Biology of parasitic protozoa IV. Apicomplexa I (SAR) Andrea Bardůnek Valigurová andreav@sci.muni.cz Notice This presentation contains some material available on the web without the permission of the creator or the copyright owner. This presentation is to be used for educational purposes only. Educational purposes are defined as a communicating material for a particular course of instruction or for the administration of such course. Educational purposes do not cover the use of copyright material in PowerPoint for public lectures or other purposes. 5 supergroups = megagroups cortical alveolae Tetrahymena (Ciliophora) SAR (Harosa) - Alveolata • SAR supergroup: Stramenopiles (heterokonts), Alveolata, and Rhizaria • Alveolata: monophyletic group; 3 main subgroups: ciliates, dinoflagellates and apicomplexans • extremely diverse modes of nutrition, such as predation, photoautotroph and intracellular parasitism • cortical alveolae = system of flattened vesicles packed into a continuous layer supporting the membrane, sometimes secondarily lost • ciliary pit or micropore • mitochondrial cristae tubular or ampulliform Cortical alveolae Ciliata Dinozoa Apicomplexa Apicomplexa • monophyletic taxon • 1.2 -10 million apicomplexan species, only about 0.1% have been named and described to date • all parasitic = obligate parasites of vertebrates and invertebrates • monoxenic or heteroxenous parasites • at least 1 stage of the life cycle with flattened cortical alveoles and apical complex • apical complex = 1 or more polar rings, conoid, rhoptries, micronemes, subpellicular microtubules • cell movement / locomotion by gliding, body flexion, longitudinal folds, and/or cilia • complicated life cycle usually comprising sexual (gametogony) and asexual (merogony also known as schizogony, sporogony) part Apicomplexan life cycle (general scheme) Apicomplexan life cycle (general scheme) Environment (exogenous part) Intermediate host Asexual reproduction Reproduction in Apicomplexa • Asexual reproduction of haploid stages via merogony, endodyogeny, endopolyogeny and/or binary fission ❑ merogony (schizogony) = multiple fission in which nuclei and other organelles in the meront (schizont) divide repeatedly and migrate to the cell periphery before internal membranes develop around them to produce a large number of daughter cells (merozoites) = budding takes place at the plasma membrane, nuclear division occurs before daughter assembly ❑ endopolyogeny = internal budding resembling merogony, but budding takes place internally within the cytoplasm, daughter assembly precedes nuclear division ❑ endodyogeny = only 2 daughter cells (endozoites) are assembled within the mother ❑ sporogony = asexual division, development of zygote (motile zygote = ookinete) producing sporozoites within oocysts (usually inside sporocysts); reductive division of the zygote forms haploid nuclei which (along with other organelles) undergo multiple divisions to form a sporoblast; sporozoites arise by cytoplasmic division of the sporoblast (sporulation) https://doi.org/10.3389/fcimb.2020.00269 Sexual reproduction Reproduction in Apicomplexa • sexual reproduction (where known) by syngamy followed by immediate meiosis to produce haploid progeny ❑ gametogony (gamogony) = transformation into a gamonts that produce male and female gametes ▪ syzygy and gametocyst formation (gregarines) ▪ isogamy (similar gametes) x anisogamy (dissimilar gametes, flagelatted microgametes in some taxa) ▪ syngamy (fusion of gametes) Fertilisation in T. gondii Oocyst = resistant stage of apicomplexans Apicoplast ✓ relict, non-photosyntethic plastid ✓ secondary endosymbiotic chloroplast limited by 4 membranes ✓ 35 kb long circular DNA strand that codes for approximately 30 proteins ✓ heme and amino acid syntheses and likely involved in lipid metabolism ✓ sensitive to herbicides ✓ promising drug target Zoite organisation • standard equipment of the eukaryotic cell • organelles characteristic of Apicomplexa: apical complex and apicoplast • conoid = hollow cone of spirally arranged fibers located within the polar rings • polar ring(s) = 1-2 electron dense structures located just behind the zoite‘s apical tip and directly under the plasma membrane • micronemes = convoluted, elongate bodies in zoite's anterior end whose contents are released during host cell invasion, likely involved in affecting plasma membrane fluidity • rhoptries - elongated, club-shaped organelles in apical region of zoite whose contents are released during host cell invasion, likely involved in forming the membranous scaffolding supporting the zoite within the parasitophorous vacuole (PV) Tachyzoite of Toxoplasma gondii Bradyzoite of T. gondii Apical complex • micropores = usually present at some stage which are morphologically similar to clathrin-coated pits and are involved in endocytosis • subpellicular microtubules = microtubules under the pellicle running posteriorly (parallel to the cell