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Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic...
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Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Public Health Microbiology".

Deadline for manuscript submissions: 15 March 2025 | Viewed by 13113

Special Issue Editor

Dr. Érica S. Martins-Duarte

E-Mail Website
Guest Editor
Department of Parasitology, Federal University, Rio de Janeiro, Brazil.
Interests: T. gondii

Special Issue Information

Dear Colleagues,

Parasitism is an ecological relation in which an organism, a parasite, lives inside or on another organism, the host, with the former depending on this organism to acquire essential nutrients and survive. Thus, it is a relationship that benefits the parasite at the expense of the host but not necessarily killing the latter. In humans, parasitic diseases hamper development and still cause high mortality, especially in children in developing countries. In addition, such infections in poultry, cattle, or swine, for example, are responsible for economic losses in livestock. In this Special Issue, we hope to receive original and review papers highlighting new advances in the diagnosis, control, and treatment of parasitic diseases that affect humans and animals of veterinary importance.

Dr. Érica S. Martins-Duarte
Guest Editor

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Keywords

  • protozoan
  • ectoparasites
  • helminths

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Published Papers (11 papers)

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23 pages, 23820 KiB  
Article
Antiproliferative and Morphological Analysis Triggered by Drugs Contained in the Medicines for Malaria Venture COVID-Box Against Toxoplasma gondii Tachyzoites
by Andréia Luiza Oliveira Costa, Mike dos Santos, Giulia Caroline Dantas-Vieira, Rosálida Estevam Nazar Lopes, Rossiane Claudia Vommaro and Érica S. Martins-Duarte
Microorganisms 2024, 12(12), 2602; https://doi.org/10.3390/microorganisms12122602 - 16 Dec 2024
Viewed by 708
Abstract
Toxoplasma gondii is a protozoan, and the etiologic agent of toxoplasmosis, a disease that causes high mortality in immunocompromised individuals and newborns. Despite the medical importance of toxoplasmosis, few drugs, which are associated with side effects and parasite resistance, are available for its [...] Read more.
Toxoplasma gondii is a protozoan, and the etiologic agent of toxoplasmosis, a disease that causes high mortality in immunocompromised individuals and newborns. Despite the medical importance of toxoplasmosis, few drugs, which are associated with side effects and parasite resistance, are available for its treatment. Here, we show a screening of molecules present in COVID-Box to discover new hits with anti-T. gondii activity. COVID-Box contains 160 molecules with known or predicted activity against SARS-CoV-2. Our analysis selected 23 COVID-Box molecules that can inhibit the tachyzoite forms of the RH strain of T. gondii in vitro by more than 70% at 1 µM after seven days of treatment. The inhibitory curves showed that most of these molecules inhibited the proliferation of tachyzoites with IC50 values below 0.80 µM; Cycloheximide and (-)-anisomycin were the most active drugs, with IC50 values of 0.02 μM. Cell viability assays showed that the compounds are not toxic at active concentrations, and most are highly selective for parasites. Overall, all 23 compounds were selective, and for two of them (apilimod and midostaurin), this is the first report of activity against T. gondii. To better understand the effect of the drugs, we analyzed the effect of nine of them on the ultrastructure of T. gondii using transmission electron microscopy. After treatment with the selected drugs, the main changes observed in parasite morphology were the arrestment of cell division and organelle alterations. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

