Chloromyxum inexpectatum n. sp. and Sphaerospora colomani n. sp. (Myxozoa: Myxosporea), parasites of the urinary system of the sterlet,Acipenser ruthenus L.

During the parasitological examination of sterlets, Acipenser ruthenus L., caught for breeding, the developmental stages of two new coelozoic species of Myxosporea were demonstrated in the urinary passages. Chloromyxum inexpectatum n. sp. occurs attached to the epithelial cells of the ureters and urinary bladder. Sphaerospora colomani n. sp. forms bisporoblastic pseudoplasmodia in the lumen of the renal tubules. A further myxosporean species, Zschokkella sturionis Tripathi, 1948, was found in the biliary ducts. The occurrence of this parasite in sterlet has not been reported so far. Examination of blood smears revealed Cryptobia acipenseris and Haemogregarina acipenseris infection. Developmental stages of the coccidia Goussia acipenseris Molnár, 1986 and Goussia vargai Molnár, 1986 were frequently seen in the intestine of the dissected fish.     

Another chloromyxid lineage: molecular phylogeny and redescription of Chloromyxum careni from the Asian horned frog Megophrys nasuta

Infection with Chloromyxum careni Mutschmann, 1999 was found in the Asian horned frog Megophrys nasuta from Malaysia and Indonesia. Kidney was the only organ infected. Coelozoic plasmodia up to 300 μm were localized in Bowman's space, embracing the glomerulus from all sides, or rarely in lumina of renal tubules. Plasmodia are polysporic, containing disporic pansporoblasts. Myxospores observed by light microscopy are colorless, variable in shape and size, measuring 6.0-8.5 × 5.0-6.5 μm, composed of two symmetrical valves joined by a meridian suture, containing four pyriform polar capsules 3.0-4.0 × 2.5-3.0 μm and a single sporoplasm. Each valve possesses 14-24 (median 21) fine longitudinal ridges clearly visible only in scanning electron microscopy. Rarely, atypical spores with a markedly pointed posterior pole and only 6-10 surface ridges are present in plasmodia together with typical spores. Both small subunit (SSU) and large subunit (LSU) rRNA gene sequences possess extremely long GU-rich inserts. In all SSU and LSU rDNA-based phylogenetic analyses, C. careni clustered as a distinct basal branch to the Myxobolus+Myxidium lieberkuehni clade, out of the marine Chloromyxum clade containing Chloromyxum leydigi, the type species of the genus. These morphological and phylogenetic data suggest erection of a new genus for the C. careni lineage, but we conservatively treat it as a Chloromyxum sensu lato until more information is available.     

Site preference of myxozoans in the kidneys of Hungarian fishes

Myxozoans are common parasites of fish kidneys, with most having specific sites of development. Five specific sites of development include (1) the lumen of renal tubules, (2) the renal corpuscles followed by location in renal tubules, (3) intracellular location within the tubular epithelium followed by a stage in the lumen of the ducts, (4) haematopoietic tissue with dispersed trophozoites, and (5) haematopoietic tissue with large, localized plasmodia. A coelozoic development preceded by presporogonic multiplication characterises most Sphaerospora spp. Early plasmodial stages of Myxidium and Chloromyxum spp. are frequently found in the renal glomerules, while spores develop in the urinary channels in plasmodia released from the renal corpuscles. In Hoferellus and Myxobilatus spp., spores are formed in small plasmodia inside the lumen of the urinary ducts after several internal cleavages in the epithelium of renal tubules. The presence of dispersed trophozoites among haematopoietic tissue cells of the renal interstitium characterises the development of Sphaerospora tincae and Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD). Spores of S. tincae are formed at the place of plasmodial development, while spore formation of PKD is in the renal tubules. A large mass of spores, often surrounded by a connective tissue capsule, can appear in the renal interstitium during infections by several Myxobolus spp.; furthermore, a large number of these spores formed in plasmodia in distant tissues can also accumulate in melano-macrophage centres.     

