ABSTRACT
Knowledge of circadian rhythm clock gene expression outside the suprachiasmatic nucleus is increasing. The purpose of this study was to determine whether expression of circadian clock genes differed within or among the bovine stress axis tissues (e.g., amygdala, hypothalamus, pituitary, adrenal cortex, and adrenal medulla). Tissues were obtained at an abattoir from eight mature nonpregnant Brahman cows that had been maintained in the same pasture and nutritional conditions. Sample tissues were stored in RNase-free sterile cryovials at -80 °C until the total RNA was extracted, quantified, assessed, and sequenced (NovaSeq 6000 system; paired-end 150 bp cycles). The trimmed reads were then mapped to a Bos taurus (B. taurus) reference genome (Umd3.1). Further analysis used the edgeR package. Raw gene count tables were read into RStudio, and low-expression genes were filtered out using the criteria of three minimum reads per gene in at least five samples. Normalization factors were then calculated using the trimmed mean of M values method to produce normalized gene counts within each sample tissue. The normalized gene counts important for a circadian rhythm were analyzed within and between each tissue of the stress axis using the GLM and CORR procedures of the Statistical Analysis System (SAS). The relative expression profiles of circadian clock genes differed (p < 0.01) within each tissue, with neuronal PAS domain protein 2 (NPAS2) having greater expression in the amygdala (p < 0.01) and period circadian regulator (PER1) having greater expression in all other tissues (p < 0.01). The expression among tissues also differed (p < 0.01) for individual circadian clock genes, with circadian locomotor output cycles protein kaput (CLOCK) expression being greater within the adrenal tissues and nuclear receptor subfamily 1 group D member 1 (NR1D1) expression being greater within the other tissues (p < 0.01). Overall, the results indicate that within each tissue, the various circadian clock genes were differentially expressed, in addition to being differentially expressed among the stress tissues of mature Brahman cows. Future use of these findings may assist in improving livestock husbandry and welfare by understanding interactions of the environment, stress responsiveness, and peripheral circadian rhythms.
Subject(s)
Circadian Clocks , Female , Cattle/genetics , Animals , Circadian Clocks/genetics , Period Circadian Proteins , Circadian Rhythm/genetics , Hypothalamus , Adrenal GlandsABSTRACT
Sanitizer neutralizers can assist foodborne pathogen detection during routine testing by counteracting sanitizer residues carried over into fluids collected and tested from food samples. This study tested sanitizer-matched neutralizers applied at increasing concentrations to facilitate Salmonella enterica survival following exposure to cetylpyridinium chloride (CPC) or peracetic acid (PAA), identifying minimum required concentrations of neutralizers to facilitate pathogen survival. Salmonella isolates were individually inoculated into a non-selective medium followed immediately by CPC (0.1 to 0.8% v/v) or PAA (0.0125 to 0.2% v/v) application, followed by neutralizers application. CPC was neutralized by lecithin and polysorbate 80, each supplemented into buffered peptone water (BPW) at 0.125 to 2.0X its respective content in Dey-Engley (D/E) neutralizing buffer. PAA was neutralized in BPW supplemented with disodium phosphate, potassium monophosphate, and sodium thiosulfate, each at 0.25 to 3.0X its respective concentration in BPW (phosphates) or D/E buffer (thiosulfate). Addition of neutralizers at 1X their respective concentrations in D/E buffer was required to allow Salmonella growth at the maximum CPC concentration (0.8%), while 2X neutralizer addition was required for Salmonella growth at the maximum PAA level (0.2%). Sanitizer neutralizers can assist pathogen survival and detection during routine food product testing.
ABSTRACT
Thrips-transmitted Iris yellow spot virus (IYSV) is an important economic constraint to the production of bulb and seed onion crops in the United States and many other parts of the world. Because the virus is exclusively spread by thrips, the ability to rapidly detect the virus in thrips vectors would facilitate studies on the role of thrips in virus epidemiology, and thus formulation of better vector management strategies. Using a polyclonal antiserum produced against the recombinant, Escherichia coli-expressed nonstructural protein coded by the small (S) RNA of IYSV, an enzyme linked immunosorbent assay was developed for detecting IYSV in individual as well as groups of adult thrips. The approach enabled estimating the proportion of potential thrips transmitters in a large number of field-collected thrips collected from field-grown onion plants. Availability of a practical and inexpensive test to identify viruliferous thrips would be useful in epidemiological studies to better understand the role of thrips vectors in outbreaks of this economically important virus of onion.
