Leishmania infantum (syn. L. chagasi) is the causative agent of visceral leishmaniasis (VL) in the New World (NW) with endemic regions extending from southern USA to northern Argentina. The two hypotheses about the origin of VL in the NW suggest (1) recent importation of L. infantum from the Old World (OW), or (2) an indigenous origin and a distinct taxonomic rank for the NW parasite. Multilocus microsatellite typing was applied in a survey of 98 L. infantum isolates from different NW foci. The microsatellite profiles obtained were compared to those of 308 L. infantum and 20 L. donovani strains from OW countries previously assigned to well-defined populations. Two main populations were identified for both NW and OW L. infantum. Most of the NW strains belonged to population 1, which corresponded to the OW MON-1 population. However, the NW population was much more homogeneous. A second, more heterogeneous, population comprised most Caribbean strains and corresponded to the OW non-MON-1 population. All Brazilian L. infantum strains belonged to population 1, although they represented 61% of the sample and originated from 9 states. Population analysis including the OW L. infantum populations indicated that the NW strains were more similar to MON-1 and non-MON-1 sub-populations of L. infantum from southwest Europe, than to any other OW sub-population. Moreover, similarity between NW and Southwest European L. infantum was higher than between OW L. infantum from distinct parts of the Mediterranean region, Middle East and Central Asia. No correlation was found between NW L. infantum genotypes and clinical picture or host background. This study represents the first continent-wide analysis of NW L. infantum population structure. It confirmed that the agent of VL in the NW is L. infantum and that the parasite has been recently imported multiple times to the NW from southwest Europe.

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Our present study is the first attempt to characterize Leishmania parasites from foci in Uzbekistan and Tajikistan endemic for visceral leishmaniasis (VL). PCR-sequencing of the ribosomal internal transcribed spacer 1 and multilocus microsatellite typing (MLMT) were applied to DNA extracted from preparations of Giemsa-stained bone marrow aspirates from 13 cases of VL. L. infantum was shown to cause VL currently occurring in this area. MLMT applying 14 microsatellite markers, previously shown to be polymorphic for strains of the L. donovani complex, revealed that microsatellite profiles of parasites causing human VL in the Namangan and Jizzakh regions in Uzbekistan, and Penjikent region in Tajikistan, basically coincide with those of strains of L. infantum MON-1. Furthermore, these parasites were assigned to a distinct cluster genetically clearly separated from the populations of L. infantum MON-1 from Europe, the Middle East and North Africa. The existence of a genetically homogeneous but distinct group of L. infantum MON-1 indicates that the parasites circulating in the Uzbeki and Tajiki foci studied have been restricted there for a long time rather than having been recently introduced from elsewhere by human or animal reservoir migration.

d Surotomycin (CB-183,315), a cyclic lipopeptide, is in phase 3 clinical development for the treatment of Clostridium difficile infection. We report here the further characterization of the in vitro mode of action of surotomycin, including its activity against growing and nongrowing C. difficile. This was assessed through time-kill kinetics, allowing a determination of the effects on the membrane potential and permeability and macromolecular synthesis in C. difficile. Against representative strains of C. difficile, surotomycin displayed concentration-dependent killing of both logarithmic-phase and stationary-phase cultures at a concentration that was <16؋ the MIC. Exposure resulted in the inhibition of macromolecular synthesis (in DNA, RNA, proteins, and cell wall). At bactericidal concentrations, surotomycin dissipated the membrane potential of C. difficile without changes to the permeability of propidium iodide. These observations are consistent with surotomycin acting as a membrane-active antibiotic, exhibiting rapid bactericidal activities against growing and nongrowing C. difficile. The Gram-positive spore-forming anaerobic bacterium Clostridium difficile is the leading cause of hospital-acquired diarrhea in North America and Europe (1, 2). Elderly hospitalized patients on broad-spectrum antibiotics are the main target population, but recent observations indicate there is an increase in the incidence of C. difficile infection (CDI) in the community without known risk factors (3, 4). In the United States in 2011, there were an estimated 500,000 cases of CDI resulting in 29,300 deaths (5), which reflects the devastating impact of CDI since the turn of the last century. Furthermore, the number of cases of severe CDI has escalated, coinciding with the emergence of epidemic ribotypes, such as BI/NAP1/027 (2, 6). BI/NAP1/027 is now responsible for a significant number of cases of hospital-acquired CDI in North America (5, 6).For Ͼ30 years, vancomycin and metronidazole have been the first-line treatment choices for CDI (7). Metronidazole is prescribed for mild to moderate CDI, while vancomycin is recommended for severe CDI (6,8). However, rates of recurrence of 20 to Ն25% in severe CDI are common following treatment with metronidazole or vancomycin (6, 9, 10). The mode of action of vancomycin is well established, involving inhibition of the later stages of peptidoglycan biosynthesis, which primarily kill rapidly growing C. difficile (11). Metronidazole undergoes biochemical reduction to form reactive species that target DNA and is potent in vitro, but only low concentrations reside in the gastrointestinal tract (12-16). Fidaxomicin, which targets the bacterial RNA polymerase inhibitor, has a narrower spectrum of activity than that of metronidazole and vancomycin and is superior in the prevention of CDI recurrence (17, 18). However, additional novel therapeutics are required to effectively treat CDI and reduce the rates of recurrence following initial therapy.Surotomycin is a minimally absorbed narrow-spectrum cycl...

