A PCR diagnostic for cyclodiene insecticide resistance in the red flour beetle Tribolium castaneum

A molecular diagnostic was used to examine the conservation of cyclodiene resistance associated mutations between different strains of Tribolium castaneum (Herbst.). An improved insecticide bioassay for discrimination between resistant genotypes was developed and seven resistant strains were established from five different continents. In order to develop a molecular diagnostic a partial cDNA of the cyclodiene insecticide resistance gene Rdl, a γ-aminobutyric-acid-gated chloride-ion channel, was cloned and sequenced. This cDNA spans exon 7, the region containing the resistance-associated mutation, and part of exon 8. An ‘allele-specific’ oligonucleotide primer, carrying the resistance-associated mutation at its 3′ end, was used in combination with a flanking ‘allele-independent’ primer in the polymerase chain reaction to selectively amplify a single resistance-associated mutation from all seven strains collected worldwide. The implications of these findings for the population genetics of insecticide resistance and its management in pest insects via quarantine are discussed.     

Total intravenous anesthesia with ketamine–medetomidine–propofol in horses

Propofol is a potentially useful intravenous anesthetic agent for total intravenous anesthesia (TIVA) in horses. The purpose of this study was to compare the anesthetic and cardiorespiratory effects of TIVA following the administration of propofol alone(P–TIVA) and ketamine–medetomidine–propofol (KM–P–TIVA) in adult horses. The carotid artery was translocated to a subcutaneous position during TIVA with P–TIVA (n = 6) or KM–P–TIVA (n = 6). All horses were premedicated with medetomidine [0.005 mg kg^–1, intravenously (IV)]. Anesthesia was induced with midazolam (0.04 mg kg^–1 IV) and ketamine (2.5 mg kg IV). All horses were orotracheally intubated and breathed 100% oxygen. The KM drug combination (ketamine 40 mg mL^–1 and medetomidine 0.05 mg mL^–1) was infused at a rate of 0.025 mL kg^–1 hour^–1. Subsequently, a loading dose of propofol (0.5 mg kg^–1, bolus IV) was administered to all horses; surgical anesthesia (determined by horse response to incision and surgical manipulation, positive response being purposeful or spontaneous movement of limbs or head) was maintained by varying the propofol infusion rate as needed. Arterial blood pressure and HR were also monitored. Both methods of producing TIVA provided excellent general anesthesia for the surgical procedure. Anesthesia time was 115 ± 17 (mean ± SD) and 112 ± 11 minutes in horses anesthetized with KM–P–TIVA and P–TIVA, respectively. The infusion rate of propofol required to maintain surgical anesthesia with KM–P–TIVA was significantly less than for P–TIVA (mean infusion rate of propofol during anesthesia; KM–P–TIVA 0.15 0.02 P–TIVA 0.23 ± 0.03 mg kg^–1 minute^–1, p = 0.004). Apnea occurred in all horses lasting 1–2 minutes and intermittent positive pressure ventilation was started. Cardiovascular function was maintained during both methods of producing TIVA. There were no differences in the time to standing after the cessation of anesthesia (KM–P–TIVA 62 ± 10 minutes versus P–TIVA 87 ± 36 minutes, p = 0.150). The quality of recovery was good in KM–P–TIVA and satisfactory in P–TIVA. KM–P–TIVA and P–TIVA produced clinically useful general anesthesia with minimum cardiovascular depression. Positive pressure ventilation was required to treat respiratory depression. Respiratory depression and apnea must be considered prior to the use of propofol in the horse.     

Astaxanthin Normalizes Epigenetic Modifications of Bovine Somatic Cell Cloned Embryos and Decreases the Generation of Lipid Peroxidation

