Streptococcus pneumoniae and Streptococcus pyogenes.Community-acquired meningitis is a monomicrobial infection caused by either viruses or bacteria in the vast majority of patients. We report here one exceptional case of a patient with mixed bacterial meningitis due to Streptococcus pneumoniae and Streptococcus pyogenes. Bacteria were undistinguishable by direct microscopic examination of the cerebrospinal fluid. He responded well to treatment with cefotaxime and dexamethasone, with no sequelae observed at the 4-month follow-up.We report the case of a 68-year-old immunocompromised Caucasian man suffering from otitis and then meningitis caused by S. pneumoniae and S. pyogenes meningitis illustrates the life-threatening consequences of barotrauma in immunocompromised patients suffering from otorhinolaryngeal infections.This first reported case of mixed  Streptococcus pneumoniae is declining, although this organism is still responsible for more than 50 % of bacterial meningitis cases , ceftriaxone , rifampicin , clindamycin  and doxycycline , but resistant to erythromycin . S. pneumoniae was not cultured from the CSF, though both S. pneumoniae and S. pyogenes were cultured from the ear pus after 1-day and 2-day incubations at 37 °C and 5 % CO2, respectively. Colonies were identified by MALDI-TOF-MS with identification scores of 2.22 and 2.35, respectively. The ear pus S. pyogenes isolate exhibited the same antibiotic susceptibility pattern as the CSF S. pyogenes isolate. The antibiotic susceptibility of the ear pus S. pneumoniae isolate, tested by using the E-test method (BioMérieux), indicated in vitro susceptibility to penicillin , amoxicillin (0.016 mg/L), ceftriaxone (0.016 mg/L), imipenem (0.004 mg/L) and vancomycin (0.250 mg/L). Susceptibility testing to oxacillin, gentamicin, erythromycin, rifampicin, clindamycin and doxycycline by using the disk diffusion method found the isolate to be susceptible to all these antibiotics.Direct microscopic examination of the Gram-stained CSF revealed 930 polymorphonuclear cells and 170 red cells per cubic millimeter, along with numerous Gram-positive cocci. Our patient’s CSF contained 3.63 g/L total protein and 1.31 mmol/L glucose.  the CSF . CSF greOne day after admission, our patient suffered epileptic seizures resistant to 1 mg clonazepam. An electroencephalogram confirmed status epilepticus and our patient was given sodium valproate and levetiracetam and was admitted to the intensive care unit. Cefotaxime (18 gr/day) was intravenously administered with a syringe pump for 14 days in association with dexamethasone the first day. Our patient eventually recovered after 4 weeks of hospitalization. A follow-up at 4 months postdischarge found no sequelae. The otolaryngologist prescribed long-term treatment with amoxicillin to prevent any further otitis.S. pneumoniae and S. pyogenes was not initially suspected after direct microscopic examination of the CSF disclosed Gram-positive cocci. Indeed, although subtle microscopic differences have been reported that distinguish both streptococci, they are unreliable for prospectively distinguishing between both pathogens in routine practice. Detection of S. pneumoniae antigen by using immunochromatographic tests and real-time PCR, however, enables rapid detection of S. pneumoniae in the CSF at the point of care [In this patient, mixed meningitis due to both  of care .S. pneumoniae is the leading bacterial etiology of purulent meningitis in adults [S. pyogenes meningitis is by itself a rare condition, and is reported in a total of 29 adult patients, including the one reported here [S. pyogenes meningitis has a poor prognosis; six patients died and most of the other patients showed neurological complications (Table Moreover, the case here reported of a mixed infection is exceptional as no such case has ever been previously reported. Indeed, while n adults , S. pyogted here –11. S. pS. pneumoniae and S. pyogenes exhibiting the same antibiotic susceptibility pattern as the CSF isolate. We hypothesized that in this patient with purulent otitis, a 12-hour flight led to a barotraumatic breach of the tympanus and central nervous system infection. Barotrauma following flight has been previously reported [S. pneumoniae meningitis following pneumococcal otitis after scuba diving has been described [S. pneumoniae. In such patients, pancytopenia at the time of admission is an additional risk factor [In our patient, purulent otitis was confirmed as the portal of entry by culture of both escribed . Patientk factor .In the patient reported here, both streptococci tested susceptible to cefotaxime, the empirical treatment administered to our patient and successfully continued for 14 days. This case indicates that immunocompromised patients should avoid barotrauma when presenting with acute uncontrolled bacterial otitis. Concerning their sensitivity to pneumococcal infections, patients should be offered the 13-valent pneumococcal conjugate vaccine at the time of their chronic lymphoid leukemia diagnosis .Written informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.


To the Editor:Dermacentor-borne necrosis erythema lymphadenopathy/tickborne lymphadenopathy (DEBONEL/TIBOLA) is an apparently benign, self-limiting rickettsial disease transmitted by Dermacentor ticks  should be considered within the differential diagnosis of a similar syndrome recently named scalp eschar associated with neck lymphadenopathy after a tick bite (SENLAT)  on November 2, 2014. Three days later, he noticed a mild ache on the occipital area of his scalp and found an attached tick that he removed with his fingers. A week later, he sought care from an infectious disease specialist because of itchy discomfort at the area of the tick bite. We recently saw a patient in whom acute myopericarditis developed after he was bitten by a large tick on the scalp and showed clinical signs of DEBONEL/TIBOLA/SENLAT, most likely attributable to Rickettsia (SFGR) was 1:128. Four days later, the patient sought care at an emergency department, reporting retrosternal chest pain. Electrocardiogram revealed a diffuse ST-segment elevation with PR-segment depression; serum creatine phosphokinase and troponin T levels were 327 IU/L (reference range 10–190 IU/L) and 420 ng/mL (reference <14 ng/mL), respectively. Myopericarditis was diagnosed. A transthoracic echocardiogram ruled out pericardial effusion, valve vegetations, and left ventricular dysfunction; cardiovascular magnetic resonance imaging performed 4 days later showed myocardial inflammation. Blood cultures were sterile, pneumococcal urinary antigen test result was negative, and IgM against coxsackievirus and Mycoplasma pneumoniae were not detected. Nonsteroidal antiinflammatory drugs were prescribed. The patient improved clinically, and electrocardiogram findings resolved. The patient received doxycycline for 4 weeks.Examination revealed an erythematous and elevated punctiform lesion with mild fluctuation in the occipital region accompanied by tender, small lymph node enlargement of both occipital lymphatic chains . No wideR. conorii, R. slovaca, and R. raoultii antibody testing showed an IgG titer of 1:512 against the 3 species. A subsequent cross-adsorption assay using R. slovaca, R. raoultii, and R. conorii antigens prepared on the basis of strains from the collection at Hospital San Pedro-CIBIR showed a decrease in IgG titers against R. conorii and R. raoultii to 1:64 and 1:256, respectively, whereas titer against R. slovaca remained at 512. IFA against Bartonella spp. and C. burnetii (Focus Diagnostics), chemiluminescence immunoassay for B. burgdorferi , and in-house microagglutination assay for F. tularensis were not reactive. The patient recovered, with only a residual scarring alopecia on the occipital region of the scalp and without cardiac dysfunction after 9-month follow-up.On a convalescent-phase serum specimen collected after 8 weeks, indirect immunofluorescence assays (IFA) for IgG against SFGR were performed in Spain’s national reference center for rickettsioses . Commercial  and in-house R. rickettsii and R. conorii , we have further epidemiologic evidence for attributing the infection to SFGR causing DEBONEL/TIBOLA/SENLAT. After serum adsorption, IFA titer against R. slovaca was 3-fold higher than that against R. conorii. R. slovaca and Candidatus R. rioja are the species most commonly found in D. marginatus ticks and in cases of DEBONEL/TIBOLA/SENLAT in Spain (Myopericarditis is a rare complication of rickettsiosis, usually associated with Rickettsia spp. with negative test results for other possible causative agents and the clinical response to doxycycline, rickettsiosis caused by R. slovaca or Candidatus R. rioja remains the most probable diagnosis. Because DEBONEL/TIBOLA/SENLAT is an emerging disease, physicians should consider that this entity may be associated with systemic complications similar to those of other tickborne rickettsioses.In view of the seroconversion to


Geocoris spp. Fallén, Hemiptera: Lygaeidae) are ubiquitous, omnivorous insect predators whose plant feeding behavior raises the question of whether they benefit or harm plants. However, several studies have investigated both the potential of Geocoris spp. to serve as biological control agents in agriculture and their importance as agents of plant indirect defense in nature. These studies have demonstrated that Geocoris spp. effectively reduce herbivore populations and increase plant yield. Previous work has also indicated that Geocoris spp. respond to visual and olfactory cues when foraging and choosing their prey and that associative learning of prey and plant cues informs their foraging strategies. For these reasons, Geocoris spp. have become models for the study of tritrophic plant-herbivore-predator interactions. Here, we present detailed images and ecological observations of G. pallens Stål and G. punctipes (Say) native to the Great Basin Desert of southwestern Utah, including observations of their life histories and color morphs, dynamics of their predatory feeding behavior and prey choice over space and time, and novel aspects of Geocoris spp.’s relationships to their host plants. These observations open up new areas to be explored regarding the behavior of Geocoris spp. and their interactions with plant and herbivore populations.Big-eyed bugs ( Geocoris spp. Fallén (Hemiptera: Lygaeidae), commonly known as big-eyed bugs, are generalist insect omnivores which occur naturally worldwide [Geocoris spp. are well known to prey on a variety of insects, including several economically important agricultural pests [Geocoris spp.—including G. bullatus (Say) [G. ochropterus (Fieber) [G. pallens Stål [G. proteus Distant [G. punctipes (Say) [G. uliginosus (Say) [G. varius (Uhler) [Geocoris spp. accept a variety of insect prey, and the field studies have also shown that Geocoris spp. reduce herbivore populations [Geocoris spp.