Figure 1
<p>Proliferation index of the best drugs of the COVID-Box after 24 h of treatment with different concentrations of (<b>A</b>) Pyrimethamine, (<b>B</b>) Cycloheximide, Anisomycin, (<b>C</b>) Merimepodib, (<b>D</b>) Midostaurin, (<b>E</b>) Salimomycin, (<b>F</b>) Bortezomib, Mycophenolic acid, Apilimod, Almitrine, and Ivermectin. Values represent mean ± SD of three experiments, except for merimepodib, salinomycin, and pyrimethamine (two experiments).</p> Full article ">Figure 2
<p>(<b>A</b>–<b>F</b>). Transmission electron microscopy and analysis of the ultrastructure of tachyzoites after treatment with cycloheximide and bortezomib. The parasites were treated with the compounds for 48 h. (<b>A</b>,<b>B</b>) Untreated parasites showed typical morphology (<b>A</b>,<b>B</b>) and division process by endodyogeny (arrows in (<b>B</b>)). (<b>C</b>) Parasites treated with the drug 62.5 nM cycloheximide showed an increase in the endoplasmic reticulum area (stars) and alterations in the structure of the plasma membrane, with regions with a lack of inner membrane complex (black arrowhead). (<b>D</b>) Parasites treated with 125 nM cycloheximide were destroyed; it is possible to observe parasite content spread through the PV. (<b>E</b>,<b>F</b>) Parasites treated with 62.5 nM bortezomib showed cell division alterations, as seen by the Golgi complex surrounded by the nucleus envelope (arrowhead) and a parasite presenting three nucleus profiles without constructing new daughter cells. Mitochondrial swelling (M) and regions of parasite devoid IMC were also observed (arrows). A—apicoplast, Ac—acidocalcisome, C—conoid; DG—dense granules, GC—Golgi complex, Lb—lipid body, M—mitochondrion, m—micronemes, N—nucleus, Rp—rhoptries, PV—parasitophorous vacuole, V—vacuolar compartment.</p> Full article ">Figure 3
<p>Fluorescence microscopy of untreated parasites (<b>A</b>) or after treatment with 31.2 nM (<b>B</b>) and 62.5 nM (<b>C</b>) of bortezomib. Parasites were labeled with anti-IMC1 for inner membrane complex (IMC, green) and DAPI for DNA (blue). (<b>A</b>) Untreated parasites showed typical morphology (arrow) and division process (arrowhead). (<b>B</b>,<b>C</b>) treated parasites showed an aberrant cell division process with large parasites harboring two or more nuclei (arrow), daughter cells without nuclei (arrowheads), and regions of the cells without IMC coverage (asterisks). (<b>D</b>) Quantitative analysis of the number of PVs presenting parasites with aberrant cell division. Results in (<b>D</b>) are the mean ± SD of two independent experiments. * <span class="html-italic">p</span> &lt; 0.05; **<span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001. Bars = 2.5 µm.</p> Full article ">Figure 4
<p>Transmission electron microscopy of <span class="html-italic">T. gondii</span> after treatment for 48 h with (-)-anisomycin (<b>A</b>,<b>B</b>) and ivermectin (<b>C</b>,<b>D</b>). (<b>A</b>) Treatment with 100 nM (-)-anisomycin induced changes in the parasite’s endoplasmic reticulum (star in inset) and (<b>B</b>) 100 nM (-)-anisomycin also induced impairment of the cell division, making it possible to observe a single parasite with two nuclei and causing discontinuation of the inner membrane complex (black arrows). (<b>C</b>) Parasites treated with 1 µM ivermectin induced the formation of myelin-like figures (inset—white arrowhead). (<b>D</b>) In this figure, it is also possible to observe an intense vacuolization process in parasites treated with 1 μM ivermectin (asterisks). M—mitochondria; N—nucleus; GC—Golgi complex; ER—endoplasmic reticulum.</p> Full article ">Figure 5
<p>Fluorescence microscopy analysis of tachyzoites treated with 62.5 nM and 125 nM (-)-anisomycin (<b>A</b>–<b>C</b>) and 1 µM ivermectin (<b>D</b>,<b>E</b>). Parasites were labeled with anti-IMC1 for inner membrane complex (IMC, green) and DAPI for DNA (blue). (<b>A</b>) Parasites treated with 62.5 nM (-)-anisomycin showed daughter cells’ budding arrestment, forming a large mass of tethered daughter cells (arrow). (<b>B</b>) Treatment with 125 nM (-)-anisomycin led to a large round mass of cells with a nucleus of increased size and disorganized profiles of IMC (arrowheads). The arrow points to a parasite region without the IMC coverage. (<b>C</b>) Quantitative analysis of the number of PVs presenting parasites with aberrant cell division after treatment with (-)-anisomycin. (<b>D</b>) Parasites treated with 1 µM ivermectin showed a divided nucleus without the construction of daughter cells (arrows). (<b>E</b>) Quantitative analysis of the number of PVs presenting parasites with aberrant cell division after treatment with ivermectin. Results in (<b>C</b>,<b>E</b>) are the mean ± SD of two independent experiments. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01. Bars = 2 µm.</p> Full article ">Figure 6
<p>Transmission electron microscopy of <span class="html-italic">T. gondii</span> tachyzoites after treatment with almitrine and midostaurin for 48 h. (<b>A</b>) Parasites treated with 1 µM almitrine showed myelin-like structures (arrowhead in inset). (<b>B</b>) Treatment with one µM almitrine also induced the formation of large vacuoles containing membranous material (asterisks) and disruption of cell division, as seen by a large mother mass harboring two non-budded daughter cells (asterisks). (<b>C</b>,<b>D</b>) Parasites treated with 250 nM midostaurin for 48 h. (<b>C</b>) Vacuole containing a mass of tachyzoite with several arrested daughter cells and IMC profiles through the cytoplasm (arrowheads). A parasite presenting a fragmented nucleus (large arrow), and a process similar to autophagy (asterisks) was observed too.</p> Full article ">Figure 7
<p>Fluorescence microscopy analysis of tachyzoites treated with almitrine and midostaurin. Parasites were labeled with anti-IMC1 for inner membrane complex (IMC, green) and DAPI for DNA (blue). (<b>A</b>) Parasites treated with 1 µM almitrine showed cell division alteration with tachyzoites presenting large nuclei (asterisks) and masses with incomplete division process (arrow). (<b>B</b>) Treatment with 0.25 µM midostaurine caused a large round mass of cells with a nucleus of increased size (asterisk), tachyzoites showing regions without the IMC cover (arrows), and daughter cells without a nucleus (arrowheads). (<b>C</b>) Quantitative analysis of the number of PVs presenting parasites with aberrant cell division after treatment with almitrine. (<b>D</b>) Quantitative analysis of the number of PVs presenting aberrant parasites after treatment with midostaurin. Results in (<b>C</b>,<b>D</b>) are the mean ± SD of two independent experiments. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01. Bars = 2.5 μm.</p> Full article ">Figure 8
<p>Morphological analysis of tachyzoites of <span class="html-italic">T. gondii</span> after treatment with 1.5 µM merimepodib. (<b>A</b>) Treatment with 1.5 µM induced Golgi complex fragmentation (vesiculation) and rhoptry disorganization, which can be seen at higher magnification in the inset. (<b>B</b>) Tachyzoites treated with 1.5 µM merimepodib also presented large vacuoles containing membranous material (asterisks). (<b>C</b>) Fluorescence microscopy analysis of tachyzoites treated with 1.5 µM merimepodib for 24 h. Parasites were labeled with anti-ARO for rhoptries (green), anti-SAG1 for parasite plasma membrane (red), and DAPI for DNA (blue). Images represent the projection of different Z focal planes. (<b>D</b>) Quantitative analysis of the number of PVs presenting parasites with rhoptry- altered morphology (arrowheads in (<b>C</b>)). Results are the mean ± SD of two independent experiments. * <span class="html-italic">p</span> &lt; 0.05. M—mitochondria; N—nucleus; GC—Golgi complex; Rp—rhoptries. Bars = 2 µm.</p> Full article ">Figure 9
<p>Transmission electron microscopy of <span class="html-italic">T. gondii</span> tachyzoites after treatment with 250 nM mycophenolic acid for 48 h. (<b>A</b>) Tachyzoites in the division process present multiple lobules (arrowheads) or mitotic nuclei (horseshoe shape) without the construction of new daughter cells (arrow). (<b>B</b>) Daughter cells without the completion of the division process with mitotic nuclei. A—apicoplast; M—mitochondria; N—nucleus.</p> Full article ">Figure 10
<p>Morphological analysis of <span class="html-italic">T. gondii</span> tachyzoites after treatment with salinomycin for 24 h. (<b>A</b>) Treatment with 125 nM caused an extensive vacuolization process (asterisks) on the parasite. (<b>B</b>) Tachyzoites treated with 250 nM showed an extensive vacuolization process (asterisks) and cell lysis (arrow). (<b>C</b>,<b>C’</b>) Fluorescence microscopy analysis of tachyzoites treated with 1.5 µM merimepodib for 24 h. Parasites were labeled with anti-LAMP1 for host cell lysosomes (green), anti-SAG1 for parasite plasma membrane (red), and DAPI for DNA (blue).</p> Full article ">
9 pages, 748 KiB  
Article
Determinants of Anemia in Schoolchildren in the Highland Bolivia
by Washington R. Cuna, Ivonne Contreras, Armando Rodriguez, Roberto Passera and Celeste Rodriguez
Microorganisms 2024, 12(12), 2491; https://doi.org/10.3390/microorganisms12122491 - 3 Dec 2024
Viewed by 621
Abstract
Anemia is a health problem of concern among schoolchildren in underprivileged rural regions, where recurrent parasitic infections are common. A cross-sectional study was conducted in 229 schoolchildren in rural highland Bolivia in the department of La Paz, an area with a high prevalence [...] Read more.
Anemia is a health problem of concern among schoolchildren in underprivileged rural regions, where recurrent parasitic infections are common. A cross-sectional study was conducted in 229 schoolchildren in rural highland Bolivia in the department of La Paz, an area with a high prevalence of protozoan and helminth infections, to determine the types and mechanisms of anemia. A substantial proportion of children (40.2%) were found to be anemic based on hemoglobin measurements. No associations were found between low hemoglobin levels and helminth or protozoan infections when evaluating infectious causes of anemia, nor with Giardia lamblia or Blastocystis hominis, which are associated with iron deficiency and nutrient malabsorption and were highly prevalent in this study. The significant association between anemia and hypochromia suggests iron deficiency, aligned with low hemoglobin levels. A total of 39 out of 150 children (26%) had markers consistent with iron deficiency anemia (IDA), 26 out of 127 children (20%) met the criteria for anemia of inflammation (AI). Furthermore, 12 of the 127 tested children (9.4%) met the criteria for mixed AI with IDA according to the soluble transferrin receptor (sTfR)/log ferritin levels, which increased significantly due to overall infections by Hymenolepis nana and Ascaris lumbricoides helminths. The findings highlight the need for integrated public health interventions to address iron nutrition and parasitic infections to effectively prevent anemia in this vulnerable population. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

