Diversity Of Protozoa Research Articles

Auranofin is lethal against feline Tritrichomonas foetus in vitro but ineffective in cats with naturally occurring infection

Protozoal diarrhea caused by Tritrichomonas foetus (blagburni) is a prevalent, lifelong, and globally distributed burden in domestic cats. Treatment is limited to the use of 5-nitroimidazoles and treatment failure is common. The repurposed gold salt compound auranofin has killing activity against diverse protozoa in vitro but evidence of efficacy in naturally occurring protozoal infections is lacking. This exploratory study investigated the efficacy and safety of auranofin for treatment of cats with naturally occurring, 5-nitroimidazole-resistant, T. foetus infection. The minimum lethal concentration (MLC) of auranofin against 5 isolates of feline T. foetus was determined under aerobic conditions in vitro. Healthy cats and cats with T. foetus infection were treated with immediate release auranofin (range, 0.5–3 mg/cat for 7 days) or guar gum-coated auranofin capsules (0.5 or 3 mg/cat for 7 days). Adverse effects were monitored by clinical signs and clinicopathologic testing. Efficacy was determined by fecal consistency score, bowel movement frequency, and single-tube nested PCR of feces for T. foetus rDNA. Fecal samples were assayed for concentrations of auranofin, known and predicted metabolites of auranofin, gold containing molecules, and total gold content using HPLC, LC-MS, ion mobility-MS, and ICP-MS, respectively. Auranofin was effective at killing isolates of feline T. foetus at MLC ≥ 1 μg/ml. Treatment of cats with T. foetus infection with either immediate release auranofin or a colon-targeted guar gum-coated tablet of auranofin did not eradicate infection. Treatment failure occurred despite fecal concentrations of gold that met or exceeded the equivalent MLC of auranofin. Neither auranofin, known or predicted metabolites of auranofin, nor any gold-containing molecules >100 Da could be detected in fecal samples of treated cats. Adverse effects associated with auranofin treatment were common but minor. These studies identify that in vitro susceptibility test results of auranofin may not translate to treatment effectiveness in vivo even when achieving gold concentrations equivalent to the MLC of auranofin in the target environment. These studies further establish the absence of any predicted or unpredicted gold containing metabolites in feces after oral administration of auranofin.

Literatur review : protozoa parasit di air sungai

River water is potentially unsafe and polluted, which can cause serious infections. Waterborne infections are a major cause of global mortality and morbidity. In cases of diarrhea, this research aims to identify the types of parasitic protozoa and their distribution in waterways in Japan and Indonesia. This research reviews literature by collecting data from various sources, such as books, national and international scientific journals. The articles analyzed are original articles relating to protozoa in river water from both countries. In Japan, protozoa such as Henneguya were found in the Omoi River and Cryptosporidium parvum in several rivers in Hokkaido. In Indonesia, species such as Codonellopsis sp., Favella sp., Tintinnopsis sp., and Codonella sp. are found. in Sorong Regency, as well as various other types in West Bandung, Central Sulawesi and South Kalimantan. The diversity and distribution of protozoa is influenced by water quality, physical conditions, water chemistry, and human activities. This research can provide information regarding protozoa which can act as bioindicators of water quality. The presence and diversity of protozoa can provide an indication of the health condition of aquatic ecosystems. As a result, conservation efforts and management of aquatic ecosystems can be improved.