axis) Apical complex Invasion process in Toxoplasma gondii Host cell invasion Parasite's superficial antigen ▪ zoite attachment, orientation and adhesion by its apical end  moving junction ▪ host cell penetration and formation of parasitophorous vacuole ▪ shedding of the zoite antigenic surface completion of PV and modification of the parasite's niche https://doi.org/10.1371/journal.ppat.0010017 Parasite niche in host cell or tissue ✓ intercellular ✓ extracellular ✓ epicellular ✓ intracellular epiplasmatic ✓ intracellular Hypothesis on evolution of parasitism in Apicomplexa: Myzocytotic predation  myzocytotic extracellular parasitism + origin of epicellular parasitism (modification of attachment apparatus and motility in trophozoite)  intracellular parasitism (lost of cell polarity and motility in trophozoite) Host-parasite interactions in Apicomplexa Valigurová et al. 2015 in https://doi.org/10.1371/journal.pone.0125063 Phylogenetic distribution of diverse organism-environment interactions in Myzoza https://doi.org/10.1016/j.pt.2020.06.002 Co-occurrence of predation and parasitism. Colpodellids (A) are predators of algae and protists. They penetrate the cell surface to access the cytosol of the predated cell. The mode of action is, in principle, quite similar to that of early-branching apicomplexan parasites such as gregarines (B) living in the gut of various animals. Apicomplexa Multiprotein phylogeny of Apicomplexa and related taxa ➢ apicomplexans are polyphyletic ➢ similar morphological features appeared convergently at least 3x ➢ gregarines and eugregarines are monophyletic ➢ Eleutheroschizon is related to Eucoccidia and Rhytidocystis has emerged as the basal group of Coccidiomorpha (the most medically important representatives) ➢ plastids are common in basal Apicomplexa; in eugregarines they are abnormally reduced or absent https://doi.org/10.7554/eLife.49662 Apicomplexa Aconoidasida••• …………………….next lecture • apical complex lacking conoid in asexual motile stages; some diploid motile zygotes (ookinetes), with conoid; macrogametes and microgametes forming independently; heteroxenous • Haemospororida•••• • Piroplasmorida•••• Conoidasida•••………………………this lecture • Gregarinasina•••• • Cryptosporidium•••• • Coccidia•••• • Adeleorina••••• • Eimeriorina••••• Apicomplexa Conoidasida••• • complete apical complex, including a conoid in all or most asexual motile stages Gregarinasina•••• • mature gamonts usually develop extracellularly; syzygy of gamonts generally occurring with production of gametocyst; similar numbers of macrogametes and microgametes maturing from paired gamonts in syzygy within the gametocyst • syngamy of mature gametes leading to gametocyst that contains few to many oocysts Cryptosporidium•••• • endogenous stages with attachment “feeder” organelle; microgametes non ciliated; oocysts without sporocysts, with 4 naked sporozoites • extracytoplasmic localisation in host cell (based on Adl et al. 2012) vs. epicellular (Valigurová et al. 2008, 2015) Coccidia•••• • mature gametes develop intracellularly; microgamont typically produces numerous microgametes • syzygy absent; zygote rarely motile; sporocysts usually form within oocysts Apicomplexa Gregarinasina • gregarines are highly diverse - intestine and body cavities of invertebrates (arthropods, annelids, molluscs, …) and hemichordates • considered the most ancestral Apicomplexa • trophozoites mucron or epimerite • large and motile trophozoites and gamonts (usually extracellular) • syzygy - pairing of 2 gamonts before gametocyst formation • syngamy of gametes within the gametocyst  zygotes  gametocyst containing few to many oocysts with sporozoites ▪ Archigregarinorida ▪ Eugregarinorida ▪ Neogregarinorida Archigregarinorida • marine invertebrates (annelids, ascidians, hemichordates, sipunculids) • ancestral to the other gregarines and probably all Apicomplexa • epicellular development at brush border of enterocytes • apical complex preserved at trophozoite/gamont stage • aseptate trophozoite, merogony • bending, rolling motility Selenidium pendula from polychaete https://doi.org/10.1016/j.protis.2016.06.001 Host-parasite interactions in archigregarines Schematic view of anterior region of Selenidium trophozoite Apical pole of Selenidium pendula • myzocytosis via mucron - conoid, rhoptries + subpellicular microtubules = plesiomorphic features present in trophozoite/gamonts stage („hypersporozoite“) Host-parasite interactions in archigregarines S. hollandei Eugregarinorida • arthropods (mostly insects), polychaetes • epicellular at brush border of enterocytes • aseptate or septate trophozoites (epimerite, protomerite, deutomerite) • apical complex present only in sporozoites • merogony absent • gliding motility, metaboly Gregarina polymorpha Life cycle of G. cuneata in mealworm larvae https://doi.org/10.1371/journal.pone.0042606 https://doi.org/10.1016/j.ijpara.2009.04.009 Host-parasite interactions in eugregarines: simple