Infections by the Yeast<Emphasis Type="Italic"> Kodomaea</Emphasis> (<Emphasis Type="Italic">Pichia</Emphasis>)<Emphasis Type="Italic"> ohmeri</Emphasis>: Two Cases and Literature Review

A 14-year-old boy who was neutropenic following chemotherapy for leukemia developed fungemia caused by the yeast Kodomaea ohmeri (Pichia ohmeri). The infection was cured by catheter removal and the use of fluconazole. A 74-year-old man who had undergone surgeries for a subcutaneous tumor developed polymicrobic cellulitis involving Kodomaea ohmeri. Despite surgical debridement and antibiotic therapy, the patient died of complications. Including these 2 cases, there have been 10 Kodomaea ohmeri infections reported thus far, all occurring in patients with pre-existing conditions. There have been seven cases of fungemia and one case each of peritonitis, funguria, and cellulitis. The treatment employed varied depending on the site/source of infection. Seven patients recovered and three died. The microbiological data available suggest that Kodomaea ohmeri can be identified definitively by biochemical tests and is susceptible to amphotericin B and either susceptible to or dose dependently susceptible to itraconazole and fluconazole.     

A foodborne outbreak of <Emphasis Type="Italic">Salmonella enterica</Emphasis> subsp. <Emphasis Type="Italic">enterica</Emphasis> serovar Madelia at a silver anniversary reception

Reported here is an outbreak of Salmonella enterica subsp. enterica serovar Madelia infection that occurred among 44 persons attending a silver anniversary reception in Hesse, Germany. Isolates of Salmonella Madelia are extremely unusual, and no outbreaks associated with this serotype have been reported previously. Forty-two attendees were interviewed and information was obtained from each of them regarding demographic and clinical characteristics and food consumed at the reception. Twenty-four attendees submitted stool samples for microbiological testing, and 10 of these were culture-positive for S. Madelia. Twenty-three attendees met the case definition of infection, while 18 met the clinical case definition (i.e. vomiting or diarrhoea within 3 days of consuming food at the reception) and five had asymptomatic infection with S. Madelia. The most likely vehicles of infection were tortellini and a red pesto sauce.     

A case of cellulitis complicating <Emphasis Type="Italic">Campylobacter jejuni</Emphasis> subspecies <Emphasis Type="Italic">jejuni</Emphasis> bacteremia and review of the literature

Infection with Campylobacter species is a predominant cause of food-borne gastroenteritis in the industrialized world. Bacteremia is detected in <1% of patients with diarrhea, mainly in immunocompromised hosts or those in the extremes of age. Reported here is the case of a 78-year-old, immunocompromised male patient with Campylobacter jejuni subsp. jejuni bacteremia complicated by cellulitis. The infection was characterized by a protracted course with several recurrences and refractoriness to multiple antibiotic regimens, responding only to a prolonged course of meropenem treatment. The frequency of cellulitis as reflected in previously reported series of Campylobacter bacteremia and the clinical characteristics of this difficult-to-treat infection are reviewed.     

Functional and structural characterisation of <Emphasis Type="Italic">Ag</Emphasis>MNPV <Emphasis Type="Italic">ie1</Emphasis>

We have located and cloned the Anticarsia gemmatalis multicapsid nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genomic DNA fragment containing the immediate early 1 ORF and its flanking regions. Computer assisted analysis of the complete ie1 locus nucleotide sequence information was used to locate regulatory signals in the upstream region and conserved nucleotide and amino acid sequences. Comparative studies led to the identification of several characteristic protein motifs and to the conclusion that AgMNPV-2D is more closely related to Choristoneura fumiferana defective NPV than to other Group I nucleopolyhedrovirus. We have also shown that the AgMNPV IE1 protein was able to transactivate an early Autographa californica MNPV promoter and its own promoter in transient expression assays. In order to investigate the biological functionality of the ie1 promoter, the ie1 upstream activating region (UAR) was molecularly dissected and cloned upstream of the E. coli lacZ ORF. The results obtained, after transfection of UFL-AG-286 insect cells, leading us to find that the −492 and −357 versions contains sequence motifs important for the level of the lacZ reporter gene expression. Electronic supplementary material The online version of this article (doi: ) contains supplementary material, which is available to authorized users. The GenBank accession number of the sequence reported in this paper is AF368905.     

