Flagellated Protozoan Parasite Research Articles

Osmotic stress and regulatory volume decrease in Crithidia fasciculataparasites

Regulatory volume decrease (RVD) describes the response to hypoosmotic stress in many cells including the parasitic flagellate Crithidia fasciculata. Hypoosmotic stress causes initial swelling, the cell expels excess liquid and returns to a normal volume. This process is not able to withstand excessive osmotic stress. We use the MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide) Assay is used to assess cell viability, while microscopy images and spectroscopic light scatter used to follow changes in cell shape and size. It was found that after being placed in 70% hypotonic (30% isotonic) solution, cells did not recover from hypoosmotic stress. We are currently developing protocols for this little‐studied parasite and investigating the mechanisms of RVD in Crithidia fasciculata including the role of the osmoregulatory contractile vacuole.

Helicobacter pylori Pathogenicity Islands and Giardia lamblia Cysteine Proteases in Role of Coinfection and Pathogenesis.

Helicobacter pylori is a well-known human-specific stomach pathogen that infects more than half of the world’s population. The infection with this bacterium can cause a variety of gastrointestinal problems, including chronic gastritis, peptic ulcers, and even cancer. H. pylori is a highly infectious bacterium. H. pylori causes an increase in gastric mucosa pH or gastric mucosa intestinal metaplasia. These modifications in the stomach environment are necessary for G. lamblia colonization to occur. Giardia lamblia is a flagellate protozoan parasite that can cause giardiasis in humans and other mammals. It dwells in the duodenum and upper jejunum. Globally, over 280 million cases of human giardiasis are predicted to occur each year. Simultaneous human colonization by G. lamblia and H. pylori is a typical occurrence since the viruses’ predisposing factors are similar in both groups. Giardiasis is a parasitic infection that affects both children and adults worldwide. Infection with Giardia is more common in underdeveloped countries. Globally, more than 200 million cases of giardiasis are detected each year. In contrast, the presence of G. lamblia in the host body triggers an immunological response comparable to that of H. pylori, with lymphocytes strongly polarized towards Th1. As a result, their combined presence exacerbates host tissue damage. The major goal of this seminar is to describe the pathophysiology, immunology, and clinical aspects of G. lamblia and H. pylori coinfection using a comprehensive search of PubMed, Lancet, and Google Scholar sources. Upper gastrointestinal problems such as upper abdominal pain, abdominal bloating, nausea, vomiting, epigastric pain/burning, and belching are all caused by both organisms. Differentiation by physical examination is impossible in people infected with both bacteria. For this coinfection distinction, a laboratory diagnosis is required. G. lamblia and H. pylori, when present together, have a synergistic effect on the host and can cause serious damage. As a result, researchers should delve deeper into the mechanics underlying this potential microbial interaction.

Diarrhea associated with Pentatrichomonas hominis in an infant

Pentatrichomonas hominis is a flagellated protozoan parasite that resides in the digestive tract of humans and is generally nonpathogenic. Although parasites of diarrheal etiology are widespread, neonatal infection with P. hominis is an uncommon incident. This case report depicts an instance of active P. hominis in an 8-month-old child with complaints of fever, vomiting, and loose stools. Stool examination was done with saline and iodine wet mount that showed motile trophozoites of P. hominis with a jerky movement. Treatment with metronidazole was effective as no parasite was found on repeated stool examinations. Pentatrichomonas hominis should be recognized as a cause of diarrhea in neonates, despite being rare.

X-ray diffraction and in vivo studies reveal the quinary structure of Trypanosoma cruzi nucleoside diphosphate kinase 1: a novel helical oligomer structure.

