Endogenous stages of Eimeria sigmodontis (Protozoa: Eimeriidae) in the cotton rat, Sigmodon hispidus

Endogenous stages of Eimeria sigmodontis were studied in experimentally infected cotton rats, Sigmodon hispidus. The parasites were located in mucosal epithelial cells of the cecum and colon. E. sigmodontis had a typical coccidian endogenous cycle that consisted of three asexual schizogonous stages and sexual stages composed of the macrogametes and microgametes. The first-, second-, and third-generation schizonts contained 10–17, 5–8 and 10–21 merozoites, respectively. Only one type of wall-forming body was present in the macrogamete. The microgametocyte had a monocentric type of development.     

A coccidian infection of the kidney of the native littleneck clam, Protothaca staminea

During the summer of 1980, numerous exposed native littleneck clams (Protothaca staminea) were observed in Sequim Bay, Washington. The kidneys of these clams were found to be parasitized by a coccidian. Heavily parasitized kidney was characterized by massive epithelial destruction and an intense inflammatory response. In addition, large numbers of “brown cells” were present throughout the tissues of infected clams.Infected kidneys harbored several stages of the life history of the parasite. The present coccidian differs markedly in several respects from other coccidians in bivalves. First, the entire life history apparently occurs in the kidney of the clam, in contrast to the presumed involvement of an alternative host in other known coccidian parasites of bivalves. Second, a thick-walled spore is produced in addition to a thin-walled oocyst. The apparent presence of two types of dissemination stages is unknown in the Coccidia. Finally, the sporocysts are tetrazoic as opposed to dizoic or multiple sporocysts in previously described species. This parasite, therefore, appears to represent an undescribed genus and species of Coccidia.     

Discovery of three novel coccidian parasites infecting California sea lions (Zalophus californianus), with evidence of sexual replication and interspecies pathogenicity

Enteric protozoal infection was identified in 5 stranded California sea lions (Zalophus californianus). Microscopically, the apical cytoplasm of distal jejunal enterocytes contained multiple stages of coccidian parasites, including schizonts with merozoites and spherical gametocytes, which were morphologically similar to coccidians. By histopathology, organisms appeared to be confined to the intestine and accompanied by only mild enteritis. Using electron microscopy, both sexual (microgametocytes, macrogamonts) and asexual (schizonts, merozoites) coccidian stages were identified in enterocytes within parasitophorous vacuoles, consistent with apicomplexan development in a definitive host. Serology was negative for tissue cyst-forming coccidians, and immunohistochemistry for Toxoplasma gondii was inconclusive and negative for Neospora caninum and Sarcocystis neurona. Analysis of ITS-1 gene sequences amplified from frozen or formalin-fixed paraffin-embedded intestinal sections identified DNA sequences with closest homology to Neospora sp. (80%); these novel sequences were referred to as belonging to coccidian parasites "A," "B," and "C." Subsequent molecular analyses completed on a neonatal harbor seal (Phoca vitulina) with protozoal lymphadenitis, hepatitis, myocarditis, and encephalitis showed that it was infected with a coccidian parasite bearing the "C" sequence type. Our results indicate that sea lions likely serve as definitive hosts for 3 newly described coccidian parasites, at least 1 of which is pathogenic in a marine mammal intermediate host species.     

Role of proteasomes in non-specific immune response of marine annelids

We investigated functioning of proteasomes and chaperones in Arenicola marina coelomocytes in conditions of lipopolysaccharide-induced inflammation. We observed the increase of chymotrypsin-like proteasome activity in coelomocytes 1 h after induction. Amount of proteasome subunits alpha- and beta-5 types increased as well. We also detected appearance of a new form of Hsp70 chaperone in infected coelomocytes. Our results allow us to consider the changes in proteasome structure and induction of chaperones as principle mechanisms in stress adaptation and defensive reactions development in annelids.     

