Laboratory diagnostics of Acanthamoeba spp. infections

2018 ◽  
Vol 54 (3) ◽  
pp. 173-178
Author(s):  
Aleksandra Bojanowska ◽  
Anna Bożym ◽  
Olga M. Koper-Lenkiewicz ◽  
Joanna Kamińska ◽  
Halina Kemona ◽  
...  

Protozoa <i>Acanthamoeba</i> castellanii are single-celled, free-living, cosmopolitan amoebae that enter the body mainly through the mucous membrane of the mouth, nasal cavity, cornea, respiratory system, damaged skin and intestinal mucosa. In addition, they can be vectors for microorganisms such as bacteria, viruses, fungi and protozoa. In the life cycle A. castellanii may take the form of trophozoites and cyst tenacity to environmental conditions. This amoeba is an etiologic agent of dangerous human diseases: <i>Acanthamoeba</i> keratitis (AK), granulomatous amoebic encephalitis (GAE), pneumonia or changes in other organs, such as the liver, kidneys and skin. The aim of the article is to present current knowledge about laboratory diagnosis of <i>Acanthamoeba</i> spp. infection, which is based on the search for trophozoites and cysts in the material collected from the patient. In the case of suspected GAE, it is also advisable to study cerebrospinal fluid sediment and perform imaging tests such as computed tomography or magnetic resonance imaging. In the case of <i>Acanthamoeba</i> keratitis, an amoeba culture (obtained from a biopsy or corneal scrapings) is established in vitro, and then identified by light microscopy. The methods of molecular biology are also useful in detecting, identifying, and determining the potential pathogenic abilities of the amoebas. After the detection of <i>Acanthamoeba</i> spp., it is important to differentiate pathogenic isolates from non-pathogenic ones. From a clinical point of view, to diagnose <i>Acanthamoeba</i> spp. infection it is necessary only to identify the type of organism and to determine the pathogenicity of the isolate.

Author(s):  
Caio Mauricio Mendes de Cordova ◽  
Caroline Galgowski ◽  
Leonardo Lange

Background. Mycoplasmas are known to cause various infections in humans, mainly in the respiratory and urogenital tracts. The different species are usually host-specific and cause diseases in well-defined sites. New species have been isolated, including those from HIV-infected persons.Summary. Itsin vitroproperties, combined with clinical findings, have led to the hypothesis that these microorganisms may act as cofactors of HIV in AIDS development. Even today this point of view is quite polemic among infectious disease specialists and many aspects remain to be clarified, in contrast to what happens, for instance, with HIV/Mycobacterium tuberculosiscoinfection. Dozens of papers have been published covering aspects of Mollicutes/HIV coinfection, but they add little to no information about the putative contribution of Mollicutes to the evolution of AIDS. Very few researchers have devoted their efforts to trying to answer this question, which remains open. In this review, we discuss the evidences that may support this statement in the light of current knowledge in the field of mycoplasmology.


2012 ◽  
Vol 12 (2) ◽  
pp. 151-157 ◽  
Author(s):  
Ewa Chronowska

Stem Cell Characteristics of Ovarian Granulosa Cells - ReviewRecently increasing interest in stem cells of mammalian ovary has been observed. Potential somatic stem cells for the follicular theca and ovarian surface epithelium have been demonstated. On the other hand, despite intensive research, difinitive evidence for stem cell characteristics of granulosa cells is still to be found. Elucidation of stem cell properties of follicular granulosa cells may have important implications both from scientific and clinical point of view. The aim of this work is to review the current knowledge about stem cell properties of cells constituting main somatic compartment of the mammalian ovary, namely granulosa cells.


2012 ◽  
Vol 57 (1) ◽  
pp. 375-381 ◽  
Author(s):  
Carmen María Martín-Navarro ◽  
Jacob Lorenzo-Morales ◽  
Rubén P. Machin ◽  
Atteneri López-Arencibia ◽  
José Manuel García-Castellano ◽  
...  