Estimation of protozoal diversity among stray cats in Malaysia

Abstract. Abdulwahab MH, Al-Talib H. 2024. Estimation of protozoal diversity among stray cats in Malaysia. Biodiversitas 25: 2115-2120. This cross sectional study was carried out to estimate the current prevalence and protozoa species in stray cats feces in Malaysia. A total of 320 fresh fecal samples were collected from S.I Home Shelter-Selangor, EJ CAT Home- Melaka, and KB-Cat Shelter - Kelantan in Malaysia. The collected fecal samples were concentrated for the presence of intestinal protozoa using a ready-made commercial Parasep tube floatation method. The fecal samples were stained with iodine and examined for protozoa by microscopic examination. The overall prevalence of intestinal protozoa in cat feces was 73.1% (n = 234). Isospora felis was most frequently detected (n = 145; 62%), followed by Toxoplasma gondii (n = 75; 32%), Giardia intestinalis (n = 9; 3.8%), Entamoeba histolytica (n = 9; 3.8%), and Tritrichomonas foetus (n = 6; 12.5%). Monoparasitism was detected in 65.8%, which was more frequent than polyparasitism (34.2%). Therefore, the local population is at risk of exposure to a wide range of zoonotic protozoa through environmental contamination with cat feces, and the current findings should be used to mitigate public health risks. Prevention and control measures need to be taken to reduce prevalence rates, especially in socioeconomically disadvantaged communities where animals live in close proximity to humans, where poor sanitation and overcrowding exist along with a lack of veterinary care and awareness of zoonotic diseases.

Responses of Protozoan Communities to Multiple Environmental Stresses (Warming, Eutrophication, and Pesticide Pollution).

To explore the impacts of multiple environmental stressors on animal communities in aquatic ecosystems, we selected protozoa-a highly sensitive group of organisms-to assess the effect of environmental change. To conduct this simulation we conducted a three-factor, outdoor, mesocosm experiment from March to November 2021. Changes in the community structure and functional group composition of protozoan communities under the separate and combined effects of these three environmental stressors were investigated by warming and the addition of nitrogen, phosphorus, and pesticides. The results were as follows: (1) Both eutrophication and pesticides had a considerable promotional effect on the abundance and biomass of protozoa; the effect of warming was not considerable. When warming was combined with eutrophication and pesticides, there was a synergistic effect and antagonistic effect, respectively. (2) Eutrophication promoted α diversity of protozoa and affected their species richness and dominant species composition; the combination of warming and pesticides remarkably reduced the α diversity of protozoa. (3) Warming, eutrophication, and pesticides were important factors affecting the functional groups of protozoa. Interaction among different environmental factors could complicate changes in the aquatic ecological environment and its protozoan communities. Indeed, in the context of climate change, it might be more difficult to predict future trends in the protozoan community. Therefore, our results provide a scientific basis for the protection and restoration of shallow lake ecosystems; they also offer valuable insights in predicting changes in shallow lakes.

A comparative study of supervised and unsupervised machine learning algorithms applied to human microbiome.

The human microbiome, consisting of diverse bacte-rial, fungal, protozoan and viral species, exerts a profound influence on various physiological processes and disease susceptibility. However, the complexity of microbiome data has presented significant challenges in the analysis and interpretation of these intricate datasets, leading to the development of specialized software that employs machine learning algorithms for these aims. In this paper, we analyze raw data taken from 16S rRNA gene sequencing from three studies, including stool samples from healthy control, patients with adenoma, and patients with colorectal cancer. Firstly, we use network-based methods to reduce dimensions of the dataset and consider only the most important features. In addition, we employ supervised machine learning algorithms to make prediction. Results show that graph-based techniques reduces dimen-sion from 255 up to 78 features with modularity score 0.73 based on different centrality measures. On the other hand, projection methods (non-negative matrix factorization and principal component analysis) reduce dimensions to 7 features. Furthermore, we apply supervised machine learning algorithms on the most important features obtained from centrality measures and on the ones obtained from projection methods, founding that the evaluation metrics have approximately the same scores when applying the algorithms on the entire dataset, on 78 feature and on 7 features. This study demonstrates the efficacy of graph-based and projection methods in the interpretation for 16S rRNA gene sequencing data. Supervised machine learning on refined features from both approaches yields comparable predictive performance, emphasizing specific microbial features-bacteroides, prevotella, fusobacterium, lysinibacillus, blautia, sphingomonas, and faecalibacterium-as key in predicting patient conditions from raw data.

