Evidence of sheep and abattoir environment as important reservoirs of multidrug resistant Salmonella and extended-spectrum beta-lactamase Escherichia coli.

The increase in antimicrobial-resistant (AMR) foodborne pathogens, including E. coli and Salmonella in animals, humans, and the environment, is a growing public health concern. Among animals, cattle, pigs, and chicken are reservoirs of these pathogens worldwide. There is a knowledge gap on the prevalence and AMR of foodborne pathogens in small ruminants (i.e., sheep and goats). This study investigates the prevalence and antimicrobial resistance of extended-spectrum beta-lactamase (ESBL) E. coli and Salmonella from sheep and their abattoir environment in North Carolina. We conducted a year-round serial cross-sectional study and collected a total of 1128 samples from sheep (n = 780) and their abattoir environment (n = 348). Sheep samples consisted of feces, cecal contents, carcass swabs, and abattoir resting area feces. Environmental samples consisted of soil samples, lairage swab, animal feed, and drinking water for animals. We used CHROMAgar EEC with 4 µg/ml of Cefotaxime for isolating ESBL E. coli, and ESBL production was confirmed by double-disk diffusion test. Salmonella was isolated and confirmed using standard methods. All of the confirmed isolates were tested against a panel of 14 antimicrobials to elucidate susceptibility profiles. The prevalence of ESBL E. coli and Salmonella was significantly higher in environmental samples (47.7% and 65.5%) compared to the sheep samples (19.5% and 17.9%), respectively (P < 0.0001). We recovered 318 ESBL E. coli and 368 Salmonella isolates from sheep and environmental samples. More than 97% (310/318) of ESBL E. coli were multidrug-resistant (MDR; resistant to ≥3 classes of antimicrobials). Most Salmonella isolates (77.2%, 284/368) were pansusceptible, and 10.1% (37/368) were MDR. We identified a total of 24 different Salmonella serotypes by whole genome sequencing (WGS). The most common serotypes were Agona (19.8%), Typhimurium (16.2%), Cannstatt (13.2%), Reading (13.2%), and Anatum (9.6%). Prevalence and percent resistance of ESBL E. coli and Salmonella isolates varied significantly by season and sample type (P < 0.0001). The co-existence of ESBL E. coli in the same sample was associated with increased percent resistance of Salmonella to Ampicillin, Chloramphenicol, Sulfisoxazole, Streptomycin, and Tetracycline. We presumed that the abattoir environment might have played a great role in the persistence and dissemination of resistant bacteria to sheep as they arrive at the abattoir. In conclusion, our study reaffirms that sheep and their abattoir environment act as important reservoirs of AMR ESBL E. coli and MDR Salmonella in the U.S. Further studies are required to determine associated public health risks.

Resistome of a carbapenemase-producing novel ST232 Klebsiella michiganensis isolate from urban hospital effluent in South Africa.

OBJECTIVES: Klebsiella michiganensis is an emerging pathogen implicated in nosocomial infections. Here we report on the resistome, virulome and mobilome of a carbapenemase-producing K. michiganensis isolate from urban hospital effluent in Pietermaritzburg, KwaZulu-Natal, South Africa. Klebsiella sp. isolate KP124 was originally isolated from the final effluent of an urban tertiary hospital in Pietermaritzburg, KwaZulu-Natal. METHODS: Following phenotypic characterisation and antibiotic susceptibility testing, the genome of carbapenemase-producing isolate KP124 was sequenced using an Illumina MiSeq platform, de novo assembled and analysed using established bioinformatics tools. RESULTS: The draft genome of strain KP124 was 6 544 586 bp in length, comprising 203 contigs >200 bp. Following confirmation of isolate KP124 as K. michiganensis using reference genomes, the blaOXA-181 carbapenemase gene as well as 11 additional genes encoding resistance against ß-lactams, aminoglycosides, fluoroquinolones and sulfonamides were detected. Virulence factors enabling iron acquisition and cell adherence, capsule locus type and plasmid replicon types were identified. CONCLUSION: This study represents the first report of an OXA-181 carbapenemase-producing K. michiganensis isolate from hospital effluent in South Africa. The presence of such a strain in the environment owing to the absence of hospital effluent treatment presents a potential risk to informal communities that may use contaminated surface water domestically.