The myxozoan fauna of spottail shiner in the Great Lakes basin: membership, richness, and geographical distribution

Spottail shiner (Notropis hudsonius) from localities in each of the Great Lakes plus some nearby waterbodies, i.e., the St. Lawrence River, and the Chester River, Maryland, were examined for myxozoan parasites. A total of 10 species was found, including 7 histozoic (Myxobolus sp.; M. algonquinensis Xiao and Desser, 1997; M. bartai Salim and Desser, 2000; M. xiaoi Salim and Desser, 2000; M. fanthami Landsberg and Lom, 1991; M. hendricksoni Mitchell, Seymour, and Gamble, 1985; Thelohanellus notatus Mavor, 1916) and 3 coelozoic (Chloromyxum sp., Zschokkella sp., Sphaerospora sp.) representatives. Infracommunity richness varied from 0 to 5 species per fish; mean infracommunity richness varied from 0 to 2.5 species. Component community richness varied from 0 to 7. Significant positive correlations were observed between mean and maximum infracommunity richness and component community richness. Similarly, maximum prevalence of each species at any 1 site was positively correlated with geographic range as measured by number of localities where a parasite species was encountered. Individual species occurred independently of each other. Representative histozoic and coelozoic species displayed similarly widespread distributions from Wisconsin to Maryland, but overall, histozoic species were dominant members within component communities. The study concludes that, under the present taxonomic paradigm, species parasitizing spottail shiner appear to be part of a larger network that cycles, in varying degrees, through certain other cyprinid and catostomid fish. The challenge of future research is to determine whether each parasite species constitutes single or multiple genetically isolated populations.     

Pathogenic mechanisms of invasive group A Streptococcus infections by influenza virus–group A Streptococcus superinfection

Group A Streptococcus (GAS) are pathogenic bacteria of the genus Streptococcus and cause severe invasive infections that comprise a wide range of diverse diseases, including acute respiratory distress syndrome, renal failure, toxic shock‐like syndrome, sepsis, cellulitis and necrotizing fasciitis. The essential virulence, infected host and external environmental factors required for invasive GAS infections have not yet been determined. Superinfection with influenza virus and GAS induced invasive GAS infections was demonstrated by our team in a mouse model, after which clinical cases of invasive GAS infections secondary to influenza virus infection were reported by other investigators in Japan, USA, Canada, UK China, and other countries. However, the pathogenic mechanisms underlying influenza virus‐GAS superinfection are not yet fully understood. The present review describes the current knowledge about invasive GAS infections by superinfection. Topics addressed include the bacteriological, virological and immunological mechanisms impacting invasion upon superinfection on top of underlying influenza virus infection by GAS and other bacteria (i.e., Streptococcus pneumoniae and Staphylococcus aureus). Future prospects are also discussed.

     

M13 Virus based detection of bacterial infections in living hosts

We report a first method for using M13 bacteriophage as a multifunctional scaffold for optically imaging bacterial infections in vivo. We demonstrate that M13 virus conjugated with hundreds of dye molecules (M13‐Dye) can target and distinguish pathogenic infections of F‐ pili expressing and F ‐negative strains of E. coli. Further, in order to tune this M13‐Dye complex suitable for targeting other strains of bacteria, we have used a 1‐step reaction for creating an anti‐bacterial antibody ‐M13‐Dye probe. As an example, we show anti‐S. aureus ‐M13‐Dye able to target and image infections of S. aureus in living hosts, with a 3.7× increase in fluorescence over background. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Phylogenetic relationships amongst Chloromyxum Mingazzini, 1890 (Myxozoa: Myxosporea), and the description of six novel species from Australian elasmobranchs