The modern methods development for microbiological water quality assessment in lakes

The functional diversity of free-living aquatic protozoans is discussed with respect to the feeding ecology, life strategies and water quality. Heterotrophic nanoflagellates (HNF) are known as most important grazers of bacteria in many aquatic ecosystems. Recent studies of flagellate feeding processes indicated that there are significant species-specific variability regarding the food uptake and food selection of bacterivorous flagellates. The Choanoflagellida have an essential role in microbial food webs contributing to a process of self-purification of water bodies as well as are used in monitoring systems. We show that diversity and relative abundance of ciliates and other protozoans can be used as indicators of toxic pollution and acidification. Multivariate statistics were used to design “the scale of toxicity” across a gradient of toxicant stress and organic compounds. A new index of periphyton flagellates (IPF) as indicator of the trophic status of a water-body and a simplified index for a sublethal toxicity assessment were developed.

Molecular detection reveals diverse tick-borne bacterial and protozoan pathogens in two tick species from Yingshan County of Hubei Province, China in 2021–2022

Molecular detection reveals diverse tick-borne bacterial and protozoan pathogens in two tick species from Yingshan County of Hubei Province, China in 2021–2022

Immunoinformatics-based multi-epitope containing fused polypeptide vaccine design against visceral leishmaniasis with high immunogenicity and TLR binding

Visceral leishmaniasis (VL) is the most lethal among all leishmaniasis diseases and remains categorized as a neglected tropical disease (NTD). This study aimed to develop a peptide-based multi-epitope vaccine construct against VL using immunoinformatics methodologies. To achieve this, four distinct proteins were screened to identify peptides consisting of 9–15 amino acids with high binding affinity to toll-like receptors (TLRs), strong antigenicity, low allergenicity, and minimal toxicity. The resulting multi-epitope vaccine construct was fused in a tandem arrangement with appropriate linker peptides and exhibited superior properties related to cytotoxic T lymphocytes (CTLs), helper T lymphocytes (HTLs), and B-cell epitopes. Subsequently, a three-dimensional (3D) model of the vaccine construct was generated, refined, and validated for structural stability and immune response capabilities. Molecular docking and simulations confirmed the vaccine construct's stability and binding affinities with TLRs, with TLR4 displaying the highest binding affinity, followed by TLR2 and TLR3. Additionally, simulations predicted robust cellular and humoral antibody-mediated immune responses elicited by the designed vaccine construct. Notably, this vaccine construct includes proteins from various pathways of Leishmania donovani (LD), which have not been previously utilized in VL vaccine design. Thus, this study opens new avenues for the development of vaccines against diverse protozoan diseases.

Diversity and distribution of ciliated protozoans on the mangrove leaf litters of Rhizophora apiculata

Diversity and distribution of ciliated protozoans on the mangrove leaf litters of Rhizophora apiculata

Investigating Bacterial and Free-Living Protozoa Diversity in Biofilms of Hot Water Pipes of Apartment Buildings in the City of Riga (Latvia)

Background: Biofilms, when formed on the surfaces of water pipes, can be responsible for a wide range of water quality and operational problems. We sought to assess the bacterial and free-living protozoa (FLP) diversity, in relation to the presence of Legionnaire's disease-causing bacteria Legionella pneumophila (L. pneumophila) in 45 biofilms of hot water distribution system pipes of apartment buildings in Riga, the capital city of Latvia.Results: 16S rRNA amplicon sequencing (metataxonomics) revealed that each biofilm contained 224 rather evenly distributed bacterial genera and that most common and most abundant were two genera, completely opposites in terms of their oxygen requirements: the obligately anaerobic Thermodesulfovibrio and the strictly aerobic Phenylobacterium. Water temperature and north-south axis (i.e., different primary water sources) displayed the most significant effect on the inter-sample variations, allowing us to re-construct three sub-networks (modules) of co-occurring genera, one involving (potentially FLP-derived) Legionella spp. Pangenome-based functional profile predictions suggested that all three may be dominated by pathways related to the development and maintenance of biofilms, including quorum sensing and nutrient transport, as well as the utilization of various energy sources, such as carbon and nitrogen. In our 18S rRNA amplicon sequencing data, potential hosts of L. pneumophila were detected in 11 out of 12 biofilm samples analyzed, however, in many cases, their relative abundance was very low (<1%). By validating our findings using culture-based methods, we detected L. pneumophila (serogroups 2, 3, 6 and 9) in nine (20%) biofilms, whereas FLP (mostly Acanthamoeba, Vahlkampfidae and Vermamoeba spp.) were present in six (~13%) biofilms. In two biofilms, L. pneumophila and its potential hosts were detected simultaneously, using culture-based methods.Conclusions: Overall, our study sheds light on the community diversity of hot water biofilms and predicts how several environmental factors, such as water temperature and source might shape it.