Community-acquired <Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis> infections: a systematic review

Stenotrophomonas maltophilia is a pathogen that causes infections mainly in immunocompromised patients. However, community-acquired S. maltophilia infections have been occasionally reported. The objective of this paper was to collect and evaluate the available published data referring to community-acquired S. maltophilia infections. We searched PubMed, the Cochrane Library, and Scopus for articles providing data for patients with community-acquired S. maltophilia infections. Eight case series and 23 case reports (involving 77 and 26 patients with community-acquired S. maltophilia infections, respectively) were regarded as eligible for inclusion in our review. Regarding the 77 patients with community-acquired S. maltophilia infections included in the identified case series, 45 had bacteremia, six ocular infections, five respiratory tract infections, four wound/soft tissue infections, two urinary tract infections, one conjunctivitis, one otitis, and one cellulitis; data were not reported for the remaining 12 patients. Comorbidity (such as malignancy, HIV infection, prior hospitalization) was common. Data included in the eight case series regarding the outcome of infection were limited. From the 26 patients with community-acquired S. maltophilia infections reported in the case reports, 22 were cured from the infection, whereas 4 of 26 patients died; one death was attributed to septic shock due to S. maltophilia. Several publications report patients with community-acquired S. maltophilia infections; the majority of them refer to patients with some kind of comorbidity. Physicians should be aware that S. maltophilia infections are not restricted to hospitalized patients.     

Epidemiology of invasive neonatal <Emphasis Type="Italic">Cronobacter</Emphasis> (<Emphasis Type="Italic">Enterobacter sakazakii</Emphasis>) infections

About 120–150 neonatal Cronobacter spp. (Enterobacter sakazakii) infections have been described. An analysis of current case numbers, epidemiological measures and risk factors is warranted. Data of microbiologically confirmed cases, published between 2000 and 2008, have been analysed statistically. More than 100 neonatal Cronobacter infections have been reported in this period. The overall lethality of the 67 invasive infections was 26.9%. The lethality of Cronobacter meningitis, bacteraemia and necrotising enterocolitis (NEC) was calculated to be 41.9% (P < 0.0001), <10% and 19.0% (P < 0.05), respectively. Logistic regression models (P < 0.0001) revealed a higher gestational age at birth and parentage not from Europe as significant factors for a higher reporting probability of neonatal Cronobacter meningitis. Neonates with Cronobacter meningitis not originating from North America have a higher risk for lethal outcome than other neonatal Cronobacter infections (P < 0.0001). Continental differences of risk factors for Cronobacter meningitis and for the lethal outcome of neonatal meningitis should be elucidated. Neonatal Cronobacter infections are mainly associated with the contamination of infant formula and of the relevant cleaning and preparation equipment. Eleven neonatal Cronobacter infections, not caused by contaminated infant formula, have been retrieved. Other environmental sources of infection should be considered. Consistent and sufficiently informative data of invasive neonatal Cronobacter infections should be recorded in a centralized reporting system.     

Tracing origin of serrated adenomas with <Emphasis Type="Italic">BRAF</Emphasis> and <Emphasis Type="Italic">KRAS</Emphasis> mutations

Serrated neoplasm of the colorectum raised many as-yet unanswered issues. To characterize serrated neoplasia pathway, we investigated BRAF and KRAS mutations in 35 traditional serrated adenomas. BRAF exons 11 and 15, and KRAS exon 2 were amplified by polymerase chain reaction and directly sequenced. BRAF V599E mutation was found in 27 serrated adenomas (77.1%), and KRAS mutations were found in 3 (8.6%) of 35 traditional serrated adenomas. In 13 cases, mixed polyps composed of traditional serrated adenomas and hyperplastic (serrated) polyps were observed, and seven of them showed the same BRAF mutations in both components. Somatic mutations of BRAF and KRAS genes were mutually exclusive. These findings suggest that BRAF mutations are early and a critical event in the serrated adenomas, and most serrated adenomas in both sides of colon may progress from microvesicular hyperplastic polyps via BRAF mutations, and some left-sided serrated adenomas develop via KRAS mutations.     

<Emphasis Type="Italic">Blastocystis hominis</Emphasis> as a cause of hypoalbuminemia and anasarca

The protozoan Blastocystis hominis has been considered nonpathogenic, but this classification has come under scrutiny in light of reports in the medical literature indicating it could be the cause of intestinal disorders and, in one case, hypoalbuminemia. Reported here is a severe case of infection with B. hominis that caused acute gastroenteritis with prolonged diarrhea, hypoalbuminemia and anasarca. The diagnosis was based on the parasitological finding, since no other pathological evidence was found. The patient responded favorably to treatment with metronidazole for 10 days. This case supports the idea that B. hominis should be considered as a cause of opportunistic infection in debilitated patients despite the controversy surrounding its pathogenicity.     

