Evaluation of steroidogenic capacity after follicle stimulating hormone stimulation in bovine granulosa cells of Revalor 200? implanted heifers

Background

Heifers not used as breeding stock are often implanted with steroids to increase growth efficiency thereby altering hormone profiles and potentially changing the environment in which ovarian follicles develop. Because bovine granulosa cell culture is a commonly used technique and often bovine ovaries are collected from abattoirs with no record of implant status, the objective of this study was to determine if the presence of an implant during bovine granulosa cell development impacts follicle stimulating hormone-regulated steroidogenic enzyme expression. Paired ovaries were collected from 16 feedlot heifers subjected to 1 of 3 treatments: non-implanted (n = 5), Revalor 200 for 28 d (n = 5), or Revalor 200 for 84 d (n = 6). Small follicle (1 to 5 mm) granulosa cells were isolated from each pair and incubated with phosphate buffered saline (n = 16) or 100 ng/mL follicle stimulating hormone (n = 16) for 24 h.

Results

Granulosa cells of implanted heifers treated with follicle stimulating hormone produced medium concentrations of progesterone similar (P = 0.22) to non-implanted heifers, while medium estradiol concentrations were increased (P < 0.10) at 28 and 84 d compared to non-implanted heifers indicating efficacy of treatment. Additionally, real-time PCR analysis in response to follicle stimulating hormone treatment demonstrated a decrease in steroidogenic acute regulatory protein (P = 0.05) mRNA expression in heifers implanted for 84 d and an increase in P450 side chain cleavage mRNA in granulosa cells of heifers implanted for 28 (P < 0.10) or 84 d (P < 0.05) compared to non-implanted females. However, no difference in expression of 3-beta-hydroxysteroid dehydrogenase (P = 0.57) and aromatase (P = 0.23) were demonstrated in implanted or non-implanted heifers.

Conclusions

These results indicate follicles which develop in the presence of high concentrations of androgenic and estrogenic steroids via an implant tend to demonstrate an altered capacity to respond to follicle stimulating hormone stimulation. Thus, efforts should be made to avoid the use of implanted heifers to study steroidogenesis in small follicle granulosa cell culture systems.     

Reducing the potential for processing contaminant formation in cereal products

Processing contaminants may be defined as substances that are produced in a food when it is cooked or processed, are not present or are present at much lower concentrations in the raw, unprocessed food, and are undesirable either because they have an adverse effect on product quality or because they are potentially harmful. The presence of very low levels of processing contaminants in common foods is becoming an increasingly important issue for the food industry, as developments in analytical techniques and equipment bring foods under closer and closer scrutiny. This review considers the formation of lipid oxidation products, hydrogenation of polyunsaturated fatty acids to prevent lipid oxidation and the associated risk of trans fatty acid formation. The formation of acrylamide in the Maillard reaction is described, as well as the genetic and agronomic approaches being taken to reduce the acrylamide-forming potential of cereal grain. The multiple routes for the formation of furan and associated chemicals, including hydroxymethylfurfuryl, are also described. The evolving regulatory and public perception situations for these processing contaminants and their implications for the cereal supply chain are discussed, emphasising the need for cereal breeders to engage with the contaminants issue.     

Allelic losses and gains during translocations of a high conservation value fish,Coregonus lavaretus

1. The use of translocations to establish new or ‘refuge’ populations for species with high conservation value is controversial but widely used in conservation management. One of the risks of this approach is that an establishing population does not adequately capture the genetic diversity of the donor gene pool. This effect, rarely examined, is tested here.
2. In this study the genetic consequences of two conservation translocations after five generations (16 years) of the European whitefish, Coregonus lavaretus, were quantified. Both translocations were made using almost the same genetic groups and thus represent a partly replicated natural study.
3. Analysis of 12 informative microsatellites showed that expected heterozygosity, the mean number of alleles per locus and allelic richness did not differ between donor and translocated populations. There was also no loss of heterozygosity in the translocated populations, nor deviations from Hardy–Weinberg equilibrium expectations, nor signs of linkage disequilibrium.
4. All populations were genetically differentiated but pairwise F_ST values were low, indicating that the magnitude of divergence was small.
5. There was no evidence of inbreeding but there were significant differences in private allelic richness between donor and translocated populations. Of 50 alleles found in the donor population, 16% of the rarer alleles were lost in one translocated population and 8% in the other.
6. Allele loss without a reduction in heterozygosity strongly points to stochastic drift effects having occurred following translocation. The evidence indicates that alleles that were not detected in the donor population have arisen de novo in the translocated populations.
7. It is concluded that conservation translocations comprising even a modest number of propagules can successfully capture a high proportion of genetic variation of the host population, and that reduced genetic variation in the translocated population may be mitigated by the emergence of new variation over short time periods.