Six novel species of Chloromyxum Mingazzini, 1890 are described using a whole evidence approach combining morphometric and molecular data, together with features of their biology. Elasmobranchs were collected in Australian waters, from the Great Barrier Reef, Queensland, off Lizard and Heron Islands; from Moreton Bay, southeast Queensland; off Hobart, Tasmania; and from the Tamar River, Launceston, Tasmania. The novel species proposed here are: Chloromyxum hemiscyllii n.sp. from Hemiscyllium ocellatum; Chloromyxum kuhlii n.sp. from Neotrygon kuhlii; Chloromyxum lesteri n.sp. from Cephaloscyllium laticeps; Chloromyxum mingazzinii n.sp. from Pristiophorus nudipinnis; Chloromyxum myliobati n.sp. from Myliobatis australis; and Chloromyxum squali n.sp. from Squalus acanthias. A seventh species from Squalus acanthias is also reported but due to limited material is not formally described. Molecular phylogenetic analyses revealed that the genus Chloromyxum is polyphyletic, and species from elasmobranchs form a well-supported sister clade, with the type species Chloromyxum leydigi, to all other congeneric species clustering within the freshwater myxosporean clade. Morphological analysis showed that elasmobranch-infecting species are predominantly pyriform shaped, have clearly thickened spore apex and possess caudal filaments, compared to other Chloromyxum species which are generally spherical or subspherical, and lack caudal filaments. These morphological and phylogenetic data provide further support for the erection of new genera, but we conservatively consider the species described in this study and other elasmobranch-infecting Chloromyxum species as Chloromyxum sensu strictu, whilst the freshwater teleost infecting and amphibian infecting species we will assign as Chloromyxum sensu lato, until more comprehensive data are available.     

The anti-biofilm and anti-virulence activities of trans-resveratrol and oxyresveratrol against uropathogenic Escherichia coli

Abstract

Uropathogenic Escherichia coli (UPEC) is the primary causative agent of urinary tract infections, which are one of the most common infectious disease types in humans. UPEC infections involve bacterial cell adhesion to bladder epithelial cells, and UPEC can also form biofilms on indwelling catheters that are often tolerant to common antibiotics. In this study, the anti-biofilm activities of t-stilbene, stilbestrol, t-resveratrol, oxyresveratrol, ε-viniferin, suffruticosol A, and vitisin A were investigated against UPEC. t-Resveratrol, oxyresveratrol, and ε-viniferin, suffruticosol A, and vitisin A significantly inhibited UPEC biofilm formation at subinhibitory concentrations (10–50?μg ml^?1). These findings were supported by observations that t-resveratrol and oxyresveratrol reduced fimbriae production and the swarming motility in UPEC. Furthermore, t-resveratrol and oxyresveratrol markedly diminished the hemagglutinating ability of UPEC, and enhanced UPEC killing by human whole blood. The findings show that t-resveratrol, oxyresveratrol, and resveratrol oligomers warrant further attention as antivirulence strategies against persistent UPEC infections.     

A frog-derived antimicrobial peptide as a potential anti-biofilm agent in combating Staphylococcus aureus skin infection

Staphylococcus aureus (S. aureus), one of the most prevalent bacteria found in atopic dermatitis lesions, can induce ongoing infections and inflammation by downregulating the expression of host defence peptides in the skin. In addition, the emergence of the ‘superbug’ Methicillin-resistant S. aureus (MRSA) has made the treatment of these infections more challenging. Antimicrobial peptides (AMPs), due to their potent antimicrobial activity, limited evidence of resistance development, and potential immunomodulatory effects, have gained increasing attention as potential therapeutic agents for atopic dermatitis. In this study, we report a novel AMP, brevinin-1E-OG9, isolated from the skin secretions of Odorrana grahami, which shows potent antibacterial activity, especially against S. aureus. Based on the characteristics of the ‘Rana Box’, we designed a set of brevinin-1E-OG9 analogues to explore its structure–activity relationship. Brevinin-1E-OG9c-De-NH₂ exhibited the most potent antimicrobial efficacy in both in vitro and ex vivo studies and attenuated inflammatory responses induced by lipoteichoic acid and heat-killed microbes. As a result, brevinin-1E-OG9c-De-NH₂ might represent a promising candidate for the treatment of S. aureus skin infections.     

Host genetic architecture in single and multiple infections

Hosts are often target to multiple simultaneous infections by genetically diverse parasite strains. The interaction among these strains and the interaction of each strain with the host was shown to have profound effects on the evolution of parasite traits. Host factors like genetic architecture of resistance have so far been largely neglected. To see whether genetic architecture differs between different kinds of infections we used joint scaling analysis to compare the genetic components of resistance in the red flour beetle Tribolium castaneum exposed to single and multiple strains of the microsporidian Nosema whitei. Our results indicate that additive, dominance and epistatic components were more important in single infections whereas maternal components play a decisive role in multiple infections. In detail, parameter estimates of additive, dominance and epistatic components correlated positively between single and multiple infections, whereas maternal components correlated negatively. These findings may suggest that specificity of host–parasite interactions are mediated by genetic and especially epistatic components whereas maternal effects constitute a more general form of resistance.     