Astaxanthin is an extremely common antioxidant scavenging reactive oxygen species (ROS) and blocking lipid peroxidation. This study was conducted to investigate the effects of astaxanthin supplementation on oocyte maturation, and development of bovine somatic cell nuclear transfer (SCNT) embryos. Cumulus–oocyte complexes were cultured in maturation medium with astaxanthin (0, 0.5, 1.0, or 1.5 mg/l), respectively. We found that 0.5 mg/l astaxanthin supplementation significantly increased the proportion of oocyte maturation. Oocytes cultured in 0.5 mg/l astaxanthin supplementation were used to construct SCNT embryos and further cultured with 0, 0.5, 1.0 or 1.5 mg/l astaxanthin. The results showed that the supplementation of 0.5 mg/l astaxanthin significantly improved the proportions of cleavage and blastulation, as well as the total cell number in blastocysts compared with the control group, yet this influence was not concentration dependent. Chromosomal analyses revealed that more blastomeres showed a normal chromosomal complement in 0.5 mg/l astaxanthin treatment group, which was similar to that in IVF embryos. The methylation levels located on the exon 1 of the imprinted gene H19 and IGF2, pluripotent gene OCT4 were normalized, and global DNA methylation, H3K9 and H4K12 acetylation were also improved significantly, which was comparable to that in vitro fertilization (IVF) embryos. Moreover, we also found that astaxanthin supplementation significantly decreased the level of lipid peroxidation. Our findings showed that the supplementation of 0.5 mg/l astaxanthin to oocyte maturation medium and embryo culture medium improved oocyte maturation, SCNT embryo development, increased chromosomal stability and normalized the epigenetic modifications, as well as inhibited overproduction of lipid peroxidation.     

Structural studies on transfer RNA: preliminary crystallographic analysis

Single-crystal diffraction patterns from Escherichia coli leucine tRNA and yeast formylmethionine tRNA show a tetragonal lattice for the former, with a = 46 angstroms and c = 137 angstroms, and a hexagonal lattice for the latter, with a = 115 angstroms and c = 137 angstroms. Initial analysis suggests a molecule with a long, double helix parallel to the c-axis for both crystals.     

Design and use of a collector for the in situ isolation of particulate trace organic species in precipitation

Extracts of particulate organic matter were examined for discrete rainfall events from metropolitan Los Angeles, California, using an in situ filtration technique. Filtration efficiency was 98 % for the collection of extractable organic C associated with particles having nominal diameters greater than 0m22 μm Organic background levels of less than 260 ng per sample were determined. Rainwater particle samples were extracted with repeated hexane and benzene: isopropanol (2: 1) solvent additions using ultrasonic agitation. Extract mixtures were quantified by high-resolution gas chromatography (HRGC) and were adjusted for component losses with perdeuterated recovery standards. Yields for the neutral fractions ranged from 130 to 669 μg with flux rates corresponding to 371 to 1097μg m^?2 day^?1. Aliquots of the neutral solvent extracts were derivatized with diazomethane to convert acidic hydroxy and carboxylic acid groups to the respective methyl ether and methyl ester analogs. This step produced increased yields of 8 to 188 %, and resulted in yields for the acid +neutral extracts that ranged from 374 to 8681μg with flux rates of 591 to 2343 μg m^?2 day^?1. Source identification was conducted by high-resolution gas chromatography/mass spectrometry (HRGC/MS) analyses. Molecular analyses indicated major anthropogenic contributions from petroleum and combustion sources, and for some samples, the significant input of microbial lipid components as well. Minor amounts of vascular plant waxes were also present in most cases. These mixed inputs of both anthropogenic and biogenic materials compared closely with previous source determinations for carbonaceous aerosol particles in the Los Angeles air basin.     

Improved efficiency for colorimetric determination of furfural in citrus juices

The operational efficiency of a method for colorimetric determination of furfural in citrus juices is improved by inclusion of a concentration-screening step prior to the color reaction. The absorbance at 277 nm may be used to determine the appropriate dilution for samples exceeding the recommended maximum furfural concentration in the reaction mixture. As much as 1 h per sample may be saved in the time required for analysis.     

Clinical evaluation of balanced anesthesia using infusion of midazolam–ketamine–medetomidine in combination with inhalation of sevoflurane (MKM–OS anesthesia) in horses

