Comparison of gross visual and microscopic assessment of four anatomic sites to monitor Besnoitia tarandi in barren-ground caribou (Rangifer tarandus)

The objective of this study was to establish a standardized protocol to monitor Besnoitia tarandi prevalence and intensity in barren-ground caribou (Rangifer tarandus) herds by: 1) calculating the relative sensitivity and specificity of the gross visual assessment of four anatomical sites compared with microscopic evaluation, and 2) determining which of four anatomical sampling sites was the most sensitive for detecting B. tarandi cysts by microscopy. Sampled tissues consisted of the conjunctiva of the left eye and skin sections from the rostrum, metatarsus, and thigh from 312 harvested caribou. Diagnosis of infection with B. tarandi was based on observation of at least one cyst by microscopic examination. For each tissue, the maximal density of cysts (number of B. tarandi cysts/mm(2) in the section examined) was calculated for a measured area consisting of the dermis extending from the epidermis of the skin to the base of the hair follicles and adnexal structures. For the conjunctiva, the entire submucosa was evaluated. Gross visual evaluation markedly underestimated B. tarandi prevalence in caribou with a relative sensitivity ranging from 0.29 in the conjunctiva to 0.13 in the skin section from the thigh, whereas relative specificities ranged from 0.98 to 1.00. The metatarsus and rostrum skin sections had the highest probabilities of cyst detection of all four anatomical sampling sites. The metatarsus harbored significantly higher densities of B. tarandi cysts than the rostrum, thigh, or conjunctiva. In conclusion, microscopic evaluation of a skin section from the anterior aspect of the mid-third portion of the metatarsal region could be used as a standardized comparative indicator of density of B. tarandi infection in Rangifer.     

Craniofacial Dermoid Cysts: Histological Analysis and Inter-site Comparison

Introduction: Dermoid cysts are common, benign, embryologically derived soft tissue cysts that can arise at a variety of craniofacial sites. It is not known whether specific histological variations exist between the different craniofacial sites. This study aims to establish whether inter-site histologic differences exist between periorbital, nasal, scalp, and postauricular dermoid cysts and analyze these in context of their distinct embryological origin and varied clinical presentation.Methods: A retrospective review of craniofacial dermoid cysts was performed. Using light microscopy with hematoxylin and eosin staining, histological appearance was directly compared between craniofacial sites.Results: All (n = 16) cysts contained keratinizing, stratified squamous epithelial lining, intraluminal keratin, and hair. Sebaceous glands were commonly present (n = 13). Eccrine (sweat) glands were less common (n = 3). Structures of mesodermal origin were seen in three periorbital cysts. Only the six ruptured cysts showed evidence of inflammation.Conclusions: Histological properties of dermoid cysts are conserved between craniofacial sites (periorbital, nasal, scalp, and postauricular). This reflects the consistency of ectodermal inclusion during early embryological development, which is independent of specific craniofacial site or surrounding anatomical structures.     

Encystation of Acanthamoeba castellanii: dye uptake for assessment by flow cytometry and confocal laser scanning microscopy

AIMS: To develop rapid means of distinguishing between cysts and trophozoites of the opportunistic pathogen, Acanthamoeba castellanii, the causative agent of keratitis. METHODS AND RESULTS: Fluorescence of Congo Red, Calcoflor White was specific for the endocyst wall; trophozoites did not become fluorescent. The anionic oxonol dye, DiBAC4(3), did not penetrate the cytoplasmic membrane after short-term (<5 min) exposure, whereas cysts are permeable and become fluorescent. Confocal scanning laser microscopy confirmed these properties and large populations of organisms were analysed by flow cytometry. CONCLUSION: These data provide a rapid alternative to traditional haemocytometer or plate counts for discrimination of trophozoites from cysts. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid and precise determination of the growth cycle of a dangerous ocular pathogen.     