Systematic review of caregiver burden in spouses and partners providing informal care to wounded, injured or sick (WIS) military personnel.

INTRODUCTION: For the purposes of this review, caregivers are individuals who provide care that is typically unpaid and usually takes place at home. This systematic review aims to identify burden among spouses/partners caring for wounded, injured or sick military personnel and the factors associated with caregiver burden. METHODS: A systematic review was undertaken using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses reporting guidelines. Five electronic databases and relevant websites were searched. Two reviewers appraised the quality of the studies and carried out data extraction. RESULTS: Ten original papers were identified, of which eight were quantitative studies and two were qualitative. These papers highlighted the potential negative impact caregiving can have on spouses/partners and also some of the positive aspects of caring that can strengthen intimate relationships. CONCLUSIONS: Caring for an injured or ill military spouse or partner is a difficult task, compounded by the complexity of dealing with potentially both their physical and mental health problems. However, research has also identified some positive aspects of caring that can strengthen intimate relationships.

Brewer's yeast is a potent inducer of fever, sickness behavior and inflammation within the brain.

Brewer's yeast, derived from the yeast species Saccharomyces cerevisiae (S. cerevisiae), is commonly used for inducing pyrexia in pharmacological studies screening antipyretics in rats. Despite its widespread use, the peripheral and central inflammatory response associated with Brewer's yeast-induced fever and sickness behavior in rats has not been investigated. Thus, we injected male Sprague-Dawley rats (150-200 g) subcutaneously with a high (4 g/kg, n = 9), medium (2 g/kg, n = 5) or low (0.4 g/kg, n = 6) dose of Brewer's yeast solution or saline (0.9%, n = 6) and measured core body temperature, cage activity, food intake and body mass for six days after injection. Blood and brain samples were collected at 2, 8, 18 and 72 h after injection; n = 5-7 per time point. Brewer's yeast administration dose-dependently induced fever, lethargy, anorexia and body mass stunting that was accompanied by increased blood plasma levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α and activation of inflammatory transcription factors (nuclear factor (NF) for interleukin-6, signal transducer and activator of transcription (STAT)-3, and NF-κB)) in the hypothalamus and circumventricular organs. The increased activation of transcription factors following Brewer's yeast administration was accompanied by increased hypothalamic mRNA expression of TNF-α, IL-1ß and IL-6 and rate-limiting enzymes for prostaglandin synthesis. Our results show that subcutaneous administration of S. cerevisae induces prolonged fever, anorexia and lethargy that is accompanied by a pronounced increase in the synthesis of pro-inflammatory cytokines, key prostaglandin synthesizing enzymes and transcription factors, in the periphery and brain.

Fever and sickness behavior: Friend or foe?

Fever has been recognized as an important symptom of disease since ancient times. For many years, fever was treated as a putative life-threatening phenomenon. More recently, it has been recognized as an important part of the body's defense mechanisms; indeed at times it has even been used as a therapeutic agent. The knowledge of the functional role of the central nervous system in the genesis of fever has greatly improved over the last decade. It is clear that the febrile process, which develops in the sick individual, is just one of many brain-controlled sickness symptoms. Not only will the sick individual appear "feverish" but they may also display a range of behavioral changes, such as anorexia, fatigue, loss of interest in usual daily activities, social withdrawal, listlessness or malaise, hyperalgesia, sleep disturbances and cognitive dysfunction, collectively termed "sickness behavior". In this review we consider the issue of whether fever and sickness behaviors are friend or foe during: a critical illness, the common cold or influenza, in pregnancy and in the newborn. Deciding whether these sickness responses are beneficial or harmful will very much shape our approach to the use of antipyretics during illness.