-plant interactions is usually beneficial to plants [Geocoris spp. can be effective biological control agents in many agricultural systems. The most important consequence of plant feeding by Geocoris spp. may be that it renders them more directly susceptible to agricultural pesticides [orldwide . Geocorial pests , 2 but hal pests –6, partial pests , 5, 7. Sal pests , 6, 8–14al pests –21, and al pests –31 have (Fieber) , G. pallens Stål , 26–31,  Distant , G. punces (Say) , 21, 33,us (Say) , 19, 33, (Uhler) —to serveulations , 26–29 (ulations ) and inculations , 31. Thuo plants , and GeoGeocoris spp. adults lay their eggs on plants in nature, or on moist cotton or paper cellulose in the laboratory. Life history traits have been characterized in laboratory colonies of G. atricolor Montandon [G. bullatus [G. lubra Kirkaldy [G. pallens [G. punctipes [G. lubra, but higher at 24°C than at 27°C for G. punctipes [ontandon , G. bullbullatus , G. lubrKirkaldy , G. pall pallens , 35, andunctipes . The speunctipes , 35, 36.unctipes , 36, 37,Geocoris spp. feed on a combination of insect prey and plant material [Geocoris spp. may even require seeds or seed pods in order to complete development [Geocoris spp. prey on many different insects of varying nutritional value. Interestingly, although Helicoverpa zea Boddie (Lepidoptera: Noctuidae) eggs are higher quality food for G. punctipes than are Acyrthosiphon pisum Harris (Hemiptera: Aphididae), G. punctipes more often preyed on A. pisum in choice tests [Geocoris spp. [material , 4, 5, 7elopment , 5, 38. Nicotiana attenuata Torr. ex S. Watson  in its native habitat, the Great Basin Desert of the southwestern USA. The postfire germination behavior of N. attenuata creates large monocultures of plants that host a diverse insect herbivore community [Corimelaena extensa Uhler (Hemiptera: Thyreocoridae) [Epitrix hirtipennis (Melsheimer) and E. subcrinita LeConte (Coleoptera: Chrysomelidae) [Manduca quinquemaculata (Haworth) and M. sexta (Linnaeus) (Lepidoptera: Sphingidae), and Tupiocoris notatus (Distant) (Hemiptera: Miridae) [Spodoptera spp. Guenée (Lepidoptera: Noctuidae) and Trimerotropis spp. (Orthoptera: Acrididae) [Empoasca spp. Walsh (Hemiptera: Cicadellidae) [Heliothis spp. Ochsenheimer (Lepidoptera: Noctuidae) [G. pallens is a common predator of herbivores on N. attenuata, and both G. pallens and G. punctipes can be found on N. attenuata or neighboring plant species during N. attenuata’s growing season [Geocoris spp. respond to volatiles emitted from N. attenuata after herbivory by removing more herbivores from emitting plants [We study the ecological interactions of the wild tobacco ommunity . This hecoridae) , Epitrixmelidae) , ManducaMiridae) ; the genrididae) ; and oppellidae) , 42 and ctuidae) . G. pallg season , 31. Geog plants , 29–31, G. pallens and G. punctipes co-occurring with N. attenuata. We have observed aspects of the life history, host plant, and insect prey preferences of both species. For G. pallens, we have also made detailed recordings of feeding behavior with a high-resolution macro lens (courtesy of A. Shillabeer with Merit Motion Pictures); quantified variation in predation activity of subpopulations with respect to the lepidopteran herbivore M. sexta; assayed the inclination of different generations of nymphs and adults from a single wild population to feed on M. sexta over a season; recorded increased occurrence of Geocoris pallens on wilting N. attenuata plants; and demonstrated that G. pallens can, in fact, survive when provided only with water and vegetative plant tissue.Here, we present quantitative and qualitative observations and high-resolution images of morphology and behavior for Geocoris pallens and G. punctipes were assayed in and collected from Lytle Preserve in the Great Basin Desert of southwestern Utah, USA , where we have annual field plantations of N. attenuata, and from a nearby location where a native N. attenuata population could be found from 2007 to 2009 after a 2006 burn . In May and June 2009, we collected adults and nymphs of G. pallens from the native N. attenuata population (four collections of 73 insects (19% adults), 31 insects (71%), 99 insects (58%), and 107 insects (95%)), allowed adults to mate and lay eggs, and observed the eggs through development to adults. These collections, together with ca. 100 insects collected from Lytle and a nearby wash, were used to start a colony of G. pallens at our institute in Jena, Germany. This colony has since received annual inputs from field collections. G. punctipes adults were also collected in June 2009 and used to start a colony in Jena. The colonies are fed a diet of Nutrimac , Manduca sexta and Spodoptera littoralis  eggs and larvae, and N. attenuata green tissue and seeds, with additional water provided using moist dental rolls in microcentrifuge tubes containing tap water. They are kept in 9 L food-quality plastic boxes (Lock & Lock) with two holes in each lid of ca. 8 cm diameter each covered with a fine mesh, and containing paper towels to provide structure for oviposition and hiding places, inside a growth chamber  with 16 h D/8 h N (06:00–22:00 D/22:00–06:00 N), 26/22°C, daylight provided by Osram L 36 W/77 fluorescent lamps (http://www.osram.com/) at 50% power, 65% RH, and ventilation by PAPST type 4656 N fans (http://www.ebmpapst.com/en/).N. attenuata population in 2009  by A. Shillabeer and kindly provided by Merit Motion Pictures . The probe lens permits the capture of HD macro images with an unusually large depth of field.Pictures were taken of insects collected from the native  in 2009  or from  in 2009 . Images M. quinquemaculata and M. sexta moths oviposit in native N. attenuata populations, the number of eggs is usually not sufficient for experiments except in outbreak years. Thus, we use M. sexta eggs and larvae from lab colonies for many field assays. In June 2007, M. sexta eggs purchased from North Carolina State University were frozen to kill developing larvae and thus prevent hatching, then thawed and used to assay native Geocoris spp. predation activity in a wild population of Nicotiana attenuata growing on a recent burn —which does not damage plants, induce volatile emission, or prevent egg predation—to the underside of a similarly sized, intact lower stem leaf in a standardized position (as in [N. attenuata population (Geocoris feeding (Although burn see . Five egn (as in ) on 35 ppulation . After 3 feeding  were couG. pallens prey on small bugs from invasive stork’s bill ground cover plants (Erodium cicutarium (L.) L’Hér. ex Aiton, Geraniales: Geraniaceae) in April and May and move to N. attenuata plants later in the spring, as E. cicutarium plants are drying up. On N. attenuata, G. pallens prey on flea beetles (Epitrix spp.), which are usually the first herbivores on N. attenuata, and mirids (T. notatus) which arrive on N. attenuata as plants begin to elongate. If Manduca spp. moths oviposit on N. attenuata (which they often do when pollinating flowers), G. pallens will begin to eat Manduca spp. eggs and young larvae [G. pallens nymphs and adults to feed on M. sexta eggs . Each individual was put with a piece of damp cotton and a single M. sexta egg into a 30 mL Dixie plastic cup (http://www.dixie.com/) with a lid containing small air holes and left for 72 h; cups were kept by a window in a shaded travel trailer at the field station (May assays) or in a laboratory (June assays), and water from an underground spring (May assays) or from a tap (June assays) was added to the cotton daily. G. pallens individuals which had eaten the egg within 72 h were counted as feeders, and those which had not were counted as nonfeeders. From the June 15th collection, two of the M. sexta eggs hatched and the larvae were eaten; these G. pallens were also counted as feeders. Native Manduca spp. oviposition in the field at this time was not sufficient for feeding assays, and the M. sexta for the May assays in 2009 were kindly provided by C. Miles of the State University of New York at Binghamton; the M. sexta for June assays came from an in-house colony of the same original stock as the Binghamton colony.We observed in many years that g larvae , 30. We xta eggs . On fourtion see ; Table 1Geocoris spp. individuals associated with diseased or damaged N. attenuata plants which began to wilt. In 2012, when a massive disease outbreak occurred which killed a huge number of plants, we investigated this phenomenon by counting the presence of Geocoris spp. on dying plants versus the two nearest healthy plants  and the second nearest neighboring healthy plant (approximately 2-3 m away) for Geocoris spp. presence. Except for one plant, only single G. pallens individuals were found on plants.In several years we observed y plants . On two Geocoris spp. have been reported to feed on seeds and insects. In 2006, to test the potential of G. pallens to survive on leaf tissue, we conducted a feeding assay in which G. pallens adults collected in Lytle were offered either water from an underground spring, or spring water and an N. attenuata leaf (n = 12 collected immediately prior to the start of the assay) was caged in a 50 mL food-quality plastic container  secured with miniature claw-style hair clips and padded on the rim with foam to avoid damaging plant leaves. These “clip cages” contained a cotton ball moistened with spring water or the moist cotton ball and part of an N. attenuata leaf. The leaf was still attached to a living plant and thus did not have to be replaced for the duration of the experiment. The plant did not harbor any insect herbivores. The cotton ball with water was exchanged every second day. Mortality was monitored once a day at noon.ata leaf . Each inhttp://udel.edu/~mcdonald/statfishers.html) for Excel (Microsoft) [M. sexta-feeding G. pallens, plants harboring G. pallens individuals, and G. pallens individuals surviving on a water-only versus water and live leaf diet. When necessary, Bonferroni post hoc corrections were calculated using Excel to correct for multiple testing.Fisher’s exact tests conducted using a spreadsheet   were useG. pallens and G. punctipes can be easily distinguished by differences in the coloration of their eggs and by the size and coloration of their nymph and adult stages  L’Hér. ex Aiton  plants and alfalfa  plantations in the early spring (mid-April to mid-May) and with N. attenuata plants in the late spring to summer (from the end of May). In contrast, we have observed G. punctipes primarily on Cucurbita foetidissima Kunth in Humb.  and Datura wrightii Regel  plants. We have not observed Geocoris spp. in areas where ants are abundant.t stages  and 2. IG. punctipes on C. foetidissima is Empoasca spp., and on D. wrightii, Lema trilineata (Olivier) (Coleoptera: Chrysomelidae) eggs and Manduca spp. eggs and larvae. The main foods for G. pallens on N. attenuata appear to be Epitrix spp., T. notatus, Manduca spp. eggs and young larvae depending on their abundance, and, when plants are setting seed, C. extensa, the seed-feeding negro bug. On N. attenuata, G. pallens begin by eating primarily flea beetles (Epitrix spp.), which are usually the first herbivores on N. attenuata, and mirids (T. notatus) which arrive on N. attenuata as plants begin to elongate but will switch to eating M. sexta and M. quinquemaculata eggs and young larvae when Manduca spp. are abundant, usually after N. attenuata begins to flower and attract Manduca spp. as pollinators [G. pallens sheltering in open N. attenuata seed capsules overnight and eating ripe seed . In 2008–2010 we observed that the number of Manduca spp. eggs preyed on Geocoris spp. increased in locations which received oviposition from native Manduca spp. moths  (In the 2009 field collections of instars)  occasioninstars)  but domiinstars) . Adults G. pallens feeding on M. sexta eggs and larvae in high-resolution macro focus . This difference was driven by total Geocoris spp. predation activity and not necessarily by the attractiveness of plants for Geocoris spp. in each site: between 91% and 100% of plants at each site had at least one egg predated. There were no significant differences among sites in the numbers of plants from which eggs were predated (P > 0.4).We found that G. pallens from a native N. attenuata population in 2009 for their tendency to eat M. sexta eggs or larvae  ate M. sexta. Collections from the same N. attenuata population were tested again in June, within 48 h after collection and transport to the lab in Jena. In a collection from June 1st, 32% of individuals ate M. sexta, and this increased slightly (but not significantly) to 37% in a collection from June 15th . G. pallens individuals from collections made within the same month did not significantly differ in their tendency to eat M. sexta (P > 0.2). Mortality over the course of the 72 h assay was less than 15% and did not differ significantly among collections .We tested field collections of r larvae , Table 1M. sexta eggs (P = 1), although in the case of second-instar nymphs, which tended to eat fewer