Figure 1
<p>Parasite prevalence by age group. Vertical bars indicate the prevalence of specific parasite infections, helminth, protozoa, and coinfections. Percentages are shown for the 5–7, 8–10, and 11–13 year subgroups.</p> Full article ">Figure 2
<p>Boxplot comparing the soluble transferrin receptor (sTfR) to log ferritin ratios, defining combined anemia of inflammation with iron-deficiency anemia among study children with and without detectable infections. The distributions of sTfR/log SF ratios are presented for sTfR-tested children who either have (N = 12) or do not have (N = 127) helminth infections. The <span class="html-italic">p</span>-value for group differences is 0.001, determined via the Mann–Whitney U test.</p> Full article ">
17 pages, 10663 KiB  
Article
Recombinant SAG2A Protein from Toxoplasma gondii Modulates Immune Profile and Induces Metabolic Changes Associated with Reduced Tachyzoite Infection in Peritoneal Exudate Cells from Susceptible C57BL/6 Mice
by Thaíse Anne Rocha dos Santos, Mário Cézar de Oliveira, Edson Mario de Andrade Silva, Uener Ribeiro dos Santos, Monaliza Macêdo Ferreira, Ana Luísa Corrêa Soares, Neide Maria Silva, Tiago Antônio de Oliveira Mendes, Jamilly Azevedo Leal-Sena, Jair Pereira da Cunha-Júnior, Tiago Wilson Patriarca Mineo, José Roberto Mineo, Érica Araújo Mendes, Jane Lima-Santos and Carlos Priminho Pirovani
Microorganisms 2024, 12(11), 2366; https://doi.org/10.3390/microorganisms12112366 - 20 Nov 2024
Viewed by 874
Abstract
Toxoplasmosis is a neglected disease that represents a significant public health problem. The antigenic profile of T. gondii is complex, and the immune response can lead to either susceptibility or resistance. Some antigens, such as surface antigen glycoprotein (SAG), are expressed on the [...] Read more.
Toxoplasmosis is a neglected disease that represents a significant public health problem. The antigenic profile of T. gondii is complex, and the immune response can lead to either susceptibility or resistance. Some antigens, such as surface antigen glycoprotein (SAG), are expressed on the surface of tachyzoite stages and interact with the host immune cells. In this study, we investigated the potential of the recombinant SAG2A protein of T. gondii to control parasitism and modulate the immune response in the peritoneal exudate cells (PECs) of both susceptible (C57BL/6) and resistant (BALB/c) mice using an in vitro infection model, gene expression, proteomic analysis, and bioinformatic tools. Our results showed that rSAG2A-treated PECs presented a lower parasitism in C57BL/6 mice but not in the PECs from BALB/c mice, and induced a pro-inflammatory cytokine profile in C57BL/6 mice (iNOS, TNF-α, and IL-6). rSAG2A modulated different exclusive proteins in each mouse lineage, with PECs from the C57BL/6 mice being more sensitive to modulation by rSAG2A. Additionally, biological processes crucial to parasite survival and immune response were modulated by rSAG2A in the C57BL/6 PECs, including fatty acid beta-oxidation, reactive oxygen species metabolism, interferon production, and cytokine-mediated signaling pathways. Together, our study indicates that rSAG2A controls T. gondii parasitism in susceptible C57BL/6 PECs through the modulation of pro-inflammatory cytokines and enhanced expression of proteins involved in the cytotoxic response. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

