Multistate Outbreak of Human Salmonella Typhimurium Infections Linked to Pet Hedgehogs - United States, 2011-2013.

Zoonotic Salmonella infections cause approximately 130 000 illnesses annually in the United States. Of 72.9 million US households owning at least one pet, five million own small mammals; 3000 hedgehogs were documented by USDA in USDA-licensed breeding facilities and pet stores in 2012. State health department collaborators and PulseNet, the national bacterial subtyping network, identified human infections of a Salmonella Typhimurium outbreak strain, which were investigated by CDC, USDA-APHIS and state public and animal health officials. A case was defined as an illness in a person infected with the outbreak strain identified between 1 December 2011 and 3 June 2013. Investigators collected information on patient exposures, cultured animal and environmental specimens for Salmonella, and conducted traceback investigations of USDA-licensed hedgehog facilities. There were 26 cases in 12 states. Illness onset dates ranged from 26 December 2011 to 8 April 2013. The median patient age was 15 years (range = <1-91 years); 58% were female. Among 23 persons with available information, 8 (35%) were hospitalized and one outbreak strain-associated death was reported. Of 25 patients with available information, 20 (80%) reported pet hedgehog contact in the week before illness onset. The outbreak strain was isolated from animal and environmental samples collected from three ill persons' homes in three states. Hedgehogs were purchased in geographically distant states from USDA-licensed breeders (10/17, 59%); a USDA-licensed pet store (1/17, 6%); unlicensed or unknown status breeders (3/17, 18%); and private individuals (3/17, 18%). Traceback investigations of USDA-licensed facilities did not reveal a single source of infection. Public and animal health collaboration linked pet hedgehog contact to human infections of Salmonella Typhimurium, highlighting the importance of a One Health investigative approach to zoonotic salmonellosis outbreaks. More efforts are needed to increase awareness among multiple stakeholders on the risk of illness associated with pet hedgehogs.

'One Health' investigation: outbreak of human Salmonella Braenderup infections traced to a mail-order hatchery - United States, 2012-2013.

Human salmonellosis linked to contact with live poultry is an increasing public health concern. In 2012, eight unrelated outbreaks of human salmonellosis linked to live poultry contact resulted in 517 illnesses. In July 2012, PulseNet, a national molecular surveillance network, reported a multistate cluster of a rare strain of Salmonella Braenderup infections which we investigated. We defined a case as infection with the outbreak strain, determined by pulsed-field gel electrophoresis, with illness onset from 25 July 2012-27 February 2013. Ill persons and mail-order hatchery (MOH) owners were interviewed using standardized questionnaires. Traceback and environmental investigations were conducted. We identified 48 cases in 24 states. Twenty-six (81%) of 32 ill persons reported live poultry contact in the week before illness; case-patients named 12 different MOHs from eight states. The investigation identified hatchery D as the ultimate poultry source. Sampling at hatchery D yielded the outbreak strain. Hatchery D improved sanitation procedures and pest control; subsequent sampling failed to yield Salmonella. This outbreak highlights the interconnectedness of humans, animals, and the environment and the importance of industry knowledge and involvement in solving complex outbreaks. Preventing these infections requires a 'One Health' approach that leverages expertise in human, animal, and environmental health.

Changes in ventral respiratory column GABAaR ε- and δ-subunits during hibernation mediate resistance to depression by EtOH and pentobarbital.