Trypanosoma cruzi is a flagellated protozoan parasite that causes Chagas disease, which represents a serious health problem in the Americas. Nucleoside diphosphate kinases (NDPKs) are key enzymes that are implicated in cellular energy management. TcNDPK1 is the canonical isoform in the T. cruzi parasite. TcNDPK1 has a cytosolic, perinuclear and nuclear distribution. It is also found in non-membrane-bound filaments adjacent to the nucleus. In the present work, X-ray diffraction and in vivo studies of TcNDPK1 are described. The structure reveals a novel, multi-hexameric, left-handed helical oligomer structure. The results of directed mutagenesis studies led to the conclusion that the microscopic TcNDPK1 granules observed in vivo in T. cruzi parasites are made up by the association of TcNDPK1 oligomers. In the absence of experimental data, analysis of the interactions in the X-ray structure of the TcNDPK1 oligomer suggests the probable assembly and disassembly steps: dimerization, assembly of the hexamer as a trimer of dimers, hexamer association to generate the left-handed helical oligomer structure and finally oligomer association in a parallel manner to form the microscopic TcNDPK1 filaments that are observed in vivo in T. cruzi parasites. Oligomer disassembly takes place on the binding of substrate in the active site of TcNDPK1, leading to dissociation of the hexamers. This study constitutes the first report of such a protein arrangement, which has never previously been seen for any protein or NDPK. Further studies are needed to determine its physiological role. However, it may suggest a paradigm for protein storage reflecting the complex mechanism of action of TcNDPK1.

Status of Giardia Infection with Other Intestinal Parasites among Paediatric Patients in Tertiary Medical College Hospitals

Background: Paediatric diarrhoea, the burning issue of nowadays, drastically affects the child mortality and morbidity. Apart from virus and bacteria, parasites also play an important role in causing diarrhoea at that age group. Giardiasis, the most prevalent among the parasitic infections, though neglected yet carries significance because of its long term health issues. The aim of the study was to identify the intestinal parasites by wet mount preparation and further detection of the flagellated protozoan parasite, Giardia by special staining - Trichrome staining technique.
 Materials and methods: It was an observational cross sectional study which was carried between the periods of July 2019 to June 2020, where a total of 200 diarrhoeic patients up to 18 years were enrolled. Direct microscopic method was done where parasites were detected by wet mount preparation and further Trichrome stainning was performed for the detection of Giardia. 
 Results: Out of 200 faecal samples, cysts of Giardia, ova of Ascaris Lumbricoides (AL) Trichuris Trichiura (TT) and both AL and TT were identified in 2.5%, 1.5%, 0.5% and 1.0% samples respectively by wet mount preparation. By Trichrome staining method we detected 3.5% of Giardia cysts.
 Conclusion: Staining technique could be applied in addition to wet mount preparation of stool samples. So that we could recover the undetected Giardia cyst in the aforesaid samples.
 JCMCTA 2021 ; 32 (2) : 111-114

Study of haemoflagellates Trypanosoma sp. infection in some fish of Iraq marshes and relationship of leukocytes with inflammatory response

Fresh water fishes infected with different species of genus Trypanosoma parasite that is similar to that same genus in blood of mammals. Trypanosomiasis or sleeping sickness disease that causes by protozoa flagellate parasites which live inside cell called Trypanosoma. Fish parasites play an important role in regulating the population of their hosts. This study was implemented to verify the infection with Trypanosoma sp. report the cellular intervening through inflammatory response in blood smears of infected fishes. A total of 190 fishes of three species Carasobarbus luteus, Aspius vorax and Liza abu were collected from Al-Hamare and Al-Chibayish marsh south of Iraq. The flagellate parasite Trypanosoma sp. observed in blood of Carasobarbus luteus 41.42% and Aspius vorax 12.28% fishes, while no infection was recorded with Trypanosoma sp. in Liza abu fishes. Prevalence and mean intensity of infection were calculated. The different counts and various types of inflammatory cells were monitored, there are increased in lymphocytes in the infected fishes. Statistically, there are significant differences (p<0.05) among species of infected fish with Trypanosoma sp.

