No evidence for condition-dependent expression of male genitalia in the dung beetle Onthophagus taurus

We examine the condition-dependence of male genitalia in the dung beetle Onthophagus taurus by manipulating the quality of dung provided for larval growth and development. We show that the influence of larval nutrition differed considerably across three different trait classes (sexual, nonsexual and genital). The size of all nonsexual traits varied with dung quality but their allometric slopes remained unchanged. Relative horn length and allometry, but not absolute horn length, showed a high degree of plasticity with differences in dung quality. In contrast, both absolute size and allometry of genitalia were largely unresponsive to changes in dung quality. Male genitalia exhibited intermediate levels of phenotypic variation and lower allometric slopes than both horns and nonsexual traits. Thus, our findings provide little support for good genes hypotheses of genital evolution. We use our findings to discuss a developmental mechanism and selection pressures that may prevent the condition-dependent expression of genitalia.     

In-depth analysis of low abundant proteins in bovine colostrum using different fractionation techniques

Bovine colostrum is well known for its large content of bioactive components and its importance for neonatal survival. Unfortunately, the colostrum proteome is complicated by a wide dynamic range, because of a few dominating proteins that hamper sensitivity and proteome coverage achieved on low abundant proteins. Moreover, the composition of colostrum is complex and the proteins are located within different physical fractions that make up the colostrum. To gain a more exhaustive picture of the bovine colostrum proteome and gather information on protein location, we performed an extensive pre-analysis fractionation of colostrum prior to 2D-LC-MS/MS analysis. Physical and chemical properties of the proteins and colostrum were used alone or in combination for the separation of proteins. ELISA was used to quantify and verify the presence of proteins in colostrum. In total, 403 proteins were identified in the nonfractionated colostrum (NF) and seven fractions (F1-F7) using six different fractionation techniques. Fractionation contributed with 69 additional proteins in the fluid phase compared with NF. Different fractionation techniques each resulted in detection of unique subsets of proteins. Whey production by high-speed centrifugation contributed most to detection of low abundant proteins. Hence, prefractionation of colostrum prior to 2D-LC-MS/MS analysis expanded our knowledge on the presence and location of low abundant proteins in bovine colostrum.     

Confidence of paternity and paternal care: covariation revealed through the experimental manipulation of the mating system in the beetle Onthophagus taurus

Theoretical models of paternal care predict that facultative reductions in male care may occur under certain conditions. One important parameter that has been shown to influence the outcome of these models is a male's confidence of paternity. In this study, we tested whether the amount of care provided by horned males in the dimorphic beetle, Onthophagus taurus, varied with his confidence of paternity. Male care results in an increased weight of dung provided in the brood masses produced by the pair. Using the sterile male technique we showed that a horned male's paternity declined with the number of sneak males in the population. The relationship was nonlinear, with paternity declining most rapidly between a frequency of one and three sneaks, and stabilizing thereafter at about 50%. A horned male's paternity was directly related to the number of copulations with the female, relative to the number of copulations achieved by sneaks. Horned males were shown to reduce their care in relation to their declining paternity. Video analysis demonstrated that reductions in male care occurred through a combination of male desertion and a trade‐off between caring and paternity assurance behaviours. The number of fights with sneak males was negatively related to the amount of care provided by a horned male. These results suggest that by gauging his expected paternity through the number of fights with sneaks, a horned male is able to assess his paternity and reduce his investment accordingly. Our data thus provide strong empirical support for the proposed link between paternity and paternal care.     

A naturally occurring αs1‐casein‐derived peptide in bovine milk inhibits apoptosis of granulosa cells induced by serum‐free conditions

Several naturally occurring peptides in bovine milk were characterized by tandem mass spectrometry and Edman degradation. Chromatograms of peptide fractions (passed through an ultra‐filtration membrane, nominal molecular weight limit 3000) prepared from colostrum (collected immediately after parturition) and transitional milk (collected 5 days postpartum) showed that they were almost identical. In total, six peptides, α_s1‐CN (f16‐23) (RPKHPIKH), α_s1‐CN (f16‐24) (RPKHPIKHQ), α_s1‐CN (f17‐25) (PKHPIKHQG), α_s1‐CN (f46‐52) (VFGKEKV), α_s1‐CN (f94‐105) (HIQKEDVPSER), and β‐CN (f121‐128) (HKEMPFPK), were identified. One of the major peptides, the N‐terminal fragment of α_s1‐casein, varied structurally during early lactation: α_s1‐CN (f17‐25) (PKHPIKHQG) and α_s1‐CN (f16‐23) (RPKHPIKH)/α_s1‐CN (f16‐24) (RPKHPIKHQ) were found in colostrum and transitional milk, respectively. A chemically synthesized peptide, α_s1‐CN (f16‐23) (RPKHPIKH), inhibited apoptosis of bovine granulosa cells induced by serum‐free conditions in a dose‐dependent manner, in consequence of caspase‐3 and caspase‐9 suppressions. The physiological function of the peptide remains unclear, but it may have potential use as pharmaceutical agent and as an anti‐apoptotic agent in cell culture medium. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.     