Nosocomial Infections and Obesity in Surgical Patients

Objective: There is an increased morbidity and mortality associated with surgery in the obese patient. This study was conducted to determine risk factors and compare the nosocomial infection rate in obese and nonobese surgical patients. Research Methods and Procedures: A total of 395 surgical patients were evaluated. BMI was calculated for each patient. Various conventional risk factors for nosocomial infections were recorded. Biochemical parameters with plasma total cholesterol and high‐density lipoprotein‐cholesterol levels were measured. The diagnosis of infection was made according to the Centers for Disease Control and Prevention criteria. Univariate and two‐step multivariate logistic regression methods were used for determination of nosocomial infection risk factors. Results: There were 117 nosocomial infections identified in 96 of 395 surgically operated patients. A significant increase in the total number of nosocomial infections was determined in obese patients compared with the normal‐weight patients (p < 0.05). High‐density lipoprotein‐cholesterol below the 10th percentile increased risk of surgical site infection. Discussion: Our results suggest that obesity is an important risk factor for postoperative nosocomial infection.     

Widespread Emergence of Multidrug Resistant <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> Isolated from CSF Samples

Bacterial infections of the central nervous system, especially acute infections such as bacterial meningitis require immediate, invariably empiric antibiotic therapy due to the widespread emergence of resistance among bacterial species. Nosocomial infections by Pseudomonas aeruginosa have been described with an increasing trend towards multidrug resistance. P. aeruginosa isolates n = 53 (66%) isolated from the cerebrospinal fluid (CSF) were used for this study. Antibiotic resistance in 53 P. aeruginosa clinical isolates from 80 CSF samples were evaluated. Of these, n = 42 (80%) of the isolates showed multidrug resistance to more than eight antibiotics and n = 17 (32%) isolates were found to be imipenem resistant P. aeruginosa (IMPR-Pa). Genotypical examination by ERIC based PCR revealed minor genetic variations. Polymicrobial infections are common in the CSF samples. However, high prevalence of P. aeruginosa as an opportunistic pathogen has been developing with increased resistance to antimicrobial agents and thus becoming a significant threat.     

Human cathelicidin peptide LL37 inhibits both attachment capability and biofilm formation of Staphylococcus epidermidis

Aims: The aim of this work was to investigate the possible effect of human cathelicidin antimicrobial peptide LL37 on biofilm formation of Staphylococcus epidermidis, a major causative agent of indwelling device‐related infections. Methods and Results: We performed initial attachment assay and biofilm formation solid surface assay in microtitre plates, as well as growth experiment in liquid medium using laboratory strain Staph. epidermidis ATCC35984. We found that already a low concentration of the peptide LL37 (1 mg l^?1) significantly decreased both the attachment of bacteria to the surface and also the biofilm mass. No growth inhibition was observed even at 16 mg l^?1 concentration of LL37, indicating a direct effect of the peptide on biofilm production. Conclusions: As biofilm protects bacteria during infections in humans and allows their survival in a hostile environment, inhibition of biofilm formation by LL37 may have a key role to prevent bacterial colonization on indwelling devices. Significance and Impact of the Study: Our findings suggest that this host defence factor can be a potential candidate in prevention and treatment strategies of Staph. epidermidis infections in humans.     

Plant teratologies as a result of phytoplasma infections

The direct correlation between teratological cases and phytoplasma infections was ascertained in spontaneous and cultivated plant species. Plants, belonging to 31 species and 12 families, showing symptoms of growth abnormalities were collected and analysed. Attempted detection of Rhodococcus fascians by isolation, PCR indexing and 16S rRNA sequencing from fasciated tissues allowed to exclude its presence. Nested PCR by universal primers and 16S rRNA sequence analyses indicated the presence of phytoplasmas, belonging to six groups, in the 44% of symptomatic samples. Among the infected species, Austrocylindropuntia exaltata, Opuntia subulata, Euphorbia characias, Euphorbia dendroides, Euphorbia linifolia, Euphorbia myrsinites, Rumex buchephalophorus, Linaria multicaulis and Fedia cornucopiae represent new phytoplasma hosts world-wide. Moreover this is the first report of phytoplasma belonging to subgroup 16SrRNA II-I in Italy. These findings together with the known erratic distribution in plant tissues of these phloem-restricted prokaryotes indicate a close correlation between fasciation and similar growth disorders and phytoplasma infections.     