Haematophagous parasites of some fish species in the Lower Irtysh

The purpose of the research is study of four fish species infected with haematophagous parasites in the rivers of the Lower Irtysh basin. Materials and methods. The work was performed in the rivers of the Lower Irtysh basin in 2017–2020. A total of 390 fish of different sex and age were examined including 47 roach (Rutilus rutilus lacustris, (Pallas)), 41 ide (Leuciscus idus, (Linnaeus, 1758)), 47 pike (Esox lucius, (Linnaeus, 1758)) and 255 Siberian sterlet (Acipenser ruthenus marsiglii (Brandt, 1833)). We took blood from the tail artery, and examined its fresh drop under a microscope, and prepared a thin smear, then dried it in the air and fixed in a mixture of 70% ethyl alcohol and diethyl ether (1 : 1) for 30 minutes. The fixed smears were dried in the air at room temperature and stained with azure and eosin at a dilution of 1 : 10 for 40 minutes. We identified species of the found parasites and calculated the prevalence of infection with standard error of the mean.Results and discussion. We found representatives of the Coccidea and Kinetoplastea classes parasitizing in fish blood. Trypanoplasma acipenseris and Haemogregarina acipenseris were recorded in the Siberian sterlet. The pike was infected with T. schulmani and H. esoci. T. schulmani was found in the blood of the ide and roach. Piscicola geometra is the main host of flagellates and sporozoans that parasitize in fish blood in the basin of the Lower Irtysh. The roach and the Siberian sterlet become infected with parasitic flagellates from the age of 1+. All examined fish were infected in age groups 2+ – 4+. Haemogregarina parasitize in the pike aged 4+ and 5+, and in the Siberian sterlet aged 1+ – 4+ and 6+ – 7+.

A New Model Trypanosomatid, Novymonas esmeraldas: Genomic Perception of Its "Candidatus Pandoraea novymonadis" Endosymbiont.

ABSTRACTThe closest relative of human pathogen Leishmania, the trypanosomatid Novymonas esmeraldas, harbors a bacterial endosymbiont “Candidatus Pandoraea novymonadis.” Based on genomic data, we performed a detailed characterization of the metabolic interactions of both partners. While in many respects the metabolism of N. esmeraldas resembles that of other Leishmaniinae, the endosymbiont provides the trypanosomatid with heme, essential amino acids, purines, some coenzymes, and vitamins. In return, N. esmeraldas shares with the bacterium several nonessential amino acids and phospholipids. Moreover, it complements its carbohydrate metabolism and urea cycle with enzymes missing from the “Ca. Pandoraea novymonadis” genome. The removal of the endosymbiont from N. esmeraldas results in a significant reduction of the overall translation rate, reduced expression of genes involved in lipid metabolism and mitochondrial respiratory activity, and downregulation of several aminoacyl-tRNA synthetases, enzymes involved in the synthesis of some amino acids, as well as proteins associated with autophagy. At the same time, the genes responsible for protection against reactive oxygen species and DNA repair become significantly upregulated in the aposymbiotic strain of this trypanosomatid. By knocking out a component of its flagellum, we turned N. esmeraldas into a new model trypanosomatid that is amenable to genetic manipulation using both conventional and CRISPR-Cas9-mediated approaches.

Prevalence and Molecular Identification of Giardiasis Isolates from Stray Dogs in Erbil Province, Kurdistan Region, Iraq

Giardia lamblia is the intestinal, flagellated protozoan parasite. It should make a species complex and comprises eight assemblages (A-H). In the current study, out of 153 examined samples, 16 (10.46%) and 36 (23.53%) specimens were positive by concentration and polymerase chain reaction (PCR) technique for giardiasis, respectively. The highest rate of infection was found in a rural area (14.38%) than in urban areas (9.15%). As well as, the infection rate in males (25/153) was higher compared to that of female dogs (11/153). Regarding fecal consistency, the highest rate (18.3%) of giardiasis was observed in diarrheic dogs, while the lowest rate of giardiasis (5.23%) was observed among non-diarrheic dogs. The PCR products were sequenced for 20 samples and further examined by sequence analysis, 16 isolates under the accession number (MN629930), independent of the host, exhibited G. lamblia GenBank ID: M36728, while in four samples under the accession number (MN629931), nucleotide substitutions generate polymorphism at position 542 and 561 (C → T) and (A → G) at position 684. The similarity between MN629931 and AY072723 genotype A2 was 99.9% which was one nucleotide substitution at position 542 (C → T). The sequencing of the PCR products recognized two assemblages in dogs suggested the possible role of dogs as the reservoir for human giardiasis in Erbil Province which is the first records in the Kurdistan Region, Iraq.