Coccidian-like Nature of Toxoplasma gondii

Specific pathogen-free domestic cats were fed with tissue cysts containing Toxoplasma gondii. In two infected cats large numbers of oocysts were produced in the faeces; no oocysts were observed in the faeces of the uninfected control cat. Five days after the feeding of the toxoplasms profuse schizogonic and gametogonic stages were observed in the epithelial cells of the small intestine of one infected cat. A single schizont was observed in an intestinal epithelial cell of a second cat six days after being fed the tissue cysts. There was no evidence of schizogony or gametogony in the uninfected control cat. The stages observed in the intestinal epithelium are identical with those of the well-known endogenous cycles of coccidian parasites. The appearance of these stages, together with the nature of the oocyst, indicates that T. gondii is a coccidian parasite closely related to the genus Isospora.     

In vitro cultivation methods for coccidian parasite research

The subclass Coccidia comprises a large group of protozoan parasites, including important pathogens of humans and animals such as Toxoplasma gondii, Neospora caninum, Eimeria spp., and Cystoisospora spp. Their life cycle includes a switch from asexual to sexual stages and is often restricted to a single host species. Current research on coccidian parasites focuses on cell biology and the underlying mechanisms of protein expression and trafficking in different life stages, host cell invasion and host-parasite interactions. Furthermore, novel anticoccidial drug targets are evaluated. Given the variety of research questions and the requirement to reduce and replace animal experimentation, in vitro cultivation of Coccidia needs to be further developed and refined to meet these requirements. For these purposes, established culture systems are constantly improved. In addition, new in vitro culture systems lately gained considerable importance in research on Coccidia. Well established and optimized in vitro cultures of monolayer cells can support the viability and development of parasite stages and even allow completion of the life cycle in vitro, as shown for Cystoisospora suis and Eimeria tenella. Furthermore, new three-dimensional cell culture models are used for propagation of Cryptosporidium spp. (close relatives of the coccidians), and the infection of three-dimensional organoids with T. gondii also gained popularity as the interaction between the parasite and host tissue can be studied in more detail. The latest advances in three-dimensional culture systems are organ-on-a-chip models, that to date have only been tested for T. gondii but promise to accelerate research in other coccidians. Lastly, the completion of the life cycle of C. suis and Cryptosporidium parvum was reported to continue in a host cell-free environment following the first occurrence of asexual stages. Such axenic cultures are becoming increasingly available and open new avenues for research on parasite life cycle stages and novel intervention strategies.     

球虫感染与捕食对根田鼠繁殖性能的影响

在自然界,捕食者和寄生物是两种主要的种群外部调节因子,二者的交互作用会对猎物和宿主种群波动产生深远影响。较低的球虫感染强度与捕食对根田鼠（Alexandromys oeconomus）繁殖无显著的交互作用。自然界球虫感染存在季节性变化,秋季感染强度最高。为了探究较高感染强度下,球虫与捕食对根田鼠繁殖的主效应及交互作用,本研究采用2×2析因实验设计,在野外围栏中测定了根田鼠种群肠道内寄生物的感染率和感染强度、雄性睾丸指数、睾酮水平、精子密度、精子活力以及雌性卵巢指数。结果表明,较高的感染强度下,球虫能显著抑制根田鼠的繁殖性能,但球虫感染与捕食对根田鼠的繁殖无显著的交互作用,这可能与球虫感染和捕食效应在时间上的错配有关。本研究认为,球虫感染对繁殖期小哺乳动物种群的调节作用虽有限,但其可通过与捕食者的耦合来降低宿主越冬时的存活率,进而影响宿主种群波动。     

Observations on Ichthyosporidium giganteum (Microsporida) with particular reference to the host-parasite relations during merogony

Connective tissue cells, particularly fibroblasts, of the fish Leiostomus xanthurus Lacépède respond to the invading microsporidian parasite Ichthyosporidium sp. [assumed to be identical with Ichthyosporidium giganteum (Thélohan)] by proliferating themselves, coalescing into a syncytium, synthesizing copious amounts of cytoplasm around the parasites, and walling off the parasitized islands of cytoplasm with fibrous capsules. The resulting cysts are xenoparasitic complexes of the syncytial xenoma type, clearly different from the cell hypertrophy tumor (xenoma sensu Weissenberg) exemplified by the Glugea cyst. These findings involve a new concept of the structure and host-parasite relations of Ichthyosporidium. Formerly, the parasitized masses of cytoplasm were interpreted as extracellular plasmodial stages of the parasite (stages uncharacteristic of the microsporidia), while the parasites themselves were interpreted as nuclei of the "plasmodia." Actually, the parasite undergoes merogony in parasitophorous vacuoles which coalesce before sporogony begins. The nuclei of the mermonts are very small chromatin granules, becoming transformed into large basophilic diplokarya of the sporonts. Sporulation is diplokaryotic throughout, the diplokarya becoming reduced in size through 2 steps during sporogony.     