ABSTRACTAcanthamoebais an opportunistic pathogen in humans, whose infections most commonly manifest asAcanthamoebakeratitis or, more rarely, granulomatous amoebic encephalitis. Although there are many therapeutic options for the treatment ofAcanthamoeba, they are generally lengthy and/or have limited efficacy. Therefore, there is a requirement for the identification, validation, and development of novel therapeutic targets against these pathogens. Recently, RNA interference (RNAi) has been widely used for these validation purposes and has proven to be a powerful tool forAcanthamoebatherapeutics. Ergosterol is one of the major sterols in the membrane ofAcanthamoeba. 3-Hydroxy-3-methylglutaryl–coenzyme A (HMG-CoA) reductase is an enzyme that catalyzes the conversion of HMG-CoA to mevalonate, one of the precursors for the production of cholesterol in humans and ergosterol in plants, fungi, and protozoa. Statins are compounds which inhibit this enzyme and so are promising as chemotherapeutics. In order to validate whether this enzyme could be an interesting therapeutic target inAcanthamoeba, small interfering RNAs (siRNAs) against HMG-CoA were developed and used to evaluate the effects induced by the inhibition ofAcanthamoebaHMG-CoA. It was found that HMG-CoA is a potential drug target in these pathogenic free-living amoebae, and various statins were evaluatedin vitroagainst three clinical strains ofAcanthamoebaby using a colorimetric assay, showing important activities against the tested strains. We conclude that the targeting of HMG-CoA andAcanthamoebatreatment using statins is a novel powerful treatment option againstAcanthamoebaspecies in human disease.


2002 ◽  
Vol 46 (3) ◽  
pp. 695-701 ◽  
Author(s):  
Julia Walochnik ◽  
Michael Duchêne ◽  
Karin Seifert ◽  
Andreas Obwaller ◽  
Thomas Hottkowitz ◽  
...  

ABSTRACT Free-living amoebae of the genus Acanthamoeba are causing serious chronic conditions such as destructive keratitis in contact lens wearers or granulomatous amoebic encephalitis in individuals with compromised immune systems. Both are characterized by the lack of availability of sufficiently effective and uncomplicated, manageable treatments. Hexadecylphosphocholine (miltefosine) is licensed for use as a topical antineoplastic agent, but it is also active in vitro against several protozoan parasites, and it was applied very successfully for the treatment of human visceral leishmaniasis. The aim of our study was to evaluate the efficacy of hexadecylphosphocholine and other alkylphosphocholines (APCs) against Acanthamoeba spp. The in vitro activities of eight different APCs against three Acanthamoeba strains of various pathogenicities were determined. All substances showed at least amoebostatic effects, and some of them disrupted the amoebae, as shown by the release of cytoplasmic enzyme activity. Hexadecylphosphocholine exhibited the highest degree of cytotoxicity against trophozoites, resulting in complete cell death at a concentration as low as 40 μM, and also displayed significant cysticidal activity. Hexadecylphosphocholine may be a promising new candidate for the topical treatment of Acanthamoeba keratitis and, conceivably, even for the oral treatment of granulomatous amoebic encephalitis.


