Liquid chromatographic determination of intermediates in D&C Yellow No. 7 and D&C Yellow No. 8

A sensitive, reproducible method that uses liquid chromatography in the reverse phase mode has been developed for the determination of phthalic acid, resorcinol, and 2-(2',4'-dihydroxybenzoyl)benzoic acid in D&C Yellow No. 7 and D&C Yellow No. 8. The method uses a 10 micron, C-8 column, a 1% acetic acid-methanol gradient, and UV absorption detection at 280 nm. Average recoveries of phthalic acid, resorcinol, and 2-(2',4'-dihydroxybenzoyl)benzoic acid were 100, 98, and 102%, respectively, from fluorescein standard (certifiable as D&C Yellow No. 7) spike with each compound at levels ranging from 0.13 to 1.3%.     

Further investigation of lead exposure as a potential threatening process for a scavenging marsupial species

There is a growing recognition of the harmful effects of lead exposure on avian and mammalian scavengers. This can lead to both lethal and non-lethal effects which may negatively impact wildlife populations. Our objective was to assess medium-term lead exposure in wild Tasmanian devils (Sarcophilus harrisii). Frozen liver samples (n = 41), opportunistically collected in 2017–2022, were analysed using inductively coupled plasma mass spectrometry (ICP-MS) to determine liver lead concentrations. These results were then used to calculate the proportion of animals with elevated lead levels (>5 mg/kg dry weight) and examine the role of explanatory variables that may have influenced the results. The majority of samples analysed were from the south-east corner of Tasmania, within 50 km of Hobart. No Tasmanian devil samples were found to have elevated lead levels. The median liver lead concentration was 0.17 mg/kg (range 0.05–1.32 mg/kg). Female devils were found to have significantly higher liver lead concentrations than males (P = 0.013), which was likely related to lactation, but other variables (age, location, body mass) were not significant. These results suggest that wild Tasmanian devil populations currently show minimal medium-term evidence of exposure to lead pollution, although samples were concentrated in peri-urban areas. The results provide a baseline level which can be used to assess the impact of any future changes in lead use in Tasmania. Furthermore, these data can be used as a comparison for lead exposure studies in other mammalian scavengers, including other carnivorous marsupial species.     

Comparison of the Level(s) of DNA Damage Using Comet Assay in Bovine Oocytes Subjected to Selected Vitrification Methods

It was suggested that the cryodamage to oocytes' DNA has been responsible for the compromised developmental competence of cryopreserved oocytes. Vitrification of bovine oocytes affected not only cellular components, but also nuclear material. A significant rate of DNA fragmentation was found in bovine frozen or vitrified oocytes analysed by Comet assay regardless of cryopreservation method. Our method of vitrification using droplet system after gentle pre-equilibration treatment is one of the most effective cryopreservation methods employed for bovine oocytes so far, making it possible to develop 30% blastocyst stage embryos. In this study, the extent of DNA damage in bovine oocytes vitrified using three vitrification methods (droplet system, Open Pulled Straw and traditional vitrification in 0.25 ml insemination straws) was compared using Comet assay. Vitrification in droplet system and Open Pull Straws vitrification did not result in detectable cryoinjuries of DNA of bovine oocytes. On the contrary, DNA fragmentation was found in four of 26 oocytes vitrified in 0.25 ml straws (15.4%, p   ≤ 0.05 in comparison with the other vitrification methods).     

Genetic engineering of plant volatile terpenoids: effects on a herbivore,a predator and a parasitoid