MKM–OS anesthesia provides general anesthesia with minimum cardiovascular depression in experimental horses. The purpose of this study was to evaluate the effect of MKM–OS anesthesia in clinical cases. Sixty‐eight horses were anesthetized with MKM–OS anesthesia for selective or emergency surgery. The horse physical status was categorized based upon the American Society of Anesthesiologists (ASA) classification scheme. Forty‐four horses were classified as ASA I or II (low‐risk; 30 soft tissue, eight ophthalmic, and six orthopedic surgeries) and 24 horses were classified as ASA III to V (high‐risk; 24 emergency colic surgeries). All horses were administered medetomidine (0.005 mg kg^–1 IV) as premedication and anesthetized with ketamine (2.5 mg kg^–1 IV) and midazolam (0.04 mg kg^–1 IV). The horses were orotracheally intubated and connected to a large animal breathing circuit that delivered oxygen‐sevoflurane and administered the midazolam (0.8 mg mL^–1)‐ketamine (40 mg mL^–1)‐medetomidine (0.05 mg mL^–1) drug combination at a rate of 0.025 mL kg^–1 hour^–1. Surgical anesthesia was maintained by controlling the dial setting of the sevoflurane vaporizer and achieved by delivering 1.6–1.8% of end‐tidal sevoflurane concentration. All horses were mechanically ventilated during anesthesia. Hypercapnia and hypoxia were not sufficiently improved in high‐risk horses (PaCO₂; low‐risk 45–53 mm Hg versus high‐risk 56–60 mm Hg, p < 0.01: PaO₂ low‐risk 248–388 mm Hg versus high‐risk 95–180 mm Hg, p < 0.01). Heart rate was significantly higher in high‐risk horses (low‐risk 37–42 bpm versus high‐risk 44–73 bpm, p < 0.01). Dobutamine infusion was required in five low‐risk (11%) and 17 high‐risk horses (68%) to maintain mean arterial blood pressure >70 mm Hg. Eleven high‐risk horses died during the perioperative period (three euthanized during surgery, two died during recovery, six died after recovery). The quality of recovery was good in low‐risk horses and good to satisfactory in high‐risk horses. MKM–OS anesthesia provided excellent surgical anesthesia with minimal to mild cardiovascular depression in low risk‐horses and mild to moderate cardiovascular depression in high risk‐horses. The possibility of preserve cardiovascular function could be the advantage of MKM–OS anesthesia in high‐risk horses.     

Long Chain Fatty Acids of Diet as Factors Influencing Reproduction in Cattle

Cattle are fed moderate amounts of long chain fatty acids (FA) with the objective to enhance lactation and growth; however, recent interest on lipid feeding to cows has focused on reproduction, immunity and health. Increasing the caloric density of the ration by fat feeding has generally improved measures of cow reproduction, but when milk yield and body weight losses were increased by fat supplementation, positive effects on reproduction were not always observed. Feeding fat has influenced reproduction by altering the size of the dominant follicle, hastening the interval to first postpartum ovulation in beef cows, increasing progesterone concentrations during the luteal phase of the oestrous cycle, modulating uterine prostaglandin (PG) synthesis, and improving oocyte and embryo quality and developmental competence. Some of these effects were altered by the type of FA fed. The polyunsaturated FA of the n-6 and n-3 families seem to have the most remarkable effects on reproductive responses of cattle, but it is not completely clear whether these effects are mediated only by them or by other potential intermediates produced during rumen biohydrogenation. Generally, feeding fat sources rich in n-6 FA during late gestation and early lactation enhanced follicle growth, uterine PG secretion, embryo quality and pregnancy in cows. Similarly, feeding n-3 FA during lactation suppressed uterine PG release, and improved embryo quality and maintenance of pregnancy. Future research ought to focus on methods to improve the delivery of specific FA and adequately powered studies should be designed to critically evaluate their effects on establishment and maintenance of pregnancy in cattle.     

Acoustic monitoring system to quantify ingestive behavior of free-grazing cattle

Estimating forage intake by free-grazing livestock is difficult and expensive. Previous approaches include behavioral observation, ratio techniques using indigestible markers, mechanical recording of ingestive jaw motion, and acoustic recording of ingestive behaviors. Acoustic recording shows great potential but has been limited by the difficulty and time required to manually identify and classify ingestive events. We present an acoustic recording and analysis system that automatically detects, classifies, and quantifies ingestive events in free-grazing beef cattle. The system utilizes a wide-frequency acoustic microphone close to the animal's mouth, mathematical signal analysis to detect and measure ingestive events, and streaming data analysis capable of handling an unlimited amount of data. Analysis parameters can be reconfigured for different animals, forages and other changing conditions. The system measures the acoustic parameters of ingestive events, such as duration, amplitude, spectrum and energy, which can support further event classification and become the inputs to a forage intake model. We validated our detection and classification technique against the results of trained human observers based on field studies with grazing steer. The software detected 95% of manually identified bites in an event-by-event comparison. Field observations and sound attenuation analysis indicate that sounds from adjacent livestock and ambient pastoral environments have an insignificant effect upon the integrity of the recorded acoustic data set. We conclude that wideband acoustic analysis allows us to identify ingestive events accurately and automatically over extended periods of time.