2021 ◽  
Author(s):  
◽  
Rosemary Jennifer Hurst

<p>The life cycle of Anisakis simplex in New Zealand waters is described from observations on the morphology, distribution and behaviour of free-living and parasitic stages. Comparison with the life cyles of two other anisakids, Phocanema decipiens Myers 1959 and Thynnascaris adunca Rudolphi 1802 shows differences in distribution, degrees of host specificity, the status of invertebrate hosts, the factors influencing infestation levels of teleost hosts, and the location and pathological effects of infestation. Larval stages occurring in intermediate and paratenic hosts were identified by comparison of larval and adult morphometrics. A. simplex larvae were also positively identified by in vitro cultivation through to adults. Some morphometric variations compared to overseas descriptions are apparent. The ventriculus of A. simplex larvae is shorter relative to body length and the intestinal caecum of P. decipiens is longer relative to ventriculus length. Egg and free-living larval stages were obtained from in vitro cultivation of (A. simplex) and collection of eggs from mature adults from definitive hosts (T. adunca). Eggs of P. decipiens were not obtained. Eggs of A. simplex and T. adunca hatch in 8-11 days at 15 [degrees] C. A. simplex eggs hatch in 6 days at a temperature of 22 [degrees] C and did not hatch in 16 days at 10 [degrees] C. Eggs and free-living stage III larvae of A. simplex and T. adunca are similar in morphology with little differentiation of internal structures. Examination of the stomach contents of pelagic fish infested with anisakids indicated that possible intermediate hosts of A. simplex are the euphausiid Nyctiphanes australis and the decapod Munida gregaria. Possible hosts of T. adunca and M. gregaria are a wide variety of smaller zooplanktonic groups, e.g. decapod larvae and copepods. Larvae of A. simplex were found in one of 8850 N. australis; larvae of T. adunca were found in 69 of 3999 chaetognaths (Sagitta spp.) a medusa and a decapod larva. These larvae are morphologically similar to Stage III larvae from teleosts. No anisakids were found in 3956 Euphausia spp., 1147 M. gregaria and 740 prawns. Twenty five T. adunca larvae and adults were found in 818 freshly eaten M. gregaria in teleost stomachs, indicating that this invertebrate may act as a paratenic and a definitive host. Experimental infection of N. australis and M. gregaria with stage II larvae of A. simplex and T. adunca was unsuccessful. The location of anisakid infestation in three pelagic teleost species, Thyrsites atun, Trachurus novaezelandiae and Trachurus declivis is described. A. simplex larvae are found mainly in the body cavity of all species, at the posterior end of the stomach, with less than one percent occurring in the musculature. Distribution of A. simplex larvae does not change with increasing size of the host or increasing total worm burden. Thyrsites atun have a higher proportion of larvae in the stomach wall (8-13%) compared to Trachurus spp. (< 4%). T. adunca larvae are found infrequently in the body cavity of all three species, on the pyloric caeca and in the stomach wall. Adults and larvae of T. adunca are found more commonly in the alimentary canal, indicating that these teleosts are more important as definitive hosts in the life cycle of this anisakid. P. decipiens larvae are found only in Thyrsites atun and occur mainly in the muscles (98.5%). No quantitative pathogenic effects of anisakid infestation on these teleosts hosts were detected. The main factors influencing the infestation of the three teleost species are age of the host, locality and season. Sex of the host and depth (over the continental shelf, 0-250 m) are not important. A. simplex infestation increased with age in all host species examined, and was higher in Trachurus declivis from the southern-most locality, suggesting the existence of at least two distinct populations of this species. Significant differences in infestation of Thyrsites atun with P. decipiens suggests that this anisakid may be more common in southern localities also. The infestation of Thyrsites atun by larval and adult T. adunca in the alimentary canal is most influenced by season and closely related to diet. Nematode samples were obtained from the marine mammals Arctocephalus forsteri, Kogia breviceps and Phocarctos hookeri. Adult A. simplex were recorded from A. forsteri (a new host record) and Kogia breviceps; preadults from Phocarctos hookeri. Adult P. decipiens were recorded from Phocarctos hookeri; preadults from Arctocephalus forsteri and K. breviceps. Other anisakids found were Anisakis physeteris (Baylis 1923), Contracaecum osculatum Rudolphi 1802 and Pseudoterranova kogiae (Johnston and Mawson 1939) Mosgovoi 1951. These records are all new for the New Zealand region except P. decipiens from P. hookeri and C. osculatum from Arctocephalus forsteri. A. simplex and C. osculatum were found associated with gastric ulcers in Arctocephalus forsteri.</p>