eggs, this was likely due to low replicate numbers.All collections comprised both adults and nymphs, and the May 15th collection had a particularly good representation of most nymphal stages and adults . There wN. attenuata plants wilted in the field due to various stresses, for example, uprooting by wind, cattle damage, or disease, we often observed Geocoris spp. around the dying plants. When a fungal disease outbreak killed a large number of N. attenuata plants in 2012, we found G. pallens more frequently on wilting plants , and no Geocoris spp. could be found on the second-nearest healthy plants (approximately 2-3 m away from wilting plants) in any of 28 replicates .When g plants . G. pallG. pallens adults survived significantly longer if reared on N. attenuata leaves and a water-soaked cotton ball than only on the wet cotton ball. After six days without any insect prey, twice as many G. pallens individuals died if given only water than if given leaf material and water , and after eight days all animals reared only on water had died, while six individuals given N. attenuata leaves remained alive (P = 0.0069).G. pallens and G. punctipes that co-occur with the native tobacco N. attenuata in the Great Basin Desert of southwestern Utah. For G. pallens, we have also captured images of feeding behavior with a high-resolution macro lens (courtesy of A. Shillabeer and Merit Motion Pictures), quantified variation in the predation of one insect prey species, M. sexta, in space and time, recorded increased occurrence around wilting or sick N. attenuata plants, and demonstrated the ability to survive on only water and vegetative plant tissue, which has not otherwise been demonstrated for any species of Geocoris. Furthermore, we describe how we have maintained laboratory colonies of both Geocoris species from field collections.We have observed aspects of the life history, host plant, and insect prey preferences of G. pallens and G. punctipes can be easily distinguished based on size and morphology at all life stages  and also tested after transport to the laboratory comprised 95% adults, all of which were likely offspring of the May collections. In the June 15th collection, the number of egg feeders had increased slightly to 37%.The June 1st collection made two weeks later comprised 68% adults; in this collection, the nymphs were certainly the offspring of the May collections, and the adults may have been a mix of May-nymphs and May-offspring. This collection was transported to the lab in Jena for testing, and although they were allowed to adapt for 24–48 h, transport and laboratory conditions may have negatively affected feeding rates. Only 32% of these Manduca spp. oviposition events which influenced their propensity to eat M. sexta eggs. There are typically two Manduca spp. oviposition peaks in the Lytle area and surroundings: one at the end of April to the first week of May (mainly on D. wrightii) and one in the middle of June (D. wrightii and N. attenuata).  Given that lepidopteran eggs are more nutritious for Geocoris spp. than aphids [T. notatus, it is interesting that G. pallens does not always readily eat M. sexta eggs but might need to learn to prey on them. G. punctipes seem to be strongly influenced by prey mobility rather than nutritional quality [G. pallens does not always seem to recognize M. sexta eggs as prey [G. pallens must first learn to associate Manduca spp. eggs with feeding larvae and the associated herbivore-induced plant volatiles.The May and June generations may have experienced separate n aphids , 38 and  as prey , 30. PerG. pallens individuals to be significantly (5-fold) more abundant on dying, wilting N. attenuata plants than on nearby healthy plants. In fact, in a 2012 plant disease outbreak, G. pallens were not found on healthy plants unless they were next to wilting plants. This could be due to greater foraging success for G. pallens when hunting insects fleeing from dying plants, or it might be that dying plants are a better nutritional supplement to G. pallens’s insect diet, which may also be more nutritious when herbivores feed on dying plants. It has long been known that nutrients, including amino acids, are mobilized from water-stressed and senescing plant tissue, although some reports indicate that the lack of turgor pressure in wilting plants reduces sap flow so that phloem feeders may not be able to access these increased resources [G. pallens are feeding directly from cells or the apoplastic space, they may be able to harvest the products of cellular senescence and degradation from dying plants. A more speculative hypothesis would be that G. pallens itself transfers disease when feeding on plant tissue, as is known for herbivores . If T. Geocoris spp. feed from vegetative plant tissue, all prior reports indicated that Geocoris individuals could not survive any better on vegetative tissue than on water alone [G. pallens individuals offered water and living leaf tissue on planta is 50% over 8 d, but for individuals given only water is 100%. The discrepancy between our results and previous results could be due to the appropriateness of the plant tissue for the particular Geocoris spp.; differences in nutritional quality of cut leaves [G. pallens individuals drinking from N. attenuata leaves in wild populations without leaving visible leaf damage .Although it has been reported that er alone . Here, wt leaves  versus lG. pallens change plant hosts and adapt to changes in host quality and herbivore prey abundance over their lifetimes. G. punctipes, though co-occurring with G. pallens, uses different host plant and herbivore resources than does G. pallens in southwestern Utah. Geocoris spp. are phenotypically plastic generalists which, though omnivorous, benefit plants by reducing their herbivore loads. These insects have become a model system to study the development of plant-herbivore-predator tritrophic interactions, and how predators learn plant cues, and have great promise as effective biological control agents for agriculture.Wild populations of


Mesorhizobium opportunistum strain WSM2075T was isolated in Western Australia in 2000 from root nodules of the pasture legume Biserrula pelecinus that had been inoculated with M. ciceri bv. biserrulae WSM1271. WSM2075T is an aerobic, motile, Gram negative, non-spore-forming rod that has gained the ability to nodulate B. pelecinus but is completely ineffective in N2 fixation with this host. This report reveals that the genome of M. opportunistum strain WSM2075T contains a chromosome of size 6,884,444 bp, encoding 6,685 protein-coding genes and 62 RNA-only encoding genes. The genome contains no plasmids, but does harbor a 455.7 kb genomic island from Mesorhizobium ciceri bv. biserrulae WSM1271 that has been integrated into a phenylalanine-tRNA gene. Biserrula pelecinus L. is an autogamous annual legume species that is common, though never dominant, on coarse textured and acidic Mediterranean soils [Trifolium subterraneum) and serradella (Ornithopus) [B. pelecinus [Mesorhizobium ciceri bv. biserrulae WSM1271 had to be used as an inoculant to establish an effective nitrogen fixing symbiosis. After 6 years of cultivation of B. pelecinus under field conditions, an isolate (designated WSM2075) was recovered from root nodules of plants grown near Northam, Western Australia that displayed an ineffective symbiotic phenotype [B. pelecinus by acquiring symbiotic genes from the original inoculant strain Mesorhizobium ciceri bv. biserrulae WSM1271 following a lateral gene transfer event [T (= LMG 24607 = HAMBI 3007) and is the type strain for a new species described as Mesorhizobium opportunistum [M. opportunistum WSM2075T is competitive for nodulation of B. pelecinus but cannot fix nitrogen [an soils  and can ithopus) . This reithopus) . At the elecinus ,3 and a henotype . AccumulM. opportunistum strain WSM2075T together with the description of the complete genome sequence and annotation. Here we reveal that a 455.7 kb genomic island from the inoculant Mesorhizobium ciceri bv. biserrulae WSM1271 has been horizontally transferred into M. opportunistum strain WSM2075T and integrated into the phenylalanine-tRNA gene.Here we present a summary classification and a set of general features for M. opportunistum strain WSM2075T is a motile, Gram-negative, non-spore-forming rod  broth [Strains of this organism are able to tolerate a pH range between 5.5 and 9.0. Carbon source utilization and fatty acid profiles have been described previously . MinimumMesorhizobium opportunistum strain WSM2075T in a 16S rRNA sequence based tree. This strain clusters in a tight group which included M. amorphae, M. huakuii, M. plurifarium and M. septentrionale and has >99% sequence identity with all four type strains. However, based on a polyphasic taxonomic study we have identified that this strain belongs to a new species [M. opportumistum strain WSM2075T forms an ineffective (non-N fixing) symbiosis with its original host of isolation, B. pelecinus L., as well as with Astragalus adsurgens, A. membranaceus, Lotus peregrinus and Macroptilium atropurpureum [T has a broader host range for nodulation than Mesorhizobium ciceri bv. biserrulae WSM1271 [urpureum ,6. In alThis organism was selected for sequencing on the basis of its environmental and agricultural relevance to issues in global carbon cycling, alternative energy production, and biogeochemical importance, and is part of the Community Sequencing Program at the U.S. Department of Energy, Joint Genome Institute (JGI) for projects of relevance to agency missions. The genome project is deposited in the Genomes OnLine Database  and the M. opportunistum strain WSM2075T was grown to mid logarithmic phase in TY rich medium [h medium  on a gyrh medium .Mesorhizobium opportunistum WSM2075T was sequenced at the Joint Genome Institute (JGI) using a combination of Illumina [in silico using software developed at JGI , and mis-assemblies were corrected using Dupfinisher [The genome of Illumina  and 454 Illumina . An IlluIllumina . IlluminIllumina , and theIllumina -30 in thfinisher , or sequGenes were identified using Prodigal  as part The genome is 6,884,444 nucleotides with 62.87% GC content  and comp