     

Assessment of plastic ingestion by pole-caught pelagic predatory fish from O'ahu,Hawai'i

1. Although the frequency of occurrence of plastic ingestion in the large-sized dolphinfish and tunas taken by the Hawai'i longline fishery is very low (frequency of occurrence < 5% of sampled individuals), the ingestion of plastic in smaller-sized specimens caught with pole-and-line gear by commercial and recreational fishers has not been investigated.
2. This study examined ingestion of >0.25 mm marine plastic debris (MPD) by four predatory fish species caught by commercial fishers around the Main Hawaiian Islands, and documented ingestion in three species: 85.7% of albacore tuna (n = 7), 40.0% of skipjack tuna (n = 10) and 12.5% of dolphinfish (n = 8).
3. Yellowfin tuna (n = 10) did not contain any MPD, probably owing to the high proportion of empty stomachs (60%).
4. For skipjack tuna, the frequency of occurrence of MPD ingestion was significantly higher for the smaller-sized specimens caught with pole-and-line (40%), compared with the larger-sized specimens caught with longlines (0%).
5. For dolphinfish, the frequency of occurrence of MPD ingestion was similar for the similar-sized specimens caught with pole-and-line and with longlines.
6. The ingested MPD items were micro–meso plastics, between 1 and 25 mm. While most ingested items were fragments, albacore also ingested line and skipjack also ingested sheets.
7. The predatory fishes ingested light MPD items that float in sea water, but there were species-specific differences in their polymer composition: albacore contained more polypropylene and polyethylene, and skipjack contained more elastomers, characterized by a high percentage of ester plasticizers.
8. Altogether, these results suggest that albacore and skipjack tunas ingest plastic of different types and polymers. Yet more research is needed to understand how differences in vertical distribution, foraging ecology and diet influence the MPD sampled by these predatory fish species.

     

Population isolation exacerbates conservation genetic concerns in the endangered Amargosa vole, Microtus californicus scirpensis

The ability of a taxon to maintain adaptive flexibility in a stochastic environment is a function of the genetic diversity within the population. In small, fragmented populations, genetic variation can become depleted more quickly than in larger, more contiguous populations. Characterizing the patterns of genetic variation and differentiation associated with these processes is an important step in establishing conservation priorities. The Amargosa vole, Microtus californicus scirpensis, is an endangered rodent persisting in the small, fragmented marsh complex surrounding the Amargosa River near Death Valley, California. This naturally patchy system has existed since the end of the Pleistocene (approximately 10,000 y.b.p.), however, fragmentation has been exacerbated by recent anthropogenic changes. For this study, I used five nuclear microsatellite loci and the cytochrome-b region of the mitochondrial genome to quantify levels of genetic variation, population substructure, and patterns of gene flow in M.c. scirpensis. These data were compared to a broadly distributed subspecies, Microtus californicus sanctidiegi. Overall levels of nuclear genetic variation were significantly lower in M.c. scirpensis, whether measured in terms of diversity or heterozygosity, compared to more broadly distributed conspecifics. Moreover, only two haplotypes were recovered from the mitochondrial data with over 90% of the observed haplotypes being identical. Despite low genetic diversity, significant genetic subdivision among M.c. scirpensis populations was detected using both pairwise F_ST and Bayesian clustering methods. Furthermore, isolation by distance analyses reveal that an important landscape feature, ephemeral tributaries, is critical for dispersal among population clusters. Recommendations for conservation management are presented.     