1,2,3-Triazole Derivatives as an Emerging Scaffold for Antifungal Drug Development against Candida albicans: A Comprehensive Review

Candida infections are most prominent among fungal infections majorly target immunocompromised and hospitalized patients and cause significant morbidity and mortality. Candida albicans is the notorious and most prevalent among all pathogenic Candida strains. Its emerging resistance toward available antifungal agents making it hard to tackle and emerging as global healthcare emergency. Simultaneously, 1,2,3-triazole nucleus is a privileged scaffold that is gaining importance in antifungal drug development due to being a prominent bioactive linker and isostere of triazole based antifungal class core 1,2,4-triazole. Numerous reports have been updated in scientific literature in last few decades related to utilization of 1,2,3-triazole nucleus in antifungal drug development against Candida albicans. Present review will shed light on various preclinical studies focused on development of 1,2,3-triazole derivatives targeting Candida albicans along with brief highlight on clinical trials and newly approved drugs. Structure-activity relationship has been precisely discussed for each architect along with future perspective that will help medicinal chemists in design and development of potent antifungal agents for tackling infections derived from Candida albicans.     

Studies on species of fungi associated with mycotic infections of fish in a Nigerian freshwater fish pond

A survey was carried out on the species of fungi associated with mycotic infections of fish in a Nigerian freshwater fish pond. A total of 24 fungal species belonging to 6 genera of aquatic phycomycete were isolated from the infected fishes. Achlya racemosa, Aphanomyces laevis, Dictyuchus sterile, Saprolegnia ferax, S. litoralis and S. parasitica had 100% frequency of occurrence amongst the infected fishes. There were similarities in the species of fungi isolated from the infected fishes in the fish pond and those isolated from the hatchery. Of the infected fishes, Clarias lazera had the highest number of fungal isolates. It was followed by Tilapia zilli, then by Chanos chanos, Ethmalosa fimbriata and finally by Cyprinus carpio. The implications of the results obtained on the development of fish farming in Nigeria are discussed.     

Integration of plant growth-promoting rhizobacteria and chemical elicitors for induction of systemic resistance in mulberry against multiple diseases

Abstract

In mulberry (Morus alba L.), various individual strains of plant growth-promoting rhizobacteria (PGPR) and synthetic analogs of naturally occurring plant activators have demonstrated their potential to elicit induced systemic resistance (ISR) against either brown leaf spot (Cercospora moricola) or leaf rust (Cerotelium fici) diseases. However, these biological and chemical elicitors have not been evaluated so far against multiple infections of both these diseases which commonly occur during the post-rainy season. The present study was therefore aimed to assess the capability of PGPR strains and chemical plant activators, as individual and in integration, in elicitation of ISR against multiple infections. Three PGPR strains, Azotobacter chroococcum strain Azc-3, Bacillus megaterium strain Bm-1 and Pseudomonas fluorescens strain Psf-4, and plant activators, acetyl-salicylic acid (ASA), sodium salicylate (NaS) and 4-amino-n-butyric acid (ABA) were selected for the study. Under in vitro tests, all the plant activators up to 2000 ppm concentration exhibited their compatibility with the PGPR strains tested. Upon assaying of elicitors with plant-pathosystem, disease suppression was significantly (p = 0.05) high with integrated application of PGPR strains and plant activators when compared to their individual applications. All the elicitors at individual application varied in their response to multiple infections with the plant age. However, integration of Azc-3 + ASA provided greater suppression to multiple infections of brown leaf spot and leaf rust diseases during the entire growth period of mulberry plants. Thus, this combination of biological and chemical elicitors holds great promise to provide an effective ecofriendly alternative to the toxic chemical fungicides presently recommended for the control of brown leaf spot and leaf rust diseases in mulberry.     