Resting cysts of Parentocirrus hortualis Voß, 1997 (Ciliophora,Hypotrichia), with preliminary notes on encystation and various types of excystation

Resting cysts of Parentocirrus hortualis were investigated, using live observation, SEM and TEM. Processes during encystation and excystation were observed in vivo under the light microscope. During encystation, the trophic body becomes globular, the ciliature is resorbed in an anterior direction, the macronuclear nodules fuse into an elongated mass, and finally a cyst wall develops. As typical for oxytrichids, the resting cysts of P. hortualis are of the kinetosome-resorbing type and their wall is made of four layers: ectocyst, mesocyst, endocyst, and metacyst. The beginning of excystation is indicated by the formation of an excystation vacuole that helps the regenerating specimen to break the cyst wall. The excysting specimen leaves the resting cyst in a thin membrane that is gradually resorbed in the outer environment. Also two other excystation modes were observed. During the rare mode, the excystation vacuole breaks the thin membrane instead of the cyst wall that ruptures under the pressure of the body of the regenerating specimen. During the reproduction mode, the regenerating specimen divides within the resting cyst, producing two to four tomites. This is the first report of division in resting cysts of oxytrichids, but reproduction in division cysts was already described in keronopsids.     

Characterization of Selenaion koniopes n. gen., n. sp., an Amoeba that Represents a New Major Lineage within Heterolobosea,Isolated from the Wieliczka Salt Mine

A new heterolobosean amoeba, Selenaion koniopes n. gen., n. sp., was isolated from 73‰ saline water in the Wieliczka salt mine, Poland. The amoeba had eruptive pseudopodia, a prominent uroid, and a nucleus without central nucleolus. Cysts had multiple crater‐like pore plugs. No flagellates were observed. Transmission electron microscopy revealed several typical heterolobosean features: flattened mitochondrial cristae, mitochondria associated with endoplasmic reticulum, and an absence of obvious Golgi dictyosomes. Two types of larger and smaller granules were sometimes abundant in the cytoplasm—these may be involved in cyst formation. Mature cysts had a fibrous endocyst that could be thick, plus an ectocyst that was covered with small granules. Pore plugs had a flattened dome shape, were bipartite, and penetrated only the endocyst. Phylogenies based on the 18S rRNA gene and the presence of 18S rRNA helix 17_1 strongly confirmed assignment to Heterolobosea. The organism was not closely related to any described genus, and instead formed the deepest branch within the Heterolobosea clade after Pharyngomonas, with support for this deep‐branching position being moderate (i.e. maximum likelihood bootstrap support—67%; posterior probability—0.98). Cells grew at 15–150‰ salinity. Thus, S. koniopes is a halotolerant, probably moderately halophilic heterolobosean, with a potentially pivotal evolutionary position within this large eukaryote group.     

American Association of Clinical Endocrinology Disease State Clinical Review: The Clinical Utility of Minimally Invasive Interventional Procedures in the Management of Benign and Malignant Thyroid Lesions

ObjectiveThe objective of this disease state clinical review is to provide clinicians with a summary of the nonsurgical, minimally invasive approaches to managing thyroid nodules/malignancy, including their indications, efficacy, side effects, and outcomes.MethodsA literature search was conducted using PubMed and appropriate key words. Relevant publications on minimally invasive thyroid techniques were used to create this clinical review.ResultsMinimally invasive thyroid techniques are effective and safe when performed by experienced centers. To date, percutaneous ethanol injection therapy is recommended for recurrent benign thyroid cysts. Both ultrasound-guided laser and radiofrequency ablation can be safely used for symptomatic solid nodules, both toxic and nontoxic. Microwave ablation and high-intensity focused ultrasound are newer approaches that need further clinical evaluation. Despite limited data, encouraging results suggest that minimally invasive techniques can also be used in small-size primary and locally recurrent thyroid cancer.ConclusionSurgery and radioiodine treatment remain the conventional and established treatments for nodular goiters. However, the new image-guided minimally invasive approaches appear safe and effective alternatives when used appropriately and by trained professionals to treat symptomatic or enlarging thyroid masses.     