Diversity of gastrointestinal parasites in sympatric mammals in Moukalaba-Doudou National Park, Gabon.

Background and Aim:Gastrointestinal parasites identified in the wild can negatively affect host fitness, lower performance, and growth. On the other side, sympatric mammals that share habitat and resources may also cross-transmit parasites, which are often zoonotic and can contribute to morbidity and mortality. This study aimed to characterize the diversity of gastrointestinal parasites circulating in mammalian hosts in Moukalaba-Doudou National Park.Materials and Methods:We screened a total of 25 fecal samples collected from nine wild mammalian species, namely, western gorilla (Gorilla gorilla gorilla), chimpanzee (Pan troglodytes), putty-nosed monkey (Cercopithecus nictitans), African forest elephant (Loxodonta cyclotis), African buffalo (Syncerus caffer), blue duiker (Philantomba monticola), bay duiker (Cephalophus dorsalis), and red river hog (Potamochoerus porcus) as well as people working as trackers (Homo sapiens) using direct microscopic observations following a sedimentation technique to concentrate the fecal material.Results:Of the total 25 fecal samples screened, 15 (60%) were positive for parasitic gastrointestinal infection. Based on the morphology of parasite eggs and cysts, we identified a rich diversity of nematodes, protozoans, trematodes, and cestodes, including unidentified strongyles (73%), Oesophagostomum spp. (53%), Ancylostoma spp. (27%), Trichuris spp. (13%), Ascaris spp. (13%), Mammomonogamus spp. (13%), Strongyloides spp. (47%), Balantidium coli (20%), Entamoeba coli (20%), Endolimax nana (6%), Fasciola hepatica (6%), Paramphistomum spp. (13%), and Taenia spp. (6%).Conclusion:All parasites were found at least once in one of the hosts, and most were potentially zoonotic and responsible for several diseases of public health concern. Because of the small sample size, our findings should not be considered conclusive. Nevertheless, they highlight the diversity of gastrointestinal parasites in this area.

Kinins and Their Receptors in Infectious Diseases.

Kinins and their receptors have been implicated in a series of pathological alterations, representing attractive pharmacological targets for several diseases. The present review article aims to discuss the role of the kinin system in infectious diseases. Literature data provides compelling evidence about the participation of kinins in infections caused by diverse agents, including viral, bacterial, fungal, protozoan, and helminth-related ills. It is tempting to propose that modulation of kinin actions and production might be an adjuvant strategy for management of infection-related complications.

From conifers to cognition: Microbes, brain and behavior.

A diversity of bacteria, protozoans and viruses ("endozoites") were recently uncovered within healthy tissues including the human brain. By contrast, it was already recognized a century ago that healthy plants tissues contain abundant endogenous microbes ("endophytes"). Taking endophytes as an informative precedent, we overview the nature, prevalence, and role of endozoites in mammalian tissues, centrally focusing on the brain, concluding that endozoites are ubiquitous in diverse tissues. These passengers often remain subclinical, but they are not silent. We address their routes of entry, mechanisms of persistence, tissue specificity, and potential to cause long-term behavioral changes and/or immunosuppression in mammals, where rabies virus is the exemplar. We extend the discussion to Herpesviridae, Coronaviridae, and Toxoplasma, as well as to diverse bacteria and yeasts, and debate the advantages and disadvantages that endozoite infection might afford to the host and to the ecosystem. We provide a clinical perspective in which endozoites are implicated in neurodegenerative disease, anxiety/depression, and schizophrenia. We conclude that endozoites are instrumental in the delicate balance between health and disease, including age-related brain disease, and that endozoites have played an important role in the evolution of brain function and human behavior.