Influence of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> infection on the expression of <Emphasis Type="Italic">MLH1</Emphasis> and <Emphasis Type="Italic">MGMT</Emphasis> in patients with chronic gastritis and gastric cancer

The aim of the present study was to evaluate the influence of Helicobacter pylori on MLH1 and MGMT mRNA levels in patients with chronic gastritis and gastric cancer. The study included 217 patients, of which 26 were uninfected, 127 had chronic gastritis and were H. pylori-positive, and 64 had gastric cancer. Bacterial genotypes were evaluated by polymerase chain reaction (PCR), and the expression levels of MLH1 and MGMT were determined by quantitative real-time PCR and immunohistochemistry. There was an association between infection with cagA, vacA s1m1 strains and gastric cancer development. When the gastric epithelium and associated inflammation were examined for expression of MLH1 and MGMT, an overall increase in expression was observed. On the other hand, these levels decrease significantly among gastric cancer patients. The loss of MLH1 and MGMT expression in gastric cancer patients suggests that it is not an early event in H. pylori-associated gastric carcinogenesis.     

Prevalence of <Emphasis Type="Italic">Blastocystis hominis</Emphasis> and <Emphasis Type="Italic">Strongyloides stercoralis</Emphasis> infection in Okinawa,Japan

This study was conducted to clarify the prevalence of Blastocystis hominis and Strongyloides stercoralis infection in Ryukyu University Hospital, Okinawa, Japan, between January 2004 and November 2006. Stool samples collected from 3,292 patients were examined by the direct smear method, formalin–ether sedimentation method, and agar plate culture method. The prevalence rate of B. hominis and S. stercoralis infection was 1.0 and 3.4%, respectively. The prevalence rate of B. hominis infection in patients aged >80 years old was significantly higher than that in patients <80 years old (P < 0.001). The prevalence rate of S. stercoralis infection was significantly higher in patients with B. hominis infection compared with those without (P < 0.001). This study demonstrated a prevalence rate for B. hominis and S. stercoralis infection and an association between B. hominis and S. stercoralis infection in Okinawa, Japan.     

Redescription of <Emphasis Type="Italic">Lemuricola</Emphasis> (<Emphasis Type="Italic">Madoxyuris</Emphasis>) <Emphasis Type="Italic">bauchoti</Emphasis> (Nematoda,Oxyuridae) from <Emphasis Type="Italic">Lemur catta</Emphasis> in Madagascar

Lemuricola (Madoxyuris) bauchoti Chabaud, Brygoo et Petter, 1965 is redescribed from material collected from the ring-tailed lemur, Lemur catta, from the Beza Mahafaly Special Reserve in Madagascar using the scanning electron microscope. This is a new host record and the first oxyurid reported from the ring-tailed lemur. Previously, records of each species of the subgenus Madoxyuris have been restricted to a single host species, but the close relationship between these nematodes and their Strepsirrhini hosts will only be proven when additional records fill in the gaps in their distribution.     

Anurans as paratenic hosts in the transmission of <Emphasis Type="Italic">Hepatozoon caimani</Emphasis> to caimans <Emphasis Type="Italic">Caiman yacare</Emphasis> and <Emphasis Type="Italic">Caiman latirostris</Emphasis>

Prevalence of Hepatozoon caimani has been reported in 76% of caimans Caiman yacare from the Pantanal region. Culex (Melanoconion) spp. mosquitoes were recently identified as natural vectors of this parasite. However, culicids are not typically eaten by crocodilians, suggesting that the main transmission route is through ingestion of insectivorous vertebrates, such as anurans. The susceptibility of wild frogs Leptodactylus chaquensis, Leptodactylus podicipinus and Scinax nasicus to infection by H. caimani was verified. Wild-caught anurans were force fed with sporulated oocysts from laboratory-bred Culex (Melanoconion) mosquitoes. Frogs were killed 30 days postinfection, and their internal organs were fed to caimans C. yacare and Caiman latirostris. Cystozoites were identified in fresh liver impression smears of L. chaquensis. C. yacare fed on anuran organ presented gametocytes in peripheral blood circulation between 74 and 80 days postinoculation (dpi). Gametocytes were also verified in C. latirostris fed on the internal organs of L. podicipinus and S. nasicus between 60–70 and 69–75 dpi, respectively. Since frogs used in experiment are sympatric with C. yacare and C. latirostris and may occur in the diet of these caimans, the results suggest these amphibians are paratenic hosts in the natural transmission cycle of H. caimani in Pantanal.     