2021 ◽  
Author(s):  
◽  
Rosemary Jennifer Hurst

<p>The life cycle of Anisakis simplex in New Zealand waters is described from observations on the morphology, distribution and behaviour of free-living and parasitic stages. Comparison with the life cyles of two other anisakids, Phocanema decipiens Myers 1959 and Thynnascaris adunca Rudolphi 1802 shows differences in distribution, degrees of host specificity, the status of invertebrate hosts, the factors influencing infestation levels of teleost hosts, and the location and pathological effects of infestation. Larval stages occurring in intermediate and paratenic hosts were identified by comparison of larval and adult morphometrics. A. simplex larvae were also positively identified by in vitro cultivation through to adults. Some morphometric variations compared to overseas descriptions are apparent. The ventriculus of A. simplex larvae is shorter relative to body length and the intestinal caecum of P. decipiens is longer relative to ventriculus length. Egg and free-living larval stages were obtained from in vitro cultivation of (A. simplex) and collection of eggs from mature adults from definitive hosts (T. adunca). Eggs of P. decipiens were not obtained. Eggs of A. simplex and T. adunca hatch in 8-11 days at 15 [degrees] C. A. simplex eggs hatch in 6 days at a temperature of 22 [degrees] C and did not hatch in 16 days at 10 [degrees] C. Eggs and free-living stage III larvae of A. simplex and T. adunca are similar in morphology with little differentiation of internal structures. Examination of the stomach contents of pelagic fish infested with anisakids indicated that possible intermediate hosts of A. simplex are the euphausiid Nyctiphanes australis and the decapod Munida gregaria. Possible hosts of T. adunca and M. gregaria are a wide variety of smaller zooplanktonic groups, e.g. decapod larvae and copepods. Larvae of A. simplex were found in one of 8850 N. australis; larvae of T. adunca were found in 69 of 3999 chaetognaths (Sagitta spp.) a medusa and a decapod larva. These larvae are morphologically similar to Stage III larvae from teleosts. No anisakids were found in 3956 Euphausia spp., 1147 M. gregaria and 740 prawns. Twenty five T. adunca larvae and adults were found in 818 freshly eaten M. gregaria in teleost stomachs, indicating that this invertebrate may act as a paratenic and a definitive host. Experimental infection of N. australis and M. gregaria with stage II larvae of A. simplex and T. adunca was unsuccessful. The location of anisakid infestation in three pelagic teleost species, Thyrsites atun, Trachurus novaezelandiae and Trachurus declivis is described. A. simplex larvae are found mainly in the body cavity of all species, at the posterior end of the stomach, with less than one percent occurring in the musculature. Distribution of A. simplex larvae does not change with increasing size of the host or increasing total worm burden. Thyrsites atun have a higher proportion of larvae in the stomach wall (8-13%) compared to Trachurus spp. (< 4%). T. adunca larvae are found infrequently in the body cavity of all three species, on the pyloric caeca and in the stomach wall. Adults and larvae of T. adunca are found more commonly in the alimentary canal, indicating that these teleosts are more important as definitive hosts in the life cycle of this anisakid. P. decipiens larvae are found only in Thyrsites atun and occur mainly in the muscles (98.5%). No quantitative pathogenic effects of anisakid infestation on these teleosts hosts were detected. The main factors influencing the infestation of the three teleost species are age of the host, locality and season. Sex of the host and depth (over the continental shelf, 0-250 m) are not important. A. simplex infestation increased with age in all host species examined, and was higher in Trachurus declivis from the southern-most locality, suggesting the existence of at least two distinct populations of this species. Significant differences in infestation of Thyrsites atun with P. decipiens suggests that this anisakid may be more common in southern localities also. The infestation of Thyrsites atun by larval and adult T. adunca in the alimentary canal is most influenced by season and closely related to diet. Nematode samples were obtained from the marine mammals Arctocephalus forsteri, Kogia breviceps and Phocarctos hookeri. Adult A. simplex were recorded from A. forsteri (a new host record) and Kogia breviceps; preadults from Phocarctos hookeri. Adult P. decipiens were recorded from Phocarctos hookeri; preadults from Arctocephalus forsteri and K. breviceps. Other anisakids found were Anisakis physeteris (Baylis 1923), Contracaecum osculatum Rudolphi 1802 and Pseudoterranova kogiae (Johnston and Mawson 1939) Mosgovoi 1951. These records are all new for the New Zealand region except P. decipiens from P. hookeri and C. osculatum from Arctocephalus forsteri. A. simplex and C. osculatum were found associated with gastric ulcers in Arctocephalus forsteri.</p>


2014 ◽  
Vol 7 (1) ◽  
pp. 20-24 ◽  
Author(s):  
Krassimira S. Yakimova ◽  
Rumen P. Nikolov ◽  
Ivan G. Todorov ◽  
Milen H. Hristov

Abstract Leptin inhibits feeding, reduces body weight and increases thermogenesis. Experimental data suggest involvement of GABAergic mechanisms in the regulation of feeding behavior and energy balance. The present study was set to determine the effect of combinations from leptin, GABAB-agonist baclofen and GABAB-antagonist CGP35348 on thermoregulation of male Wistar rats, using in vivo and in vitro experiments. The substances used for in vivo experiments were administered intraperitoneally (i.p.). The measurement of the body temperature was done via thermistor probes (TX8) and monitored on multichannel recorder Iso-Thermex16. In vitro experiments were conducted on rat PO/AH neurons, recorded extracellulary by conventional electrophysiological equipment, using brain slice preparations. The separate intraperitoneal injection of leptin as well as GABAB-antagonist CGP35348 produced significant hyperthermia in rats while the GABAB-agonist baclofen caused a decrease in the core body temperature. The probable synergy between the hyperthermic effects of leptin and GABAB-antagonist did not occur. On the contrary, the effect of this combination was lower as compared to the result of the separate administration of GABAB-antagonist. When leptin was applied just prior to GABAB-agonist baclofen, neither of their separate effects appeared. In vivo effects determined correlated with in vitro changes of firing rate observed in PO/AH neurons. The data from this study provide a new point of view concerning the interactions of leptin and GABA on the level of thermoregulation. These results represent a step forward in understanding the complicated mechanisms involved in thermoregulation.