The Functional Anatomy of the Neck and Forelimbs of the Cape Golden Mole,Chrysochloris Asiatica (Lipotyphla: Chrysochloridae)

The anatomy of the neck and forelimb of Chrysochloris asiatica is described and illustrated. The sequence of events during digging and modifications for fossorial action are described. Modifications include the appearance of a third bone in the forearm; the shortening and fusion of bones in the manus; enlarged processes on the scapula, humerus and ulna for greater muscle attachment; enlarged neck muscles and a dip in the spine in the cervical region to accommodate these and the enlarged shoulder muscles; an enlarged occiput for insertion of the powerful neck muscles; a greatly enlarged triceps and movement of the shoulder girdle to a position anterior to that normal in mammals. The possibility of the third forearm bone being the ossified tendon of a flexor muscle is discussed, without any conclusion being reached as to its true origin.     

A comparison of three combinations of injectable anesthetics in miniature donkeys

Objective To compare three combinations of injectable anesthetics in miniature donkeys for quality of induction, recovery, muscle relaxation, cardiopulmonary changes during anesthesia and duration of recumbency. Design Prospective, randomized experimental study. Animals Six miniature donkeys (< 90 cm in height at the withers) weighing 92–127 kg were used. Materials and methods The drug combinations were: xylazine?butorphanol?ketamine (XBK), xylazine?butorphanol?tiletamine?zolazepam (XBT) and xylazine?propofol (XP). Each miniature donkey was anesthetized with each combination at 1‐week intervals in random order. Heart and respiratory rates, indirect blood pressure and temperature were measured before and at 5‐minute intervals during recumbency. Arterial blood samples were drawn for blood‐gas analysis before and at 5, 15 and 30 minutes of anesthesia when samples could be collected. Recumbency time to sternal and time to standing were recorded and a subjective evaluation of induction, muscle relaxation and recovery were made. Results Mean recumbency time ± SD was 14.7 ± 9.4, 33.8 ± 6.3 and 14.6 ± 1.9 minutes with XBK, XBT and XP, respectively. Mean time to standing ± SD was 28.4 ± 11.3, 43.7 ± 7.2 and 26.3 ± 2.9 minutes with XBK, XBT and XP, respectively. Heart and respiratory rates and blood pressures varied from baseline but were always within normal ranges. Hemoglobin saturation, pH and PaO₂ tended to be lower with these doses of XBT and XP. Conclusions and clinical relevance Overall quality of anesthesia was poor with XBK. At the doses used this combination did not provide sufficient anesthesia compared with the combinations of XBT and XP, which appeared to provide acceptable anesthesia of short duration in miniature donkeys.     

The role of p38 mitogen-activated kinase (MAPK) in the mechanism regulating cyclooxygenase gene expression in equine leukocytes

The goal of this study was to define the role for p38 mitogen-activated kinase (MAPK) in the signaling mechanism regulating pro-inflammatory cyclooxygenase (COX) gene expression in lipopolysaccharide (LPS)-activated equine leukocytes for the purposes of identifying novel targets for anti-inflammatory therapy in endotoxemic horses. The p38 MAPK has been shown to positively regulate inflammatory gene expression in human leukocytes and can be activated by a variety of stimuli including LPS, TNF-alpha, and IL-1. Activation-associated phosphorylated p38 MAPK has been implicated in the up-regulation of several inflammatory genes, including COX-2 which ultimately results in the production of prostanoids that are responsible for the pathophysiology associated with endotoxemia. Our hypothesis is that activation of p38 MAPK is essential for LPS-induced COX-2 expression in equine peripheral blood leukocytes. We tested our hypothesis by investigating the effects of the specific p38 MAPK inhibitors SB203580 and SB202190 on LPS-induced COX-2 protein expression and PGE(2) production in equine leukocytes. LPS stimulation activated p38 MAPK and increased COX-2 expression in a dose-dependent manner with maximal activation observed after 30min and 4h, respectively, at a concentration of 10 ng/ml LPS. In contrast, LPS stimulation did not affect COX-1 protein expression. Pretreatment with SB203580 or SB202190 significantly inhibited LPS-induced activation-associated p38 MAPK phosphorylation, COX-2 mRNA and protein levels, and PGE(2) production in equine leukocytes. Maximal inhibition of LPS-induced COX-2 protein expression was achieved at a concentration of 10 microM SB203580. We concluded that p38 MAPK is essential for LPS-induced COX-2 expression suggesting that p38 MAPK is a potential target for anti-inflammatory therapy during equine endotoxemia.