Intraperitoneal and subcutaneous injections of the TLR9 agonist ODN 1668 in rats: brain inflammatory responses are related to peripheral IL-6 rather than interferons.

Subcutaneous or intraperitoneal administration of Toll-like receptor (TLR)-9 agonist, ODN 1668 caused moderate fever and anorexia. In comparison to stimulation of other intracellular TLRs, activation of TLR9 did not result in pronounced peripheral induction of interferons, but rather induced interleukin-6. Expression of cytokines (TNFα, IL-1ß) and inducible forms of enzymes for prostaglandin E2 synthesis occurred in the brain, in conjunction with a moderate activation of the transcription factors STAT3 and NF-IL6 in brain endothelial cells. The lack of a septic-like state in ODN 1668-treated rats reinforces the therapeutic value of this drug.

Factors affecting temperature variation and habitat use in free-ranging diamondback terrapins.

Measuring the thermal conditions of aquatic reptiles with temperature dataloggers is a cost-effective way to study their behavior and habitat use. Temperature dataloggers are a particularly useful and informative approach to studying organisms such as the estuarine diamondback terrapin (Malaclemys terrapin) that inhabits a dynamic environment often inaccessible to researchers. We used carapace-mounted dataloggers to measure hourly carapace temperature (Tc) of free-ranging terrapins in South Carolina from October 2007 to 2008 to examine the effects of month, sex, creek site, and tide on Tc and to determine the effects of month, sex, and time of day on terrapin basking frequency. Simultaneous measurements of environmental temperatures (Te; shallow mud, deep mud, water) allowed us to make inferences about terrapin microhabitat use. Terrapin Tc differed significantly among months and creek and between sexes. Terrapin microhabitat use also varied monthly, with shallow mud temperature being the best predictor of Tc November-March and water temperature being the best predictor of Tc April-October. Terrapins basked most frequently in spring and fall and males basked more frequently than females. Our study contributes to a fuller understanding of terrapin thermal biology and provides support for using dataloggers to investigate behavior and habitat use of aquatic ectotherms inhabiting dynamic environments.

Isolation of Campylobacter from feral swine (Sus scrofa) on the ranch associated with the 2006 Escherichia coli O157:H7 spinach outbreak investigation in California.

We report the isolation of Campylobacter species from the same population of feral swine that was investigated in San Benito County, California, during the 2006 spinach-related Escherichia coli O157:H7 outbreak. This is the first survey of Campylobacter in a free-ranging feral swine population in the United States. Campylobacter species were cultured from buccal and rectal-anal swabs, colonic faeces and tonsils using a combination of selective enrichment and antibiotic-free membrane filtration methods. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS, Bruker Daltonics, Inc., Billerica, MA, USA) was used to identify species followed by confirmatory multiplex PCR or 16S rRNA sequencing. Genetic relatedness of Campylobacter jejuni and Campylobacter coli strains was determined by multilocus sequence typing (MLST) and porA allele sequencing. Altogether, 12 (40%) of 30 feral swine gastrointestinal and oral cavity specimens were positive, and six species were isolated: Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalsis, Campylobacter jejuni, Campylobacter lanienae and Campylobacter sputorum. Campylobacter jejuni subtypes were closely related to MLST sequence type 21 (ST-21) and had identical porA sequences. Campylobacter coli subtypes were unrelated to isolates in the pubMLST/porA database. This feral swine population lived in close association with a 'grassfed' beef cattle herd adjacent to spinach and other leafy green row crop fields. The findings underscore the importance of protecting raw vegetable crops from faecal contamination by wild or feral animals. The study also illustrates a potential risk of Campylobacter exposure for hunters during handling and processing of wild swine meat.

Fever, sickness behavior, and expression of inflammatory genes in the hypothalamus after systemic and localized subcutaneous stimulation of rats with the Toll-like receptor 7 agonist imiquimod.