Metabolic evolutionary roots of the macrophage immune response in amoeba-bacteria interactions: The conserved role of hypoxia-induced Factor and AMP kinase

The bacteria Legionella, being able to infect both macrophages and protozoans, reduce oxidative phosphorylation and induce glycolysis, which allows pathogens to grow and replicate in these cells. In amoeba-like inflammatory macrophages (M1), the phagocytizing cells of the primary immune defense, an increase in the rate of glycolysis is followed by a decrease of oxidative phosphorylation. The opposite takes place in anti-inflammatory macrophages (M2). They change from glycolysis to oxidative metabolism when AMP-dependent kinase (AMPK) is activated by a high ratio of AMP/ATP. Stimulation of macrophages with anti-inflammatory cytokines causes activation of AMPK. Infection of macrophages with the parasitic flagellate Leishmania infantum induces a switch from an initial glycolytic phase to oxidative phase with the essential role of AMPK in this change. Activated AMPK induces catabolic pathways effectively producing ATP as well as processes requiring the energy supply. AMPK regulates the migration of cells and enhances the phagocytic activity of macrophages. In macrophages, bacterial products activate TLRs and NF-κB signaling, causing an increase of transcription of hypoxia-induced factor HIF-1α (a subunit of HIF-1). This brings about induction of the enzyme and transporter expression essential for glycolysis and the pentose phosphate pathway to proceed and makes biosynthetic processes and ROS production in macrophages possible. Hypoxia augments macrophage phagocytosis in a HIF-1α-dependent manner. Multicellular parasites experience changes in the availability of oxygen in their life cycle. In the nematode Ascaris suum, HIF participates in the pre-adaptation to hypoxic conditions after infection of their hosts. Also, the freshwater and marine invertebrates meet changes of oxygen concentrations. In the anaerobic branch of the respiratory chain of these invertebrates, fumarate serves as the terminal electron acceptor that is reduced to succinate in complex II of the ETC. In mammalian cells, accumulation of succinate under hypoxic conditions suggests that the mammalian complex II may reduce fumarate to succinate, too. The data reviewed here show that the ability to shift the cell metabolism towards glycolysis observed in activated macrophages can be traced back in evolution to metabolic changes characterizing protozoans infected with bacteria. Anabolic needs of multiplying bacteria direct host metabolism to glycolysis that produces, aside from ATP, precursors of the amino acids used by the pathogen for its protein synthesis. M1-activated mammalian macrophages behave in the same way. Regulation of metabolism in M1 and M2 macrophages is further enhanced by HIF-1 and AMPK, respectively. These archaic functions of AMPK and HIF, important also to control phagocytosis and cell migration were extended to embryonic development in multicellular organisms.

The thermal proteome stability profile of Trypanosoma cruzi in epimastigote and trypomastigote life stages