Aims There is a need for novel treatments for Clostridium difficile infection (CDI). Antibacterial flavonoids are part of a large family of polyphenol phytochemicals with a long history of use in ethnomedicine, but are unexamined against C. difficile. We explored their anti-difficile properties. Methods and Results Anti-difficile activities were determined for several naturally occurring flavonoids, olympicin A and synthetic 4-chromanone and chalcone analogs. With the exception of olympicin A, most naturally occurring phytochemicals tested were poorly active. Diversified synthetic flavonoids resembling olympicin A retained anti-difficile activity, suggesting olympicin A could act as a pharmacophore to obtain novel agents. They also demonstrated concentration dependent killing of logarithmic and stationary phase cultures and reduced sporulation and toxin production. Olympicin A and some synthetic flavonoids dissipated the bacterial transmembrane potential. Interestingly, mutants could only be selected with the analog 207 at a frequency of 10-9. Conclusions Based on the potent anti-difficile properties of olympicin A and modified flavonoids, further exploration of this class of phytochemicals is warranted. Significance and Impact of the Study CDI is a major problem in developed countries. These studies point to there being an avenue for optimizing plant-derived flavonoids, and related antibacterial phytochemicals, as nature-inspired approaches to treat CDI.

To minimize fertilizer and water use, and NO3-N runoff from container culture, growth, and nutrient status of forsythia (Forsythia ×intermedia Zab. ‘Spring Glory’) in No. 2 containers were compared in response to a controlled-release fertilizer (CRF; Nutricote 18-6-8 100-day at rates of 2, 4, and 6 kg·m−3) and placements (incorporation and topdress) under three irrigation strategies [drip-irrigated low (25% or less) leaching fraction (DrLoLF), hand-sprinkled low leaching fraction (HsLoLF), and hand-sprinkled high (50% or less) leaching fraction (HsHiLF)]. In a coexperiment under drip irrigation only, forsythia response was also examined under incorporation, topdress, and dibble fertilizer placements with the same CRF rates applied as single or split dose. Dibble fertilizer placement was superior to both incorporation and topdress in this order. Maximum growth of forsythia occurred at rates of 4.7 kg·m−3 with dibble. With incorporation and topdress, maximum growth was not achieved even at the 6 kg·m−3 maximum rate tested. Forsythia grew better with incorporated than with topdressed CRF with the DrLoLF treatment. The response was reverse with HsHiLF or showed no differences with HsLoLF. Under drip irrigation, greater concentrations of NO3-N generally leached from incorporation and dibbled containers in this order than from topdress. Less nitrate was leached from the topdressed containers because less was released from the CRF prills. At the 6 kg·m−3 CRF rate, total cumulative NO3-N leachings were 76, 85, and 22 kg·ha−1 (45 × 45-cm container spacing) for dibbling, incorporation, and topdress, respectively, under drip irrigation. Split application of CRF greatly reduced NO3-N in leachate, although plant growth also was reduced as a result of less availability of and uptake of nutrients under this strategy.

Cutaneous leishmaniasis (CL) in Morocco is caused by three species, Leishmania major, L. tropica and L. infantum. CL has been known in Chichaoua province since 2000. Using DNA extracted from microscopic slides and parasite cultures, collected in the years 2006 and 2009, we identified for the first time L. tropica as the causative agent of CL in this region. Species identification was achieved by performing the ITS1-PCR-RFLP approach. By using this method it was possible to identify parasites in Giemsa stained slides containing less than five parasites per oil-immersion field even they were conserved for up to four months.

BackgroundThe Namangan Region in the Pap District, located in Eastern Uzbekistan is the main focus of visceral leishmaniasis (VL) in Uzbekistan. In total, 28 cases of human VL were registered during 2006-2008 in this region. A study on the epidemiology of VL in this area was carried out in 2007-2008 in the villages of Chodak, Oltinkan, Gulistan and Chorkesar located at elevations of 900-1200 above sea level.ResultsA total of 162 dogs were tested for Leishmania infection. Blood was drawn for serology and PCR. When clinical signs of the disease were present, aspirates from lymph nodes and the spleen were taken. Forty-two dogs (25.9%) had clinical signs suggestive of VL and 51 (31.5%) were sero-positive. ITS-1 PCR was performed for 135 dogs using blood and tissue samples and 40 (29.6%) of them were PCR-positive. Leishmanial parasites were cultured from lymph node or spleen aspirates from 10 dogs.Eight Leishmania strains isolated from dogs were typed by multi-locus microsatellite typing (MLMT) and by multilocus enzyme electrophoretic analysis (MLEE), using a 15 enzyme system. These analyses revealed that the strains belong to the most common zymodeme of L. infantum, i.e., MON-1, and form a unique group when compared to MON-1 strains from other geographical regions.ConclusionsThe data obtained through this study confirm the existence of an active focus of VL in the Namangan region of Uzbekistan. The fact that L. infantum was the causative agent of canine infection with typical clinical signs, and also of human infection affecting only infants, suggests that a zoonotic form of VL similar in epidemiology to Mediterranean VL is present in Uzbekistan.