Focus: Zoonotic Disease: Retrospective Evaluation of Hydatid Cyst Cases During Pregnancy

Aim: Zoonotic parasite infections affect many pregnant people around the world. Hydatid cystic disease is also a zoonotic disease caused by Echinococcus sp. This study aims to present the maternal-fetal results and clinical treatment of pregnant women diagnosed with liver hydatid cyst (CH). This zoonotic disease is discussed again in the light of current literature. Materials and Methods: Pregnant women with hydatid cyst monitored in a tertiary health center between 2018 and 2020 were evaluated. Seven cases were included in this study. We retrospectively collected and analyzed clinical data, which did not interfere with medical treatment. Results: Albendazole was started as medical therapy in six patients, and percutaneous drainage was applied to one patient. Three of our six patients who started medical treatment had to undergo surgery due to maternal complications that developed despite medical treatment. Two of our patients were delivered with a cesarean section due to the obstetric indications. Discussion: Hydatid cysts are most commonly caused by Echinococcus granulosus infection and most common in the liver. The diagnosis of liver hydatid cysts is not difficult, but pregnant women’s treatment methods have some problems. Although both medical and surgical treatments are available, there is no consensus. We would also like to underscore that echinococcal disease of the liver should be kept in mind in the differential diagnosis of abdominal pain, jaundice, and/or fever, especially in endemic regions. We think that when we increase awareness about this disease, we can improve fetal and maternal outcomes by making an early diagnosis and management.     

Cytochemical and Electrophoretic Studies on the Cyst Wall of a Ciliate, Histriculus muscorum Kahl

The localization of proteins and polysaccharides in the cyst wall of a ciliate, Histriculus muscorum, was examined by light and electron microscope cytochemistry and by using an enzyme digestion test on thin sections. The endocyst and ectocyst were digested by treatment with pepsin or protease VI. The endocyst was intensely PAS-positive and alcian blue-positive. The ectocyst was also PAS- and alcian blue-positive, but the reaction was weaker than that of the endocyst. At the electron microscope level, an intensely positive reaction to methenamine silver was observed in the endocyst and a weak reaction in the ectocyst of the mature cyst. In the ectocyst of the encysting organism, however, the reaction to methenamine silver was more intense than that of the mature cyst. These results demonstrate the possible presence of glycoproteins in the ectocyst and endocyst. The mesocyst was negative to all cytochemical and enzyme digestion tests examined. The cyst wall, isolated by sonication, was analyzed by SDS-polyacrylamide gel electrophoresis. Two bands, 190 and 140 kilodaltons, were specific for the cyst wall. The 190 kilodalton band was the only PAS-positive band and its localization in the cyst was was discussed.     

The Excystment Process in the Ciliate Nassula ornata Ehrbg.

SYNOPSIS. The ciliate was cultured in natural lake water on Oscillatoria curviceps. Encystment was induced by lack of food. The resting cyst has 2 membranes—a faceted, lamellate ectocyst and a thin endocyst. The kinetosomal rows and the cytopharynx are retained in the cyst, and the position of the cytopharynx with respect to the rows remains unchanged. Thus, the usual polarity of the organism is preserved. Distilled water and plant infusions were relatively ineffective excystment-inducing agents, but a 0.1% (w/v) aqueous solution of peptone, pre-inoculated with wild bacteria, was highly effective. At 21 C it induced the excystment of 97% of the cysts within 2-3.5 hr. During excystment, the contractile vacuole enlarges, thereby producing pressure which ruptures the membranes. The ectocyst tears first—always at the posterior end of the cyst—the endocyst somewhat later. Emergence is largely by cytoplasmic streaming through a relatively small slit in the ectocyst, and the posterior end of the ciliate always emerges first.     

Influence of temperature and darkness on embryonic diapause termination in dormant Artemia cysts that have never been desiccated

Environmental cues for embryonic diapause termination (EDT) were investigated in the laboratory-produced encysted dormant embryos of the brine shrimp, Artemia franciscana. The cysts were spawned and kept throughout in a 2% sea salt solution. They were activated by various temperatures of the temperate zones or by continuous dark condition (DD), resulting in a state of EDT, and were thereafter able to resume their subsequent development and hatch under appropriate conditions. The level of EDT was conveniently assayed by a hatch test observed within 2 days at 28 degrees C under continuous light condition (LL). A cold treatment of the newly spawned dormant cysts, at 4 degrees C under DD for more than 100 days, resulted in more than 95% hatch of the dormant cysts. Similar treatments of the dormant cysts but at room temperature or 28 degrees C led to significantly different results (30-40% hatch). Almost all the residual non-hatched cysts derived from the above could hatch after an additional cold treatment (at 4 degrees C under DD for about 100 days). This might prove to be latent partial bivoltine in Artemia. Meanwhile, a rearing condition (28 degrees C under LL) induced the newly spawned cysts to hatch scatteredly at and after 1 month, resulting in 22% cumulative hatch on the 92nd day after spawning. When the newly spawned dormant cysts were pretreated at 28 degrees C under DD for 5 or 14 days and then reared at 28 degrees C under LL, the cumulative hatch significantly increased (60%). These results are discussed with respect to probable diapause regulator(s) involved in EDT.     