Trypanosoma cruzi is a flagellate protozoa being the etiological agent of Chagas disease, a neglected tropical disease, which still poses a public health problem worldwide. The intricate molecular changes during T. cruzi-host interaction have been explored using different largescale omics techniques. However, protein stability is largely unknown. Thermal proteome profiling (TPP) methodology has the potential to characterize proteome-wide stability highlighting key proteins during T. cruzi infection and life stage transition from the invertebrate to the mammalian host. In the present work, T. cruzi epimastigotes and trypomastigotes cell lysates were subjected to TPP workflow and analyzed by quantitative large-scale mass spectrometry-based proteomics to fit a melting profile for each protein. A total of 2884 proteins were identified and associated to 1741 melting curves being 1370 in trypomastigotes (TmAVG 53.53 °C) and 1279 in epimastigotes (TmAVG 50.89 °C). A total of 453 proteins were identified with statistically different melting profiles between the two life stages. Proteins associated to pathogenesis and intracellular transport had regulated melting temperatures. Membrane and glycosylated proteins had a higher average Tm in trypomastigotes compared to epimastigotes. This study represents the first large-scale comparison of parasite protein stability between life stages. SignificanceTrypanosoma cruzi, a unicellular flagellate parasite, is the etiological agent of Chagas disease, endemic in South America and affecting more that 7 million people worldwide. There is an intense research to identify novel chemotherapeutic and diagnostic targets of Chagas disease. Proteomic approaches have helped in elucidating the quantitative proteome and PTMs changes of T. cruzi during life cycle transition and upon different biotic and abiotic stimuli. However, a comprehensive knowledge of the protein-protein interaction and protein conformation is still missing. In order to fill this gap, this manuscript elucidates the T. cruzi Y strain proteome-wide thermal stability map in the epimastigote and trypomastigote life stages. Comparison between life stages showed a higher average melting temperature stability for trypomastigotes than epimastigotes indicating a host temperature adaptation. Both presented a selective thermal stability shift for cellular compartments, molecular functions and biological processes based on the T. cruzi life stage. Membrane and glycosylated proteins presented a higher thermal stability in trypomastigotes when compared to the epimastigotes.

Molecular detection of glutamate dehydrogenase gene of Giardia lamblia isolated from food handlers in Erbil city

Background and objective: Giardia lamblia is the intestinal flagellated protozoan parasite that infects vertebrates, including humans. Giardiasis is the major diarrheal disease found worldwide. It can be symptomatic or may be an asymptomatic carrier that led to chronic disease. This study aimed to determine the proportion of giardiasis among food handlers and evaluate the correlation between two laboratory methods for identifying the Giardia lamblia. Methods: A total of 308 stool samples were collected from food handlers that annually attend the central laboratory in Erbil City. Wetmount microscopic examination was performed for the diagnosis of cysts and trophozoites of the Giardia parasite. Molecular analysis done for positive samples, DNA extraction performed using the QIAamp Fast DNA Stool Mini Kit (Qiagen Company, Germany). Nested PCR analysis was done targeting the Glutamate dehydrogenase gene using two sets of primers for amplification of 734bp fragment. Gel electrophoresis was performed for visualizing the amplified DNA by Ultraviolet light. Results: The mean age of food handler participants was 29 years. Most (93.8%) of the food handlers were males, and the majority (98.6%) of the participants did not have any signs and symptoms. Four (7.4%) microscopy positive sample participants were highly educated. There was no association between educational level and positive rate by microscopy (P = 0.066). The majority of participants did not receive treatments, particularly most of the microscopic positive samples. The food handlers did not take any antiparasitic treatments 9 (3.4%) (P = 0.676). From 11 (3.6%) microscopically positive samples, 10 (90.9%) Giardia lamblia gdh gene 734 bp fragments were amplified by nested PCR. Conclusion: Amplification of 734bp of gdh gene by the nested PCR is the most specific and sensitive method for identifying Giardia lamblia. Food handlers were important people to care about sanitation and preparing food, particularly for avoiding diseases transmitted by food. Keywords: Giardiasis; Genetic characterization; gdh gene; Nested PCR.