Experimental transmission of<Emphasis Type="Italic"> Cryptosporidium molnari</Emphasis> (Apicomplexa: Coccidia) to gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis> L.) and European sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis> L.)

Cryptosporidium molnari was experimentally transmitted to gilthead sea bream (Sparus aurata) and European sea bass (Dicentrachus labrax) by oral infection with infected stomach scrapings. The infection was also cross-transmitted from infected gilthead sea bream to sea bass by cohabitation. The course of the infection was assessed after necropsy by three microscopic diagnostic methods and their sensitivity was compared. At the end of all the experiments the prevalence of infection reached 100%. In the oral experiments, both fish hosts appeared infected as early as 7 days post exposure (p.e.), but gilthead sea bream exhibited a higher intensity of infection and infection proceeded at a faster rate than in sea bass. The cellular host reaction was stronger in sea bass than in sea bream, whereas the histopathological effect was lower in the former. Transmission could be favoured by cannibalism among cohabiting fish. This is the first report on piscine Cryptosporidium transmission. The implications for the aquaculture industry are discussed.     

Repeat induced point mutation in two asexual fungi, <Emphasis Type="Italic">Aspergillus niger</Emphasis> and <Emphasis Type="Italic">Penicillium </Emphasis><Emphasis Type="Italic">chrysogenum</Emphasis>

Repeat induced point mutation (RIP) is a gene silencing mechanism present in fungal genomes. During RIP, duplicated sequences are efficiently and irreversibly mutated by transitions from C:G to T:A. For the first time, we have identified traces of RIP in transposable elements of Aspergillus niger and Penicillium chrysogenum, two biotechnologically relevant fungi. We found that RIP in P. chrysogenum has affected a large set of sequences, which also contain other mutations. On the other hand, RIP in A. niger is limited to only few sequences, but literally all mutations are RIP-like. Surprisingly, RIP occurred only in transposon sequences that have disrupted open reading frames in A. niger, a phenomenon not yet reported for other fungi. In both fungal species, we identified two sequences with strong sequence similarity to Neurospora crassa RID. RID is a putative DNA methyltransferase and the only known enzyme involved in the RIP process. Our findings suggest that both A. niger and P. chrysogenum either had a sexual past or have a sexual potential. These findings have important implications for future strain development of these fungi.     

Agroinfection of cloned Sri Lankan cassava mosaic virus DNA to <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>, <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> and cassava

Sri Lankan cassava mosaic virus (SLCMV) is a bipartite begomovirus infecting cassava in India and Sri Lanka. We have used Agrobacterium-mediated inoculation (agroinoculation) of cloned SLCMV DNA to inoculate additional hosts, Nicotiana tabacum and Arabidopsis. Although SLCMV infection in these hosts caused stunting, leaf deformation and developmental abnormalities, accumulation levels of viral DNA in the infected plants suggested that this virus was poorly adapted to them. In the natural host, cassava, agroinoculation produced infection at a low frequency. The monopartite nature of SLCMV, reported earlier in N. benthamiana, was maintained in the new hosts as well as in cassava.     

Horses naturally infected by <Emphasis Type="Italic">Trypanosoma vivax</Emphasis> in southern Brazil

In this study, we reported the first outbreak of the infection by Trypanosoma vivax in horses in southern Brazil, a non-endemic region where bovines have only recently been found infected by this trypanosome species. We evaluated 12 horses from a farm in southern Brazil, where four horses displayed pale mucous membranes, fever, weight loss, and swelling of abdomen, prepuce, or vulva. The diagnosis of T. vivax was confirmed in four horses by morphological parameters of trypomastigotes in blood smears and species-specific PCR. All T. vivax-infected animals showed anemia, and most showed increased levels of beta-1, beta-2, and gamma globulins. Horses were treated with diminazene aceturate, but cure was not achieved, and the disease relapsed after therapy. These findings demonstrated that Brazilian T. vivax isolates, which were already reported infecting cattle, buffaloes, goats, and sheep, can be highly pathogenic for horses, causing severe disease and even death of the animals due to the recurrence of the infection.     

