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European Journal of Microbiology and Immunology
Authors: Hans Kollenda, Ralf Matthias Hagen, Miriam Hanke, Sandra Rojak, Rebecca Hinz, Lars Wassill, Sven Poppert, Egbert Tannich, and Hagen Frickmann

Background: The objective of this study was to assess an in-house loop-mediated isothermal amplification (LAMP) platform for malaria parasite detection and identification on species level.

Methods: LAMP primers specific for the human Plasmodium spp., namely, P. falciparum, P. vivax, P. ovale, P. malariae, and P. knowlesi, as well as genus-specific primers, were tested against a composite gold standard comprising microscopy from thick and thin blood films, commercial genus-specific Meridian illumigene Malaria LAMP, in-house real-time polymerase chain reaction (PCR), and commercial fast-track diagnostics (FTD) Malaria differentiation PCR.

Results: Of the 523 blood samples analyzed, the composite gold standard indicated 243 Plasmodium-species-DNA-containing samples (46.5%). Sensitivity and specificity of the analyzed genus- and species-specific LAMP primers were 71.0%–100.0% and 90.8%–100.0%, respectively. The influence of parasitemia was best documented for P. falciparum-specific LAMP with sensitivity values of 35.5% (22/62) for microscopically negative samples containing P. falciparum DNA, 50% (19/38) for parasitemia ≤50/μL, 84% (21/25) for parasitemia ≤500/μL, and 100% (92/92) for parasitemia >500/μL.

Conclusions: In our hands, performance characteristics of species-specific in-house LAMP for malaria lack reliability required for diagnostic laboratories. The use of the easy-to-apply technique for surveillance purposes may be considered.

Open access
Community Ecology
Authors: A.F.S. Garcia, A.M. Garcia, S.R. Vollrath, F. Schneck, C.F.M. Silva, Í.J. Marchetti, and J.P. Vieira

Food partitioning among coexisting species in different habitats remains an important research topic in trophic ecology. In this work, we combined carbon and nitrogen stable isotope ratios and stomach content analyses to investigate differences in diet and niche overlap of two congeneric juvenile mullet species (Mugil curema and Mugil liza) coexisting in a marine surf-zone and an estuarine zone in southern Brazil (29oS). These habitats have contrasting levels of food availability, especially in terms of prey diversity, with higher microalgae diversity in the estuary than in the marine surf-zone. In these contrasting conditions, we predicted that both mullet species will have (a) higher niche overlap and smaller niche breadth at the marine surf-zone due to the common exploration of highly abundant surf-zone diatoms and (b) lower niche overlap and higher niche breadth inside the estuary due to selective feeding on more diverse food resources. Isotope niche areas (measured as standard ellipse areas) were higher in the estuary (6.10 and 6.18) than in the marine surf-zone (3.68 and 3.37) for both M. curema and M. liza, respectively. We observed an overlap of 52% in isotopic niches of both species in the marine surf-zone and none in the estuary. We also found contrasting patterns in the diet composition between species according to the habitat. At the marine surfzone, diatoms of the classes Bacillariophyceae and Coscinodiscophyceae dominated (> 99%) the food content of both mullet species. In contrast, green algae, cyanobacteria, dinoflagellates and flagellates comprised the diet of both species in the estuary. These results could be explained by spatial differences in food availability (especially regarding diversity of microalgae) between both habitats. At the marine site, both species explored the most abundant microalgae available (mostly the surf-zone diatom Asterionellopsis cf. guyunusae and fragments of Coscinodiscus), whereas in the estuary both species shifted their diets to explore the greater diversity of microalgae resources. Overall, our findings revealed that niche partitioning theory could not fully predict changes in breadth and overlap of food niches of estuarine dependent fish species with complex life cycles encompassing marine to estuarine systems with contrasting food availabilities.