Natural infections of Pintomyia verrucarum and Pintomyia maranonensis by Leishmania (Viannia) peruviana in the Eastern Andes of northern Peru

The natural infection of sand flies by Leishmania was investigated in Andean areas located between the Central and Eastern Cordilleras of northern Peru where cutaneous leishmaniasis caused by Leishmania (Viannia) peruviana is endemic. Sand flies were captured at five locations along the Utcubamba River in the Department of Amazonas, and morphologically identified under a microscope. Among 422 female sand flies dissected, the most dominant species was Pintomyia verrucarum (320 flies), followed by Pi. maranonensis (83 flies), Pi. robusta (13 flies), and Lutzomyia castanea (6 flies). Genetic analysis of sand flies from these areas together with those from other areas revealed that individuals of Pi. verrucarum were closely related regardless of morphological variation of their spermathecae. On the other hand, individuals of Pi. maranonensis collected in the study area were distant from those of other areas with genetic distances over the intraspecific level but mostly below the interspecific level, suggesting the unique characteristics of sand flies in this area. The natural infection of sand flies by flagellate parasites was detected mainly in the hindgut of each one of Pi. verrucarum and Pi. maranonensis. Both parasite species were identified as L. (V.) peruviana based on cytochrome b and mannose phosphate isomerase gene analyses. In addition, parasite species obtained from the lesion of a patient with cutaneous leishmaniasis in the study area in this period was identified as L. (V.) peruviana. These results strongly suggest that Pi. verrucarum and Pi. maranonensis are responsible for the transmission of L. (V.) peruviana in these areas. This is the first report of the natural infection of Pi. maranonensis by L. (V.) peruviana.

Genetic and immunological basis of human African trypanosomiasis.

Human African trypanosomiasis, or sleeping sickness, results from infection by two subspecies of the protozoan flagellate parasite Trypanosoma brucei, termed Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense, prevalent in western and eastern Africa respectively. These subspecies escape the trypanolytic potential of human serum, which efficiently acts against the prototype species Trypanosoma brucei brucei, responsible for the Nagana disease in cattle. We review the various strategies and components used by trypanosomes to counteract the immune defences of their host, highlighting the adaptive genomic evolution that occurred in both parasite and host to take the lead in this battle. The main parasite surface antigen, named Variant Surface Glycoprotein or VSG, appears to play a key role in different processes involved in the dialogue with the host.

Naphthoquinone derivatives exhibit apoptosis-like effect and anti-trypanosomal activity against Trypanosoma evansi

Trypanosoma evansi is an extracellular flagellate blood protozoan parasite and an etiological agent of animal trypanosomosis. Presently, only a few drugs are registered and have been used for the treatment of animal trypanosomosis, but they show severe toxic effects and also face the problem of drug resistance. Naphthoquinones (NTQ) are considered as fortunate structures in the field of medicinal chemistry as they have been reported for their antitrypanosomal potential against other trypanosomes-T. brucei and T. cruzi. In the present study, six naphthoquinones (NTQ1-NTQ6) derivatives were evaluated for anti-trypanosomal activity by demonstrating their growth inhibitory effect against T. evansi. All NTQs significantly (p < 0.001) exhibited activity against parasite growth and multiplication with IC50 values of 11.48 μM, 373.6 μM, 12.97 μM, 21.97 μM, 18.19 μM and 5.758 μM but NTQ1, NTQ3 and NTQ6 were selected based on their IC50 value for further studies. The dose-and time-dependent morphological effect on parasite was evaluated including the measurement of reactive oxygen species (ROS) by spectrofluorometery and measurement of apoptosis by flow cytometry. The selected NTQs exhibited a significant production of ROS and displayed a significant AV+ and PI+ labelled cells in the axenic culture of T. evansi than quinapyramine methyl sulphate (QPS), as reference control. NTQs also showed more cytotoxic effect on horse peripheral blood mononuclear cells as compare to QPS. Therefore, we confirmed the antitrypanosomal activity and apoptotic-like mechanism of NTQs in an axenic culture of T. evansi.

Bacteria cultured from the gut of Meccus pallidipennis (Hemiptera: Reduviidae), a triatomine species endemic to Mexico.