Figure 1
<p>rSAG2A decreases the parasitism in PECs infected with tachyzoites of <span class="html-italic">T. gondii</span>. (<b>A</b>) PECs from BALB/c and C57BL/6 mice were infected with tachyzoites of <span class="html-italic">T. gondii</span> (RH-2FI strain; MOI = 1) for 3 h, followed or not by addition of rSAG2A. (<b>B</b>) The number of intracellular tachyzoites was measured by the β -galactosidase colorimetric assay after incubation for 24 h with rSAG2A. Data represent mean + SD of two independent experiments. *** <span class="html-italic">p</span> &lt; 0.001 by one-way ANOVA followed by the Tukey post-test.</p> Full article ">Figure 2
<p>rSAG2A alters the pro-inflammatory and anti-inflammatory cytokines expression in PECs infected with <span class="html-italic">T. gondii</span>. Cytokine transcript accumulation was evaluated during the presence of parasites for 3 h in PECs from both BALB/c and C57BL/6 mice or treated with rSAG2A during 24 h, through qPCR. (<b>A</b>) INOS, (<b>B</b>) Arginase, (<b>C</b>) TNF-α, (<b>D</b>) IL-10, (<b>E</b>) IL-1β, (<b>F</b>) TGF-β, and (<b>G</b>) IL-6. (<b>H</b>) Hierarchical clustering for the same mean relative quantification (RQ) values for the same genes, represented in z score scale, in the different conditions studied. Data represent mean + SD of two independent experiments using a pool of PECs n = 4 animals/experiment. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001 between control and rSAG2A of 6 replicates/group by ANOVA followed by the Tukey post-test.</p> Full article ">Figure 3
<p>Venn diagram of proteins shared by PECs of mice infected with <span class="html-italic">T. gondii</span> or treated with rSAG2A. Venn diagram showing the number of proteins in common and not in common between the different groups. PECs of the BALB/c mice treated with rSAG2A (BALB/c_rSAG2A), BALB/c mice infected with <span class="html-italic">T. gondii</span> (BALB/c_<span class="html-italic">T. gondii</span>), C57BL/6 mice treated with rSAG2A (C57BL/6_rSAG2A), and C57BL/6 mice infected with <span class="html-italic">T. gondii</span> (C57BL/6_<span class="html-italic">T. gondii</span>). The asterisk values represent proteins augmented with fold change &gt; 2 exclusively in PECs of C57BL/6 mice treated with rSAG2A.</p> Full article ">Figure 4
<p>Accumulation profile and hierarchical clustering for exclusive/up-modulated or down-modulated proteins in C57BL/6 mice PECs treated with rSAG2A (<b>A</b>) or infected with <span class="html-italic">T. gondii</span> (<b>B</b>), compared to BALB/c PECs in the same conditions. Proteins in red are those exclusive or that had a Log2FC (compared to the other conditions) at least twice as high in C57BL/6 PECs treated with rSAG2A (red names in <b>A</b>) or in C57BL/6 PECs infected with <span class="html-italic">T. gondii</span> (red names in <b>B</b>). Proteins in blue are those that were repressed only in C57BL/6 PECs treated with rSAG2A (blue names in <b>A</b>) or in C57BL/6 PECs infected with <span class="html-italic">T. gondii</span> (blue names in <b>B</b>). Heatmap colors are represented by Log2FC scale, which indicates the accumulation of proteins in Log2 fold change scale; blue shades indicate the repressed proteins and red shades indicate proteins that increased in relation to their respective controls. Black arrows represent proteins present in the cytokine network.</p> Full article ">Figure 5
<p>Protein–protein interaction network for exclusive and augmented proteins from PECs of C57BL/6 mice treated with rSAG2A (<b>A</b>) or infected with <span class="html-italic">T. gondii</span> (<b>B</b>). The bigger nodes represent the proteins identified through proteomic analysis. Smaller nodes were aggregated with networks using the STRING database. The different colors represent different clusters identified in the module analysis. The biological processes enriched in each cluster are described with the respective colors around the networks.</p> Full article ">Figure 6
<p>Network of protein–protein interactions for down-modulated proteins in PECs of C57BL/6 mice treated with rSAG2A (<b>A</b>) or infected with <span class="html-italic">T. gondii</span> (<b>B</b>). The bigger nodes represent the proteins identified through proteomic analysis. Smaller nodes were aggregated with networks using the STRING database. The different colors represent different clusters identified in the module analysis. The biological processes enriched in each cluster are described with the respective colors around the networks.</p> Full article ">Figure 7
<p>Protein interaction network related with inflammatory and regulatory cytokines. The large circles represent proteins whose abundance was altered in PECs treated with rSAG2A. Diamonds represent the inflammatory and regulatory cytokines, besides INOs. Small circles represent proteins not identified during the proteomic analysis. The fat connectors highlight first-degree interaction with the cytokines and the slender connectors the interactions in distinct degrees. The arrows indicate the most relevant proteins in the study. The red and blue borders represent up-modulated and down-modulated proteins, respectively.</p> Full article ">
11 pages, 1145 KiB  
Article
Cryptosporidium Infections in Neonatal Calves on a Dairy Farm
by Michaela Kaduková, Andrea Schreiberová, Pavol Mudroň, Csilla Tóthová, Pavel Gomulec and Gabriela Štrkolcová
Microorganisms 2024, 12(7), 1416; https://doi.org/10.3390/microorganisms12071416 - 12 Jul 2024
Viewed by 1212
Abstract
This study was conducted with the aim of the molecular identification of the protozoan parasite Cryptosporidium spp. in calves in the early stage of their development on a dairy farm in Eastern Slovakia. Twenty-five Holstein and Holstein cross calves were included in the [...] Read more.
This study was conducted with the aim of the molecular identification of the protozoan parasite Cryptosporidium spp. in calves in the early stage of their development on a dairy farm in Eastern Slovakia. Twenty-five Holstein and Holstein cross calves were included in the study and monitored from their birth to the fifth week of life (1–5 weeks). Fresh fecal samples were collected from the same group of calves each week, except during the fourth week, and with the exception of Sample 8. All samples were analyzed using the Ziehl–Neelsen staining method and coproantigen was tested using the ELISA test as the screening method. Using the ELISA method, the highest incidence of cryptosporidiosis was observed in the second week of life of the calves, while the antigen was detected in 21 (91.6%) calves. Using the Ziehl–Neelsen staining method, the highest incidence was also observed in the second week, with an incidence rate of 62.5%. Positive isolates confirmed by the ELISA test were molecularly characterized. The species and subtypes of Cryptosporidium in the positive isolates were identified using PCR and the sequence analysis of the small subunit of the ribosomal 18S RNA (ssu rRNA) and the 60 kDa glycoprotein (gp60) genes of the parasite. The sequence analysis of 29 isolates at the 18S rRNA loci confirmed the presence of two species—Cryptosporidium parvum and Cryptosporidium ryanae. Out of 29 isolates, 25 were assigned to the species C. parvum, with the gp60 locus identified as genotype IIaA17G1R1. Among the individual animal groups, calves are the most common reservoirs of the C. parvum zoonotic species. This disease has significant public health implications as contact with livestock and their feces and working with barn manure are major sources of infection, not only for other animals but also for humans. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