During hibernation in the 13-lined ground squirrel, Ictidomys tridecemlineatus, the cerebral cortex is electrically silent, yet the brainstem continues to regulate cardiorespiratory function. Previous work showed that neurons in slices through the medullary ventral respiratory column (VRC) but not the cortex are insensitive to high doses of pentobarbital during hibernation, leading to the hypothesis that GABA(A) receptors (GABA(A)R) in the VRC undergo a seasonal modification in subunit composition. To test whether alteration of GABA(A)R subunits are responsible for hibernation-associated pentobarbital insensitivity, we examined an array of subunits using RT-PCR and Western blots and identified changes in ε- and δ-subunits in the medulla but not the cortex. Using immunohistochemistry, we confirmed that during hibernation, the expression of ε-subunit-containing GABA(A)Rs nearly doubles in the VRC. We also identified a population of δ-subunit-containing GABA(A)Rs adjacent to the VRC that were differentially expressed during hibernation. As δ-subunit-containing GABA(A)Rs are particularly sensitive to ethanol (EtOH), multichannel electrodes were inserted in slices of medulla and cortex from hibernating squirrels and EtOH was applied. EtOH, which normally inhibits neuronal activity, excited VRC but not cortical neurons during hibernation. This excitation was prevented by bicuculline pretreatment, indicating the involvement of GABA(A)Rs. We propose that neuronal activity in the VRC during hibernation is unaffected by pentobarbital due to upregulation of ε-subunit-containing GABA(A)Rs on VRC neurons. Synaptic input from adjacent inhibitory interneurons that express δ-subunit-containing GABA(A)Rs is responsible for the excitatory effects of EtOH on VRC neurons during hibernation.

Up-regulation of site-specific remodeling without accumulation of microcracking and loss of osteocytes.

Functional adaptation of bone normally protects the skeleton from fracture during daily activity. Accumulation of microcracking and loss of osteocytes have been implicated in the regulation and initiation of targeted (reparative) remodeling of bone and, in certain situations, the development of fatigue or stress fracture. We performed a histologic study of the dorsal cortex of the mid-diaphysis of the third metacarpal (Mc-III) bone of Thoroughbred racehorses after bones were bulk-stained in basic fuchsin and transverse calcified sections were prepared. The Thoroughbred racehorse is an extreme athlete whose Mc-III bone experiences particularly high cyclic strains during training and racing. A group of non-athletic horses was also included in the experiment. The following variables were quantified: activation frequency (Ac.f); bone formation rate (BFR); resorption space density (Rs.N/T.Ar); microcrack density (Cr.Dn); microcrack mean length (Cr.Le); microcrack surface density (Cr.S.Dn); osteocyte density (Ot.N/T.Ar; Ot.N/B.Ar); and bone volume fraction (B.Ar/T.Ar). Ac.f and BFR were estimated using a mathematical algorithm. Using confocal microscopy, bones were examined for fine microcracks, diffuse matrix injury, and disruption of the osteocyte syncytium. Low values for Cr.Dn (#/mm2) were found in both groups (0.022+/-0.008 and 0.013+/-0.006 for racing Thoroughbreds and non-athletic horses, respectively). There was no significant relationship between Cr.Dn and Ot.N/T.Ar; Ot.N/B.Ar, B.Ar/T.Ar, and Ot.N/T.Ar; Ot.N/B.Ar, and remodeling (Ac.f, Rs.N/T.Ar) and Ot.N/T.Ar; Ot.N/B.Ar. Intense remodeling of the Mc-III dorsal cortex was found in the racing Thoroughbreds (Ac.f 12.8+/-7.4 #/mm2/year; BFR 31.5+/-15.6%; Rs.N/T.Ar 0.19+/-0.09 #/mm2) and was significantly increased compared with non-athletic horses. Overall, remodeling was weakly correlated with Cr.Dn (r2=0.15, P<0.05). Subtle matrix injury, not detectable by bright-field microscopy, was particularly evident adjacent to resorption spaces in Thoroughbred bone. In non-athletic horses, disruption of the dendritic cell processes of osteocytes associated with cement lines and interstitial fragments was more evident. Taken together, these findings suggest that site-specific (targeted) induction of remodeling during functional adaptation of bone in a high-strain skeletal site is not dependent on accumulation of microcracking or loss of osteocytes. We hypothesize that athleticism can directly influence bone turnover in this extreme athlete through pathways that do not involve classical linear microcracks.

Filopodial calcium transients promote substrate-dependent growth cone turning.

Filopodia that extend from neuronal growth cones sample the environment for extracellular guidance cues, but the signals they transmit to growth cones are unknown. Filopodia were observed generating localized transient elevations of intracellular calcium ([Ca2+]i) that propagate back to the growth cone and stimulate global Ca2+ elevations. The frequency of filopodial Ca2+ transients was substrate-dependent and may be due in part to influx of Ca2+ through channels activated by integrin receptors. These transients slowed neurite outgrowth by reducing filopodial motility and promoted turning when stimulated differentially within filopodia on one side of the growth cone. These rapid signals appear to serve both as autonomous regulators of filopodial movement and as frequency-coded signals integrated within the growth cone and could be a common signaling process for many motile cells.