Morphological study of cysts of Pelomyxa palustris Greeff, 1874

Pelomyxa palustris Greeff, 1874, is the only representative of pelomyxoid amoebae with rest cysts in its life cycle. The morphology of the P. palustris cysts was studied using light and electron microscopy. The encystation of P. palustris under the climatic conditions of northwestern Russia occurs in August through September. The rest of the cysts have complex, trilaminar walls. The most developed are the two inner layers, i.e., the electron-dense structureless endocyst and the laminated mesocyst; the thickness of each layer can reach 0.6–0.7 μm. The thickness of the superficial electron-dense ectocyst lamina usually does not exceed 0.1–0.2 μm. The encysted cell of P. palustris has a unique structure. About 60% of its cell volume is occupied by a giant central vacuole filled with prokaryotic cytobionts. This vacuole has also been found to contain vacuoles and vesicles of different natures, restricted by vacuole membranes, autophagosomes, and lipid droplets. The amoeba cytoplasm occupies the space between the endocyst inner surface and the central vacuole. It contains no inclusions, prokaryotic cytobionts and most of cell organelles. In the cytoplasm there are 4 large nuclei filled with relatively homogeneous karyoplasm. The nuclear envelope forms numerous long tubular outgrowths, piercing the cytoplasm and underlining the central vacuole membrane. In this state the encysted pelomyxoids survive until the beginning of excystation. The excystation of P. palustris in the studied region occurs in spring, during the second half of April through the beginning of May. The cysts undergo complex morphofunctional changes due to reorganization of the wall and formation of young multinucleate amoebae. Out of the three initial lamina of the wall, only one persists until the moment of encystation. The central vacuole is destroyed and its content penetrates into the cytoplasm. Pelomyxoid nuclei divide twice. Prokaryotic cytobionts are localized in the cytoplasm and in the perinuclear area. Young multinuclear P. palustris individuals exiting cysts do not differ from the adult forms by their organization.     

有害甲藻孢囊的分类鉴定研究进展

有害甲藻孢囊主要是指能产生毒素和(或)能引起有害藻华发生并对水生态系统产生各种危害效应的甲藻孢囊。我国沿海共记录了10属18种,占全球有害甲藻孢囊的3/4。这些有害甲藻孢囊广泛分布于我国沿海,会对水产养殖业造成严重的经济损失,甚至会威胁人类的身体健康。因此,有害甲藻孢囊的多样性及分布越来越受到人们的关注。对有害甲藻孢囊的准确判断不仅对研究其多样性及分布至关重要,而且有助于水产品的安全检验和有害藻华的早期预警。对有害甲藻孢囊的分类主要存在鉴定困难、鉴定不准确等问题。本文综述了有害甲藻孢囊的危害、中国沿海有害甲藻孢囊的种类和分布,以及有害甲藻孢囊的鉴定等3个方面的研究进展,并提出利用孢囊及营养细胞的形态学特征、分子生物学、毒理学等多学科研究手段准确鉴定有害甲藻孢囊的建议。     

Effects of various physical and chemical factors on excystation of the encysted metacercariae of Echinostoma caproni

Various physical and chemical factors were studied to determine their effects on the viability of encysted metacercariae of Echinostoma caproni. Viability was equated with chemical excystation in an alkaline trypsin-bile salts (TB) medium. Control cysts showed excystation percentages of > 90% in TB. Excystation proved to be a more reliable criterion of cyst viability than observations by light microscopy. Isolated cysts and cysts left in the snail (in situ cysts) were studied. Generally, in situ cysts proved more resistant to various physical and chemical treatments than did isolated cysts. Cysts stored for 7 days at 28 C in a Locke's 1:1 solution showed 97% excystation, suggesting that cysts of this species would survive postal delays during shipment. Of numerous marinades tested, the one that was most harmful to isolated and in situ cysts was vinegar. Isolated and in situ cysts were killed by boiling (100 C) for 1 or 3 min, but freezing at -10 C did not kill all isolated or in situ cysts after 24 hr. Concentrations of potassium permanganate ranging from 300 to 1,200 mg/L killed most isolated cysts within 5 min, but in situ cysts survived these concentrations for 24 hr. Concentrated solutions of NaCl and sucrose had no effect on the viability of isolated and in situ cysts, suggesting that their use in food preparations for molluscs would not be effective in killing echinostomatid cysts in tainted snail tissues.     