Open access
European Journal of Microbiology and Immunology
Authors: Cosme Alvarado-Esquivel, Luis Francisco Sánchez-Anguiano, Jesús Hernández-Tinoco, Agar Ramos-Nevarez, Sergio Estrada-Martínez, Sandra Margarita Cerrillo-Soto, Miriam Alejandra Mijarez-Hernández, Carlos Alberto Guido-Arreola, Alma Rosa Pérez-Álamos, Isabel Beristain-Garcia, and Elizabeth Rábago-Sánchez

We determined the association between having a history of surgery and the seroreactivity to T. gondii. An age- and gender-matched case-control study of 391 subjects with a history of surgery and 391 subjects without this history was performed. Sera of subjects were analyzed for detection of anti-T. gondii immunoglobulin G (IgG) and M (IgM) antibodies using enzyme-linked immunoassays. Anti-T. gondii IgG antibodies were found in 25 (6.4%) of the 391 cases and in 21 (5.4%) of the 391 controls (odds ratio [OR] = 1.29; 95% confidence interval [CI]: 0.66–2.18; P = 0.54). The frequency of cases with high IgG antibody levels (10/25: 40.0%) was equal to that found in controls (8/21: 38.1%) (OR = 1.08; 95% CI: 0.32–3.56; P = 0.89). Of the 25 anti-T. gondii IgG antibody seropositive cases, 5 (16.0%) were also positive for anti-T. gondii IgM antibodies. Meanwhile, of the 21 anti-T. gondii IgG antibody seropositive controls, 4 (19.0%) were also positive for anti-T. gondii IgM antibodies (OR = 0.81; 95% CI: 0.17–3.72; P = 0.80). Logistic regression showed that only the variable “hysterectomy” was associated with T. gondii seropositivity (OR = 4.6; 95% CI: 1.6–13.4; P = 0.005). Results suggest that having a history of surgery is not an important risk factor for infection with T. gondii. However, the link between T. gondii infection and hysterectomy should be further investigated.

Open access

One of the main goals of community ecology is to measure the relative importance of environmental filters to understand patterns of species distribution at different temporal and spatial scales. Likewise, the identification of factors that shape symbiont metacommunity structures is important in disease ecology because resulting structures drive disease transmission. We tested the hypothesis that distributions of virus species and viral families from rodents and bats are defined by shared responses to host phylogeny and host functional characteristics, shaping the viral metacommunity structures at four spatial scales (Continental, Biogeographical, Zoogeographical, and Regional). The contribution of host phylogeny and host traits to the metacommunity of viruses at each spatial scale was calculated using a redundant analysis of canonical ordering (RDA). For rodents, at American Continental scale the coherence of viral species metacommunity increased while the spatial scale decreased and Quasi-Clementsian structures were observed. This pattern suggests a restricted distribution of viruses through their hosts, while in the Big Mass (Europe, Africa, and Asia), the coherence decreased as spatial scale decreased. Viral species metacommunities associated with bats was dominated by random structures along all spatial scales. We suggest that this random pattern is a result of the presence of viruses with high occupancy range such as rabies (73%) and coronavirus (27%), that disrupt such structures. At viral family scale, viral metacommunities associated with bats showed coherent structures, with the emergence of Quasi- Clementsian and Checkerboard structures. RDA analysis indicates that the assemblage of viral diversity associated with rodents and bats responds to phylogenetic and functional characteristics, which alternate between spatial scales. Several of these variations could be subject to the spatial scale, in spite of this, we could identify patterns at macro ecological scale. The application of metacommunity theory at symbiont scales is particularly useful for large-scale ecological analysis. Understanding the rules of host-virus association can be useful to take better decisions in epidemiological surveillance, control and even predictions of viral distribution and dissemination.

Open access
European Journal of Microbiology and Immunology
Authors: Ahmed Elfiky, Agnes Bonifacius, Joern Pezoldt, Maria Pasztoi, Paweena Chaoprasid, Pooja Sadana, Nagla El-Sherbeeny, Magda Hagras, Andrea Scrima, Petra Dersch, and Jochen Huehn