Salmonellae (NTS) cause a large burden of invasive and gastrointestinal disease among young children in sub-Saharan Africa. No vaccine is currently available. Previous reports indicate the importance of the O-antigen of Salmonella lipopolysaccharide for virulence and resistance to antibody-mediated killing. We hypothesised that isolates with more O-antigen have increased resistance to antibody-mediated killing and are more likely to be invasive than gastrointestinal.Nontyphoidal S. Enteritidis were less susceptible and expressed more O-antigen than Typhimurium (p<0.0001 for both comparisons). For S. Typhimurium, but not Enteritidis, O-antigen expression correlated with reduced sensitivity to killing . Both serovars expressed O-antigen populations ranging 21–33 kDa average molecular weight. O-antigen from most Typhimurium were O-acetylated on rhamnose and abequose residues, while Enteritidis O-antigen had low or no O-acetylation. Both Typhimurium and Enteritidis O-antigen were approximately 20%–50% glucosylated. Amount of S. Typhimurium O-antigen and O-antigen glucosylation level were inversely related. There was no clear association between clinical presentation and antibody susceptibility, O-antigen level or other O-antigen features.We studied 192 NTS isolates  from blood and stools, mostly from paediatric admissions in Kenya 2000–2011. Isolates were tested for susceptibility to antibody-mediated killing, using whole adult serum. O-antigen structural characteristics, including O-acetylation and glucosylation, were investigated. Overall, isolates were susceptible to antibody-mediated killing, but S. Typhimurium and Enteritidis clinical isolates are susceptible to antibody-mediated killing, with degree of susceptibility varying with level of O-antigen for S. Typhimurium. This supports the development of an antibody-inducing vaccine against NTS for Africa. No clear differences were found in the phenotype of isolates from blood and stool, suggesting that the same isolates can cause invasive disease and gastroenteritis. Genome studies are required to understand whether invasive and gastrointestinal isolates differ at the genotypic level.Kenyan  Salmonellae (NTS) are an emerging major cause of invasive bacterial disease in African children aged less than 5 years and immunocompromised adults, with an estimated case fatality rate of 20–25%. NTS also cause diarrhoea, a killer of about 1.5 million young children annually, mainly in low- and middle-income countries. No vaccine against NTS is available, but improved understanding of the Salmonella bacteria that cause disease in Africa would help the development of new vaccines. The authors characterized a collection of 192 Kenyan NTS strains (114 S. Typhimurium and 78 S. Enteritidis) from blood and stool specimens. All strains could be killed to differing extents by antibodies present in the blood of healthy HIV-uninfected African adults, supporting the development of a vaccine that will induce protective antibodies when given to African children. Differences in killing by antibody were partly related to the amount of O-antigen on the bacterial surface. There were no clear distinction between stains causing invasive disease and diarrhoea, suggesting that the same strains may be capable of causing both forms of disease. Clarification of this will require genomic analysis.Nontyphoidal  Salmonella enterica serovars Typhimurium and Enteritidis account for nearly 80% of all human isolates reported globally [S. pneumoniae disease [NTS are a major but neglected cause of invasive disease (hence iNTS disease) in Africa –3. Salmoglobally . While iglobally –7, in Af disease  and bloo disease . No vacc disease –13.S. Typhimurium are genetically distinct from those in the rest of the world, and are characterized by a new sequence type, ST313, that has spread throughout the continent [S. Enteritidis which is more prevalent than S. Typhimurium in some parts of Africa [In Kenya, iNTS disease is particularly frequent in rural areas , with inontinent . The ST3f Africa ,19, and f Africa . InvestiSalmonella and other Gram-negative bacteria, and is key to the interaction between Salmonella and its environment. The O-antigen chain  constitutes the outermost part of LPS [Salmonella, LPS plays an important role in the interaction between the bacterium and its host and is a virulence factor required for colonization and resistance to antibody-mediated killing [Salmonella that lack OAg are avirulent and succumb readily to complement-mediated killing [Lipopolysaccharide (LPS) forms the outer layer of t of LPS . In path killing . While t killing . OAg pro killing –26, whic killing –29 and p killing .S. Typhimurium and 78 S. Enteritidis isolates derived from patients, mostly children, presenting to three main Kenyan hospitals in Nairobi and from 31 healthy carriers related to these patients. Our primary objective was to characterise NTS isolates circulating in endemic areas, with the goal of identifying bacterial features associated with either invasive or gastrointestinal pathology. We hypothesised that invasive isolates would, by nature, be more resistant to survival in the blood than gastrointestinal isolates, and focused our analysis on the susceptibility of isolates to antibody-mediated killing and characterization of the OAg expressed by the various isolates. We investigated the total amount of OAg expressed by the various isolates and OAg specific characteristics such as molecular weight (MW), O-acetylation and glucosylation levels, parameters which can influence OAg immunogenicity. Additionally, we tested the sensitivity of the isolates to antibody killing, using whole human serum from HIV-uninfected adults [In this study we analysed a bacterial collection of 114 S. Typhimurium and 78 S. Enteritidis) collected between 2000 and 2011 at the Centre for Microbiology Research (CMR), Kenya Medical Research Institute (KEMRI) in Nairobi, Kenya. Study isolates were selected from the archived bacterial culture collection based on availability of unequivocal serovar identification and availability of clinical records and metadata. The isolates were stored in Tryptic Soy Broth with 15% glycerol at-80°C. The isolates were from blood and stools with a few additional samples from urine (3 samples), cerebrospinal fluid  and environmental sources (2 samples). The isolates were mostly from children admitted to three main hospitals in Nairobi county, namely: Kenyatta National Referral hospital, Gertrude’s Children’s Hospital and Aga Khan University Hospital. This collection also included isolates from healthy carriers (31 samples). Before use in the present study, the archived isolates were re-cultured and the serotype was confirmed by antibody-based agglutination .The study culture collection contained 192 archived NTS isolates  agar after 0 and 180 min. As a negative control, reference sera were heat-inactivated at 56°C for 45 min to inactivate endogenous complement and included in each SBA experiment. Additional internal controls included testing the representative endemic African S. Typhimurium isolate D23580, previously shown to be susceptible to serum killing [Salmonella concentration.The study isolates were tested for their susceptibility/resistance to killing against the serum pool described above, using a protocol involving whole serum and endogenous complement activity as previously described . Briefly killing  and hype killing . SusceptOAg extraction was performed by acid hydrolysis . Bacteri1H NMR analysis was performed as a confirmation of the identity of the OAg samples and to verify the presence of O-acetyl groups along the OAg chain as previously described [escribed .Glucosylation level of OAg samples was estimated by HPAEC—PAD after acid hydrolysis of the OAg to release the monosaccharides constituting the sugar chain as described previously. Commercial glucose was used for building the calibration curve (0.5–10 μg/ml). Glucose (Glc) level is calculated as molar ratio of Glc to rhamnose (Rha), sugar present in each OAg repeating unit. The analysis was also used as confirmation of the OAg samples identity verifying the correct monosaccharides ratios in the repeating unit ,36.2PO4, 5% CH3CN, pH 7.2, at the flow rate of 0.5 ml/min (isocratic method for 30 min). OAg peaks were detected by differential refractive index (dRI). Void and bed volume calibration was performed with λ-DNA (λ-DNA molecular weight (MW) Marker III 0.12–21.2 kb; Roche) and sodium azide , respectively. OAg average MW was estimated on standard dextrans (Sigma) calibration curve.HPLC—SEC analysis was used to estimate the molecular size distribution of OAg populations ,36. SampSalmonella serovars examined (S. Typhimurium and S. Enteritidis) was compared using the Mann-Whitney U test. Possible correlations  were analysed by Spearman rank.Serum susceptibility and OAg amounts of the two S. Typhimurium and 78 (41%) were S. Enteritidis. This study collection included isolates from blood, stools , CSF, urine and environmental/animal sources as described in S. Typhimurium, 57 with S. Enteritidis). The age ranged from 1 month to 59 years for S. Typhimurium isolates and 0 months to 64 years for S. Enteritidis isolates, the majority  of these patients being below 5 years . Overall, the median age of subjects from whom S. Typhimurium and S. Enteritidis strains were isolated was 2 and 3.5 years, respectively. Ages were not available for the healthy carriers.Of the 192 NTS isolates in the study, 114 (59%) were S. Typhimurium and S. Enteritidis isolates  underwent a reduction in viable bacterial counts over the 180 minute time course of the assay, and so were designated ‘sensitive’ to antibody-mediated killing. Overall median Log10 reduction in viable bacteria after 180 minutes was 2.9 for S. Typhimurium. Indeed 50% (57/114) S. Typhimurium were highly susceptible to killing, undergoing a 3.0 Log10 reduction. In contrast S. Enteritidis as a group were less susceptible to antibody-mediated killing than S. Typhimurium, being killed by a median of 1.2 Log10 . However, all S. Enteritidis were sensitive to killing. A small proportion  was killed by a full 3.0 Log10.SBA results using the serum pool demonstrated a marked difference in susceptibility to antibody-mediated killing among individual isolates and a clear difference in susceptibility between S. Enteritidis isolates expressed more OAg than S. Typhimurium, with 70.9% of the isolates producing more than 15 μg/ml/OD OAg compared to only 42.1% S. Typhimurium. Median OAg production for S. Enteritidis isolates was 16.8 μg/ml/OD compared to 14.4 μg/ml/OD of S. Typhimurium (p<0.0001 by Mann Whitney test)  . All stuey test) ; <5% isoS. Typhimurium and 45 S. Enteritidis) showed that only 6.8% of S. Typhimurium OAg were not O-acetylated, compared with 35.6% and 42.2% of S. Enteritidis OAg with no or traces of O-acetyl groups, respectively. For S. Typhimurium, OAg, O-acetylation was detected on both rhamnose (Rha) and abequose (Abe) sugar residues in the majority of isolates (63%), but also on Rha only (9.6%) and Abe only (17.8%)  in accordance with our initial hypothesis, but this was not present for S. Enteritidis   and either antibody susceptibility or polysaccharide production, for either isolates . There we<0.001) .NTS infections are a major problem in sub-Saharan Africa and can either present as bacteraemia, with symptoms of gastroenteritis in under half of patients, or as diarrheal disease without bloodstream infection ,11,37. BS. Typhimurium, were susceptible to antibody-dependent complement-mediated killing, using a serum pool from an endemic region. Previously it has been shown that the antibody response specific to NTS is acquired with age, corresponding with a decline in cases of NTS bacteraemia [S. Typhimurium and S. Enteritidis) antibodies as expected among healthy adults in a Salmonella endemic region of sub-Saharan Africa such as Kenya, Malawi or Tanzania [Salmonella antibodies against the vast majority of endemic NTS isolates in the current study. These findings support an important role for antibodies in protective vaccines against iNTS disease [The main finding of the study is that all isolates studied, apart from one teraemia ,38. The Tanzania . The stu disease ,33,39.Salmonella serovars studied. According to our study hypothesis, decreased susceptibility of S. Enteritidis compared with S. Typhimurium could be the result of higher levels of OAg, not excluding the contributions of other pathogen features, not investigated here. A previous report from Malaysia found that S. Enteritidis isolates were more often isolated from the blood than stool of patients compared with other Salmonella serovars [S. Enteritidis isolates to evade host first line defence mechanisms and gain access to the bloodstream. It has also been reported that, when exposed to serum, S. Typhimurium upregulates genes for increased production of very long chain OAg to escape complement-dependent killing [S. Typhimurium isolates is consistent with the hypothesis that high OAg expression protects against such killing. It is also well know that strains lacking OAg are rapidly eliminated from the host [S. Enteritidis isolates and susceptibility to antibody-mediated killing does not support our hypothesis for this serovar concerning OAg levels and resistance to antibody killing. This finding suggests that other inherent factors of Salmonella, particularly S. Enteritidis, have a role in determining loss of sensitivity. In other words, quantity of OAg is more important for resistance to antibody-killing of S. Typhimurium than S. Enteritidis. Further studies are required to better understand the factors contributing to the inherent resistance of S. Enteritidis.Another key finding is the difference in relationship