Frequency and Molecular Detection of Giardia intestinalis in Children Attending Pediatrics of Punjab, Pakistan

Background: Giardia intestinalis is the most common protozoan infecting the small intestine of human beings and a major cause of enteric infection, especially in children throughout the world. It is a highly diverse protozoan, which comprises a complex of eight genetic assemblages that are further differentiated into sub-assemblages. Objectives: A cross-sectional study was conducted to detect the frequency, molecular detection and assemblage identification of G. intestinalis in children of Punjab, Pakistan. Methods: A total of 800 stool samples were collected from children ranging 0 - 10 years of age with gastrointestinal disturbances and subjected to direct microscopy, enzyme-linked immunosorbent assay and polymerase chain reaction targeting small-subunit ribosomal RNA (SSU rRNA) and triosephosphate isomerase (tpi) genes. A predesigned questionnaire was filled prior to sampling from the guardian of each child to collect information. Results: The results indicated that the prevalence of 9.5% (76/800) was achieved by microscopy, ELISA and PCR targeting SSU-rRNA gene. The genetic DNA from 69 out of 76 (90.80%) was successfully amplified by tpi gene. Among these tpi gene-positive samples, 38 were successfully typed in assemblage B (55.07%) followed by 20 (28.98%) in assemblage A and 11 (15.94%) in mixed type assemblages (A & B). Residency and socioeconomic status were statistically associated with giardiasis. Among the clinical presentations, abdominal pain is prominent in assemblage B (57.89%) and vomiting in assemblage A (40%) type infections. Conclusions: Advanced molecular tools for giardiasis are well-adapted to get true prevalence, better discrimination of assemblages and their correlation with clinical signs.

Cascade dam impoundments restrain the trophic transfer efficiencies in benthic microbial food web

Determination of the effects of cascade dams on benthic microbial ecosystem is essential for dam regulation and ecological function protection. However, no comprehensive investigation has yet shown the ecosystem-level responses of microbiota to dam impoundments. This study conducted DNA metabarcoding and microbial food web analysis for multiple species and their interrelationships along a cascade dam-affected river. The composition, distribution and diversity of bacteria, protozoans and metazoans were obviously different between river and reservoirs, mainly controlled by hydrological (P < 0.01) and nutrient parameters (P < 0.05). Those three groups make up a co-occurrence network, with most edges direct from higher to lower trophic levels or vice versa and more than 50% keystones participate in the food web, indicating the significant role of predator-prey relationships. Based on the microbial food web analysis, the predator biomass, especially at higher trophic levels, decreased by about 10% from the riverine to the lacustrine system. The structural equation model illustrates that both bottom-up forces (environmental factors particularly velocity and nutrient concentrations) and top-down forces (higher trophic levels) critically control microbial food web patterns (P < 0.05). As a result of dam impoundments, the lower velocity in the reservoirs has direct negative effects on trophic transfer efficiencies that may be further magnified by nutrient accumulation, probably leading to an increase of eutrophication and posing a risk to water quality. The results suggest the potential ecological risk in the reservoirs and highlight the need to consider from the perspective of ecosystem during the operation of cascade dams.