Toxins ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 297
Author(s):  
Franjo Banović ◽  
Horst Schroten ◽  
Christian Schwerk

Although it rarely induces disease in humans, Listeria monocytogenes (Lm) is important due to the frequency of serious pathological conditions—such as sepsis and meningitis—it causes in those few people that do get infected. Virulence factors (VF) of Lm—especially those involved in the passage through multiple cellular barriers of the body, including internalin (Inl) family members and listeriolysin O (LLO)—have been investigated both in vitro and in vivo, but the majority of work was focused on the mechanisms utilized during penetration of the gut and fetoplacental barriers. The role of listerial VF during entry into other organs remain as only partially solved puzzles. Here, we review the current knowledge on the entry of Lm into one of its more significant destinations, the brain, with a specific focus on the role of various VF in cellular adhesion and invasion.


2015 ◽  
Vol 24 (135) ◽  
pp. 115-131 ◽  
Author(s):  
Oluf Dimitri Røe ◽  
Giulia Maria Stella

Asbestos is the term for a family of naturally occurring minerals that have been used on a small scale since ancient times. Industrialisation demanded increased mining and refining in the 20th century, and in 1960, Wagner, Sleggs and Marchand from South Africa linked asbestos to mesothelioma, paving the way to the current knowledge of the aetiology, epidemiology and biology of malignant pleural mesothelioma. Pleural mesothelioma is one of the most lethal cancers, with increasing incidence worldwide. This review will give some snapshots of the history of pleural mesothelioma discovery, and the body of epidemiological and biological research, including some of the controversies and unresolved questions. Translational research is currently unravelling novel circulating biomarkers for earlier diagnosis and novel treatment targets. Current breakthrough discoveries of clinically promising noninvasive biomarkers, such as the 13-protein signature, microRNAs and theBAP1mesothelioma/cancer syndrome, are highlighted. The asbestos history is a lesson to not be repeated, but here we also review recentin vivoandin vitrostudies showing that manmade carbon nanofibres could pose a similar danger to human health. This should be taken seriously by regulatory bodies to ensure thorough testing of novel materials before release in the society.


2018 ◽  
Vol 9 (1) ◽  
pp. 75-79
Author(s):  
Y. A. Gugosyan ◽  
V. A. Yevstafyeva ◽  
О. А. Gorb ◽  
V. V. Melnychuk ◽  
I. О. Yasnolob ◽  
...  

Strongyloides westeri (Ihle, 1917), a parasitic horse nematode, has an unusual lifecycle, which allows it to exist for a long time in the environment. Morphometric features of eggs, larvae and free-living S. westeri were studied in vitro under different temperature regimes. The optimal temperature for their embryonic development is 25 °С, under which 90% of the first stage rhabditiform larvae are formed and released within 7 hours of cultivation. The temperatures of 20 and 30 °С are less favorable for their development. Embryonic development of Strongyloides has four stages that differ in morphology and size. The eggs of a parthenogenetic female are 3.7% longer and 19.6% wider than eggs isolated from free-living females of S. westeri. In embryogenesis, the eggs shorten by 4.4 μm (6.5%) and widen by 5.35 μm (8.3%). New data were obtained on postembryonic development of S. westeri. The differential morphometric features of stage 1 and 2 rhabditiform larvae which grow both in length and width (33.7% and 30.4% respectively) are established. The development of filariform larvae is associated with loss of bulbous thickening and formation of cylindrical oesophagus. Simultaneously, the body elongates, and the gut becomes shorter. Differential morphometric features of free-living males and females of S. westeri are the length and width of body, length of oesophagus, gut, tail end, and size of spicules. Postembryonic development of the free-living and parasitic generations from rhabditiform larvae is temperature-dependent. Most of the free-living generations of Strongyloides (54.0%) are formed at 20 °С, and filariform larvae mostly (70.0%) develop at 30 °С. The obtained results of morphological studies improve differential diagnostics of the nematode at various stages of development and further advance the study of its intraspecific variability.


Sign in / Sign up

Export Citation Format

Share Document