Adaptive immunity is essentially required to control acute infection with enteropathogenic Yersinia pseudotuberculosis (Yptb). We have recently demonstrated that Yptb can directly modulate naïve CD4+ T cell differentiation. However, whether fully differentiated forkhead box protein P3 (Foxp3+) regulatory T cells (Tregs), fundamental key players to maintain immune homeostasis, are targeted by Yptb remains elusive. Here, we demonstrate that within the CD4+ T cell compartment Yptb preferentially targets Tregs and injects Yersinia outer proteins (Yops) in a process that depends on the type III secretion system and invasins. Remarkably, Yop-translocation into ex vivo isolated Foxp3+ Tregs resulted in a substantial downregulation of Foxp3 expression and a decreased capacity to express the immunosuppressive cytokine interleukin-10 (IL-10). Together, these findings highlight that invasins are critically required to mediate Yptb attachment to Foxp3+ Tregs, which allows efficient Yop-translocation and finally enables the modulation of the Foxp3+ Tregs' suppressive phenotype.

Open access
European Journal of Microbiology and Immunology
Authors: Manja Boehm, Daniel Simson, Ulrike Escher, Anna-Maria Schmidt, Stefan Bereswill, Nicole Tegtmeyer, Steffen Backert, and Markus M. Heimesaat

Campylobacter jejuni is a major food-borne zoonotic pathogen, responsible for a large proportion of bacterial gastroenteritis cases, as well as Guillian-Barré and Miller-Fisher syndromes. During infection, tissue damage is mainly caused by bacteria invading epithelial cells and traversing the intestinal barrier. C. jejuni is able to enter the lamina propria and the bloodstream and may move into other organs, such as spleen, liver, or mesenteric lymph nodes. However, the involved molecular mechanisms are not fully understood. C. jejuni can transmigrate effectively across polarized intestinal epithelial cells mainly by the paracellular route using the serine protease high-temperature requirement A (HtrA). However, it appears that HtrA has a dual function, as it also acts as a chaperone, interacting with denatured or misfolded periplasmic proteins under stress conditions. Here, we review recent progress on the role of HtrA in C. jejuni pathogenesis. HtrA can be transported into the extracellular space and cleaves cell-to-cell junction factors, such as E-cadherin and probably others, disrupting the epithelial barrier and enabling paracellular transmigration of the bacteria. The secretion of HtrA is a newly discovered strategy also utilized by other pathogens. Thus, secreted HtrA proteases represent highly attractive targets for anti-bacterial treatment and may provide a suitable candidate for vaccine development.

Open access
European Journal of Microbiology and Immunology
Authors: Markus Krohn, Thomas Wanek, Marie-Claude Menet, Andreas Noack, Xavier Declèves, Oliver Langer, Wolfgang Löscher, and Jens Pahnke

ATP-binding cassette (ABC) transporters are of major importance for the restricted access of toxins and drugs to the human body. At the body's barrier tissues like the blood–brain barrier, these transporters are highly represented. Especially, ABCB1 (P-glycoprotein) has been a priority target of pharmaceutical research, for instance, to aid chemotherapy of cancers, therapy resistant epilepsy, and lately even neurodegenerative diseases. To improve translational research, the humanization of mouse genes has become a popular tool although, like recently seen for Abcb1, not all approaches were successful. Here, we report the characterization of another unsuccessful commercially available ABCB1 humanized mouse strain. In vivo assessment of transporter activity using positron emission tomography imaging revealed a severe reduction of ABCB1 function in the brain of these mice. Analyses of brain mRNA and protein expression showed that the murine Abcb1a gene is still expressed in homozygous humanized animals while expression of the human gene is minimal. Promoter region analyses underpinned that the introduced human gene might dysregulate normal expression and provided insights into the regulation of both transcription and translation of Abcb1a. We conclude that insertion of the human coding DNA sequence (CDS) into exon 3 instead of exon 2 most probably represents a more promising strategy for Abcb1a humanization.