Characterization of Encephalitozoon (Septata) intestinalis Isolates Cultured from Nasal Mucosa and Bronchoalveolar Lavage Fluids of Two AIDS Patients

ABSTRACT. Microsporidia are obligate intracellular protozoan parasites that can cause opportunistic infections in AIDS patients. Species from five genera of microsporidia are presently known to infect man. One species, Septata intestinalis originally was detected in stool specimens of individuals with chronic diarrhea and subsequently was found to disseminate to the kidneys, lungs, and nasal sinuses. This organism has since been reclassified as Encephalitozoon and in this study, we report the culture of Encephalitozoon intestinalis from a bronchoalveolar lavage specimen and a nasal mucus aspirate of two AIDS patients living in the USA. The bronchoalveolar and nasal microsporidian isolates grew in several continuous cell lines including RK-13, MDCK, HT-29, Caco-2, Vero, and 1047. Transmission electron microscopy of the clinical and cell culture specimens revealed that the new isolates appeared to be E. intestinalis based on morphology and growth of organisms in septated membrane-bound parasitophorous vacuoles. The new E. intestinalis isolates were characterized and compared with the first isolated E. intestinalis that was cultured from stool to confirm their identity and to determine if there existed any minor differences, as seen in the closely related Encephalitozoon cuniculi strains. By the methods of sodium dodecyl sulfate-polyacrylamide gel electrophoresis staining for proteins and carbohydrates, Western blot immunodetection, and polymerase chain reaction-based methods with restriction endonuclease digestion, double-stranded DNA heteroduplex mobility shift analysis, and DNA sequencing of the ribosomal DNA intergenic spacer region, the new isolates were identical to each other and to the reference isolate of E. intestinalis. In addition, with any of these methods, the E. intestinalis organisms could be distinguished from the three E. cuniculi strains, Encephalitozoon hellem, and Vittaforma corneae, which is important for diagnostics, therapeutic strategies, and epidemiology.     

Detection of latent infections caused by Colletotrichum sp. in olive fruit

Aims

To set up a practical method to detect latent infections of Colletotrichum sp., the causal agent of olive anthracnose, on olives before the onset of disease symptoms.

Methods and Results

Freezing, sodium hydroxide (NaOH), ethanol and ethylene treatments were evaluated to detect latent infections on inoculated and naturally infected olive fruit by Colletotrichum sp. as non‐hazardous alternatives to paraquat. Treatments were conducted using fruit of cultivars Arbequina and Hojiblanca. The disease incidence and T₅₀ were calculated. Dipping in NaOH 0·05% solution and the paraquat method were the most effective treatments on both inoculated and naturally infected fruit, although the value of T₅₀ was lower for the NaOH method than for the paraquat method in one of the experiments. Subsequently, the dipping time in NaOH 0·05% was evaluated. Longer dipping times in NaOH 0·05% were better than shorter ones in cultivar Arbequina, with 72 h being the most effective in cultivar Hojiblanca.

Conclusions

NaOH solution is a practical method to detect latent infections of Colletotrichum sp. on immature olive fruit.

Significance and Impact of the Study

This study is relevant because we set up a viable, non‐hazardous alternative to paraquat to detect latent infections of Colletotrichum sp. using NaOH. The use of NaOH is a simple and eco‐friendly tool that allows the determination of the level of latent infections by Colletotrichum in olives. Therefore, our method will be useful in decision‐making processes for disease management before the appearance of the first visible symptoms.     

Prevalence of integrons and a new dfrA17 variant in Gram‐negative bacilli which cause community‐acquired infections

A hundred and eleven Gram‐negative bacilli from community‐acquired infections were characterized by antimicrobial susceptibility testing, screened for class 1 and 2 integrons, and statistically evaluated for the association between antibiotic profile and the presence of integrons. The frequency with which integrons were harbored was 28.8%. Three E. coli strains contained a dfrA17 variant inserted in a class 1 integron. Results of PFGE indicated that some E. coli strains carrying integrons were clonally related. Carriage of gene cassettes was significantly associated with resistance to certain antibiotics (P < 0.05).