Cervical transfer of deep frozen cattle embryos

Cattle blastocysts were collected from 29 donors 7-8 days after estrus and frozen and stored in liquid nitrogen up to several months. Two procedures were used for freezing and thawing: After thawing, the embryos were cultured from 8 to 12 hours before transfer; 36% of the embryos continued normal development during culture; both procedures resulted in a high pregnancy rate (procedure A: 10 15 ; procedure B: 11 15 ) after single cervical transfer of the frozen thawed embryos which developed normaly in vitro . However the overall survival rate was low (25%) and varied between donors, indicating that progress must be made before the technique of freezing can be extended to applied conditions.     

Necines of alkaloids in Heliotropium species from Mexico and the U.S.A.

The qualitative and quantitative compositions of necines in plants of 20 Heliotropium species collected in Mexico and the U.S.A. and one species from Spain are reported. Trachelanthamidine, supinidine and retronecine were found in all species after hydrolysis of their alkaloids; lindelofidine was detected in most species, whereas heliotridine only in four. Trachelanthamidine, lindelofidine, and supinidine were dominant in four, two and one species, respectively; retronecine was dominant in 15 species, whereas heliotridine only in one. The dominant necine in H. ternatum was either retronecine or lindelofidine depending on the collection locality. Qualitative as well as quantitative differences depending on the collection locality were found in H. curassavicum. Plants from Oaxaca, Mexico, contained lindelofidine and a pyrrolizidine-diol as major necines, trachelanthamidine as minor, and traces of retronecine. Plants originating from two other localities contained trachelanthamidine (dominant), retronecine, and supinidine. The necine patterns found in the examined species differ significantly from those previously reported for 21 species mainly collected in Asia, the Middle East and Australia.     

Pyrrolizidine alkaloids of Heliotropium from Mexico and adjacent U.S.A.

The levels of alkaloids were determined in leaves and stems of 24 species of Heliotropium collected in Mexico and adjacent U.S.A. All species contained unsaturated pyrrolizidines, their content in leaves ranging from 0.003 to 0.18%. Saturated pyrrolizidine alkaloids were detected in nine species. Greenhouse-grown plants of five species had a significantly higher alkaloid content in their leaves than the corresponding field samples. In all these species, except for H. indicum, the differences were due to higher levels of saturated pyrrolizidines.     

Prion gene haplotypes of U.S. cattle

Background

Inguinal and scrotal hernias are of great concern to pig producers, and lead to poor animal welfare and severe economic loss. Selection against these conditions is highly preferable, but at this time no gene, Quantitative Trait Loci (QTL), or mode of inheritance has been identified in pigs or in any other species. Therefore, a complete genome scan was performed in order to identify genomic regions affecting inguinal and scrotal hernias in pigs. Records from seedstock breeding farms were collected. No clinical examinations were executed on the pigs and there was therefore no distinction between inguinal and scrotal hernias. The genome scan utilised affected sib pairs (ASP), and the data was analysed using both an ASP test based on Non-parametric Linkage (NPL) analysis, and a Transmission Disequilibrium Test (TDT).

Results

Significant QTLs (p < 0.01) were detected on 8 out of 19 porcine chromosomes. The most promising QTLs, however, were detected in SSC1, SSC2, SSC5, SSC6, SSC15, SSC17 and SSCX; all of these regions showed either statistical significance with both statistical methods, or convincing significance with one of the methods. Haplotypes from these suggestive QTL regions were constructed and analysed with TDT. Of these, six different haplotypes were found to be differently transmitted (p < 0.01) to healthy and affected pigs. The most interesting result was one haplotype on SSC5 that was found to be transmitted to hernia pigs with four times higher frequency than to healthy pigs (p < 0.00005).

Conclusion

For the first time in any species, a genome scan has revealed suggestive QTLs for inguinal and scrotal hernias. While this study permitted the detection of chromosomal regions only, it is interesting to note that several promising candidate genes, including INSL3, MIS, and CGRP, are located within the highly significant QTL regions. Further studies are required in order to narrow down the suggestive QTL regions, investigate the candidate genes, and to confirm the suggestive QTLs in other populations. The haplotype associated with inguinal and scrotal hernias may help in achieving selection against the disorder.     

U.S. fishery negotiations with Canada and Mexico

Abstract

In the last decade, the world has witnessed a fundamental reorientation of posture toward marine resources as evidenced by consensus in the Third United Nations Conference on the Law of the Sea (UNCLOS III) and the near universal move toward fishery jurisdictions of 200 nautical miles (370 km). These and other non‐marine‐specific economic and political pressures impose a new constellation of constraints on North American fishery relations. This has resulted in disharmonies on two borders.