Empirical validation of SAR values predicted by FDTD modeling.

Rapid increase in the use of numerical techniques to predict current density or specific absorption rate (SAR) in sophisticated three dimensional anatomical computer models of man and animals has resulted in the need to understand how numerical solutions of the complex electrodynamics equations match with empirical measurements. This aspect is particularly important because different numerical codes and computer models are used in research settings as a guide in designing clinical devices, telecommunication systems, and safety standards. To ensure compliance with safety guidelines during equipment design, manufacturing and maintenance, realistic and accurate models could be used as a bridge between empirical data and actual exposure conditions. Before these tools are transitioned into the hands of health safety officers and system designers, their accuracy and limitations must be verified under a variety of exposure conditions using available analytical and empirical dosimetry techniques. In this paper, empirical validation of SAR values predicted by finite difference time domain (FDTD) numerical code on sphere and rat is presented. The results of this study show a good agreement between empirical and theoretical methods and, thus, offer a relatively high confidence in SAR predictions obtained from digital anatomical models based on the FDTD numerical code.     

Enhanced affinity bifunctional bisphosphonates for targeted delivery of therapeutic agents to bone

Skeletal diseases have a major impact on the worldwide population and economy. Although several therapeutic agents and treatments are available for addressing bone diseases, they are not being fully utilized because of their uptake in nontargeted sites and related side effects. Active targeting with controlled delivery is an ideal approach for treatment of such diseases. Because bisphosphonates are known to have high affinity to bone and are being widely used in treatment of osteoporosis, they are well-suited for drug targeting to bone. In this study, a targeted delivery of therapeutic agent to resorption sites and wound healing sites of bone was explored. Toward this goal, bifunctional hydrazine-bisphosphonates (HBPs), with spacers of various lengths, were synthesized and studied for their enhanced affinity to bone. Crystal growth inhibition studies showed that these HBPs have high affinity to hydroxyapatite, and HBPs with shorter spacers bind more strongly than alendronate to hydroxyapatite. The HBPs did not affect proliferation of MC3T3-E1 preosteoblasts, did not induce apoptosis, and were not cytotoxic at the concentration range tested (10(-6)-10(-4) M). Furthermore, drugs can be linked to the HBPs through a hydrazone linkage that is cleavable at the low pH of bone resorption and wound healing sites, leading to release of the drug. This was demonstrated using hydroxyapatite as a model material of bone and 4-nitrobenzaldehyde as a model drug. This study suggests that these HBPs could be used for targeted delivery of therapeutic agents to bone.     

Justified concern or exaggerated fear: the risk of anaphylaxis in percutaneous treatment of cystic echinococcosis-a systematic literature review

Percutaneous treatment (PT) emerged in the mid-1980s as an alternative to surgery for selected cases of abdominal cystic echinococcosis (CE). Despite its efficacy and widespread use, the puncture of echinococcal cysts is still far from being universally accepted. One of the main reasons for this reluctance is the perceived risk of anaphylaxis linked to PTs. To quantify the risk of anaphylactic reactions and lethal anaphylaxis with PT, we systematically searched MEDLINE for publications on PT of CE and reviewed the PT-related complications. After including 124 publications published between 1980 and 2010, we collected a total number of 5943 PT procedures on 5517 hepatic and non-hepatic echinococcal cysts. Overall, two cases of lethal anaphylaxis and 99 reversible anaphylactic reactions were reported. Lethal anaphylaxis occurred in 0.03% of PT procedures, corresponding to 0.04% of treated cysts, while reversible allergic reactions complicated 1.7% of PTs, corresponding to 1.8% of treated echinococcal cysts. Analysis of the literature shows that lethal anaphylaxis related to percutaneous treatment of CE is an extremely rare event and is observed no more frequently than drug-related anaphylactic side effects.     