Open access

The fever-inducing effect of lipopolysaccharides (LPS) is well known, and human blood is extremely responsive to this pyrogen. Recently, the safety of LPS-containing food supplements and probiotic drugs as immune-stimulants has been questioned, although these products are orally taken and do not reach the bloodstream undigested. The concerns are understandable, as endotoxaemia is a pathological condition, but the oral uptake of probiotic products containing LPS or Gram-negative bacteria does not pose a health risk, based on the available scientific evidence, as is reviewed here. The available methods developed to detect LPS and other pyrogens are mostly used for quality control of parentally applied therapeuticals. Their outcome varies considerably when applied to food supplements, as demonstrated in a simple comparative experiment. Products containing different Escherichia coli strains can result in vastly different results on their LPS content, depending on the method of testing. This is an inherent complication to pyrogen testing, which hampers the communication that the LPS content of food supplements is not a safety concern.

Open access
European Journal of Microbiology and Immunology
Authors: Kouassi N’Guessan, Timothée Ouassa, Anna S. Dean, Riccardo Alagna, Guy Damien Adagra, Valeri Ibode, Daniela M. Cirillo, and Jacquemin Kouakou

Setting: Tuberculosis (TB) drug resistance survey was conducted in 2016–2017 to estimate the burden of drug-resistant TB in Côte d'Ivoire.

Design: A cross-sectional cluster-based survey was conducted. All eligible smear positive patients were interviewed using a structured questionnaire to collect clinical and sociodemographic information and tested by the Xpert Mycobacterium tuberculosis/rifampicin (MTB/RIF) assay. If resistant to rifampicin, solid and liquid cultures were performed. Phenotypic drug susceptibility testing (DST) was conducted in liquid medium for rifampicin, isoniazid, ethambutol, streptomycin, ofloxacin, and amikacin.

Results: Of the 1105 sputum smear positive patients enrolled, 995 new and 100 previously treated patients were positive for Mycobacterium tuberculosis complex by Xpert. Proportion of patients with rifampicin resistance was 4.6% (95% CI: 2.4–6.7) and 22% (95% CI: 13.7–30.3), respectively, for new and previously treated patients. Second-line DST results were available for most rifampicin-resistant patients. None were resistant to amikacin, only two were ofloxacin-resistant. Apart from the antecedent of previously treatment for TB, no other risk factors for rifampicin resistance were detected.

Conclusion: Prevalence of rifampicin resistance among TB patients in Côte d'Ivoire is higher than that in other countries in the region. Surveillance of drug resistance, through an expanded GeneXpert network, and programmatic management of drug-resistant TB (PMDT) must be strengthened in Côte d'Ivoire.

Open access
European Journal of Microbiology and Immunology
Authors: Anabel Cruz-Romero, Cosme Alvarado-Esquivel, Dora Romero-Salas, Ángel Osvaldo Alvarado-Félix, Sokani Sánchez-Montes, Jesús Hernández-Tinoco, and Luis Francisco Sánchez-Anguiano

Purpose: This study aimed to determine the seroprevalence and correlates of Leptospira IgG antibodies in backyard pigs in the northern Mexican state of Durango. We performed a cross-sectional study of 305 backyard pigs. Anti-Leptospira IgG antibodies were detected using microscopic agglutination assay (MAT) with a panel of 12 Leptospira antigens.

Results: Overall, antibodies against Leptospira (any of the 12 Leptospira serovars examined) were found in 186 (61.0%) of the 305 pigs studied. Seropositive pigs were found on 80 (70.2%) of the 114 properties surveyed. The predominant serovar was Leptospira interrogans Pomona (n = 55); followed by Leptospira noguchii Lousiana and Leptospira santarosai Tarassovi (n = 53 each); L. interrogans Bataviae (n = 47); Leptospira biflexa Semaranga and L. interrogans Hebdomadis (n = 36 each); L. interrogans Pyrogenes (n = 30); L. interrogans Djasiman (n = 20); Leptospira borgpetersenii Ballum (n = 11); L. noguchii Panama and L. interrogans Canicola (n = 5 each); and L. borgpetersenii Mini (n = 2). Logistic regression showed that seropositivity was associated with low (<1000 m above sea level) altitude (odds ratio [OR] = 3.24; 95% confidence interval [CI]: 2.01–5.20; P < 0.001).

Conclusions: This is the first report of Leptospira exposure in backyard pigs in Mexico and of an association between Leptospira exposure in pigs and altitude. Backyard pigs represent a high-risk group for Leptospira exposure.

Open access