Minimally invasive physiological correlates of social behaviour in belugas (Delphinapterus leucas) under human care

Simultaneous observations of reproductive physiology and behaviour were conducted on a group of two male and two female belugas under professionally managed care for one year to explore potential mating strategies. Weekly blow sampling for progesterone in females was used to define the breeding season by detecting three oestrous cycles in one female. Twice weekly blow sampling for testosterone and twice monthly testes measurements via ultrasonography were used to detect reproductive seasonality in both males. Female–male association frequency varied longitudinally, with 70% of all interactions occurring during the 16-week breeding season. Male–male associations did not vary seasonally. Male display behaviours towards the female occurred 14.8 times more frequently during the breeding season (0.164 ± 0.188 behaviours/min) than outside of the breeding season (0.011 ± 0.042 behaviours/min). The cycling female responded variably to male display behaviours by altering swim speed or body orientation towards the male. Although this small sample size limits broad conclusions, the frequent display behaviours, low copulation rate and lack of serious male–male aggression are consistent with predictions for pre-copulatory female mate choice developed from the current knowledge of beluga reproductive physiology. These observations, which are not feasible for wild belugas, provide important management considerations because reduced opportunities for mate choice could limit the reproductive rate, especially in small populations

The Green Eating Project: web-based intervention to promote environmentally conscious eating behaviours in US university students

Objective: To investigate the effectiveness of an online, interactive intervention, referred to as the Green Eating (GE) Project, to motivate university students to adopt GE behaviours. Design: The study was quasi-experimental and integrated into courses for credit/extra credit. Courses were randomly stratified into experimental or non-treatment control. The 5-week intervention consisted of four modules based on different GE topics. Participants completed the GE survey at baseline (experimental, n 241; control, n 367) and post (experimental, n 187; control, n 304). The GE survey has been previously validated and consists of Transtheoretical Model constructs including stage of change (SOC), decisional balance (DB: Pros and Cons) and self-efficacy (SE: School and Home) as well as behaviours for GE. Modules contained basic information regarding each topic and knowledge items to assess content learning. Setting: The GE Project took place at a public university in the north-eastern USA. Subjects: Participants were full-time students between the ages of 18 and 24 years. Results: The GE Project was effective in significantly increasing GE behaviours, DB Pros, SE School and knowledge in experimental compared with control, but did not reduce DB Cons or increase SE Home. Experimental participants were also more likely to be in later SOC for GE at post testing. Conclusions: The GE Project was effective in increasing GE behaviours in university students. Motivating consumers towards adopting GE could assist in potentially mitigating negative consequences of the food system on the environment. Future research could tailor the intervention to participant SOC to further increase the effects or design the modules for other participants

Evaluating beluga (Delphinapterus leucas) blow samples as a potential diagnostic for immune function gene expression within the respiratory system

Evaluating respiratory health is important in the management of cetaceans, which are vulnerable to respiratory diseases. Quantifying the expression of genes related to immune function within the respiratory tract could be a valuable tool for directly assessing respiratory health. Blow (exhale) samples allow DNA analysis, and we hypothesized that RNA could also be isolated from blow samples for gene expression studies of immune function. We evaluated the potential to extract RNA from beluga blow samples and tested whether transcripts associated with immune function could be detected with endpoint polymerase chain reaction. A total of 54 blow samples were collected from clinically healthy aquarium belugas (n = 3), and 15 were collected from wild belugas temporarily restrained for health assessment in Bristol Bay, Alaska (n = 9). Although RNA yield varied widely (range, 0–265.2 ng; mean = 85.8; SD = 71.3), measurable RNA was extracted from 97% of the samples. Extracted RNA was assessed in 1–6 PCR reactions targeting housekeeping genes (Rpl8, Gapdh or ActB) or genes associated with immune function (TNFα, IL-12p40 or Cox-2). Fifty of the aquarium samples (93%) amplified at least one transcript; overall PCR success for housekeeping genes (96/110, 87%) and genes associated with immune function (90/104, 87%) were similarly high. Both RNA yield and overall PCR success (27%) were lower for wild beluga samples, which is most likely due to the reduced forcefulness of the exhale when compared with trained or free-swimming belugas. Overall, the high detection rate with PCR suggests measuring gene expression in blow samples could provide diagnostic information about immune responses within the respiratory tract. While further study is required to determine if quantitative gene expression data from blow samples is associated with disease states, the non-invasive nature of this approach may prove valuable for belugas, which face increasing anthropogenic disturbances