between susceptibility to antibody-mediated killing and levels of OAg expression for the two serovars , but theserovars , increas killing . Our finthe host ,42, and S. Typhimurium were more heterogeneous than S. Enteritidis. OAg O-acetylation occurred in almost all S. Typhimurium isolates, and most often at two different sites: C-2 Abe (known to provide factor O:5 specificity) and C-2/C-3 Rha. Rha O-acetylation was first reported by Wollin et al [S. Typhimurium isolate [S. Typhimurium isolates with either Abe- or Rha-O-acetylation only, shows that O-acetylation at both sites can occur independently. A S. Typhimurium OAg-based vaccine for Africa would need to induce antibodies capable of binding to both OAg specificities. In contrast, O-acetylation of S. Enteritidis OAg was not as common, with more than two-thirds of the isolates lacking or having very low levels of O-acetylation.In relation to OAg fine specificity (O-acetylation) of the isolates in our collection, in et al  and we h isolate . The extS. Typhimurium and can occur on the O:12 antigen galactose (C4 position) generating the 12–2 variant, and on the O:1 antigen galactose (C6 position) (phage-determined) [S. Typhimurium isolates in our collection were glucosylated, with less than 30% of them at levels < 20%, showing that glucosylation is a very common feature among endemic strains. Similarly, 86% S. Enteritidis isolates investigated showed a glucosylation level >20%. Glucosylation in S. Enteritidis is not associated with specific OAg factors and it has not been extensively studied. A report on S. Enteritidis food-borne salmonellosis strains showed glucosylated LPS in some mouse isolates, closely related to strains obtained from eggs [S. Enteritidis strains.Glucosylation is another OAg modification for which variation has been described for ermined) . It has ermined) . All S. rom eggs , but notShigella, LPS glucosylation promotes bacterial invasion by enhancing type III secretion system (T3SS) function and is associated with reduced LPS chain length [Salmonella, T3SS is necessary for invasion of intestinal epithelium and survival within macrophages and further investigation into different glucosylation levels and T3SS-mediated invasion is still to be performed. The inverse correlation between S. Typhimurium OAg glucosylation and OAg expression observed in the current study may be related to T3SS function, as previously reported for Shigella [S. Typhimurium [Salmonella isolates defective for the synthesis of long and very long OAg species have been shown to have increased translocation of a Salmonella pathogenicity island 1 (SPI1)-T3SS effector protein promoting invasion. Therefore, also for Salmonella, there may be the need to balance T3SS activity, possibly enhanced by shorter and glucosylated LPS, and protection against innate immune effectors, which is favoured by a thicker LPS layer [In n length . In SalmShigella  and S. Thimurium . SalmonePS layer ,28.Salmonella in the blood of bacteraemic patients can be as low as 1 CFU/ml [The reasons for NTS disease presenting as invasive or enteric disease are not well understood and could relate to host immunity and/or pathogen virulence. Epidemiological data indicate a frequent presentation of the invasive syndrome, without diarrhoea symptoms, in younger age groups , often a1 CFU/ml  and bloo1 CFU/ml .The lack of obvious difference between phenotype of NTS isolates from invasive and gastrointestinal disease episodes is consistent with the concept that invasive NTS isolates are able to cause diarrhoea and vice versa. If true, this would implicate host immunity as the major determinant of NTS disease presentation in endemic sub-Saharan African countries. Other previous studies  have notThere are several limitations to the current study: categorization of isolates as “invasive” or “gastroenteric” was based on clinical presentation, as confirmed by microbiological analysis. Blood and stool samples belonging to the same patient were rarely collected, making it impossible to exclude that bacteria from bacteremic patients were not also present in stool and vice versa . AnotherS. Enteritidis were generally less susceptible to killing than S. Typhimurium isolates and expressed higher levels of OAg, but while OAg expression correlated with antibody resistance for S. Typhimurium isolates, no correlation could be found for S. Enteritidis, supporting a role for other inherent bacterial factors in conferring resistance to antibody killing for this Salmonella serogroup. Serovar- and strain-specific differences in OAg expression and fine specificity were found. Those features could not be correlated to clinical presentation, so the same strains may be able to cause both invasive disease and diarrhoea. Whole genome analysis of invasive and gastrointestinal NTS isolates from Africa will provide additional insights regarding whether these strains are the same or different [In summary, Kenyan NTS isolates from patients with bacteraemia and gastroenteritis were susceptible to antibody-mediated killing, supporting the development of an antibody-inducing vaccine against NTS for Africa. ifferent .S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 TableS. Typhimurium and S. Enteritidis isolates.b. Glucosylation results for (XLSX)Click here for additional data file.


Oryza sativa) is a staple crop that supports half the world's population and an important monocot model system. Monocot leaf matures in a basipetal manner, and has a well-defined developmental gradient along the longitudinal axis. However, little is known about its transcriptional dynamics after leaf maturation. In this study, we have reconstructed a high spatial resolution transcriptome for the matured rice leaf by sectioning the leaf into seven 3-cm fragments. We have performed strand-specific Illumina sequencing to generate gene expression profiles for each fragment. We found that the matured leaf contains a longitudinal gene expression gradient, with 6.97%  of the expressed genes showing differentially expression along the seven sections. The leaf transcriptome showed a gradual transition from accumulating transcripts related to primary cell wall and basic cellular metabolism at the base to those involved in photosynthesis and energy production in the middle, and catabolic metabolism process toward the tip.Rice ( Monocot or cereal crops such as rice, maize, wheat and barley provide the majority of the calories in human diets, and their leaf tissues are the most important source of the photosynthetic energy. Unlike dicot, monocot has distinct leaf architecture with a basipetal developmental gradient (maturation from tip to base) along the leaf bladeIt has been shown that a gene expression gradient exists in the developing rice and maize leaves and the transcriptome inside a developing leaf is highly dynamic23http://137.189.43.55/JBrowse/). Our data revealed the existence of a dynamic gene expression gradient within the matured leaf and could provide a valuable genetic resource for C3 monocot research and future crop improvement programs.In this study, we examined the gene expression patterns within the matured rice leaf by dividing the fully expanded rice leaf blade it into seven 3 cm consecutive sections, and performed strand-specific RNA sequencing to generate a comprehensive transcriptome database for the matured rice leaf  in at least one of the seven sections, and 18,992 genes (33.92%) in all seven sections. The numbers of expressed genes are similar in all sections, with 30,321, 29,649, 29,628, 28,986, 30,200, 29,552 and 30,340 expressed genes identified in sections 1 to 7, respectively. In total, 2,603 differentially expressed genes (DEGs)  were identified in at least one pairwise comparison between the sections, and 142 out of 2,603 DEGs have ORGO annotation . DEG anaGO, AgriGO and KEGG enrichment analysis indicated that the 2,603 DEGs we identified are associated with various biological processes, molecular functions, cellular components and pathways . We clusLeaf blade is the major organ for photosynthesis, and photosystem differentiation is tightly linked to the leaf development. In term of sheer transcript abundance, the middle sector of leaf blade accumulated more photosynthesis related transcripts. Our KEGG enrichment analysis of DEGs revealed that many enriched pathways in the middle sector (cluster group 3) are related to photosynthesis . The expWe further examined the expression pattern of several key genes involved in photosynthesis, such as the genes encoding the antenna proteins that are responsible for light-harvestingOur data also revealed a longitudinal dynamics of transcription factor gene expression. According to the annotation of PlantTFDB7Using a comprehensive transcriptome dataset, we have demonstrated that a nonuniform gene expression pattern exists along longitudinal axis in matured rice leaf. Gene expression analysis showed that highly expressed genes near the base are associated with primary cell wall formation, basic cellular metabolism and secondary metabolites. Many enriched pathways in the leaf base are associated with well-defined plant secondary metabolism processes, such as starch and sucrose metabolism, phenylpropanoid and flavonoid production and linoleic acid metabolism . In contRBCS (LOC_Os12g17600) is the gene with the highest expression level in all sections along the mature rice leaf longitudinal axis  is most abundant in the tip sections, and a similar tip to base gene expression gradient is observed for the PHOTOSYSTEM II SUBUNIT R gene910Previous studies have uncovered photosynthetic differences within the developing maize and rice leaves2nal axis , which rCS LOC_Osg17600 isOryza sativa l. japonica) seedlings were grown in the greenhouse, with temperature at 25–28°C. About 20 days after planting, the fully opened 4th leaves was cut it into seven 3-cm segments, from bottom to tip and labeled as sections 1 to 7, respectively. The tissues were immediately frozen in liquid nitrogen for total RNA extraction. Two biological replicates were collected for each section.The rice Nipponbare . Sequence quality was examined using FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/). Reads were mapped to the Oryza sativa genome (MSU Release 7.0) using TophatData processing and analysis were performed as previously describedp values from the hypergeometric test were corrected by FDR. Only GOslims with q < 0.05 were considered significantly enriched. AgriGO enrichment analysis was performed using http://bioinfo.cau.edu.cn/agriGO/q < 0.05 were considered statistically significant. The OGRO annotation was retrieved from the http://qtaro.abr.affrc.go.jp/ogro database and DEGs with OGRO annotation is listed in Enrichment analyses were carried out as previously describedeEF-1a)To verify RNA-seq results, real-time PCR was performed using SYBR green (Applied Biosystems) and ABI Step One Plus Realtime, quantiutative (q) PCR System (Applied Biosystems). A set of genes selected on the basis of their relatively broad distribution of expression among all sections . Expresshttp://137.189.43.55/JBrowse/The data has been deposited in NCBI SRA under the accession number SRP045512 and the BAM alignment results are available at B.L., C.X., P.L., S.Z., Z.F. and J.J.G. conceived and supervised the project. W.Y., A.Z. and N.L. performed the experiments, Y.C., W.Z. and J.X. performed the Illumina sequencing. N.L., S.Y., D.Z. and C.X. analyzed the data.sup fig 1-4Dataset