Figure 1
<p>Oocysts of <span class="html-italic">Cryptosporidium</span> spp. according to Ziehl–Neelsen staining method; bar—20 µm.</p> Full article ">Figure 2
<p>Phylogenetic tree constructed using the maximum likelihood method and Tamura–Nei model and depicting the relationships among <span class="html-italic">C. parvum</span> and <span class="html-italic">C. ryanae</span> based on the small subunit 18S rRNA gene sequence data available in the GenBank database.</p> Full article ">
10 pages, 2182 KiB  
Article
Parasitic Effects on the Congenital Transmission of Trypanosoma cruzi in Mother–Newborn Pairs
by Ana Gabriela Herrera Choque, Washington R. Cuna, Simona Gabrielli, Simonetta Mattiucci, Roberto Passera and Celeste Rodriguez
Microorganisms 2024, 12(6), 1243; https://doi.org/10.3390/microorganisms12061243 - 20 Jun 2024
Viewed by 1008
Abstract
Maternal parasitemia and placental parasite load were examined in mother–newborn pairs to determine their effect on the congenital transmission of Trypanosoma cruzi. Parasitemia was qualitatively assessed in mothers and newborns by the microhematocrit test; parasite load was determined in the placental tissues [...] Read more.
Maternal parasitemia and placental parasite load were examined in mother–newborn pairs to determine their effect on the congenital transmission of Trypanosoma cruzi. Parasitemia was qualitatively assessed in mothers and newborns by the microhematocrit test; parasite load was determined in the placental tissues of transmitting and non-transmitting mothers by the detection of T. cruzi DNA and by histology. Compared to transmitter mothers, the frequency and prevalence of parasitemia were found to be increased in non-transmitter mothers; however, the frequency and prevalence of parasite load were higher among the transmitter mothers than among their non-transmitter counterparts. Additionally, serum levels of interferon (IFN)-γ were measured by an enzyme-linked immunosorbent assay (ELISA) in peripheral, placental, and cord blood samples. Median values of IFN-γ were significantly increased in the cord blood of uninfected newborns. The median IFN-γ values of transmitter and non-transmitter mothers were not significantly different; however, non-transmitter mothers had the highest total IFN-γ production among the group of mothers. Collectively, the results of this study suggest that the anti-T. cruzi immune response occurring in the placenta and cord is under the influence of the cytokines from the mother’s blood and results in the control of parasitemia in uninfected newborns. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

Figure 1
<p>Results of N-PCR amplifications with <span class="html-italic">T. cruzi</span> DNA nuclear, electrophoresed on a 2% agarose gel and visualized by ethidium bromide staining. The 149 base pairs (bp) were amplified through N-PCR with primers TCZ3 and TCZ4. M: molecular weight marker (50 bp); C+: positive control (<span class="html-italic">T. cruzi</span> II of Y strain); S1–S7: representative amplicons of positive patients, from transmitter (S2–S4) and non-transmitter (S1, S5–S7) mothers; C−: negative control from a patient with negative serology for <span class="html-italic">T. cruzi</span>.</p> Full article ">Figure 2
<p>Histological section of the placenta from a non-transmitter mother. Arrows point to (<b>A</b>,<b>B</b>) amastigote nests; and (<b>C</b>) released parasites (H&amp;E). Scale bar: 25 μm.</p> Full article ">Figure 3
<p>Histological section of the placenta from a transmitter mother. Arrows point to (<b>A</b>) amastigote nest; and (<b>B</b>,<b>C</b>) released parasites (H&amp;E). Scale bar: 25 μm.</p> Full article ">
16 pages, 919 KiB  
Article
Subtype Distribution of Blastocystis spp. in Patients with Gastrointestinal Symptoms in Northern Spain
by Cristina Matovelle, Joaquín Quílez, María Teresa Tejedor, Antonio Beltrán, Patricia Chueca and Luis Vicente Monteagudo
Microorganisms 2024, 12(6), 1084; https://doi.org/10.3390/microorganisms12061084 - 27 May 2024
Cited by 3 | Viewed by 1174
Abstract
Limited molecular data exist on the prevalence and subtype distribution of Blastocystis spp., the most prevalent parasite in human and animal feces worldwide. A total of 44 different subtypes (STs) of Blastocystis are currently recognized based on the sequence of the small subunit [...] Read more.
Limited molecular data exist on the prevalence and subtype distribution of Blastocystis spp., the most prevalent parasite in human and animal feces worldwide. A total of 44 different subtypes (STs) of Blastocystis are currently recognized based on the sequence of the small subunit ribosomal RNA (SSU-rRNA) gene. This is a molecular study of Blastocystis spp. in hospitalized patients with gastrointestinal symptoms in northern Spain. We analyzed 173 Blastocystis-positive patients with gastrointestinal symptoms by using nested PCR for molecular detection, subtype identification, phylogenetic analyses, and genetic diversity assessment. ST2 (34.1%) and ST3 (34.7%) predominated, followed by ST1 (15.6%) and ST4 (15.6%). Mixed infections with different subtypes were observed in some patients. Sequence analysis revealed for the first time in European humans the allele 88 (a variant of ST1). In other cases, alleles commonly found in animal samples were detected (allele 9 in ST2, allele 34 in ST3, and allele 42 in ST4). Phylogenetic analysis showed high variability in ST1 and ST2, suggesting a polyphyletic origin, while both ST3 and ST4 exhibited higher genetic homogeneity, indicating a possible monophyletic origin and recent transmission to humans. These data confirm Blastocystis spp. subtype diversity and may help in understanding the evolutionary processes and potential zoonotic transmission of this parasite. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

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<p>Neighbor joining analysis of the partial sequences of the <span class="html-italic">SSU-rRNA</span> gene of <span class="html-italic">Blastocystis</span> and reference sequences representative of different subtypes. A sequence of <span class="html-italic">Proteromonas lacertae</span> was used as the outgroup and sequences of other <span class="html-italic">Blastocystis</span> subtypes were obtained from GenBank<sup>®</sup> (LC414134.1 and MK874786.1: ST1 <span class="html-italic">Homo sapiens</span>, EU445491.1: ST2 Monkey (sic.), M25.1 and MK874818.1: ST2 <span class="html-italic">Homo sapiens</span>, MN914073.1: ST3 <span class="html-italic">Homo sapiens</span>, and MH127478.1: ST4 <span class="html-italic">Rattus exulans</span>). Genetic distances were calculated using the Kimura 2 model (own image). The length of the branch connecting the outgroup sample was reduced by 61% to simplify the image. The symbol * indicates the sequences obtained in the present work.</p> Full article ">
15 pages, 4162 KiB  
Article
Synthetic Peptides Selected by Immunoinformatics as Potential Tools for the Specific Diagnosis of Canine Visceral Leishmaniasis
by Gabriel Moreira, Rodrigo Maia, Nathália Soares, Thais Ostolin, Wendel Coura-Vital, Rodrigo Aguiar-Soares, Jeronimo Ruiz, Daniela Resende, Rory de Brito, Alexandre Reis and Bruno Roatt
Microorganisms 2024, 12(5), 906; https://doi.org/10.3390/microorganisms12050906 - 30 Apr 2024
Cited by 2 | Viewed by 1170
Abstract
Diagnosing canine visceral leishmaniasis (CVL) in Brazil faces challenges due to the limitations regarding the sensitivity and specificity of the current diagnostic protocol. Therefore, it is urgent to map new antigens or enhance the existing ones for future diagnostic techniques. Immunoinformatic tools are [...] Read more.
Diagnosing canine visceral leishmaniasis (CVL) in Brazil faces challenges due to the limitations regarding the sensitivity and specificity of the current diagnostic protocol. Therefore, it is urgent to map new antigens or enhance the existing ones for future diagnostic techniques. Immunoinformatic tools are promising in the identification of new potential epitopes or antigen candidates. In this study, we evaluated peptides selected by epitope prediction for CVL serodiagnosis in ELISA assays. Ten B-cell epitopes were immunogenic in silico, but two peptides (peptides No. 45 and No. 48) showed the best performance in vitro. The selected peptides, both individually and in combination, were highly diagnostically accurate, with sensitivities ranging from 86.4% to 100% and with a specificity of approximately 90%. We observed that the combination of peptides showed better performance when compared to peptide alone, by detecting all asymptomatic dogs, showing lower cross-reactivity in sera from dogs with other canine infections, and did not detect vaccinated animals. Moreover, our data indicate the potential use of immunoinformatic tools associated with ELISA assays for the selection and evaluation of potential new targets, such as peptides, applied to the diagnosis of CVL. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