Trans-cinnamaldehyde nanoemulsion wash inactivates Salmonella Enteritidis on shelled eggs without affecting egg color

Salmonella Enteritidis is a major foodborne pathogen that causes enteric illnesses in humans, primarily through the consumption of contaminated poultry meat and eggs. Despite implementation of traditional disinfection approaches to reduce S. Enteritidis contamination, egg-borne outbreaks continue to occur, raising public health concerns and adversely affecting the popularity and profitability for the poultry industry. Generally Recognized as Safe (GRAS) status phytochemicals such as Trans-cinnamaldehyde (TC) have previously shown to exhibit anti-Salmonella efficacy, however, the low solubility of TC is a major hurdle in its adoption as an egg wash treatment. Therefore, the present study investigated the efficacy of Trans-cinnamaldehyde nanoemulsions (TCNE) prepared with emulsifiers Tween 80 (Tw.80) or Gum Arabic and lecithin (GAL) as dip treatments, at 34°C, for reducing S. Enteritidis on shelled eggs in presence or absence of 5% chicken litter. In addition, the efficacy of TCNE dip treatments in reducing trans-shell migration of S. Enteritidis across shell barrier was investigated. The effect of wash treatments on shell color were evaluated on d 0, 1, 7, and 14 of refrigerated storage. TCNE-Tw.80 or GAL treatments (0.06, 0.12, 0.24, 0.48%) were effective in inactivating S. Enteritidis by at least 2 to 2.5 log cfu/egg as early as 1 min of washing time (P \u3c 0.05). In presence of organic matter, nanoemulsions (0.48%) reduced S. Enteritidis counts by ∼ 2 to 2.5 log cfu/egg as early as 1 min, (P \u3c 0.05). Nanoemulsion wash also inhibited trans-shell migration of S. Enteritidis, as compared to control (P \u3c 0.05). The nanoemulsion wash treatments did not affect shell color (P \u3e 0.05). Results suggest that TCNE could potentially be used as an antimicrobial wash to reduce S. Enteritidis on shelled eggs, although further studies investigating the effect of TCNE wash treatments on organoleptic properties of eggs are necessary

A developmental switch in transcription factor isoforms during spermatogenesis controlled by alternative messenger RNA 3\u27-end formation

Spermatogenic cells elaborate a highly specialized differentiation program that is mediated in part by germ cell-enriched transcription factors. This includes a novel member of the sterol response element-binding factor family, SREBF2_v1/SREBP2gc. Somatic SREBFs are predominantly synthesized as precursor proteins and are critical regulators of cholesterol and fatty acid synthesis. In contrast, SREBF2_v1 bypasses the precursor pathway and has been directly implicated in spermatogenic cell-specific gene expression. During spermatogenesis, SREBF2 precursor transcripts predominate in premeiotic stages, while SREBF2_v1 is highly upregulated specifically in pachytene spermatocytes and round spermatids. In the present study, we demonstrate thatSrebf2_v1mRNAs are present in the testis of several mammalian species, including humans. The basis for the stage-dependent transition in SREBF2 isoforms was also investigated. A 3\u27 rapid amplification of cDNA ends (RACE)-PCR analysis of the rat and human revealed thatSrebf2_v1transcripts are generated by alternative pre-mRNA cleavage/polyadenylation. This involves the use of an intronic, A(A/U)UAAA-independent poly(A) signal within intron 7 of theSrebf2gene. Developmentally regulated competition between germ cell factors that control RNA splicing and pre-mRNA cleavage/polyadenylation may underlie this process. These results define an important role for alternative polyadenylation in male germ cell gene expression and development by controlling a stage-dependent switch in transcription factor structure and function during spermatogenesis. TheSrebf2gene thus provides a useful model to explore the role of alternative polyadenylation in regulating stage-dependent functions of important protein regulators in spermatogenic cells

Minimally invasive physiological correlates of social behaviour in belugas (Delphinapterus leucas) under human care