First molecular detection of Anaplasma phagocytophilum in the hard tick Rhipicephalus haemaphysaloides in Taiwan

Anaplasma phagocytophilum is transmitted mainly by hard ticks and can cause potentially fatal granulocytic anaplasmosis in humans, but its occurrence in ticks in Taiwan has never been investigated although this pathogen has been detected in Taiwanese rodents before. Ticks collected from small mammals in Hualien, eastern Taiwan, were assayed for Anaplasma infections; infections of Rickettsia and Apicomplexa protozoans were also studied. Of the 270 individually assayed Rhipicephalus haemaphysaloides ticks, A. phagocytophilum was identified in a nymphal tick. Parasites most similar to Anaplasma bovis, Rickettsia rickettsii, Rickettsia sp. TwKM01, and at least seven apicomplexan species (including genera Cryptosporidium, Hepatozoon, and Theileria) were also identified. This study shows that A. phagocytophilum does occur in the hard tick in Taiwan, although whether R. haemaphysaloides can vector this pathogen remains to be determined. This work also reveals a high diversity of tick-borne bacteria and protozoans circulating in a small region and calls for further research on their potential risks for human health.

Recent Advances in Halophilic Protozoa Research.

Most research on microorganisms adapted to hypersaline habitats has focused on Archaea and Bacteria, with microbial eukaryotes receiving much less attention. Over the past 15yr, our knowledge of phagotrophic microbial eukaryotes, i.e. protozoa, from hypersaline habitats has greatly improved through combinations of microscopy, molecular phylogenetics, environmental sequencing, transcriptomics and growth experiments. High salinity waters from salterns, other landlocked water masses and deep hypersaline anoxic basins contain unique and diverse halophilic protozoan assemblages. These have the potential to exert substantial grazing pressure on prokaryotes and other eukaryotes. They represent many separate evolutionary lineages; species of Heterolobosea, Bicosoecida, and Ciliophora have been most intensively characterized, with several proven to be extreme (or borderline extreme) halophiles. Transcriptomic examinations of the bicosoecid Halocafeteria (and the heteroloboseid Pharyngomonas) indicate that high-salt adaptation is associated with a subtle shift in protein amino acid composition, and involves the differential expression of genes participating in ion homeostasis, signal transduction, stress management, and lipid remodeling. Instances of gene duplication and lateral transfer possibly conferring adaptation have been documented. Indirect evidence suggests that these protozoa use "salt-out" osmoadaptive strategies.

Disentangling food webs interactions in the littoral of temperate shallow lakes

Elucidation of food web interactions provides better understanding of ecosystem functioning, indicates anthropogenic impacts which often cause alterations in environmental conditions and detoriorations in feeding network in terrestrial and aquatic ecosystems. The importance of microfauna and macroinvertebrates in littoral zones of shallow waters has poorly been studied regarding their trophic interactions. This study compares invertebrate assemblage and food web interactions among epiphyton, zooplankton and benthic macroinvertebrates across structural heterogeneity in the littoral zone of three temperate shallow water bodies. Submerged and structurally-complex stands of Ceratophyllum demersum inhabited higher abundance of cladocerans and supported higher diversity and biomass of epiphytic protozoans and invertebrates in comparison to floating-leaved Nuphar lutea stands. Analysis of the ecosystem functioning illustrated the increased biomass of macroinvertebrate predators and a preference of predation over planktonic crustaceans in more complex macrophyte stands. Food webs displayed higher complexity and size with habitat heterogeneity, while epiphyton and zooplankton shared the important fraction in food resources among the invertebrate trophic network. Results of food web modelling indicated zooplankton and epiphyton as more vulnerable to invertebrate predation in the complex submerged macrophytes than in floating-leaved macrophyte stands. Integrated community, ecosystem and food web approaches in explanation of complex trophic interactions in the littoral zones justified an increase in diversity and food-web functional complexity with structural heterogeneity of microhabitats.

An Investigation into Rumen Fungal and Protozoal Diversity in Three Rumen Fractions, during High-Fiber or Grain-Induced Sub-Acute Ruminal Acidosis Conditions, with or without Active Dry Yeast Supplementation.