Clostridium perfringens bacteremia is associated with adverse outcomes. Known risk factors include chronic kidney disease, malignancy, diabetes mellitus, and gastrointestinal disease. We present a 74-year-old man admitted with confusion, vomiting, and abdominal pain. Exam revealed tachycardia, hypotension, lethargy, distended abdomen, and cold extremities. He required intubation and aggressive resuscitation for septic shock. Laboratory data showed leukocytosis, metabolic acidosis, acute kidney injury, and elevated lipase. CT scan of abdomen revealed acute pancreatitis and small bowel ileus. He was started on vancomycin and piperacillin-tazobactam. Initial blood cultures were positive for C. perfringens on day five. Metronidazole and clindamycin were added to the regimen. Repeat CT (day 7) revealed pancreatic necrosis. The patient developed profound circulatory shock requiring multiple vasopressors, renal failure requiring dialysis, and bacteremia with vancomycin-resistant enterococci. Hemodynamic instability precluded surgical intervention and he succumbed to multiorgan failure. Interestingly, our isolate was beta lactamase producing. We review the epidemiology, risk factors, presentation, and management of C. perfringens bacteremia. This case indicates a need for high clinical suspicion for clostridial sepsis and that extended spectrum beta lactam antibiotic coverage may be inadequate and should be supplemented with use of clindamycin or metronidazole if culture is positive, until sensitivities are known.  Clostridium species are Gram-positive, spore-forming, obligate anaerobic bacilli. Malignancies, renal insufficiency, and other chronic illnesses have been associated with Clostridium perfringens (C. perfringens) bacteremia [ C. perfringens bacteremia associated with acute pancreatitis. The purpose of this case report is to alert physicians to suspect C. perfringens bacteremia in elderly patients presenting with abdominal symptoms and sepsis. Additionally, providers should be aware that although rare, C. perfringens can produce beta lactamase which can complicate antibiotic management.cteremia . This encteremia . We presA 74-year-old man was brought to our emergency room with altered mental status, abdominal pain, and multiple episodes of vomiting and diarrhea for one day. Review of systems was negative for any other systemic complaints including fever. His medical history was significant for hypertension. He had no prior history of diabetes or surgery and had no pertinent family history. Personal history included prior alcohol use for fifty years with a reported intake of approximately 3-4 beers daily.On physical exam, he was tachycardic, hypotensive, and obtunded with cold extremities. His abdomen was tense and distended with sluggish bowel sounds. He was emergently intubated and aggressive resuscitation initiated for presumptive septic shock.3/μL), anion gap metabolic acidosis (pH 7.10 with anion gap of 31), acute kidney injury (creatinine level 2.6 mg/dL), elevated lipase (1088 U/L), and mildly elevated transaminases . Mean corpuscular volume (MCV) was 98 fL and there was no evidence of hemolysis. Of note, his HbA1c was 8.4% suggesting undiagnosed diabetes mellitus. Urine toxicology screen was negative and serum ethanol level was less than 10 mg/dL. Computerized tomography (CT) scan of the abdomen  exist and can produce as many as 12 different toxins. The 4 principal toxins of C. perfringens are alpha, beta, epsilon, and iota [ C. perfringens bacteremia has been associated with intravascular hemolysis and death [Five subtypes ofand iota –10. Alphand iota  as well and iota . C. perfnd death , 16. Lownd death . Epsilonnd death . When innd death .C. perfringens bacteremia, especially with a penicillinase producing strain, is a rare clinical entity. Epidemiological studies examining Clostridium bacteremia have been conducted in Taiwan, Japan, Canada, and the United States. C. perfringens bacteremia of 0.97 per 100,000 population. Elderly patients with comorbid illnesses, especially renal insufficiency or malignancy, were at increased risk. The 30-day and attributed mortalities were 26.9% and 8.6%, respectively. Nosocomial acquired C. perfringens infection was a significant predictor of 30-day mortality. Most C. perfringens blood isolates were susceptible to the antibiotics tested. Resistance was observed in only seven out of ninety-three isolates, primarily to penicillin and clindamycin.A study from northern Taiwan  demonstr C. perfringens bacteremia. Overall 30-day mortality was 27%. Septic shock at initial presentation was significantly associated with mortality [ Clostridium bacteremia. C. perfringens was the most common species isolated [A review of all blood cultures drawn in a Japanese tertiary center from 2001 to 2009 demonstrated only 18 patients withortality . A popul Clostridium infection in 0.12% with C. perfringens again, being the most common isolate (21.7%) [Review of blood cultures drawn in a rural hospital in Wisconsin from 1990 to 1997 yielded (21.7%) . Clostridium species and have demonstrated that failure to institute early, appropriate antimicrobial therapy may be associated with a poor outcome [Several studies have identified other conditions associated with the pathogenicity of outcome , 21, 22. C. perfringens (77%), Clostridium bifermentans (9%), and Clostridium septicum (4%). Clostridium bacteremia in patients with underlying liver cirrhosis and septic shock on initial presentation were poor prognostic factors [In another Taiwanese study, a review of 73 patients with clostridial bacteremia in an 11-year period identified diabetes mellitus and liver cirrhosis as the most common underlying comorbidities. Etiological species identified were factors . Clostridium was also studied in Israel. They found that growth of Clostridium species in blood cultures, even in the absence of one of the histotoxic syndromes, is often of clinical significance. Patients with Clostridium bacteremia were older, had a higher frequency of gastrointestinal disease, especially colorectal tumors, were more frequently associated with polymicrobial bacteremia, and had a higher mortality rate [The significance of positive blood culture fority rate . Clostridium infection were readily apparent. Although C. perfringens bacteremia was not initially suspected in our patient, he was treated with appropriate antibiotics.Our patient had several known risk factors for clostridial infection and subsequent mortality. He was older, apparently diabetic, had gastrointestinal disease, and presented in shock. However, none of the histotoxic syndromes associated withOur isolate was reported to be beta lactamase producing. Based on the susceptibility report see , our pat C. perfringens in vitro may predict cephalosporinase activity in vivo. Beta lactam resistance has been well studied in many pathogenic bacteria. In vitro susceptibility does not necessarily produce in vivo activity of an apparently appropriate antibiotic [ C. perfringens. Early studies have demonstrated in vitro susceptibility of C. perfringens strains to vancomycin [ C. perfringens [The patient was initiated on piperacillin-tazobactam and vancomycin, both of which could be expected to have efficacy towards our isolate. Nevertheless, our patient deteriorated, requiring the addition of metronidazole and clindamycin. It is conceivable that penicillinasetibiotic . Unfortuncomycin . Howeverfringens . C. perfringens bacteremia, clinicians should be aware of the risk factors and presentation associated with this pathogen. We recommend immediate initiation of additional antibiotic coverage, for example, clindamycin or metronidazole, as soon as C. perfringens is isolated in culture, pending sensitivity. Data regarding clinical outcomes in beta lactamase producing Clostridia are scarce and may warrant further subgroup analysis. Furthermore, more microbiologic studies are required exploring in vitro and in vivo susceptibility patterns.Although rare, given the significant mortality of


Campylobacter jejuni is one of the leading bacterial causes of human gastroenteritis. Due to the increasing rates of human campylobacteriosis, C. jejuni is considered as a serious public health concern worldwide. C. jejuni is a microaerophilic, fastidious bacterium. C. jejuni must overcome a wide range of stress conditions during foodborne transmission to humans, such as food preservation and processing conditions, and even in infection of the gastrointestinal tracts of humans. Particularly, this microaerophilic foodborne pathogen must survive in the atmospheric conditions prior to the initiation of infection. C. jejuni possesses unique regulatory mechanisms for oxidative stress resistance. Lacking OxyR and SoxRS that are highly conserved in other Gram-negative foodborne pathogens, C. jejuni modulates the expression of genes involved in oxidative stress resistance mainly via the peroxide resistance regulator and Campylobacter oxidative stress regulator. Based on recent findings of ours and others, in this review, we described how C. jejuni regulates the expression of oxidative stress defense. Campylobacter jejuni is considered as the most common bacterial cause of acute gastroenteritis in humans  resistance is found in approximately 92% C. jejuni isolates from raw chicken in South Korea (C. jejuni isolates from children in Thailand (The Centers for Disease Control and Prevention (CDC) estimated that  country . In Canaper year . Worldwiper year . Despiteng trend . In addi concern . In someth Korea , and 100Thailand .Campylobacter sp. are isolated from a wide range of animals, such as poultry, cattle, sheep, and dogs, and most of them are associated with various disease symptoms in animals and humans  and hydrogen peroxide (H2O2). Subsequently, intracellular ferrous iron (Fe2+) and H2O2 produce a highly reactive hydroxyl radical (HO⋅) via the Fenton reaction  in the atmosphere , such as the superoxide anion  and peroxide stress   render C. jejuni significantly sensitive to menadione, a superoxide generator, and slightly to H2O2 (C. jejuni (Superoxide dismutase (SOD) is the primary enzyme that is responsible for the detoxification of superoxide . Whereasectively , C. jejured SodB , and a seroxide) . Accordiparaquat . In S. fate SDS; ; however to H2O2 . It woul. jejuni .2O2 to H2O and O2, plays an important role in oxidative defense mechanism , and two putative peroxidases  H as an electron donor to transfer electrons to AhpC , but not to H2O2  is a periplasmic protein that reduces H2O2 to water. C. jejuni possesses two putative CCP genes; docA and Cjj0382  would affect oxidative stress defense strain-dependently. Also, the autoinducer 2-depdendent quorum sensing affects oxidative stress depending on the strain. A luxS mutant in C. jejuni NCTC 11168 exhibits comparable resistance to oxidative stress to the wild type  and H2O2 ] . In Salm to AhpC . Unlike e genome . AhpC exe genome . An ahpC to H2O2 , possiblhpC does . Two Tpxperoxide . Cytochr Cjj0382 . Even thi 81–176 . In C. jsistance , suggestild type ; howeverand H2O2 .C. jejuni from oxidative stress by scavenging free intracellular irons associated with ROS generation or by repairing cellular damages caused by ROS. The E. coli Dps (DNA binding protein from starved cells) is known to protect DNA by sequestrating free Fe2+ and by reducing the formation of ROS , a recent extensive mutagenesis and colonization study done by ahpC, katA, and sodB play the primary role in the oxidative stress defense of C. jejuni.Although a number of genes and proteins have been reported to contribute to oxidative stress defense in E. coli, oxidative stress is involved in bacterial response to a variety of stress conditions, such as heat, salinity, heavy metals, and lactic acid bacteria  by about twofold  than low temperatures , suggesting that temperature affects oxidative stress resistance in C. jejuni  and severe (5% NaCl) salt stress also increases expression of oxidative stress defense genes, such as ahpC, katA, and katE, along with genes associated with iron homeostatis, such as ferrochelatase  state. Biofilm formation is one of extensively studied survival mechanisms in pathogenic bacteria in stress environments  conditions than in oxygen-limited  conditions  and the formation of a disulfide bond , the negative auto-regulation of perR involves Mn2+ in B. subtilis  by H2O2 in B. subtilis PerR results in the dissociation of Fe2+ from PerR, and the demetallated PerR cannot bind