The study of intestinal microbiota in vector insects like triatomines is paramount in parasitology because many parasitic species inhabit the vector's gut. Although knowledge on the gut microbiota in various vectors of the parasitic flagellate Trypanosoma cruzi has grown, research efforts have focused on South American triatomines. This study reports the isolation of bacterial microbiota in the anterior and posterior gut of Meccus pallidipennis (a triatomine species endemic to Mexico) by culture, as well as its identification by phenotypic and biochemical tests and its quantification by counting colony-forming units. The study was performed on fifth-instar nymph and adult specimens of M. pallidipennis, either laboratory-bred or collected in the field and either infected or not with T. cruzi. Overall, 17 bacterial species were identified, with the genera Bacillus and Staphylococcus being the most prevalent regardless of the origin of the insects. No differences were observed in the number of bacterial species in the gut of laboratory-bred and field-collected insects, neither with respect to life stage or infection status. In general, the Shannon-Weaver diversity index was higher in non-infected insects than in infected ones. Further studies using non-culture methods are required to determine whether bacterial species diversity is modified by laboratory breeding.

Lophomonas blattarum - A blunder!

Background: Lophomonas blattarum is a parasite colonising gut of cockroaches. There are 136 case reports from immunocompromised patients and children suffering from respiratory tract infections. Here we report a case where broncho alveolar lavage fluid from a child showed structures which convincingly appeared like a flagellate parasite but was later confirmed to be ciliocytophthoria. Case Description: A 5 y old female child hailing from rural southern India came with complaints of cough, cold and abdominal pain since 6 m. In view of increased severity of symptoms, high grade fever and cavitatory lesions in the lung, the child was referred to our centre. We considered the differential diagnoses - Hydatid cyst, Tuberculosis, non infectious causes of lung cyst. Investigations showed Eosinophilia and raised ESR. CT scan of chest showed consolidation with cavities involving apical segment of right lower lobe, anterior segment of left upper lobe suggestive of infective etiology. Broncho alveolar lavage fluid was negative for acid fast bacilli and TB PCR. On wet mount examination, motile structures appearing to have tuft of flagella were seen. Modified Trichrome stain showed numerous inflammatory cells and pyriform shaped structures with tuft of flagella arranged irregularly at one pole. Inner structures were not appreciated very clearly and a presumptive diagnosis of the parasite Lophomonas blattarum was made. The patient was treated with metronidazole 7.5 mg/kg body weight for 10 d awaiting further results. Child improved symptomatically and discharged. Discussion: Since it was difficult to do the molecular tests, we sent the micrscopic photographs and videos to CDC Diagnostic Assistance team to help us solve the case. They confirmed it to be ciliocytophthoria. Bronchial epithelial cells are cylindrical with basal nuclei and a visible terminal bar just beneath the regularly arranged cilia at the other end. This was a learning experience for us to differenciate the cells from the parasite. We now routinely perform special stains on samples found to have motile structures like these. Conclusion: Lophomonas blattarum rarely causes human disease more often misdiagnosed because the movement of the flagellate parasite resembles the motion of detached ciliated bronchoepithelial cells when brochoalveolar lavage fluid is examined under the microscope.

Vickermania gen. nov., trypanosomatids that use two joined flagella to resist midgut peristaltic flow within the fly host

BackgroundThe family Trypanosomatidae encompasses parasitic flagellates, some of which cause serious vector-transmitted diseases of humans and domestic animals. However, insect-restricted parasites represent the ancestral and most diverse group within the family. They display a range of unusual features and their study can provide insights into the biology of human pathogens. Here we describe Vickermania, a new genus of fly midgut-dwelling parasites that bear two flagella in contrast to other trypanosomatids, which are unambiguously uniflagellate.ResultsVickermania has an odd cell cycle, in which shortly after the division the uniflagellate cell starts growing a new flagellum attached to the old one and preserves their contact until the late cytokinesis. The flagella connect to each other throughout their whole length and carry a peculiar seizing structure with a paddle-like apex and two lateral extensions at their tip. In contrast to typical trypanosomatids, which attach to the insect host’s intestinal wall, Vickermania is separated from it by a continuous peritrophic membrane and resides freely in the fly midgut lumen.ConclusionsWe propose that Vickermania developed a survival strategy that relies on constant movement preventing discharge from the host gut due to intestinal peristalsis. Since these parasites cannot attach to the midgut wall, they were forced to shorten the period of impaired motility when two separate flagella in dividing cells interfere with each other. The connection between the flagella ensures their coordinate movement until the separation of the daughter cells. We propose that Trypanosoma brucei, a severe human pathogen, during its development in the tsetse fly midgut faces the same conditions and follows the same strategy as Vickermania by employing an analogous adaptation, the flagellar connector.