Coding of neuronal differentiation by calcium transients.

Excitability has long been recognized as the basis for rapid signaling in the mature nervous system, but roles of channels and receptors in controlling slower processes of differentiation have been identified only more recently. Voltage-dependent and transmitter-activated channels are often expressed at early stages of development prior to synaptogenesis, and allow influx of Ca(2+). Here we examine the functions of spontaneous transient elevations of intracellular Ca(2+) in embryonic neurons. These Ca(2+) transients abruptly raise levels of Ca(2+) as much as tenfold, for brief periods, repeatedly, and can be highly localized. Like cloudbursts on the developing landscape, Ca(2+) transients modulate growth and stimulate differentiation, in a frequency-dependent manner, probably by changes in phosphorylation or proteolysis of regulatory and structural proteins in local regions. We review the mechanisms by which Ca(2+) transients are generated and their effects in regulating motility via the cytoskeleton and differentiation via transcription.

In vivo regulation of axon extension and pathfinding by growth-cone calcium transients.

Growth cones at the tips of extending neurites migrate through complex environments in the developing nervous system and guide axons to appropriate target regions using local cues. The intracellular calcium concentration ([Ca2+]i) of growth cones correlates with motility in vitro, but the physiological links between environmental cues and axon growth in vivo are unknown. Here we report that growth cones generate transient elevations of [Ca2+]i as they migrate within the embryonic spinal cord and that the rate of axon outgrowth is inversely proportional to the frequency of transients. Suppressing Ca2+ transients by photorelease of a Ca2+ chelator accelerates axon extension, whereas mimicking transients with photorelease of Ca2+ slows otherwise rapid axonal growth. The frequency of Ca2+ transients is cell-type specific and depends on the position of growth cones along their pathway. Furthermore, growth-cone stalling and axon retraction, which are two important aspects of pathfinding, are associated with high frequencies of Ca2+ transients. Our results indicate that environmentally regulated growth-cone Ca2+ transients control axon growth in the developing spinal cord.

A mixed foodborne outbreak with Salmonella heidelberg and Campylobacter jejuni in a nursing home.

OBJECTIVE: To investigate a mixed Salmonella heidelberg and Campylobacter jejuni foodborne outbreak in a nursing home. DESIGN: Retrospective cohort study with a nested case-control design. Cases were defined by positive stool-culture results. Controls needed to be both asymptomatic and culture-negative. SETTING AND PATIENTS: Residents of a 580-bed nursing home in Brooklyn, New York. RESULTS: Of the 580 residents, 119 (21%) developed illness. Of the 93 symptomatic patients who submitted specimens, cultures were positive for S heidelberg in 24 (26%), C jejuni in 14 (15%), and both microorganisms in 25 (27%). Only the pureed diet was associated highly with infection by either Salmonella (odds ratio [OR], 17.6; 95% confidence interval [CI95], 4.8-68.7; P < .001), Campylobacter (OR, 13.3; CI95, 3.2-59.2; P < .001), or both organisms (OR, 8.9; CI95, 2.7-30.3; P < .001). Among the 42 pureed foods served during the 5 days before the outbreak, five meat or poultry items were associated most strongly with culture positivity. Of these five meat items, only a chopped-liver salad was implicated by the two employees reporting illness. A reported food-handling error occurred when ground, cooked chicken livers were placed in a bowl containing raw chicken-liver juices. INTERVENTION: Recommendations for proper cleaning and sanitizing of kitchen equipment to prevent cross-contamination between raw and cooked foods. CONCLUSIONS: Mixed foodborne outbreaks occur rarely. During this outbreak, contamination of a single food item with multiple bacterial pathogens was the likely source of transmission. Improper food-handling techniques that promote growth of one microorganism also allow growth of other pathogens that may be present. Because different sources and routes of transmission may be implicated for different pathogens, specific preventive measures may vary depending on the organisms involved.

Foodborne salmonellosis.

