these cells.