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<p>Flow chart with an experimental design for the selection and application of synthetic peptides, based on the epitope prediction approach, in the serological diagnosis of CVL [<a href="#B14-microorganisms-12-00906" class="html-bibr">14</a>].</p> Full article ">Figure 2
<p>The experimental design employed in the serological test for peptides No. 45, No. 48, and mix. The control noninfected group (CNI), dogs from the endemic area and nonendemic area. The <span class="html-italic">L. infantum</span>-infected dogs (CVL) were stratified according to their statuses as asymptomatic dogs (AD), oligosymptomatic dogs (OD), and symptomatic dogs (SD). Vaccinated dogs (LBSap and Leish-Tec<sup>®</sup>) and dogs infected with other pathogens (<span class="html-italic">Trypanosoma cruzi</span>, <span class="html-italic">Ehrlichia canis</span>, and <span class="html-italic">Babesia canis</span>) constitute the other two groups evaluated.</p> Full article ">Figure 3
<p>Performance of the peptides No. 45, No. 48, and mix in the discrimination of noninfected and <span class="html-italic">L. infantum</span>-infected dogs presenting different clinical forms. (<b>a</b>) Distribution of the individual optical density results of the control noninfected dogs (CNI) and <span class="html-italic">L. infantum</span>-infected dogs (CVL) tested by the peptides No. 45, No. 48, mix, and soluble <span class="html-italic">Leishmania infantum</span> antigen (SLiA). (<b>b</b>) Distribution of the individual optical density results of the control noninfected dogs (CNI) and <span class="html-italic">L. infantum</span>-infected dogs according to clinical forms. The CVL dogs were stratified according to their clinical statuses as asymptomatic dogs (AD), oligosymptomatic dogs (OD), and symptomatic dogs (SD). The dotted lines within the graphs represent the cut-offs calculated by the ROC curve between the negative and positive results of SLiA (OD = 0.273), Pep45 (OD = 0.41), Pep48 (OD = 0.431), and mix (OD = 0.331). (<b>c</b>) The ROC curves were constructed with the results of the control noninfected, and <span class="html-italic">L. infantum</span>-infected control serum samples tested by each assay. (<b>d</b>) Results of sensitivity, specificity, negative predictive values (NPV), positive predictive values (PPV), and accuracy from the canine visceral leishmaniasis positive and negative animals tested by the peptides No. 45, No. 48, the mix, and SLiA.</p> Full article ">Figure 4
<p>Cross-reactivity of the peptides No. 45, No. 48, and mix with samples from dogs infected with other pathogens of medical and diagnostic importance. (<b>a</b>) Distribution of the individual optical density results from serum samples from dogs infected with <span class="html-italic">Leishmania infantum</span>, <span class="html-italic">Trypanosoma cruzi</span>, <span class="html-italic">Ehrlichia canis</span>, and <span class="html-italic">Babesia canis</span> using the peptides No. 45, No. 48, mix, and soluble <span class="html-italic">Leishmania infantum</span> antigen (SLiA). The dotted lines within the graphs represent the cut-offs calculated by the ROC curve between the negative and positive results of SLiA (OD = 0.273), Pep45 (OD = 0.41), Pep48 (OD = 0.431), and mix (OD = 0.331). (<b>b</b>) The ROC curves were constructed with the results of serum samples from control noninfected dogs, <span class="html-italic">L. infantum</span>-infected dogs, and dogs infected with other pathogens tested by each assay. (<b>c</b>) Results of sensitivity, specificity, negative predictive values (NPV), positive predictive values (PPV), and accuracy from noninfected control dogs, <span class="html-italic">L.infantum</span>-infected dogs, and dogs with other canine pathogens tested by the peptides No. 45, No. 48, mix, and SLiA.</p> Full article ">Figure 5
<p>Performance of the peptides No. 45, No. 48, and mix in the discrimination of <span class="html-italic">L. infantum</span> infected dogs from vaccinated dogs. (<b>a</b>) Distribution of the individual optical density results from serum samples from dogs infected with <span class="html-italic">L. infantum</span> and vaccinated dogs using the peptides No. 45, No. 48, mix, and soluble <span class="html-italic">Leishmania infantum</span> antigen (SLiA). The dotted lines within the graphs represent the cut-offs calculated by the ROC curve between the negative and positive results of SLiA (OD = 0.273), Pep45 (OD = 0.41), Pep48 (OD = 0.431), and mix (OD = 0.331). (<b>b</b>) Results of sensitivity, specificity, negative predictive values (NPV), positive predictive values (PPV), and accuracy from noninfected control dogs, <span class="html-italic">L. infantum</span>-infected dogs, and vaccinated dogs tested by the peptides No. 45, No. 48, mix, and SLiA.</p> Full article ">