野外围栏条件下根田鼠肠道寄生物的感染率

寄生物是调节小哺乳动物种群波动的因子之一。许多生物和非生物因子可影响寄生物的感染。本研究以根田鼠为对象,在野外围栏条件下测定了根田鼠肠道寄生物种类及其感染率,并分析了根田鼠建群者数量、性别、年龄及时间对寄生物感染率的影响。结果表明,根田鼠肠道寄生物有球虫、线虫和绦虫,球虫是主要寄生物, Eimeria wenrichi 为球虫中的优势种;其感染率随根田鼠建群者数量增加而增加,且随时间而显著变化。在根田鼠幼体中,高密度处理组E. wenrichi 的感染率显著高于低密度处理组,而根田鼠幼体的E. ochrogasteri 感染率显著高于成体。本研究未发现根田鼠性别间的球虫感染率差异。上述结果说明,在根田鼠种群中,球虫感染率有时间变化,且受宿主密度的制约,高密度种群引起的生前应激与当前密度制约性感染可能对球虫感染率具有耦合效应。     

Coccidian infection may explain the differences in the life history of octopus host populations

The prevalence of coccidian parasites in three Octopus tehuelchus populations from San Matías Gulf (Patagonia, Argentina) is compared. The prevalence was similar between sexes, but varied between seasons (being highest during cold months) and sites. Islote Lobos had the highest prevalence (42.7–100%) followed by San Antonio Bay (0–66%) and El Fuerte (0–24.5%). Octopuses under 27 mm of dorsal mantle length showed a low prevalence (less than 50%), which increased with size. We hypothesize that the high prevalence of parasites, which affect the three populations differentially, could account for the observed variability in life-span and growth, size–frequency distributions, reproduction and densities of O. tehuelchus populations.     

Occurrence of Apicomplexa-like structures in the digestive gland of Strombus gigas throughout the Caribbean

The queen conch, Strombus gigas, is a marine resource of ecological and economical importance in the Caribbean region. Given its importance in this region, and the critical status of most populations, the reproductive biology of this species has been studied to support management decisions. It was from these studies that a generalized sporozoan infection was detected. This study describes the geographic distribution of a coccidian (Apicomplexa) parasite infecting the digestive gland of S. gigas throughout the Caribbean. The parasite was present in every location sampled. Based on histological analysis, the parasites from all locations are similar and appear to complete their life cycle within the digestive gland. The highest occurrence of the parasites was registered in samples from Puerto Rico (54 parasites per field) and Martinique (45 parasites per field). The lowest incidence was registered on the Mexican coast of Yucatan peninsula, at Alacranes and Chinchorro with 17 parasites per field. Data showed significant differences among sites (Kruskal Wallis H = 106.957; p ? 0.05). The abundance of parasites found in the digestive ducts and in the faeces suggests the liberation of parasites to the environment. A gradual decrease in abundance was found from East to West of the Caribbean sea.     

Disruption of a mitochondrial protease machinery in Plasmodium falciparum is an intrinsic signal for parasite cell death

The ATP-dependent ClpQY protease system in Plasmodium falciparum is a prokaryotic machinery in the parasite. In the present study, we have identified the complete ClpQY system in P. falciparum and elucidated its functional importance in survival and growth of asexual stage parasites. We characterized the interaction of P. falciparum ClpQ protease (PfClpQ) and PfClpY ATPase components, and showed that a short stretch of residues at the C terminus of PfClpY has an important role in this interaction; a synthetic peptide corresponding to this region antagonizes this interaction and interferes with the functioning of this machinery in the parasite. Disruption of ClpQY function by this peptide caused hindrance in the parasite growth and maturation of asexual stages of parasites. Detailed analyses of cellular effects in these parasites showed features of apoptosis-like cell death. The peptide-treated parasites showed mitochondrial dysfunction and loss of mitochondrial membrane potential. Dysfunctioning of mitochondria initiated a cascade of reactions in parasites, including activation of VAD–FMK-binding proteases and nucleases, which resulted in apoptosis-like cell death. These results show functional importance of mitochondrial proteases in the parasite and involvement of mitochondria in programmed cell death in the malaria parasites.     