Foodborne diseases caused by non-typhoid Salmonella are a very important public health problem and an economic burden in many parts of the world. Salmonellosis data from the WHO Global Databank on Foodborne Disease, from the literature and from the WHO Surveillance Programme in Europe were reviewed for the years 1985-1995, showing an apparent increase in the incidence of salmonellosis in many parts of the world. In industrialized countries, this increase may be due to the emergence and increase of S. enteritidis and S. typhimurium DT104. In order to reduce the incidence of human foodborne salmonellosis, measures should be taken simultaneously during the production, processing, distribution, retail marketing and handling/preparation of food to prevent the introduction of Salmonella and its multiplication. These control measures need to be supported by effective foodborne disease surveillance programmes which make it possible to recognize and investigate outbreaks and emerging pathogens, and to assess the need for and evaluate interventions by monitoring longer term trends.

Properties of a projection pathway from the medial preoptic nucleus to the midbrain periaqueductal gray of the rat and its role in the regulation of cardiovascular function.

In this study we examined (1) the effect of stimulation of the MPO on the firing activity of neurons in the PAG, (2) the role of glutamic acid in this interaction and, (3) whether reversible blockade of neuronal activity in the PAG by lidocaine can alter the effect of stimulation of the MPO on arterial blood pressure. Single pulse stimulation of the MPO produced a biphasic response in 2/32 cells and inhibited 3/32 cells. Train electrical stimulation excited 21/54 cells and inhibited 12/54 cells. The latencies to the onset of the excitatory and the inhibitory effects were not different, but the duration of the excitatory effect was slightly longer than that of the inhibitory effect. Chemical stimulation of the MPO excited 17/97 cells and inhibited 16/97 cells. The latency to onset of the excitatory response to stimulation of the MPO was longer but the duration was shorter than that of the inhibitory response. In 83% of the animals (29/35), stimulation of the MPO produced a decrease in mean arterial pressure (MAP). The duration of the response was 196.9 +/- 20.9 s and the average decrease in the MAP was 18.2 +/- 1.4 mmHg. Application of KYN blocked the excitatory response to stimulation of the MPO in 8/16 cells and the inhibitory response of 3/10 cells. Injection of lidocaine into the PAG by itself had no effect on the arterial blood pressure. However, in all animals (n = 10) lidocaine totally or significantly reduced the magnitude of the blood pressure change produced by stimulation of the MPO in a reversible manner. These studies electrophysiologically confirm a pathway between the MPO and the PAG that is, in part, under glutamatergic control. In addition, our results demonstrate that stimulation of the MPO produces a distinctive depressor effect that is mediated through the PAG.

[Experimental and pathological models that explain the regulation of cellular migration and axon's orientations]. / Modelos experimental y patológico que explican cómo se regula la migración celular y se orientan los axones.

Since Cajal discovery of nerve growth cones and their role in the growth, metabolism and destination of the axon in 1890 numerous studies have corroborated with more sophisticated techniques his original findings and ideas. The use of modified Golgi staining, drawing from the camera lucida in 2 combination with electron microscopy and t retrograde biochemical markers have helped to identify the neuronal sites of synapsis. Dynamic studies of the nerve growth cones using time-lapsed video microscopic images of living neuroblast in t culture or the intact animal have demonstrated their protactil and retractile membranes and exploratory filipodia properties. Interaction between growth cone membrane receptors and different molecules along the migrating axon determine also its final destination. Finally the role that genes play on neuronal migration and in guiding the axons to reach their targets have been identified in patients with Kallmann syndrome, a migrational disorder of olfactory axons and hypothalamic gonadotropic hormone-releasing hormone with absent gene at Xp22.3 locus manifested by anosmia and hypogonadotropic hypogonadism.

Recurrent outbreaks of Salmonella Enteritidis infections in a Texas restaurant: phage type 4 arrives in the United States.