The potential for dispersal of microalgal resting cysts by migratory birds

Most microalgal species are geographically widespread, but little is known about how they are dispersed. One potential mechanism for long‐distance dispersal is through birds, which may transport cells internally (endozoochory) and deposit them during, or in‐between, their migratory stopovers. We hypothesize that dinoflagellates, in particular resting stages, can tolerate bird digestion; that bird temperature, acidity, and retention time negatively affect dinoflagellate viability; and that recovered cysts can germinate after passage through the birds’ gut, contributing to species‐specific dispersal of the dinoflagellates across scales. Tolerance of two dinoflagellate species (Peridiniopsis borgei, a warm‐water species and Apocalathium malmogiense, a cold‐water species) to Mallard gut passage was investigated using in vitro experiments simulating the gizzard and caeca conditions. The effect of in vitro digestion and retention time on cell integrity, cell viability, and germination capacity of the dinoflagellate species was examined targeting both their vegetative and resting stages. Resting stages (cysts) of both species were able to survive simulated bird gut passage, even if their survival rate and germination were negatively affected by exposure to acidic condition and bird internal temperature. Cysts of A. malmogiense were more sensitive than P. borgei to treatments and to the presence of digestive enzymes. Vegetative cells did not survive conditions of bird internal temperature and formed pellicle cysts when exposed to gizzard‐like acid conditions. We show that dinoflagellate resting cysts serve as dispersal propagules through migratory birds. Assuming a retention time of viable cysts of 2–12 h to duck stomach conditions, cysts could be dispersed 150–800 km and beyond.     

Anaerobes in the upper respiratory tract in infancy

Development of the indigenous microbiota begins on the surfaces of the human body after birth when infants are exposed to continuous person-to-person and environmental contacts with microbes. Anaerobes constitute a significant part of indigenous bacterial communities at different body sites. Pioneering anaerobic commensals are able to colonize and survive in the oral cavity during the first months of life. After teeth emerge, more attachment sites and potential niches are available for anaerobic bacterial colonization. Specific partner relationships influence the composition and stability of forming multigeneric communities, biofilms, where Fusobacterium nucleatum is of specific interest. In infancy, the oral colonization seems to be rather stable at species level, though not at clonal level. The colonization pattern in the nasopharynx is different from that in the oral cavity; anaerobes are absent from healthy nasopharynges but transiently colonize this anatomical site during infection. The most plausible origin for nasopharyngeal anaerobes is the oral cavity and, conceivably, saliva is the most likely transmission vehicle. Whether anaerobic bacteria colonize the nasopharynx just because of ecological changes favoring their growth or whether they could play an active role in the pathogenesis of respiratory infections is not known.     

Determination of biological and physicochemical parameters of Artemia franciscana strains in hypersaline environments for aquaculture in the Colombian Caribbean

Background

Artemia (Crustacea, Anostraca), also known as brine shrimp, are typical inhabitants of extreme environments. These hypersaline environments vary considerably in their physicochemical composition, and even their climatic conditions and elevation. Several thalassohaline (marine) environments along the Colombian Caribbean coast were surveyed in order to contribute to the knowledge of brine shrimp biotopes in South America by determining some vital biological and physicochemical parameters for Artemia survival. Additionally, cyst quality tests, biometrical and essential fatty acids analysis were performed to evaluate the economic viability of some of these strains for the aquaculture industry.