to DNA, and this conformational changes in PerR induce gene expression .Unlike other Gram-negative foodborne pathogenic bacteria, such as i genome . InsteadPerR: The C. jejuni genome contains two genes encoding Fur homologs. Whereas Fur regulates iron uptake genes  ke genes , anotherke genes . As PerRess PerR . Similarlis PerR , a perR peroxide . A microlis PerR  and showpromoter . Similarcription . Whereassubtilis , perR auanganese .C. jejuni survival under aerobic conditions  are conserved in C. jejuni PerR  . jejuni . In addisistance . A recenegulator . In fact studies . Given tion does .CosR: CosR (Campylobacter oxidative stress regulator) is an OmpR-type essential response regulator that plays an important role in the control of oxidative stress resistance in C. jejuni , and cprR (Cj1227c) mutants could not be generated, suggesting that these two genes are essential in C. jejuni. C. jejuni to paraquat, a superoxide generator, reduces the protein level of CosR (Cj0355c) in a 2DGE analysis, suggesting a potential role of CosR in oxidative stress defense. In the two studies above, the essentiality of cosR was an obstacle to functional characterization of CosR, because it is not possible to generate its knock-out mutant. Thus, the function of CosR has been characterized in an alternative way by using antisense-mediated gene knockdown  is direct based on the results of electrophoretic mobility shift assays and DNase I footprinting assays  . jejuni , 2012. Tor CheA; . In an enockdown , which wactivity . CosR reg assays , 2012. A. jejuni . The Cosuni  and the sensor kinase CprS  and functionally complements a csrA mutation in E. coli in the regulation of biofilm formation and motility , RpoN (σ54), and FliA (σ28), and lacks stress-related sigma factors found in other bacteria  ase CprS . Due to e stress . Further and gne . Cj1556 y Cj1556 . A Cj155rophages . Cj1103 motility . Inactivc stress . C. jeju. jejuni . Althoug. jejuni .C. jejuni may encounter during foodborne transmission to humans, and (2) critical survival mechanisms, such as aerotolerance, biofilm and VBNC formation. C. jejuni’s resistance to and survival in stress conditions that are associated with oxidative stress defense may eventually increase the possibility of C. jejuni transmission to humans via food, resulting in food safety problems. Additionally, studies thus far have demonstrated that oxidative stress response in C. jejuni is controlled by many different regulatory mechanisms that are distinctly different from mechanisms reported in other foodborne pathogens. Investigation of molecular mechanisms for oxidative stress resistance in C. jejuni has been greatly facilitated by the availability of improved experimental techniques. Further characterization of the mechanisms for oxidative stress resistance and regulation will tell us how this fastidious microaerophilic pathogen survives in stress conditions and causes such huge problems in food safety.In summary, oxidative stress resistance affects: (1) bacterial response to various stress conditions that The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Brucella spp. and Mycobacterium bovis (M. bovis), respectively, as comparative models to outline the merits of LAV platforms with emphasis on molecular strategies that have been pursued to generate LAVs with enhanced vaccine safety and efficacy profiles. Finally, we discuss the prospects of LAV platforms in the fight against brucellosis and BTb and outline new avenues for future research towards developing effective vaccines using LAV platforms.Vaccination of humans and animals with live attenuated organisms has proven to be an effective means of combatting some important infectious diseases. In fact, the 20th century witnessed tremendous improvements in human and animal health worldwide as a consequence of large-scale vaccination programs with live attenuated vaccines (LAVs). Here, we use the neglected zoonotic diseases brucellosis and bovine tuberculosis (BTb) caused by  Vaccination provides the most effective means of preventing and eradicating infectious diseases, and historically, live attenuated vaccines (LAVs) have proven effective in protecting humans and animals from infection. LAVs are weakened versions of the pathogen, obtained by disrupting or mutating one or more genes, which renders the pathogen incapable of causing disease; however, the pathogen remains sufficiently potent to educate the immune system to initiate and establish highly specific short-term or lifelong immunity . LAVs ofBrucellosis, also called Bang’s disease, is a worldwide zoonosis of profound importance . GloballBrucella has the potential to render patients severely debilitated, which can perpetuate poverty and tax health care resources [Brucella infection, typical treatment regimens are prolonged and are often accompanied by unwanted side effects [In humans, brucellosis typically presents as an undulant fever but gradually becomes systemic, affecting practically every organ system of the body with protracted symptoms of arthopathy, myalgia, and debilitation . In rumiesources ,52. Alth effects .Mycobacterium bovis induces both pulmonary and extrapulmonary symptoms in humans and animals [M. bovis shares 99.95% genome sequence similarity with M. tuberculosis and a live attenuated version of the pathogen, Bacillus Calmette–Guérin (BCG), provides the current and only widely used vaccine against tuberculosis [M. bovis in humans as well as a wide variety of animal species is increasing. More than 50 million cattle are estimated to be infected with the pathogen worldwide [M. bovis infection of humans presents with similar symptoms as M. tuberculosis; however, human infection with virulent M. bovis infection is not responsive to the antibiotic treatment commonly used to treat M. tuberculosis [The intracellular bacteria  animals . M. bovirculosis . The gloorldwide , resultiorldwide . M. bovirculosis . Althougrculosis , wildlifrculosis , especiarculosis .Brucella and M. bovis share similar features that support LAV development. First, the unavailability of highly effective subunit vaccines that protect humans against these pathogens can be largely attributed to their intricate immune evasion strategies [Brucella spp. and M. bovis are intracellular vacuolar pathogens that establish replicative niches within acidic compartments of professional phagocytic cells [M. bovis and Brucella spp. and their respective host proteins will reveal novel avenues for engineering next-generation LAVs. The purpose of this article is not to comprehensively review the development of vaccines for M. bovis or Brucella spp., which can be found elsewhere [The pathogenic programs of rategies –67. Howerategies . Therefoic cells  and theric cells  includinic cells , phagosoic cells . Therefoic cells  or stimulsewhere –76, but M. bovis resulted in attenuation and subsequent establishment of M. bovis BCG [M. bovis. Serial passage has also been used to generate Brucella vaccines strains. For example, RB51 is a spontaneous rough mutant derived using repeated passage of Brucella abortus strain 2308 in vitro [B. abortus strains 45/20 [Several approaches to LAV generation have been described , includiovis BCG . Serial ovis BCG . Strainsin vitro . The mains 45/20 , proved B. abortus strain 19 (S19), for example, is a smooth strain that became attenuated during prolonged cultivation under dehydrating conditions [Exposure of microorganisms to irradiation ,99 or otnditions . The molnditions , includinditions . Irradianditions ,104, an nditions , and thenditions . More renditions  and RNA-nditions . The appBrucella [M. bovis [M. bovis yielded mutants with similar efficacy to BCG in a guinea pig model of tuberculosis [Brucella spp., this strategy led to the mutation and identification of several virulence genes [Brucella infection [Transposon-mediated mutagenesis by random gene inactivation has been used to identify virulence factors and construct mutations in Brucella ,108 and M. bovis ,110. Thirculosis . In Brucce genes . For exace genes , were denfection ,33,114. nfection . This stnfection . Followinfection  or gene nfection . FinallySalmonella enterica serovar Typhimurium) that express heterologous Brucella protective antigens  have been tested [Brucella vaccine antigens L7/L12 [Brucella challenge. However, only modest protection, which was lower than reported for the LAVs RB51 or S19, was observed in these studies.Various vaccine modalities, including DNA, protein, viral vector, and live attenuated vaccines, have been developed for protecting animals or humans from brucellosis ,76. For n tested . Vaccinin tested  or Semlin tested  vectors s L7/L12 , SodC [1s L7/L12 , or tranBrucella has yet to be developed, several LAV formulations show promise. Although efficacy has been demonstrated in bovine populations with S19, this vaccine can induce abortion in pregnant animals [Brucella virulence determinants and the role of individual genes in the survival and virulence of the pathogen in vitro and in vivo. Collectively, this work has provided opportunities to build upon the merits of the LAV approach by using rational bioengineering of strains for the generation of attenuated agents that harbor deletions of key genes that are essential for virulence yet maintain efficacy in vaccine challenge experiments. However, it is often easier said than done with Brucella spp., as well as with other bacterial pathogens, which unlike their viral counterparts have larger genomes and greater genetic complexities that require defining strains with levels of attenuation that ensure both safety and protection. Continued efforts in our labs to develop attenuated Brucella mutants as vaccine candidates have yielded promising candidates, including ∆mucR [∆asp24 [∆vjbR mutants [B. melitensis ∆vjbR strains were shown to be defective for survival within macrophages and rapidly cleared from the spleen in BALB/c mice [∆vjbR strains was further revealed by the absence of splenomegaly in inoculated mice [vjbR mice were 5-fold less than wild-type controls [B. melitensis ∆vjbR strains are promising and warrant further evaluation in large animal and nonhuman primate models to develop an improved vaccine candidate for possible human use. In addition, an expanded analysis of delivery systems, including encapsulation of B. melitensis ∆vjbR or other LAV strains, is recommended [B. ovis ΔabcBA vaccine [B. ovis in rams [abcBA strain in ovine monocyte-derived macrophages revealed defects in intracellular multiplication, trafficking, and Brucella-containing vacuole (BCV) maturation compared to wild-type infection [While an effective subunit vaccine for  animals . It can  animals ,125 or i animals . The seang ∆mucR , ∆asp24  [∆asp24 , and ∆vj mutants . B. meliB/c mice . The safted mice . Even atcontrols ,128. Remcontrols . Therefocontrols ,33. Diff vaccine . Remarka in rams . An in vnfection . TherefoM. bovis [M. bovis auxotrophs displayed reduced bacterial burden and pathology following challenge with virulent M. bovis. These data confirm the efficacy of LAVs in preventing infection with this pathogen. In contrast, subunit vaccine antigens, when delivered alone, have generally proven to be less efficacious than BCG in conferring protection to cattle from M. bovis  to boost BCG have been tested. To date, field trials have demonstrated that BCG vaccination can protect cattle from natural exposure to M. bovis . Similariew, see ). MoreovM. bovis  and DNA e as BCG . Howevere as BCG  or viruse as BCG ,135 havee as BCG . In one hallenge . BCG strhallenge . Similarhallenge ,139. Thecfp10-esat6 locus is responsible for loss of virulence [M. bovis ΔRD1 displayed a reduced number and severity of TB lesions as well as reduced bacterial burden in BCG vaccinates [M. bovis mutant, Δmce2, exhibited greater immunological reactivity in response to tuberculin purified protein derivative (PPD) than BCG vaccinates [Sequence analysis of BCG has provided information that can be exploited to develop