Review

Jump to: Research, Other

19 pages, 3257 KiB  
Review
Cryptosporidium Species Infections Detected from Fecal Samples of Animal and Human Hosts in South Africa: Systematic Review and Meta-Analysis
by Mpho Tawana, ThankGod E. Onyiche, Tsepo Ramatla, Sebolelo Jane Nkhebenyane, Dennis J. Grab and Oriel Thekisoe
Microorganisms 2024, 12(12), 2426; https://doi.org/10.3390/microorganisms12122426 - 25 Nov 2024
Viewed by 587
Abstract
This study presents a systematic review and meta-analysis approach of Cryptosporidium species prevalence studies in animal and human hosts published between 1980 and 2020 in South Africa. Extensive searches were conducted on three electronic databases including PubMed, ScienceDirect and Google Scholar. The findings [...] Read more.
This study presents a systematic review and meta-analysis approach of Cryptosporidium species prevalence studies in animal and human hosts published between 1980 and 2020 in South Africa. Extensive searches were conducted on three electronic databases including PubMed, ScienceDirect and Google Scholar. The findings indicated an overall pooled prevalence estimate (PPE) of Cryptosporidium spp. infections in animals and humans at 21.5% and 18.1%, respectively. The PCR–RFLP appeared to be the most sensitive diagnostic method with a PPE of 77.8% for the detection of Cryptosporidium spp. infections followed by ELISA (66.7%); LAMP (45.4%); PCR (25.3%); qPCR (20.7%); microscopy (10.1%); IFAT (8.4%); and RDT (7.9%). In animal hosts, C. parvum had the highest PPE of 3.7%, followed by C. andersoni (1.5%), C. ubiquitum (1.4%) and C. bovis (1.0%), while in humans, C. parvum also had the highest PPE of 18.3% followed by C. meleagridis at 0.4%. The data generated in this study indicated that Cryptosporidium spp. infections were highly prevalent in both animals and humans in South Africa, especially in the KwaZulu-Natal and North West provinces. However, we further observed that there was a lack of prevalence studies for both animals and humans in some of the provinces. This study highlights the necessity for a “One Health” strategic approach promoting public hygiene, animal husbandry and regular screening for Cryptosporidium spp. infections in both animals and humans. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

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<p>Flow chart of included studies according to PRISMA guidelines.</p> Full article ">Figure 2
<p>Forest plot of the prevalence of <span class="html-italic">Cryptosporidium</span> spp. from animal studies conducted during 2001–2010. The squares demonstrate the individual point estimates. The diamond at the base indicates the pooled estimate from the overall studies [<a href="#B30-microorganisms-12-02426" class="html-bibr">30</a>,<a href="#B36-microorganisms-12-02426" class="html-bibr">36</a>,<a href="#B37-microorganisms-12-02426" class="html-bibr">37</a>,<a href="#B38-microorganisms-12-02426" class="html-bibr">38</a>].</p> Full article ">Figure 3
<p>Funnel plot with 95% confidence limits of <span class="html-italic">Cryptosporidium</span> spp. pooled prevalence estimates of cattle subgroup studies that tested positive for <span class="html-italic">Cryptosporidium</span> species. The diamond at the base indicates the pooled estimate from the studies overall.</p> Full article ">Figure 4
<p>Forest plot showing the pooled estimates of <span class="html-italic">Cryptosporidium</span> spp. from studies conducted on (<b>A</b>) males and (<b>B</b>) females. The squares demonstrate the individual point estimates. The diamonds at the base indicate the pooled estimates from the overall studies [<a href="#B40-microorganisms-12-02426" class="html-bibr">40</a>,<a href="#B47-microorganisms-12-02426" class="html-bibr">47</a>,<a href="#B52-microorganisms-12-02426" class="html-bibr">52</a>,<a href="#B53-microorganisms-12-02426" class="html-bibr">53</a>,<a href="#B57-microorganisms-12-02426" class="html-bibr">57</a>].</p> Full article ">Figure 4 Cont.
<p>Forest plot showing the pooled estimates of <span class="html-italic">Cryptosporidium</span> spp. from studies conducted on (<b>A</b>) males and (<b>B</b>) females. The squares demonstrate the individual point estimates. The diamonds at the base indicate the pooled estimates from the overall studies [<a href="#B40-microorganisms-12-02426" class="html-bibr">40</a>,<a href="#B47-microorganisms-12-02426" class="html-bibr">47</a>,<a href="#B52-microorganisms-12-02426" class="html-bibr">52</a>,<a href="#B53-microorganisms-12-02426" class="html-bibr">53</a>,<a href="#B57-microorganisms-12-02426" class="html-bibr">57</a>].</p> Full article ">Figure 5
<p>Funnel plot with 95% confidence limits of <span class="html-italic">Cryptosporidium</span> spp. pooled prevalence estimates of 1981–1990 interval subgroup studies that tested positive for <span class="html-italic">Cryptosporidium</span> spp. in humans. The diamond at the base indicates the pooled estimate from the studies overall.</p> Full article ">
13 pages, 1160 KiB  
Review
Plasmodium cynomolgi: What Should We Know?
by Fauzi Muh, Ariesta Erwina, Fadhila Fitriana, Jadidan Hada Syahada, Angga Dwi Cahya, Seongjun Choe, Hojong Jun, Triwibowo Ambar Garjito, Josephine Elizabeth Siregar and Jin-Hee Han
Microorganisms 2024, 12(8), 1607; https://doi.org/10.3390/microorganisms12081607 - 7 Aug 2024
Viewed by 2497
Abstract
Even though malaria has markedly reduced its global burden, it remains a serious threat to people living in or visiting malaria-endemic areas. The six Plasmodium species (Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale curtisi, Plasmodium ovale wallikeri [...] Read more.
Even though malaria has markedly reduced its global burden, it remains a serious threat to people living in or visiting malaria-endemic areas. The six Plasmodium species (Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale curtisi, Plasmodium ovale wallikeri and Plasmodium knowlesi) are known to associate with human malaria by the Anopheles mosquito. Highlighting the dynamic nature of malaria transmission, the simian malaria parasite Plasmodium cynomolgi has recently been transferred to humans. The first human natural infection case of P. cynomolgi was confirmed in 2011, and the number of cases is gradually increasing. It is assumed that it was probably misdiagnosed as P. vivax in the past due to its similar morphological features and genome sequences. Comprehensive perspectives that encompass the relationships within the natural environment, including parasites, vectors, humans, and reservoir hosts (macaques), are required to understand this zoonotic malaria and prevent potential unknown risks to human health. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1
<p>The morphology of <span class="html-italic">P. cynomolgi</span> K4 line is shown in each development stage cultured in 100% of rhesus macaque RBCs (Fauzi Muh et al., original unpublished data). Scale bar indicates 5 µm.</p> Full article ">Figure 2
<p>The geographical distribution of natural hosts and vectors of <span class="html-italic">P. cynomolgi</span>.</p> Full article ">