Results

In addition to the three locations (Galerazamba, Manaure, and Pozos Colorados) reported in the literature three decades ago in the Colombian Caribbean, six new locations were registered (Salina Cero, Kangaru, Tayrona, Bahía Hondita, Warrego and Pusheo). All habitats sampled showed that chloride was the prevailing anion, as expected, because of their thalassohaline origin. There were significant differences in cyst diameter grouping strains in the following manner according to this parameter: 1) San Francisco Bay (SFB-Control, USA), 2) Galerazamba and Tayrona, 3) Kangarú, 4) Manaure, and 5) Salina Cero and Pozos Colorados. Chorion thickness values were smaller in Tayrona, followed by Salina Cero, Galerazamba, Manaure, SFB, Kangarú and Pozos Colorados. There were significant differences in naupliar size, grouping strains as follows (smallest to largest): 1) Galerazamba, 2) Manaure, 3) SFB, Kangarú, and Salina Cero, 4) Pozos Colorados, and 5) Tayrona. Overall, cyst quality analysis conducted on samples from Manaure, Galerazamba, and Salina Cero revealed that all sites exhibited a relatively high number of cysts.g^-1. Essential fatty acids (EFA) analysis performed on nauplii from cyst samples from Manaure, Galerazamba, Salina Cero and Tayrona revealed that cysts from all sites exhibited high arachidonic acid:20:4(n-6) (ArA) and eicosapentaenoic acid: 20:5(n-3) (EPA) levels comparable to the control sample (SFB). In contrast, most cysts collected (including SFB) at different locations, and during different months, presented low docosahexaenoic acid: 22:6(n-3) (DHA) levels (Manaure was the only exception with high DHA levels). Some variations in EPA and ArA levels were observed in all sites, contrasting with the much lower DHA levels which remained constant for all locations, except for Manaure which exhibited variable DHA levels. DHA/EPA ratio was overall very low for all sites compared to SFB cysts. All strains had a low DHA/ArA, but a high EPA/ArA ratio, including the control.

Conclusion

The Colombian A. franciscana habitats analyzed were determined to be thalassohaline, and suitable for A. franciscana development. EFA profiles demonstrated that Tayrona, Galerazamba, Manaure and Salina Cero strains are suitable food for marine fish and crustacean culture because of their high EPA/ArA ratio, but might have to be fortified with DHA rich emulsions depending on the nutritional requirements of the species to be cultured, because of their overall low DHA content. The relatively small nauplii are appropriate for marine larvaeculture. In contrast, the strains from Tayrona, Kangarú, Salina Cero, and Pozos Colorados may be of use but limited to Artemia small biomass production quantities, because of the small surface area of their respective locations; Artemia could be exploited at these locations for local aquaculture applications. In general, cyst quality evaluation for Manaure, Salina Cero and Galerazamba cysts revealed that cysts from these three locations could improve their quality by concentrating efforts on cyst processing techniques. Finally, most locations had great A. franciscana production potential and require different degrees of water quality and/or infrastructure management.     

The morphological study of the cysts Pelomyxa palustris Greeff, 1874

Pelomyxa palustris Greeff, 1874, is the only species of pelomixoid amoebas with the rest cysts in its life cycle. The morphology of the P. palustris has been studied by the light and electronic microscopy. Encystation of P. palustris under climatic conditions of North-West of Russia occurs within August-September. Rest cysts have a complex, trilaminar wall. Two inner lamina are the dense endocyst and the laminated mesocyst, thickness of each layer runs up to 0.6-0.7 microm. Thickness of the electron-dense ectocyst usually does not exceed 0.1-0.2 microm. The encystated cell of P. palustris has the unique structure. About 60 % of the cell volume are occupied by a huge vacuole placed in the center and filled up with the prokaryotic cytobionts. Different vacuoles, small vesicles of various nature, autophagosomes and lipid drops could be found inside that huge vacuole. The amoebae cytoplasm occupies the space in between endocyst's inner surface and the central vacuole. No any inclusions, prokaryotic cytobionts and most of cell organelles are absent in the cytoplasm. There are 4 large nuclei filled with relatively homogeneous karyoplasm lying in the cytoplasm. Nuclear envelope forms a lot of long tubular channels, running through the cytoplasm and lining the membrane of the central vacuole. Encysted pelomixoid stay in this state up until the beginning of excystation. Excystation of P. palustris in the studied region occurs in spring, during the latter half of April and the beginning of May. Cysts undergo complex morphofunctional changes, related to the reorganization of the wall and formation of young multinucleate amoebas. Only one wall lamina of the 3 initial ones is left up to the moment of excystation. The central vacuole endures ruination and its content penetrates into the cytoplasm. Pelomixoid nuclei divide twice. Prokaryotic cytobionts are localized in cytoplasm and in the perinuclear area. Young multinuclear species of P. palustris coming out of the cysts do not differ in their structure from the adult forms.