This paper, then, addresses the form of binational fishery negotiations between the United States and her two neighbors. One section presents a general model of the binational negotiation process. The next section introduces the institutional and political context of U.S.‐Canada relations, and then applies the negotiation model to the evolution of West Coast salmon deliberations. In a parallel fashion, the following section focuses on U.S.‐Mexico relations and the tuna, shrimp, and anchovy fisheries. The last section concludes with remarks on future directions for North American binational fishery relations.     

Yeasts from U.S.A. soils

Summary Investigations of yeasts from 38 U.S.A. soils samples show the occurrence of 22 different species (16 sporogenous and 6 asporogenous).The most widespread species were Pichia fermentans and Hansenula anomala. The other isolated species were Saccharomyces ellipsoideus, Torulaspora delbrueckii, Pichia membranefaciens, Saccharomyces smittii, Saccharomyces carlsbergensis, Saccharomyces uvarum, Torulaspora rosei, Zygosaccharomyces rouxii, Zygosaccharomyces n. sp., Hansenula saturnus, Hansenula californica, Hansenula n. sp., Hansenula suaveolens, Debaryomyces n. sp., Torulopsis glabrata, Torulopsis n. sp. Candida tropicalis, Candida robusta, Rhodotorula glutinis, Trichosporon cutaneum and Trichosporon cutaneum var. multisporus.     

Interleukin-8 haplotype structure from nucleotide sequence variation in commercial populations of U.S. beef cattle

The aim of the present study was twofold: first, to design a panel of 96 sires that reflects the breadth of genetic diversity in U.S. beef cattle, and second, to use this panel to discover nucleotide sequence diversity and haplotype structures of interleukin (IL)-8 in commercial populations. The latter is a requisite for epidemiological studies designed to test whether IL8 alleles are risk factors for acquiring or maintaining bacterial infections in production environments. IL-8 encodes a proinflammatory cytokine that plays a central role in cell-mediated immunity by attracting and activating neutrophils in the early stages of host defense against bacterial invasion. Seven single-nucleotide polymorphism (SNP) markers were identified by sequencing two IL8 DNA segments amplified from the panel of 17 popular cattle breeds (MARC beef cattle diversity panel, version 2.1). Assays for automated genotype scoring by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were developed to independently verify the seven SNP alleles in the 96 bulls and 313 cattle from the MARC reference population. Five haplotype structures, spanning the two IL8 DNA segments, were unambiguously defined for the set of seven IL8 SNPs. Based on the breadth of germplasm in bovine diversity panel, the five haplotype structures for IL8 are estimated to represent >98% of those present in these DNA segments in commercial populations of U.S. beef cattle. The frequencies of the five respective haplotypes in the eight Angus sires of the diversity panel (0.75, 0.25, 0.00, 0.00, 0.00) were similar to those scored in 150 purebred Angus cattle from six herds in four Midwestern states (0.82, 0.18, 0.01, 0.00, 0.00), suggesting that the diversity panel may also be useful for estimating allele frequencies in commercial populations. Received: 29 August 2000 / Accepted: 17 November 2000     

Frequency and occurrence of late-gestation losses from cattle cloned embryos.

Nuclear transfer from somatic cells still has limited efficiency in terms of live calves born due to high fetal loss after transfer. In this study, we addressed the type of donor cells used for cloning in in vivo development. We used a combination of repeated ultrasonography and maternal pregnancy serum protein (PSP60) assays to monitor the evolution of pregnancy after somatic cloning in order to detect the occurrence of late-gestation losses and their frequency, compared with embryo cloning or in vitro fertilization (IVF). Incidence of loss between Day 90 of gestation and calving was 43.7% for adult somatic clones and 33.3% for fetal somatic clones, compared with 4.3% after embryo cloning and 0% in the control IVF group. Using PSP60 levels in maternal blood as a criterion for placental function, we observed that after somatic cloning, recipients that lost their pregnancy before Day 100 showed significantly higher PSP60 levels by Day 50 than those that maintained pregnancy (7.77 +/- 3.3 ng/ml vs. 2.45 +/- 0.27 ng/ml for normal pregnancies, P < 0.05). At later stages of gestation, between 4 mo and calving, mean PSP60 concentrations were significantly increased in pathologic pregnancy after somatic cloning compared with other groups (P < 0.05 by Day 150, P < 0.001 by Day 180, and P < 0.01 by Day 210). In those situations, and confirmed by ultrasonographic measurements, recipients developed severe hydroallantois together with larger placentome size. Our findings suggest that assessing placental development with PSP60 and ultrasonography will lead to better care of recipient animals in bovine somatic cloning.