泽西牛犊对热胁迫的生理应答及其作为动物福利下降指标的潜在应用性

本研究以泽西奶牛(Bostaurus)牛犊为对象,探讨温、热环境(30℃)下短暂暴露是否影响其体温、心跳和呼吸频率以及细胞和内分泌应激反应(皮质醇、甲状腺激素、牛生长激素及热应激蛋白72)。结果表明:热暴露导致呼吸频率和心率加快,体温显著升高,但幅度较小。在所研究的细胞和内分泌应激反应中,血浆甲状腺激素浓度略下降,肌肉热应激蛋白72的浓度稍增加。因此,尽管夏季饲养动物所经历的较高环境温度(但非极端温度)能影响泽西牛犊的生理,但是泽西牛犊能对温和的暂时性环境温度改变做出适应性反应。我们认为,直肠温度和细胞热应激蛋白72浓度是所研究参数中评价热胁迫降低动物福利的最适指标。     

Radiation hybrid mapping of all 10 characterized bovine Toll-like receptors

Toll-like receptors (TLRs) are cell-surface signalling molecules that recognize a range of highly conserved pathogen molecules and instigate the appropriate immune response. Here, we report the mapping of all 10 characterized bovine TLR genes using a radiation hybrid panel. The genomic organization of the bovine TLRs is similar to that of humans and mice. TLR1, TLR6 and TLR10 map closely together on Bos taurus chromosome 6 (BTA6), while TLR7 and TLR8 map to the X chromosome. TLR2, TLR3, TLR4, TLR5 and TLR9 map to BTA17, BTA27, BTA8, BTA16 and BTA22 respectively. Our increased knowledge of the genomic organization of the bovine TLR genes may promote our understanding of their evolution and help in the identification of bovine genes underlying disease-resistance traits.     

Proteomic investigation in the detection of the illicit treatment of calves with growth-promoting agents

The use of beta-agonists, sexual steroids, and corticosteroids as growth-promoting agents (GPAs) in veal calves is forbidden in the European Union (EU) and subjected to restrictions in the US because it may be potentially noxious for both treated animals and the consumer. Although official controls performed in the EU have revealed a limited number of positive samples, the analysis of seized preparations indicate that the use of illegal GPAs is far from being abandoned. The presence of these compounds in matrixes of biological origin often goes unnoticed because of the use of very low dosages and/or of molecules of unknown chemical structure. It is therefore necessary to develop screening methods based on the biological effects of these substances that allow the simultaneous screening of many components, as proteome analysis. When hepatic cytosols and microsomes from calves treated with a combination of GPAs were analyzed by 2-DE, we found changes in the expression of two proteins, which we identified as adenosine kinase and reticulocalbin. Our aim was not to speculate about molecular mechanisms, but to show the ability of the proteomic approach to find biomarkers of illicit treatments and to use it as a basis to develop large-scale screening methods.     

Considerations on the diet composition and feeding rate of dermersal sharks in 15 European public aquaria

To get an overview of the feeding practices applied to dermersal sharks in captivity, a questionnaire was sent to 15 European public aquaria. The most common dermersal shark species kept in the participating aquaria are Carcharhinus plumbeus (n=9), Carcharias taurus (n=7), and C. melanopterus (n=6). Body weight (BW) was calculated using captive estimated body length (BL) and actual weight data obtained from commercial fisheries. The estimated BW for C. plumbeus in all participating aquaria is higher than the calculated BW. The feeding rate (FR) for adult C. plumbeus varied widely within participating aquaria (2.3–11.5% BW_cal/week or 32–112 kcal/kg BW_cal/week). The FR for adult C. plumbeus was lower (1.2–7.9% BW_est/week or 17–99 kcal/kg BW_est/week) using the estimated BW. Adult Carcharias taurus were fed at 1.3–5.0% BW_cal/week or 70–170 kcal/kg BW_cal/week. Negaprion brevirostris was fed at 5.0% BW_est/week or 57 kcal/BW_est/week. The relationship between FR and estimated BW is expressed as FR=54.5 e^?0.033BW. In this study, the feeding frequency for C. plumbeus varied between 1–7 times a week, with an average of 3.3 times a week. Diet in all aquaria consisted of both lean and fatty fish, and sometimes squid or crustaceans. Supplementation of vitamins is common practice in 80% of the surveyed aquaria, although a wide variety of quantity and quality was observed. Ten aquaria (66%) add iodine, as potassium iodide (0.4–12.5 mg KI/kg BW/week) or calcium‐iodate (322–1,500 mg/kg food/week), to their food. Zoo Biol 22:203–226, 2003. © 2003 Wiley‐Liss, Inc.     

A polymorphism in the bovine gamma-S-crystallin gene revealed by allele-specific amplification

Summary
A polymorphism was detected in the 3' untranslated region of the bovine gamma-S-crystallin gene by direct sequencing of polymerase chain reaction (PCR) products from genomic DNA of an N'Dama bull and a Boran cow. A set of three PCR primers was designed to detect this difference and thus give allele-specific amplification. The two allele-specific primers differ in length by 20 nucleotides so that the allelic products may be distinguished by simple agarose gel electrophoresis following a single PCR reaction. This provides a simple and rapid assay for this polymorphism.