Molecular detection and genetic characterization of Trichomonas gallinae in falcons in Saudi Arabia.

Avian trichomonosis is primarily caused by Trichomonas gallinae, a flagellated protozoan parasite that especially infects the upper digestive tract of columbid bird species and their avian predators. However, this parasite has recently been found to be distributed worldwide in various other avian species. This parasitic disease is common in captive falcons in Saudi Arabia and the Middle East. This study aimed to examine and identify the genetic variation of T. gallinae obtained from three species of falcons in Saudi Arabia via the sequencing analysis of the internal transcribed spacer (ITS) region. Swab samples from 97 saker falcons (Falco cherrug), 24 peregrine falcons (Falco peregrinus) and 37 gyrfalcons (Falco rusticolus) were cultured and analysed for infection between 2018 and 2019. The overall prevalence of infection by T. gallinae was 26.58% (n = 42), of which 35 (83.33%) were collected from Riyadh region and seven (16.67%) were collected from Qassim region. The results indicate the presence of four genotypes of T. gallinae in Saudi falcons: A, C, II, and KSA11. This study reports for the first time genetic diversity of T. gallinae in these falcons in Saudi Arabia.

Study of Some Immunological Parameters with Level of Iron in Women Infected with Trichomonas Vaginalis

Trichomonas vaginalis is a flagellated protozoan parasite that causes a disease called trichomoniasis inhumans. It affects the urogenital tract and is one of the most important causes of non-viral genitourinarysexually transmitted infection (STI). The host’s immune response plays a very important role in the onsetof pathological symptoms and their differences between infected people. During the period from July toDecember 2019, 30 samples were collected, 20 of infected women with T. vaginalis and 10 samples as acontrol group from reviews of external women’s clinics in Kirkuk city, whose ages ranged between 18-56 yearsso this study was designed to know the effect of infection with T. vaginalis parasite on some immunologicalparameters such as the role of humoral immune response based on the production of immunoglobulins IgM,IgG, IgA against the parasite and there estimation in serum by using Enzyme Linked Immunosorbent Assay(ELISA) technique as well as estimating the C-Reactive protein concentration as an indication of systemicinflammation and of serum iron level . The results of the current study showed a “significant” increase inthe level of immunoglobulins IgM, IgG in serum of women infected with T. vaginalis compared to thecontrol group, as the level of IgM (8. 2 ± 2.9 ng/dl), (6.3 ± 0.5 ng/dl) Consequently, at the level (p <0.05),the level of IgG was (8.16 ± 2.20 ng/dl), (6.53 ± 1.69 ng/dl) for the infected women and the control group,respectively. As for the level of IgA, the results showed non-significant increase in the affected womencompared with control group (8.22 ± 3.1 ng/dl), (7.82 ± 2.8 ng/dl), respectively. The results of estimatingthe CRP concentration showed a significant increase in the infected women compared to the control group(5.76± 1.56 mg/dl), (4.61± 0.66 mg/dl), respectively. While the results of measuring the level of iron showeda significant decrease among infected women and the control group (38.79 ±8.23 mg/dl), (93.83± 34.76 mg/dl), respectively. We conclude from the above that infection with T.vaginals stimulates the humoral immuneresponse and affects on immunoglobulins levels, CRP concentration and Iron serum level.