The Toll-like receptor 7 (TLR7) agonist imiquimod is used for topical treatment of skin cancers. We studied the consequences of injections of imiquimod into a subcutaneous (s.c.) air pouch or of intraperitoneal (i.p.) injections on the manifestation of fever, sickness behavior, and the peripheral and brain-intrinsic induction of a variety of inflammatory molecules. Rats were given imiqimod s.c. or i.p. (1 or 5 mg/kg). Body temperature, motor activity, and food and water intake were recorded by telemetric devices. Peripheral and brain-intrinsic induction of inflammatory mediators was analyzed by real-time polymerase chain reaction (RT-PCR), bioassays, enzyme-linked immunosorbent assays (ELISAs), and immunohistochemistry. Imiquimod is the first TLR-agonist to produce more potent effects with s.c. than i.p. administration. Peripheral induction of interferons (IFNs) and putative circulating pyrogens corresponded to the magnitude of the illness responses. In the brain, an expression of cytokines (TNFα, IL-1ß, and IL-6) and inducible forms of enzymes for prostaglandin E2 synthesis (COX-2 and mPGES) occurred, which was accompanied by a moderate activation of the transcription factors NFκB and STAT3, and a strong activation of the transcription factor NF-IL6, in cells of specific areas with an open blood-brain barrier. These inflammatory responses noted within the brain were more marked after s.c. administration, than i.p. administration of imiquimod. At a dose of 5 mg/kg, imiquimod causes rather moderate brain-inflammatory responses, which are related to peripheral IFN-expression and possibly mediated by brain-intrinsic activation of NF-IL6 and induction of a proinflammatory cocktail. The lack of a septic-like state in imiquimod-treated rats reinforces the therapeutic use of this drug.

Visual evoked potentials, reaction times and eye dominance in cricketers.

AIM: Few studies have examined the physiology of cricket, including the difference in ability between batsmen to make controlled contact with a ball bowled at high speed. METHODS: We therefore measured visual evoked potentials and choice reaction times with dominant eyes, non-dominant eyes, and both eyes together, in 15 elite batsmen and 10 elite bowlers (aged 20.9 SD 1.9 years) and 9 control subjects (aged 20.2 SD 1.5 years). The latency and amplitude of waves N70, P100 and N145 were determined for each visual evoked potential (VEP). In addition interpeak latencies and peak to peak amplitudes were measured. The subjects also completed a choice reaction test to a visual stimulus. RESULTS: We found that cricketers were not more likely to have crossed dominance (dominant eye contralateral to dominant hand) than controls. Cricketers had a faster latency for VEP wave N70 than controls (p=0.03). However reaction time was not different between cricketers and the control group. Across all subjects, in comparison to monocular testing, binocular testing led to a faster choice reaction time (p=0.02) and larger amplitudes of VEP wave N70 (p=0.01). CONCLUSION: Visual processing during the first 100(-1)50 ms of the balls flight together with binocular vision facilitates retinal activation in talented cricketers.

Determinants of ball release speed in schoolboy fast-medium bowlers in cricket.

AIM: Studies investigating determinants of ball release speed have examined the technique and anthropometry of fast bowlers with little work being done on muscular strength. The aim of our study was to determine whether knee biomechanics during bowling and strength of the shoulder and knee could predict ball release speed. METHODS: Twelve cricketers, aged 16.6+/-0.7) years, from schools in Johannesburg, South Africa, volunteered for the study. Subjects were fast-medium bowlers (mean ball release speed of 29.2+/-1.8 m.s(-1)) and had been bowling for at least 5 years. Three accurate deliveries were filmed on an outdoor cricket pitch, in the sagittal plane with a high-speed digital camera recording at 250 frames per second. The mean ball release speed, knee angle at ball release and knee angle at front foot strike were determined using simple two-dimensional kinematics. On a separate day, peak concentric isokinetic muscle torque was measured for both knees and the dominant shoulder. RESULTS: Ball release speed was positively correlated to a straight knee at front foot strike (r=0.72, P=0.009) and at ball release (r=0.71, P=0.011). No significant correlation was found between ball release speed and any of the peak torque values (knee extension peak torque, r=-0.11, knee flexion peak torque, r=-0.08, shoulder internal rotation peak torque, r=0.21 and shoulder external rotation, r=0.29, P>0.05). A multiple regression model using knee angle at front foot strike and at ball release, and the angle at which peak torque is generated during shoulder internal and external rotation, predicted ball release speed (adjusted r2=0.85, P<0.002). CONCLUSIONS: We have confirmed that the angle of the front knee at the beginning and end of a delivery is an important correlate of ball release speed in schoolboy fast-medium bowlers. In addition we have also demonstrated that a multiple regression model based on knee kinematics and shoulder peak torque angles can be used to predict ball release speed.