Other

Jump to: Research, Review

9 pages, 1741 KiB  
Brief Report
Oxidative Stress in the Murine Model of Extraparenchymal Neurocysticercosis
by Diego Generoso, Tatiane de Camargo Martins, Camila Renata Corrêa Camacho, Manuella Pacífico de Freitas Segredo, Sabrina Setembre Batah, Alexandre Todorovic Fabro, Edda Sciutto, Agnès Fleury, Pedro Tadao Hamamoto Filho and Marco Antônio Zanini
Microorganisms 2024, 12(9), 1860; https://doi.org/10.3390/microorganisms12091860 - 8 Sep 2024
Viewed by 963
Abstract
Oxidative stress is associated with several infectious diseases, as well as the severity of inflammatory reactions. The control of inflammation during parasite destruction is a target of neurocysticercosis treatment, as inflammation is strongly related to symptom severity. In this study, we investigated the [...] Read more.
Oxidative stress is associated with several infectious diseases, as well as the severity of inflammatory reactions. The control of inflammation during parasite destruction is a target of neurocysticercosis treatment, as inflammation is strongly related to symptom severity. In this study, we investigated the presence of malondialdehyde and protein carbonyl, two by-products of reactive oxygen species (ROS), in an experimental model of extraparenchymal neurocysticercosis. Twenty male and twenty female rats were inoculated with 50 cysts of Taenia crassiceps in the subarachnoid space of the cisterna magna. Ten animals (five males and five females) were used as controls. Three months after inoculation, their brains were harvested for oxidative stress and histological assessments. Infected animals had higher scores for inflammatory cell infiltrates, malondialdehyde, and protein carbonyl. These results encourage future efforts to monitor oxidative stress status in neurocysticercosis, particularly in the context of controlling inflammation. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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Figure 1

Figure 1
<p>Schematic representation of the experimental procedures. (<b>A</b>): The cyst injection was performed at the cisterna magna, i.e., the subarachnoid space between the posterior-inferior portion of the cerebellum and the dorsal portion of the medulla. (<b>B</b>): Three months after the inoculation, the cysts could be observed throughout the subarachnoid space, located predominantly in the basal convexity of the brain and brainstem. (<b>C</b>): The right hemisphere was used for histologic analysis, and the left hemisphere for oxidative stress essays. (<b>D</b>): The dorsal and coronal views at the level of tissue histologic assessment.</p> Full article ">Figure 2
<p>Microscopic view of the basal arachnoid–pia–mater–brain interfaces for male (<b>A</b>,<b>B</b>) and female (<b>C</b>,<b>D</b>) rats. In control animals, a thin arachnoid layer (arrows) juxtaposed to the pia–mater with preserved histoarchitecture (<b>B</b>,<b>D</b>) can be observed. In infected animals (<b>A</b>,<b>C</b>), the arachnoid is thickened and lymphocytes (*) can be observed in intense (<b>A</b>) or discrete (<b>C</b>) infiltrations.</p> Full article ">Figure 3
<p>Comparisons of the malondialdehyde concentration between the experimental groups. When considering all infected animals, the mean concentration was higher than the control animals (<b>A</b>). However, the differences were no longer significant (ns) when analyzing male (<b>B</b>) and female (<b>C</b>) animals with their controls. Further, considering the infected animals, males had a higher concentration of malondialdehyde than females (<b>D</b>).</p> Full article ">Figure 4
<p>Comparisons of protein carbonyl between the experimental groups. The infected animals presented higher concentrations when comparing all animals (<b>A</b>), the males (<b>B</b>), and females (<b>C</b>) with their respective controls. There was no difference between infected males and females (<b>D</b>). ns: non-significant.</p> Full article ">
10 pages, 1028 KiB  
Brief Report
A Pilot Study for the Characterization of Bacillus spp. and Analysis of Possible B. thuringiensis/Strongyloides stercoralis Correlation
by Elena Pomari, Pierantonio Orza, Milena Bernardi, Fabio Fracchetti, Ilenia Campedelli, Patrick De Marta, Alessandra Recchia, Paola Paradies and Dora Buonfrate
Microorganisms 2024, 12(8), 1603; https://doi.org/10.3390/microorganisms12081603 - 6 Aug 2024
Cited by 1 | Viewed by 1006
Abstract
Differentiating between Bacillus species is relevant in human medicine. Bacillus thuringiensis toxins might be effective against Strongyloides stercoralis, a nematode causing relevant human morbidity. Our first objective was to evaluate genomic and MALDI-TOF identification methods for B. thuringiensis. Our secondary objective [...] Read more.
Differentiating between Bacillus species is relevant in human medicine. Bacillus thuringiensis toxins might be effective against Strongyloides stercoralis, a nematode causing relevant human morbidity. Our first objective was to evaluate genomic and MALDI-TOF identification methods for B. thuringiensis. Our secondary objective was to evaluate a possible negative selection pressure of B. thuringiensis against S. stercoralis. PCR and Sanger were compared to MALDI-TOF on a collection of 44 B. cereus group strains. B. thuringiensis toxin genes were searched on 17 stool samples from S. stercoralis-infected and uninfected dogs. Metagenomic 16S rRNA was used for microbiome composition. The inter-rate agreement between PCR, Sanger, and MALDI-TOF was 0.631 k (p-value = 6.4 × 10−10). B. thuringiensis toxins were not found in dogs’ stool. Bacteroidota and Bacillota were the major phyla in the dogs’ microbiome (both represented >20% of the total bacterial community). Prevotella was underrepresented in all Strongyloides-positive dogs. However, the general composition of bacterial communities was not significantly linked with S. stercoralis infection. The genomic methods allowed accurate differentiation between B. thuringiensis and B. cereus. There was no association between B. thuringiensis and S. stercoralis infection, but further studies are needed to confirm this finding. We provide the first descriptive results about bacterial fecal composition in dogs with S. stercoralis infection. Full article
(This article belongs to the Special Issue Parasites and Infection: Strategies to Control, Diagnose, and Treat Parasitic Diseases)
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<p>Neighbor-Joining phylogenetic tree based on the <span class="html-italic">gyrB</span> gene sequence comparison for the <span class="html-italic">Bacillus</span> strains under analysis with the corresponding sequence retrieved for the type strain <span class="html-italic">B. mycoides</span> DSM 2048<sup>T</sup>, <span class="html-italic">B. cytotoxicus</span> NVH 391-98<sup>T</sup>, and <span class="html-italic">B. subtilis</span> subsp. <span class="html-italic">subtilis</span> BCRC 10255<sup>T</sup>. The tree was reconstructed through MEGA11 with the Tamura–Nei model and complete deletion treatment for gaps. The accession numbers of the sequence deposited and/or available in NCBI database were reported in brackets.</p> Full article ">Figure 2
<p>Phylum- and order-level gut microbiota composition in the fecal samples of 10 dogs. (<b>A</b>) Data obtained with Kraken2/Bracken. (<b>B</b>) Data obtained with DADA2.</p> Full article ">
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