In recent years infection caused by Salmonella serotype Enteritidis (SE) phage type 4 has spread through Europe but has been uncommon in the USA. The first recognized outbreak of this strain in the USA occurred in a Chinese restaurant in EI Paso, Texas, in April 1993; no source was identified. In September 1993, a second outbreak caused by SE phage type 4 was associated with the same restaurant. To determine the cause of the second outbreak, we compared food exposures of the 19 patients with that of two control groups. Egg rolls were the only item significantly associated with illness in both analyses (first control group: odds ratio [OR] 8.2, 95% confidence interval [CI] 2.3-31.6; second control group: OR 13.1, 95% CI 2.1-97.0). Retrospective analysis of the April outbreak also implicated egg rolls (OR 32.4, 95% CI 9.1-126.6). Egg roll batter was made from pooled shell eggs and was left at room temperature throughout the day. These two outbreaks of SE phage type 4 likely could have been prevented by using pasteurized eggs and safe food preparation practices.

The role of the basolateral nucleus of the amygdala in the pathway between the amygdala and the midbrain periaqueductal gray in the rat.

We electrophysiologically examined the connection between the basolateral nucleus of the amygdala (BLA) and the periaqueductal gray (PAG) and examined the role of the central nucleus of the amygdala (CNA) in this pathway. Train electrical stimulation of the BLA excited 21% (7/33) and inhibited 27% (9/33) of the cells recorded in the PAG. Chemical stimulation of the BLA excited 23% (13/56) and inhibited 16% (9/56) of the cells recorded in the PAG. Injection of lidocaine into the CNA by itself had no effect on PAG cells (n = 9) or on blood pressure but blocked the effect of BLA stimulation on PAG neurons in 78% of the cells recorded. It was concluded that: (1) PAG cells respond to BLA stimulation; (2) the majority of these cells are located in the dorsolateral and lateral columns of the PAG; and (3) the CNA modulates a majority of the activities of the BLA in the PAG.

Chick sensory neuronal growth cones distinguish fibronectin from laminin by making substratum contacts that resemble focal contacts.

The adhesive interactions of nerve growth cones stabilize elongating nerve fibers and mediate transmembrane signaling to regulate growth cone behaviors. We used interference reflection microscopy and immunocytochemistry to examine the dynamics and composition of substratum contacts that growth cones of chick sensory neurons make with extracellular adhesive glycoproteins, fibronectin and laminin. Interference reflection microscopy indicated that sensory neuronal growth cones on fibronectin-treated substrata, but not on laminin, make contacts that have the appearance and immobility of fibroblastic focal contacts. Interference reflection microscopy and subsequent immunocytochemical staining showed that beta 1 integrin and phosphotyrosine residues were concentrated at growth cone sites that resemble focal contacts. Two other components of focal contacts, paxillin and zyxin, were also co-localized with concentrated phosphotyrosine residues at sites that resemble focal contacts. Such staining patterns were not observed on laminin-treated substrata. Growth cone migration on fibronectin-treated substrata was inhibited by herbimycin A, a tyrosine kinase inhibitor. We conclude that sensory neuronal growth cones distinguish fibronectin from laminin by making contacts with distinct organization and regulation of cytoskeletal components at the adhesive sites. This finding suggests that growth cone interactions with different adhesive molecules lead to distinctive transmembrane organization and signaling to regulate nerve fiber elongation.

An electrophysiological characterization of the projection from the central nucleus of the amygdala to the periaqueductal gray of the rat: the role of opioid receptors.

The midbrain periaqueductal gray (PAG) and the central nucleus of the amygdala (CNA) are both known to be involved in fear and anxiety, analgesia, vocalization, cardiovascular and respiratory changes, and freezing. Anatomical studies have shown that a connection between these two regions exists but little is known about the physiology or the neurochemical constituents of this pathway. The goals of this study were to characterize the projection from the CNA to the PAG using electrophysiological techniques and to determine whether mu- and/or delta-opioid receptors, which play a large role in a majority of the functions of the PAG, are involved in this pathway. Of the 38 PAG cells tested with single shock stimulation of the CNA, 44% responded; of those, 46% were excited and 54% were inhibited. The latency to onset of response for the inhibitory cells (12.71 +/- 6.61 ms) was shorter than that of the excitatory cells (22.33 +/- 4.04 ms). Forty-six percent of the 129 PAG cells tested with train electrical stimulation of the CNA responded; 44% were excited and 56% were inhibited. Chemical stimulation of the CNA (10 mM D,L-homocysteic acid) produced similar results; 48% (62/128) of PAG cells responded; 45% of cells were excited and 55% were inhibited. The baseline firing rate of the inhibitory cells was significantly higher compared to the excitatory cells. Chemical stimulation of the CNA produced an increase in blood pressure in 12 animals, a decrease in two animals, and had no effect on the blood pressure of 68 animals. The blood pressure changes ranged between 8.5 and 26.3 mmHg with a mean of 16.2 +/- 2.2 mmHg. The effect of naloxone (given either on site in the PAG or systemically) on the response to CNA stimulation was tested in 21 cells. Twenty-five percent of the excitatory cells (2/8) and 77% (10/13) of the inhibitory cells were blocked by naloxone with the majority of the blocked cells located in the ventrolateral PAG. It is concluded that: (1) Approximately 50% of cells in the lateral and ventrolateral columns of the PAG respond to CNA stimulation; (2) the inhibitory response is mediated by a faster conducting or a more direct pathway than the pathway that mediates the excitatory response; (3) neurons that are inhibited by CNA stimulation have a significantly higher baseline firing rate than neurons that are excited, suggesting that they may be tonically active interneurons; and (4) at least one link in the CNA-PAG pathway utilizes mu- or delta-opioid receptors.