Cinnamaldehyde content in foods determined by gas chromatography-mass spectrometry.

trans-Cinnamaldehyde, the principal component of cinnamon flavor, is a potent antimicrobial compound present in essential oils such as cinnamon. In the course of studies designed to discover its maximum microbial lethality under food-processing conditions, a gas chromatographic-mass spectrophotometric procedure was developed for the extraction and analysis of essential oil components such as cinnamaldehyde from commercial cinnamon-containing foods (several brands of cinnamon breads, cereals, cookies, puddings, applesauces, and fruit juices). The cinnamaldehyde content ranged from trace amounts in orange juice to 12.2 mg/100 g (122 ppm) in apple cinnamon cereals and 31.1 mg/100 g (311 ppm) for cinnamon swirl bread (highest value). To ascertain the heat stability of cinnamaldehyde, pure cinnamaldehyde, pure eugenol, cinnamon oil, and mixtures consisting of cinnamaldehyde plus eugenol or cinnamon oil were heated at graded temperatures up to 210 degrees C and 60 min, and then possible compositional changes were examined. Eugenol was stable to heat, as were the components of cinnamon oil: carvone, eugenol, and linalool. In contrast, starting at approximately 60 degrees C, pure cinnamaldehyde undergoes a temperature-dependent transformation to benzaldehyde under the influence of heat. Eugenol, both pure and in cinnamon oil, when added to pure cinnamaldehyde protected the aldehyde against heat destruction. The protection may due to an antioxidative action of eugenol. The possible mechanism of this effect and the significance of these findings for food chemistry and microbiology are discussed.

Identification of three triterpenoids in almond hulls.

Three triterpenoids, betulinic acid, oleanolic acid, and ursolic acid, were isolated as their methyl esters (treatment with diazomethane) from diethyl ether extracts of almond hulls (Nonpareil variety) using flash chromatography and preparative high-performance liquid chromatography. The triterpenoids, which comprised approximately 1% of the hulls, were characterized using chromatographic and spectroscopic methods. These studies demonstrate that almond hulls are a rich source of these triterpenoids, which have reported anti-inflammatory, anti-HIV, and anti-cancer activities.

Probable toxic shock syndrome without shock and multisystem involvement.

Three menstruating adolescents using tampons colonized with Staphylococcus aureus developed high fever, vomiting, diarrhea, and abdominal pain followed by conjunctival injection and a sunburn-like rash. Two girls had transient orthostatic hypotension but none developed shock or evidence of multiple organ injury. They are reported as having probably toxic shock syndrome without shock or multiple organ system involvement.

Simultaneous hemodialysis and exchange transfusion in hemolytic uremic syndrome.

We report our experience in the acute management of a seven-year-old child with severe hemolytic uremic syndrome. On three separate occasions, acute neurologic signs dramatically resolved during simultaneous exchange transfusion and hemodialysis. The ease with which exchange transfusion may be combined with acute hemodialysis is discussed. In selected cases of hemolytic uremic syndrome with marked neurologic manifestations, exchange transfusion may offer a benefit similar to that reported in adults with thrombotic thrombocytopenic purpura.