Morphology and ultrastructure of the uterus of Lineolepis scutigera (Dujardin, 1845) Karpenko, 1985 (Cestoda,Cyclophyllidea, Hymenolepididae) in formation of uterine capsules

The microanatomy and ultrastructure of the uterus in different stages of ontogenesis were studied in a cyclophyllidean cestode Lineolepis scutigera. In the early stages, developing embryos lie freely in the cavity of a single unbroken uterus, which is later fragmented into separate compartments. As a result, numerous spherical uterine capsules are formed; each of them contains one formed egg. Capsules represent a fragmented but actively functioning uterus. Muscular cells containing numerous lipid inclusions are located around them. In the final stages, placenta-like relationships are formed between eggs and the epithelium of capsules. A comparative morphofunctional analysis of uterine capsules in cestodes is presented. Attention is paid to the formation of close interactions between the uterine epithelium and developing eggs.     

Cellular and biochemical responses to environmental and experimentally induced stress in sea urchin coelomocytes

Coelomocytes are considered to be immune effectors of sea urchins. Subpopulations of coelomocytes can be purified from a total cell suspension. The proportion of each cell type can vary not only among species, but also between individuals of the same species, according to their size and physiological conditions. We tested the hypothesis that coelomocytes play a role in defense mechanisms activated by adverse external conditions. Total coelomocytes from control and stressed (temperature, pollution, and injuries) sea urchins were analyzed for their expression of the 70 kDa heat shock protein (hsp70), a well recognized stress marker. Further analysis was performed by separation of coelomocytes into subpopulations by step gradients. We demonstrated that sea urchin coelomocytes respond to temperature shock and to polluted seawater by the upregulation of hsp70. Among coelomocytes certain cells, known as red spherula cells, showed a great increase in number in animals collected from polluted seawaters or subjected to "accidental" injury. The present study confirms the immunological function of sea urchin coelomocytes, as indicated by the upregulation of the hsp70 molecular marker, and suggests that sea urchin coelomocytes can be utilized as sensitive bio-indicators of environmental stress.     

Survey of gastrointestinal parasitic infections of beef cattle in regions under Mediterranean weather in Greece

Gastrointestinal (GI) parasitic infections of beef cattle were investigated in 15 farms from 3 different regions of Greece, characterized by temperate Mediterranean climate, during two years. A total of 262 faecal samples were collected. Helminth eggs were detected in 42 (16%) samples. Strongyle-type eggs were found in 28 (10.7%) samples, Strongyloides spp. and Toxocara spp. eggs in 8 (3.1%) samples each, and Capillaria spp. and Moniezia spp. eggs in 1 (0.4%) sample each. Coccidian oocysts were found in 123 (46.9%) samples. A logistic regression model showed that the age of animals, the number of employees, and the region of animals were associated with coccidian infections. There was almost a four-fold increase in the risk of coccidian infections in calves less than 12 months old compared with animals that were more than 36 months old (p < 0.05). The risk of coccidian infections was 0.07, 0.18 and 0.27-fold lower for farms with 1, 2 and 3 employees respectively compared to farms with 4 employees; farms with less than 4 employees had a protective effect (p < 0.05 in all cases). Almost a four-fold increased risk of coccidian infections was shown for animals from Epirus compared with those from Thessaly (p < 0.05). These findings are useful to formulate appropriate control strategies for coccidian parasites of beef cattle in Greece and other areas with similar climatic conditions.     

Toxoplasma gondii: the enzymic defect of a mutant resistant to 5-fluorodeoxyuridine.