Subject(s)
Bunyaviridae/isolation & purification , Insect Vectors/virology , Thysanoptera/virology , Animals , Bunyaviridae/immunology , Enzyme-Linked Immunosorbent Assay , Onions/virology , Viral Proteins/immunologyABSTRACT
Thrips-transmitted Iris yellow spot virus (IYSV) (Family Bunyaviridae, Genus Tospovirus) affects onion production in the United States and worldwide. The presence of IYSV in Georgia was confirmed in 2003. Two important thrips species that transmit tospoviruses, the onion thrips (Thrips tabaci (Lindeman)) and the tobacco thrips (Frankliniella fusca (Hinds)) are known to infest onion in Georgia. However, T. tabaci is the only confirmed vector of IYSV. Experiments were conducted to test the vector status of F. fusca in comparison with T. tabaci. F. fusca and T. tabaci larvae and adults reared on IYSV-infected hosts were tested with antiserum specific to the nonstructural protein of IYSV through an antigen coated plate ELISA. The detection rates for F. fusca larvae and adults were 4.5 and 5.1%, respectively, and for T. tabaci larvae and adults they were 20.0 and 24.0%, respectively, indicating that both F. fusca and T. tabaci can transmit IYSV. Further, transmission efficiencies of F. fusca and T. tabaci were evaluated by using an indicator host, lisianthus (Eustoma russellianum (Salisbury)). Both F. fusca and T. tabaci transmitted IYSV at 18.3 and 76.6%, respectively. Results confirmed that F. fusca also can transmit IYSV but at a lower efficiency than T. tabaci. To attest if low vector competency of our laboratory-reared F. fusca population affected its IYSV transmission capability, a Tomato spotted wilt virus (Family Bunyaviridae, Genus Tospovirus) transmission experiment was conducted. F. fusca transmitted Tomato spotted wilt virus at a competent rate (90%) suggesting that the transmission efficiency of a competent thrips vector can widely vary between two closely related viruses.
Subject(s)
Enzyme-Linked Immunosorbent Assay/methods , Insect Vectors/virology , Onions/virology , Plant Diseases/virology , Thysanoptera/virology , Tospovirus/physiology , Agriculture , Animals , Gentianaceae/virology , Georgia , Insect Vectors/growth & development , Larva/growth & development , Larva/virology , Molecular Sequence Data , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, RNA , Species Specificity , Thysanoptera/growth & development , Tospovirus/geneticsABSTRACT
The impact of applying edible oils to corn silks on ear-feeding insects in sweet corn, Zea mays L., production was evaluated in 2006 and 2007. Six edible oils used in this experiment were canola, corn, olive, peanut, sesame, and soybean. Water and two commercial insecticidal oils (Neemix neem oil and nC21 Sunspray Ultrafine, a horticultural mineral oil) were used as the controls for the experiment. Six parameters evaluated in this experiment were corn earworm [Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)] damage rating, the number of sap beetle [Carpophilus spp. (Coleoptera: Nitidulidae)] adults and larvae, the number of corn silk fly (or picture-winged fly) (Diptera: Ulidiidae) larvae, common smut [Ustilago maydis (D.C.) Corda] infection rate, and corn husk coverage. Among the two control treatments, neem oil reduced corn earworm damage at both pre- and postpollination applications in 2006, but not in 2007, whereas the mineral oil applied at postpollination treatments reduced corn earworm damage in both years. The mineral oil also reduced the number of sap beetle adults, whereas the neem oil applied at postpollination attracted the most sap beetle adults in 2007. Among the six edible oil treatments, the corn and sesame oils applied at postpollination reduced corn earworm damage only in 2007. The application of the peanut oil at postpollination attracted more sap beetle adults in 2006, and more sap beetle larvae in 2007. Olive and neem oils significantly reduced husk coverage compared with the water control in both years. The mineral oil application consistently increased smut infection rate in both 2006 and 2007. Ramifications of using oil treatments in ear pest management also are discussed.