Characterization of spontaneous calcium transients in nerve growth cones and their effect on growth cone migration.

This study examines the mechanisms of spontaneous and induced [Ca2+]i spiking in nerve growth cones and the effect of spikes on growth cone migration. Over a 10-20 min observation period, 29% of DRG growth cones undergo spontaneous and transient elevations in physiological extracellular Ca2+ ((Ca2+)o; 2 mM), whereas 67% of growth cones exposed to 20 mM (Ca2+)o exhibit similar [Ca2+]i spikes. Spontaneous [Ca2+]i spiking was not observed in neuronal cell bodies or nonneuronal cells. Ca2+ influx through non-voltage-gated Ca2+ channels was required for spontaneous [Ca2+]i spikes in growth cones, since removal of (Ca2+)o, or addition of the general Ca2+ channel blockers La3+ or Ni2+, reversibly blocked [Ca2+]i spiking, while blockers of the voltage-gated Ca2+ channels did not. Experiments using agents that influence intracellular Ca2+ stores suggest that Ca2+ stores may buffer and release Ca2+ during growth cone [Ca2+]i spikes. Growth cone migration was immediately and transiently inhibited by [Ca2+]i spikes, but eventually returned to prespike rates.

Outbreak of brucellosis at a United States pork packing plant.

In 1992, the North Carolina Department of Environment, Health, and Natural Resources received 18 case reports of brucellosis from a county health department. All patients had potential exposure to the kill floor of one pork processing plant. A subsequent National Institute for Occupational Safety and Health health hazard evaluation surveyed 154 (99%) of 156 kill floor workers of this plant and found that 30 (19%) had evidence of recent (or persistent) brucellosis. These data show that significant exposure to Brucella is occurring among packing plant workers in North Carolina and suggest that some of the approximately 38,000 production workers in pork processing plants in the United States are at risk of contracting swine brucellosis. Additional measures may need to be taken to prevent occupational exposure to Brucella.

Filopodia initiate choices made by sensory neuron growth cones at laminin/fibronectin borders in vitro.

Localized expression of environmental cues is thought to provide directional information to migrating neuronal growth cones by enhancing or suppressing axon outgrowth over limited regions. To investigate how such a mechanism may function in vivo, we observed growth cones of embryonic chick dorsal root ganglion neurons at a substratum border between the extracellular matrix components laminin and fibronectin in vitro using time-lapse phase-contrast and interference reflection microscopy. We found that patterns of laminin and fibronectin could locally promote or suppress the direction of growth cone migration. While migrating on either laminin or fibronectin, at least 79% of growth cones changed their rate and/or direction of outgrowth upon contact with the alternative substratum, in a manner suggesting that growth cones were selecting one substratum over the other. Complex changes in growth cone behavior were initiated by filopodial contact with the alternate substratum, suggesting that filopodia were providing intracellular signals to the growth cone. Using interference reflection microscopy, we have found that selection of a substratum is independent of the degree of close contact to the substratum. We conclude that spatially localized ECM components can direct axon outgrowth by mechanisms based on intracellular signaling through growth cone filopodia.