BACKGROUND: Most insect‐resistant transgenic crops employ toxins to control pests. A novel approach is to enhance the effectiveness of natural enemies by genetic engineering of the biosynthesis of volatile organic compounds (VOCs). Before the commercialisation of such transgenic plants can be pursued, detailed fundamental studies of their effects on herbivores and their natural enemies are necessary. The linalool/nerolidol synthase gene FaNES1 was constitutively expressed from strawberry in three Arabidopsis thaliana accessions, and the behaviour of the aphid Brevicoryne brassicae L., the parasitoid Diaeretiella rapae McIntosh and the predator Episyrphus balteatus de Geer was studied. RESULTS: Transgenic FaNES1‐expressing plants emitted (E)‐nerolidol and larger amounts of (E)‐DMNT and linalool. Brevicoryne brassicae was repelled by the transgenic lines of two of the accessions, whereas its performance was not affected. Diaeretiella rapae preferred aphid‐infested transgenic plants over aphid‐infested wild‐type plants for two of the accessions. In contrast, female E. balteatus predators did not differentiate between aphid‐infested transgenic or wild‐type plants. CONCLUSION: The results indicate that the genetic engineering of plants to modify their emission of VOCs holds considerable promise for facilitating biological control of herbivores. Validation for crop plants is a necessary next step to assess the usefulness of modified volatile emission in integrated pest management. Copyright © 2012 Society of Chemical Industry     

Distinct Effects of Boar Seminal Plasma Fractions Exhibiting Different Protein Profiles on the Functionality of Highly Diluted Boar Spermatozoa

The aim of this study was to evaluate how different protein profiles of seminal plasma (SP) fractions affect sperm functionality in vitro. Ejaculates from three boars were separated into six fractions. The fractions differed from each other in their sperm content, in their total SP protein content, and their spermadhesin PSP-I/PSP-II and heparin-binding protein (HBP) concentrations. Spermatozoa were mainly recovered in fraction 2 (sperm-rich fraction, >1800 × 10⁶ spermatozoa/ml), whereas the pre-sperm fraction 1 and the post-sperm fractions 4–6 contained low numbers of spermatozoa (<500 × 10⁶/ml). Except in fraction 2, the total SP protein concentration and the concentration of both, spermadhesin PSP-I/PSP-II and the HBPs increased with fraction order. Distinct time-dependent effects were observed on motility characteristics and membrane integrity of highly diluted boar spermatozoa upon incubation with a 10% dilution of the SP from each fraction. The highest sperm viability was recorded after exposure for 5 h to fraction 2, followed by fractions 1 and 3. The percentages of motile spermatozoa also differed significantly among fractions after 5 h of incubation. Spermatozoa incubated with SP of fractions 1–3 showed the highest percentage motility. We conclude that different SP fractions exert distinct effects on the functionality of highly diluted boar spermatozoa. Fractions 1–3 appear to promote sperm survival, whereas fractions 4–6 seem to be harmful for preserving the physiological functions of highly diluted boar spermatozoa.     

Effects of Dextrose Plus Lactose in the Sows Diet on Subsequent Reproductive Performance and within Litter Birth Weight Variation

Effects of dextrose plus lactose in sow’s feed were tested on subsequent reproductive performance and within litter birth weight variation. During the last week of gestation and lactation, sows were either fed a commercial lactation diet (Control: C), or an isocaloric diet containing 25 g/kg dextrose plus 25 g/kg lactose (Treatment: T). In the subsequent weaning‐to‐oestrus interval (WEI), all sows received the same amount of a commercial feed, but T sows were supplemented with 150 g dextrose plus 150 g lactose per day. Weight and backfat changes were recorded as well as litter characteristics during the treatment period and the subsequent parity. No significant effect of treatment was found on the subsequent reproductive performance, including the number of piglets born, although the number of live born piglets was 0.51 larger (p = 0.31) and weight of the live born piglets was 84 g higher in the T sows (p = 0.07) than in the C sows. When sows were categorized in sows with 12 or less and more than 12 total born piglets in the previous litter, treatment of sows with dextrose plus lactose resulted for the group with 12 or less piglets in a strong increase in subsequent total born piglets (13.97), whereas in the untreated sows the subsequent litter size was 11.89. In the group with more than 12 total born piglets, no effect of treatment was found (interaction between previous litter size and treatment p = 0.03). The within litter variation in birth weight in the subsequent litter was numerically lower in the T sows. We concluded that the use of dextrose and lactose during lactation and WEI seems to enhance litter size in sows with low previous litter size and seems to have the potential to reduce the within litter variation in birth weight.     

Nutritional evaluation of waste date fruit as partial substitute for soybean meal in practical diets of juvenile Nile tilapia, Oreochromis niloticus L.