Simultaneous observations of reproductive physiology and behaviour were conducted on a group of two male and two female belugas under professionally managed care for one year to explore potential mating strategies. Weekly blow sampling for progesterone in females was used to define the breeding season by detecting three oestrous cycles in one female. Twice weekly blow sampling for testosterone and twice monthly testes measurements via ultrasonography were used to detect reproductive seasonality in both males. Female–male association frequency varied longitudinally, with 70% of all interactions occurring during the 16-week breeding season. Male–male associations did not vary seasonally. Male display behaviours towards the female occurred 14.8 times more frequently during the breeding season (0.164 ± 0.188 behaviours/min) than outside of the breeding season (0.011 ± 0.042 behaviours/min). The cycling female responded variably to male display behaviours by altering swim speed or body orientation towards the male. Although this small sample size limits broad conclusions, the frequent display behaviours, low copulation rate and lack of serious male–male aggression are consistent with predictions for pre-copulatory female mate choice developed from the current knowledge of beluga reproductive physiology. These observations, which are not feasible for wild belugas, provide important management considerations because reduced opportunities for mate choice could limit the reproductive rate, especially in small populations

Pre-messenger RNA cleavage factor I (CFIm): potential role in alternative polyadenylation during spermatogenesis

A hallmark of male germ cell gene expression is the generation by alternative polyadenylation of cell-specific mRNAs, many of which utilize noncanonical A(A/U)UAAA-independent polyadenylation signals. Cleavage factor I (CFIm), a component of the pre-mRNA cleavage and polyadenylation protein complex, can direct A(A/U)UAAA-independent polyadenylation site selection of somatic cell mRNAs. Here we report that the CFIm subunits NUDT21/CPSF5 and CPSF6 are highly enriched in mouse male germ cells relative to somatic cells. Both subunits are expressed from spermatogenic cell mRNAs that are shorter than the corresponding somatic transcripts. Complementary DNA sequencing and Northern blotting revealed that the shorter Nudt21 and Cpsf6 mRNAs are generated by alternative polyadenylation in male germ cells using proximal poly(A) signals. Both sets of transcripts contain CFIm binding sites within their 3\u27-untranslated regions, suggesting autoregulation of CFIm subunit formation in male germ cells. CFIm subunit mRNA and protein levels exhibit distinct developmental variation during spermatogenesis, indicating stage-dependent translational and/or posttranslational regulation. CFIm binding sites were identified near the 3\u27 ends of numerous male germ cell transcripts utilizing A(A/U)UAAA-independent sites. Together these findings suggest that CFIm complexes participate in alternative polyadenylation directed by noncanonical poly(A) signals during spermatogenesis

Assessing the quantity and downstream performance of DNA isolated from Beluga (Delphinapterus leucas) blow samples

Blow (exhale) sampling in cetaceans may provide a minimally invasive alternative to biopsy sampling for genetic analyses that may be favored in vulnerable populations. However, the utility of single-exhale blow samples has not been evaluated, and the relationship between the number of exhales collected and DNA yield and its subsequent performance during polymerase chain reaction (PCR) is unknown. DNA was extracted from 98 blow samples collected from 11 aquarium- housed and 29 wild belugas in Bristol Bay, Alaska. Blow samples consisted of one, two, or four successive exhales, with at least nine samples per type from both aquarium-housed and wild belugas. DNA concentration and purity was assessed with a spectrophotometer, and PCR performance was assessed through the amplification of a fragment of the mitochondrial DNA control region or a nuclear marker of sex. Measurable DNA was recovered from 96 samples (98%), although DNA yield varied widely, both by sample (range: 0 to 4,406 ng, mean = 701.5, SD = 1,033.7) and by number of exhalations (ng DNA/exhale) (range: 0 to 3,723, mean = 427.1, SD = 721.8). The amount of DNA extracted per exhale was greater for aquarium samples than for wild samples, but total yield was not proportional with the number of exhales for either group. Successful beluga-specific PCR amplification occurred in 56/59 of the aquarium samples (23/25 single-exhale samples) and 28/39 of the wild samples tested (7/10 of the single-exhale samples). The forcefulness of the breath and chance collection of large pieces of cellular debris likely shaped the relationship between the number of exhales and the DNA yield. Using these methods, a single, forceful exhale should yield enough DNA to perform multiple experiments. This technique is immediately applicable to live-stranded belugas such as the temporary mass strandings that occasionally occur in Cook Inlet, Alaska, and has the potential to increase genetic sampling in protected populations with less disturbance than direct tissue sampling