Sub-acute ruminal acidosis (SARA) is a gastrointestinal functional disorder in livestock characterized by low rumen pH, which reduces rumen function, microbial diversity, host performance, and host immune function. Dietary management is used to prevent SARA, often with yeast supplementation as a pH buffer. Almost nothing is known about the effect of SARA or yeast supplementation on ruminal protozoal and fungal diversity, despite their roles in fiber degradation. Dairy cows were switched from a high-fiber to high-grain diet abruptly to induce SARA, with and without active dry yeast (ADY, Saccharomyces cerevisiae) supplementation, and sampled from the rumen fluid, solids, and epimural fractions to determine microbial diversity using the protozoal 18S rRNA and the fungal ITS1 genes via Illumina MiSeq sequencing. Diet-induced SARA dramatically increased the number and abundance of rare fungal taxa, even in fluid fractions where total reads were very low, and reduced protozoal diversity. SARA selected for more lactic-acid utilizing taxa, and fewer fiber-degrading taxa. ADY treatment increased fungal richness (OTUs) but not diversity (Inverse Simpson, Shannon), but increased protozoal richness and diversity in some fractions. ADY treatment itself significantly (P < 0.05) affected the abundance of numerous fungal genera as seen in the high-fiber diet: Lewia, Neocallimastix, and Phoma were increased, while Alternaria, Candida Orpinomyces, and Piromyces spp. were decreased. Likewise, for protozoa, ADY itself increased Isotricha intestinalis but decreased Entodinium furca spp. Multivariate analyses showed diet type was most significant in driving diversity, followed by yeast treatment, for AMOVA, ANOSIM, and weighted UniFrac. Diet, ADY, and location were all significant factors for fungi (PERMANOVA, P = 0.0001, P = 0.0452, P = 0.0068, Monte Carlo correction, respectively, and location was a significant factor (P = 0.001, Monte Carlo correction) for protozoa. Diet-induced SARA shifts diversity of rumen fungi and protozoa and selects against fiber-degrading species. Supplementation with ADY mitigated this reduction in protozoa, presumptively by triggering microbial diversity shifts (as seen even in the high-fiber diet) that resulted in pH stabilization. ADY did not recover the initial community structure that was seen in pre-SARA conditions.

Molecular analysis of Giardia duodenalis isolates from symptomatic and asymptomatic children from La Habana, Cuba

Giardiasis is considered the most common intestinal parasitic disease in humans worldwide. In Cuba, this infection has particularly a strong clinical impact on the child population. Giardia duodenalis is a highly diverse protozoan, which comprises a complex of eight morphologically identical genetic assemblages, further divided into sub-assemblages. The present study used triose phosphate isomerase (tpi) and small-subunit ribosomal RNA (SSU rRNA) genes as genetic markers for the identification of G. duodenalis assemblages and sub-assemblages in correlation with clinical and epidemiological data in children attended at the Paediatric Hospital “William Soler” and at Pedro Kouri Institute, between 2015 and 2016. A prevalence of 8% of G. duodenalis infection was recorded in stool samples after concentration techniques from 68 children out of 847 analysed. A 100% detection of Giardia DNA was achieved by a SSU-rRNA PCR, whereas DNA from 63 of 68 (92.6%) was successfully amplified by tpi-PCR. By this assemblage-specific tpi-PCR 32 (50.8%) assemblage B, 17 (27.0%) assemblage A and 14 (22.2%) mixed infection (A+B) were identified. Assemblage B was significantly (P<0.02) more frequently found in children with diarrhoea. Sequence analysis of the tpi gene of Giardia isolates from symptomatic children showed that assemblage A belonged to the sub-assemblage AII, and 4 sub assemblages BIV and 1 sub assemblage BIII were also recorded. Only 2 discordant genotyping results were observed by phylogenetic comparison of SSU-rRNA and tpi sequences. Further studies with novel molecular tools for a better discrimination at the sub-assemblage level are needed to identify the dynamics of spread of giardiasis and to verify possible correlations between Giardia genetic diversity and clinical manifestation.