Persistent Colonization of Nine Cystic Fibrosis Patients with an<Emphasis Type="Italic"> Achromobacter</Emphasis> (<Emphasis Type="Italic">Alcaligenes</Emphasis>)<Emphasis Type="Italic"> xylosoxidans</Emphasis> Clone

The present study was conducted to investigate the increasing incidence of Achromobacter (previously Alcaligenes) xylosoxidans isolates being recovered from sputum samples of cystic fibrosis patients at a cystic fibrosis department for adults in Athens, Greece. During the 1-year study period, a total of 34 isolates were detected persistently in 9 of 71 cystic fibrosis patients. The isolates exhibited resistance to multiple antimicrobial agents. Isolates that were recovered repeatedly from each patient exhibited identical macrorestriction profiles with pulsed-field gel electrophoresis, indicating that the same strain persisted in the lungs of these patients. Isolates from five of the patients were genetically related, suggesting a common-source outbreak of Achromobacter xylosoxidans colonization or infection.     

Prevalence of the <Emphasis Type="Italic">ica</Emphasis> operon and insertion sequence IS<Emphasis Type="Italic">256</Emphasis> among <Emphasis Type="Italic">Staphylococcus epidermidis</Emphasis> prosthetic joint infection isolates

Joint replacement surgery has improved the quality of life for hundreds of thousands of patients. However, the infection of a joint implant is an important and serious complication, though the prevalence is low. Staphylococcus epidermidis is the most important pathogen involved in foreign-body infections. S. epidermidis is also a commensal that comprises a substantial part of the normal skin flora of humans. The possibility to demonstrate potential specific virulence markers may facilitate the interpretation of the bacteriological findings, as well as the clinical decision. The prevalence of the ica locus and insertion sequence IS256 by using polymerase chain reaction (PCR) among 32 clinical S. epidermidis isolates from prosthetic joint infections (PJIs) and 24 commensal isolates from nares and skin was investigated. Sixteen (50%) of the 32 PJI isolates harbored the ica operon compared with one-third of the commensal isolates obtained from the samples of the skin and nares of healthy individuals. The IS256 was demonstrated in 26 (81%) out of 32 PJI isolates. By contrast, IS256 was found in one of 24 commensal isolates. In conclusion, IS256 may be superior to the ica operon as a marker of the invasive capacity of S. epidermidis, since it was found in most of the PJI isolates, but rarely among commensals.     

Cysteine proteinases from promastigotes of <Emphasis Type="Italic">Leishmania</Emphasis> (<Emphasis Type="Italic">Viannia</Emphasis>) <Emphasis Type="Italic">braziliensis</Emphasis>

Leishmania (Viannia) braziliensis is the major causative agent of American tegumentary leishmaniasis, a disease that has a wide geographical distribution and is a severe public health problem. The cysteine proteinase B (CPB) from Leishmania spp. represents an important virulence factor. In this study, we characterized and localized cysteine proteinases in L. (V.) braziliensis promastigotes. By a combination of triton X-114 extraction, concanavalin A-affinity, and ion exchange chromatographies, we obtained an enriched fraction of hydrophobic proteins rich in mannose residues. This fraction contained two proteinases of 63 and 43 kDa, which were recognized by a CPB antiserum, and were partially sensitive to E-64 in enzymatic assays with the peptide Glu-Phe-Leu. In confocal microscopy, the CPB homologues localized in the peripheral region of the parasite. This data together with direct agglutination and flow cytometry assays suggest a surface localization of the CPB homologues. The incubation of intact promastigotes with phospholipase C reduced the number of CPB-positive cells, while anti-cross-reacting determinant and anti-CPB antisera recognized two polypeptides (63 and 43 kDa) derived from phospholipase C treatment, suggesting that some CPB isoforms may be glycosylphosphatidylinositol-anchored. Collectively, our results suggest the presence of CPB homologues in L. braziliensis surface and highlight the need for further studies on L. braziliensis cysteine proteinases, which require enrichment methods for enzymatic detection.     

Characterization of <Emphasis Type="Italic">p</Emphasis>24 Gene of <Emphasis Type="Italic">Spodoptera litura</Emphasis> Multicapsid Nucleopolyhedrovirus

Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) p24 gene is 753 bp long, potentially encoding 244 amino acids with a predicted molecular weight of 27.3 kDa. Homology analysis indicated that SpltMNPV P24 has 20–36% amino acid identity with that of other known baculoviruses. RT-PCR results showed that the p24 gene is transcribed actively at the late stage of infection and the mRNA start site was mapped within a consensus baculovirus late promoter sequence (ATAAG). Western blot analysis of extracts from SpltMNPV-infected S. litura cells detected a specific 28 kDa protein, and this protein was not N-glycosylated. Structural localization revealed that SpltMNPV P24 was associated with the nucleocapsid of occlusion-derived virus (ODV) as a complex form of 83 kDa.