The potential of waste date meal (WDM; low-quality date palm, Phoenix dactylifera L.) as a carbohydrate source in formulated diets for Nile tilapia was evaluated. Four isocaloric-practical diets (15.7 kJ g⁻¹) were formulated incorporating WDM at 0, 100, 200 and 300 g kg⁻¹ levels as partial substitutes for soybean meal (SBM). These were designated D₀ [284 g crude protein (CP) and 383 g carbohydrate (CHO) kg⁻¹ diet], D₁ (279 g CP and 446 g CHO kg⁻¹ diet), D₂ (207 g CP and 495 g CHO kg⁻¹ diet) and D₃ (175 g CP and 578 g CHO kg⁻¹ diet). Each diet was fed to three replicate groups of 30 fish [20.20 ± 0.09 g (±SE)] for 75 days. No feed-related mortality was observed during the entire experimental period. Final body weight (FBW) and specific growth rate (SGR) in the different treatments were statistically not significantly different ( P  > 0.05). Protein efficiency rate (PER) was lowest in diet D₀ and increased with decrease of SBM content (D₁–D₃). A significant increase in whole body lipid content was recorded in fish fed diets D₂ and D₃. Results showed that WDM could be a substitute for SBM up to 300 g kg⁻¹ in practical Nile tilapia diets without compromising growth.     

Identification of Polymorphisms in the Osteopontin Gene and Their Associations with Certain Semen Production Traits of Water Buffaloes in the Brazilian Amazon

The osteopontin gene may influence the fertility of water buffaloes because it is a protein present in sperm. The aim of this work was to identify polymorphisms in this gene and associate them with fertility parameters of animals kept under extensive grazing. A total of 306 male buffaloes older than 18 months, from two farms, one in the state of Amapá and the other in the state of Pará, Brazil were used in the study. Seven SNPs were identified in the regions studied. The polymorphisms were in gene positions 1478, 1513 and 1611 in the region 5′upstrem and positions 6690, 6737, 6925 and 6952 in the region amplified in intron 5. The SNPs were associated with the traits, namely scrotal circumference, scrotal volume, sperm motility, sperm concentration and sperm pathology. There were significant SNPs (p < 0.05) for all the traits. The SNP 6690 was significant for scrotal circumference, sperm concentration, sperm motility and sperm pathology and the SNP 6737 for scrotal volume. The genotype AA of SNP 6690 presented the highest averages for scrotal circumference, sperm concentration and motility and the lowest total number of sperm pathologies. For the scrotal volume trait, the animals with the largest volume were correlated with the presence of the genotype GG of SNP 6737. These results indicate a significance of the osteopontin gene as it seems to exert a substantial influence on the semen production traits of male buffaloes.     

Production of Oocytes of Nile Tilapia (Oreochromis niloticus) for In Vitro Fertilization via Hormonal Treatments

Only a few studies have described hormonal treatments for induction of synchronicity and gamete collection in Nile tilapia (Oreochromis niloticus), both important for assortative matings in breeding programmes and essential for polyploidy technologies. In this study, we compared the effectiveness of carp pituitary extract (CPE), Nile tilapia pituitary extract (TPE), human chorionic gonadotropin (hCG) and gonadotropin‐releasing hormone (GnRH) protocols on the induction of spawning and egg production in Nile tilapia. Among the hormonal treatments analysed, only hCG was effective for producing viable gametes for in vitro fertilization. To verify the viability of this hormonal treatment, hCG was tested using different doses (1000, 2000, 3000, 4000 and 5000 IU/kg) in a large number of females (208 animals) from two Nile tilapia lines. The results indicated that hCG doses between 1000 and 5000 IU/kg could be used to induce final oocyte maturation in Nile tilapia with collection of stripped oocytes. This is the first study to report differential reproductive responses to hormonal treatment between tilapia lines: line 1 was more efficient at producing eggs and post‐hatching larvae after hCG induction than line 2. In conclusion, we demonstrated that the hCG protocol may be applied on a large scale to induce final oocyte maturation in Nile tilapia. The development of a protocol for in vitro fertilization in Nile tilapia may aid in breeding programmes and biotechnological assays for the development of genetically modified lines of Nile tilapia.