We have previously described a mutant of Toxoplasma gondii that was 100-fold more resistant to 5-fluorodeoxyuridine, as measured by growth in human fibroblast cultures. Various pyrimidine salvage enzymes were measured in the wild type and the mutant parasites to determine the biochemical basis for resistance to fluorodeoxyuridine. Both the resistant mutant and the wild type parasite had little or no uridine kinase, an enzyme readily detectable in the human fibroblast host cells. Uridine and deoxyuridine phosphorylases were found in both parasites while human fibroblasts had much less of these enzymes. The critical difference between the mutant and the wild type parasites proved to be a 100-fold lower concentration of uracil phosphoribosyltransferase in the fluorodeoxyuridine-resistant mutant. A back mutant of the resistant strain, selected for its ability to use uracil, simultaneously regained uracil phosphoribosyltransferase and sensitivity to fluorodeoxyuridine. This enzymic evidence together with previously published data show that in wild type T. gondii, deoxyuridine is incorporated into nucleic acids through a phosphorolysis to produce uracil which is then converted to uridylic acid by uracil phosphoribosyltransferase.     

Characterisation of immune responses in the pea aphid, Acyrthosiphon pisum

The innate immune system of insects provides effective defence against a range of parasites and pathogens. The pea aphid, Acyrthosiphon pisum, is a novel study system for investigating host-parasite interactions due to its complex associations with both well-characterised bacterial symbionts and a diversity of pathogens and parasites, including several important biological control agents. However, little is known about the cellular and humoral immune responses of aphids. Here we identify three morphologically distinct types of haemocytes in circulation that we name prohemocytes, granulocytes and oenocytoids. Granulocytes avidly phagocytose Gram negative Escherechia coli and Gram positive Micrococcus luteus while oenocytoids exhibit melanotic activity. Prohaemocytes increase in abundance immediately following an immune challenge, irrespective of the source of stimulus. Pea aphids form melanotic capsules around Sephadex beads but do not form cellular capsules. We also did not detect any antimicrobial peptide activity in the haemolymph using zone of inhibition assays. We discuss these results in relation to recent findings from the pea aphid genome annotation project that suggest that aphids have a reduced immune gene repertoire compared to other insects.     

Time of day, age and feeding habits influence coccidian oocyst shedding in wild passerines

Protozoan coccidia are one of the most common intestinal parasites in birds. Ordinary coccidian detection and quantification techniques have proved to be inaccurate for wild passerines due to the existence of marked oocyst shedding rhythms throughout the day. Previous studies have suggested that these rhythms should be taken into account when analysing coccidian load and prevalence data, but their pattern and magnitude still remain poorly known. In this study we characterised shedding rhythms in the field by means of 406 samples of faeces taken from two species of passerines with different diets: the European Serin (a granivorous species), and the Garden Warbler (an insectivorous species). Both coccidian prevalence and load were two-phased, with maximums occurring in the afternoon. Oocyst elimination remained consistently high during the second half of the day, whereas prevalence peaked during the afternoon, lowering throughout the evening. This pattern was found in both species. We found a high repeatability of prevalence and intensity when differences between the morning and afternoon were statistically controlled. As a result, we suggest that sampling periods used in the analysis of coccidian prevalence and/or load studies should take into account these differences in times of shedding and be limited to the afternoon, otherwise a statistical control of this factor will be required.     

A game-theoretic model of interspecific brood parasitism with sequential decisions

The interaction between hosts and parasites in bird populations has been studied extensively. This paper uses game-theoretic methods to model this interaction. This has been done in previous papers but has not been studied taking into account the detailed sequential nature of this game. We introduce a model allowing the host and parasite to make a number of decisions which will depend on various natural factors. The sequence of events begins with the host forming a nest and laying a number of eggs, followed by the possibility that a parasite bird will arrive at the nest; if it does it can choose to destroy some of the host eggs and lay one of its own. A sequence of events follows, which is broken down into two key stages; firstly the interaction between the host and the parasite adult, and secondly that between the host and the parasite chick. The final decision involves the host choosing whether to raise or abandon the chicks that are in the nest. There are certain natural parameters and probabilities which are central to these various decisions; in particular the host is generally uncertain whether parasitism has taken place, but can assess the likelihood of parasitism based upon certain cues (e.g. how many eggs remain in its nest). We then use this methodology to model two real-world interactions, that of the Reed Warbler with the Common Cuckoo and also the Yellow Warbler with the Brown-headed Cowbird. These parasites have different methods in the way they parasitize the nests of their hosts, and the hosts can in turn have different reactions to these parasites. Our model predictions generally match the real results well, and the model also makes predictions of the effect of changes in various key parameters on the type of parasitic interactions that should occur.