novel candidate LAVs. For example, mutation of chromosomal regions of difference 1 (RD1) at the irulence . M. boviccinates . Anotherccinates . TherefoB. melitensis strains harboring mutations in the LuxR family member ∆vjbR have exhibited promise as LAVs for reducing brucellosis [M. tuberculosis strain H37Rv were shown to decrease cell survival under hypoxic and reductive stress triggered by vitamin C [M. bovis strains harboring deletions in LuxR family members may provide a useful framework for the development of LAVs to address M. bovis infection.Although cellosis ,114, simitamin C . Furtheritamin C . These sAs a result of their ability to elicit protective immune responses, LAVs against bacteria may represent a superior alternative to subunit, killed, or DNA vaccines. Although subunit vaccines have an advantage in perceived safety over live attenuated organisms, including viruses, their inability to penetrate deep lymphoid tissue results in a weak immune response lacking recall and frequently requiring the addition of toxic immune stimulants or attenuated vectors for delivery ,144. AddWe have argued that LAVs provide a compelling technology to control the neglected zoonotic diseases brucellosis and BTb; this is because of the better immune response they engender and a history of past successes in disease eradication . We noteBrucella subunit vaccines with corresponding candidate LAVs remain in their infancy. Efforts to genetically engineer a multivalent or universal Brucella vaccine that affords potent cross protection also constitutes an exciting area of future research. The development of encapsulation technologies to enhance the stability and reduce the requirement of cold chain storage for LAVs may also transform the feasibility of large-scale vaccination efforts [M. bovis and Brucella, provides an exciting avenue of future investigation. Finally, new advances in genetics, bioinformatics, and pharmaceutical technology may provide avenues for addressing these challenges and thereby promote development of next-generation vaccines for addressing neglected zoonotic diseases of global consequence.There are many exciting avenues for future research and testing of LAV vaccines. For example, the analyses of prime-boost strategies that pair promising  efforts ,147. EngLAVs against brucellosis and Btb display greater efficacy and/or safety in various model systems than competing subunit vaccine platforms against these diseases.Brucella LAVs have demonstrated excellent efficacy and safety in preclinical studies.Some candidate The potential for reversion to virulence of LAVs has been partly addressed by the use of modern techniques that remove species-defining genetic loci that are neither readily reacquired nor restore function, and the removal of multiple loci reduces the risk of reversion to infinitesimal levels.Brucella vaccine, encapsulation technologies to deliver Brucella LAVs, and engineering LAVs to protect against multiple pathogens, including M. bovis and Brucella, provides exciting avenues of future investigation.Evaluation of prime-boost strategies, a multivalent or universal High-throughput approaches for generating and screening banks of mutant bacteria, including the TraSH system and RNA-guided gene editing using CRISPR-Cas technology, provide exciting areas for future investigation.Brucella abortus S19 ΔvjbR live vaccine candidate is safer than S19 and confers protection against wild-type challenge in BALB/c mice when delivered in a sustained-release vehicle. Infect Immun. 2009;77(2): 877–84.Arenas-Gamboa AM, Ficht TA, Kahl-McDonagh MM, Gomez G, Rice-Ficht AC. The Brucella ovis lacking a putative ATP-binding cassette transporter (ΔabcBA) protects against wild type Brucella ovis in rams. PLoS ONE. 2015;10(8): e0136865.Silva APC, Macêdo AA, Costa LF, Rocha CE, Garcia LNN, Farias JRD, et al. Encapsulated Brucella abortus RB51 with a functional wboA Gene results in O-antigen synthesis and enhanced vaccine efficacy but no change in rough phenotype and attenuation. Infect Immun. 2000;68(7): 3927–32.Vemulapalli R, He Y, Buccolo LS, Boyle SM, Sriranganathan N, Schurig GG. Complementation of Mycobacterium bovis BCG and Mycobacterium microti. Mol Microbiol. 2002;46(3):709–17.Pym AS, Brodin P, Brosch R, Huerre M, Cole ST. Loss of RD1 contributed to the attenuation of the live tuberculosis vaccines Mycobacterium bovis against bovine tuberculosis. Immunology. 2003;108(4):548–55.Skinner MA, Ramsay AJ, Buchan GS, Keen DL, Ranasinghe C, Slobbe L, et al. A DNA prime-live vaccine boost strategy in mice can augment IFN-γ responses to mycobacterial antigens but does not increase the protective efficacy of two attenuated strains of


Microbacterium spp. isolated from heavy metal (HM)-contaminated environments (soil and plants) can play a role in mobilization processes and in the phytoextraction of HM. Here, we report the whole-genome sequences and annotation of 10 Microbacterium spp. isolated from both HM-contaminated and -noncontaminated compartments. Microbacterium spp. have been isolated from diverse environments, including soil, plants, water, and humans . Based on related publications (–d humans –4. They helators . Interesed soils –8. The id soils –, 10. Bacications –15, we ccations –.Microbacterium sp. genomes in similar percentages.Genomic DNA was extracted with a phenol-chloroform extraction protocol . The NexMicrobacterium genes involved in HM resistance and mobilization is currently being performed and will be published in a subsequent report.Genome annotation (summarized in The nucleotide sequences have been deposited at the DDBJ/EMBL/GenBank under the accession numbers provided in


Halomonas sp. strain MCTG39a was isolated from coastal sea surface water based on its ability to utilize n-hexadecane. During growth in marine medium the strain produces an amphiphilic exopolymeric substance (EPS) amended with glucose, which emulsifies a variety of oil hydrocarbon substrates. Here, we present the genome sequence of this strain, which is 4,979,193 bp with 4,614 genes and an average G+C content of 55.0%. Halamonas sp. strain MCTG39a from a sea surface water sample collected off the coast of California was isolated by enrichment with n-hexadecane as the sole carbon source . The strain produces a surface-active (amphiphilic) exopolymer, which emulsifies a range of hydrocarbons and increases their bioavailability for biodegradation. Strain MCTG39a is a strictly aerobic and motile, rod-shaped marine bacterium that degrades hydrocarbons. It produces an exopolymeric substance (EPS) with strong emulsifying qualities against a range of hydrocarbons.Halomonas sp. strain MCTG39a. Genomic DNA was isolated, and the sequence was generated at the Department of Energy (DOE) Joint Genome Institute (JGI) using Pacific Biosciences (PacBio) technology. A Pacbio SMRTbellTM library was constructed and sequenced on the PacBio RS platform, which generated 247,476 filtered subreads totaling 781.1 Mbp. All general aspects of library construction and sequencing performed at the JGI can be found at http://www.jgi.doe.gov. The raw reads were assembled using HGAP (version: 2.1.1) . Additional gene prediction analysis and manual functional annotation was performed within the Integrated Microbial Genomes–Expert Review (IMG ER) platform (http://img.jgi.doe.gov) developed by the Joint Genome Institute, Walnut Creek, CA, USA  with functional predictions for 3,957 of them. A total of 92 RNA genes were detected. Other genes, characteristic for the genus, are given in the IMG database . This geHalomonas sp. strain MCTG39a obtained in this study was deposited in GenBank as part of BioProject no. PRJNA224116, with individual genome sequences submitted as whole-genome shotgun projects under the accession no. JQLV00000000.The draft genome sequence of


Ananas comosus var. bracteatus (Red Pineapple) is an important ornamental plant for its colorful leaves and decorative red fruits. Because of its complex genome, it is difficult to understand the molecular mechanisms involved in the growth and development. Thus high-throughput transcriptome sequencing of Ananas comosus var. bracteatus is necessary to generate large quantities of transcript sequences for the purpose of gene discovery and functional genomic studies.Ananas comosus var. bracteatus transcriptome was sequenced by the Illumina paired-end sequencing technology. We obtained a total of 23.5 million high quality sequencing reads, 1,555,808 contigs and 41,052 unigenes. In total 41,052 unigenes of Ananas comosus var. bracteatus, 23,275 unigenes were annotated in the NCBI non-redundant protein database and 23,134 unigenes were annotated in the Swiss-Port database. Out of these, 17,748 and 8,505 unigenes were assigned to gene ontology categories and clusters of orthologous groups, respectively. Functional annotation against Kyoto Encyclopedia of Genes and Genomes Pathway database identified 5,825 unigenes which were mapped to 117 pathways. The assembly predicted many unigenes that were previously unknown. The annotated unigenes were compared against pineapple, rice, maize, Arabidopsis, and sorghum. Unigenes that did not match any of those five sequence datasets are considered to be Ananas comosus var. bracteatus unique. We predicted unigenes encoding enzymes involved in terpenoid and phenylpropanoid biosynthesis.The Ananas comosus var. bracteatus. To our knowledge; this is the first report on the de novo transcriptome sequencing of the Ananas comosus var. bracteatus. Unigenes obtained in this study, may help improve future gene expression, genetic and genomics studies in Ananas comosus var. bracteatus.The sequence data provide the most comprehensive transcriptomic resource currently available for  Bromeliaceae, sub family Bromelioideae, in the order Poales. According to the recent classification, Coppens d’Eeckenbrugge and Leal, proposed one genus, Ananas, with two species A. comosus (L.) Merr.  and A. macrodontes Morren . A. comosus is subdivided into five botanical varieties: var. comosus, var. ananassoides, var. erectifolius, var. parguazensis and var. bracteatus . Leaves with white margin and red color, roots, stem were dissected and total RNA extracted using the RNeasy plus Micro Kit . The quality and quantity of total RNA was assessed at an absorbance ratio (OD260/280and OD260/230) and 1% agarose gel electrophoresis. Equal amounts of RNA from each sample were mixed into a single large pool for cDNA synthesis.To obtain cDNA library, first stand synthesis cDNA was synthesized using random hexamer primers from purified Poly (A) mRNA. Second strand cDNA was synthesized using DNA polymerase I and RNase H. Short fragments were then separated by agarose gel electrophoresis and selected for PCR amplification as sequencing templates. Finally, cDNA library was sequenced on Illumina HiSeq 2000 sequencing platform, Beijing Biomarker Technologies, Beijing, China.http://trinityrnaseq.sourceforge.net/), which recovers more full-length transcripts across a broad range of expression levels and sequencing depths [The raw reads were cleaned by removing adapter sequences, low quality sequences (reads with ambiguous bases ‘N’), and reads with >10% Q-value<20 bases. All sequences smaller than 60 bases were eliminated based on the assumption that small reads might represent sequencing artifacts . The higg depths . Subsequ-5 [The assembled sequences were compared against the NCBI Nr, Nt and Swiss-port databases with an E-value < 10-5 . The ope-5 .We quant-5 . The RPKhttp://www.genome.jp/kegg/kaas/. All the mapped sequences were annotated to the KEGG database to obtain the enzyme commission (EC) number. The EC were then mapped to the KEGG pathway to obtain the KEGG Pathway-Maps.To annotate the assembled sequences with GO terms, the Swiss-Port BLAST results were imported into BLAST2GO, a software package that retrieves GO terms, allowing gene functions to be determined and compared . The uniS1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.