epimerite Cephaloidophora communis from crustacean Gregarina garnhami from locust Actinocephalus dujardini from centipede Beloides tenius from skin beetle Ancyrophora stelliformis from ground beetle Host-parasite interactions in eugregarines: complicated epimerite Host-parasite interactions in eugregarines: modified protomerite Urospora travisiae from polychaete Bothriopsides histrio from diving beetle Neogregarinorida • neogregarines derived from eugregarines, merogony (secondarily obtained) • parasitising insects and are usually found in the fat body, haemolymph, hypodermis, intestine or Malpighian tubules • intracellular, epicellular development • apical complex present in later stages • trophozoite with epimerite or mucron • usually pathogenic for their hosts, even fatal https://doi.org/10.3390/microorganisms9071430; https://doi.org/10.1016/j.ejop.2006.07.007 Mattesia dispora in fat body of larval Mediterranean flour moth Apicomplexa Cryptosporidium spp. • single genus Cryptosporidium, currently more than 30 species • cosmopolitan, mainly intestinal or gastric • monoxenous • epicellular niche - parasite attached to the host cell by a feeder organelle • small oocysts (4,8-5,6 x 4,2-4,8 μm) with 4 naked sporozoites (no sporocysts) • autoinfective stages - type I merozoites capable of forming new type I meronts and thin-walled oocysts that may liberate sporozoites to initiate new infections https://doi.org/10.1016/j.ijpara.2007.11.003 https://doi.org/10.1016/j.ijpara.2007.11.003 Formation of epicellular niche in cryptosporidia https://doi.org/10.3390/microorganisms9122434 Schematic diagram of host-parasite interactions in cryptosporidia. 3 colours are used to distinguish between the parasite (in green), the host cell including its parts modified due to parasitisation (in grey), and the contact zone between the host and the parasite (in white with black dots) where the interactions of the two organisms become more intimate. (A) Invading zoite (either sporozoite or merozoite). (B) An early trophozoite partially enveloped by an incomplete parasitophorous sac (PS). (C) Young trophozoite almost completely enveloped by a PS. Note the tunnel connection between the interior of the anterior vacuole and the host cell cytoplasm that developed as the result of the Y-shaped membrane junction. (D) Almost mature trophozoite. Note the folding of the anterior vacuolar membrane during its transformation into the feeder organelle (FO). (E) Mature stage with a prominent filamentous projection at the base of the PS and with a fully developed FO, the lamellae of FO formed from the anterior vacuole membrane. av - anterior vacuole, db - dense band, dl - dense line separating the feeder organelle from the filamentous projection of the PS, if - incomplete fusion of PS, mv - host microvilli, po - pore on the PS, tu - tunnel connection Life cycle of cryptosporidia https://www.youtube.com/watch?v=ee159M3wxA8 https://www.youtube.com/watch?v=pCTwMCwxudU Extracellular stages in the life cycle of cryptosporidia ??? https://doi.org/10.1016/j.pt.2008.08.002 https://doi.org/10.1016/j.watres.2016.09.013 https://doi.org/10.1016/j.ijpara.2013.07.005 Zoonotic potential of cryptosporidia https://doi.org/10.1016/j.ijpara.2013.07.005 https://doi.org/10.3390/microorganisms9122434 Cryptosporidum parvum • intestinal pathogen of mammalian hosts Pathological changes in rodents with intestinal cryptosporidiosis Healthy jejunum Jejunum with C. parvum, absorptive cells exhibiting protuberances Ileum (BALB/c) with C. parvum, villous atrophy • epithelial cells lining intestinal tract, does not invade deeper layers of the intestinal mucosa • major pathological changes: villous atrophy, shortening of microvilli and sloughing of enterocytes Pathological changes in rodents with intestinal cryptosporidiosis Colon – mucus production (BALB/c mouse)Goblet cells with mucus (BALB/c mouse) • cryptosporidia on the surface of the intestinal mucosa • villous atrophy of the small intestine, inflammatory infiltrate in the lamina propria Cryptosporidium muris https://doi.org/10.1017/S0031182013001637 sporozoite trophozoites 10 µm oocyst type II merogony type I merozoites I type II merozoitesmacrogamontsmicrogamonts zygote excystation type I merogony host cell invasion • epithelial cells within crypts of the gastric glands (glandular part of the gastric mucosa) • prepatent period in BALB/c mice 7-10 days, in Mastomys natalensis 14-21 days • later this isolate has been identified as a new species and named C. proliferans merogony Life cycle of C. muris in rodent host type II merogony 6 µm type I merogony Life cycle of C. muris in rodent host 6 µm microgamontmacrogamont fertilisation Life cycle of C. muris in rodent host thick-walled oocyst zygotes developing oocysts thin-walled oocyst Pathological changes in rodents with gastric cryptosporidiosis healthy gastric epithelium crypt of gastric gland glandular part of gastric mucosa parasitised gastric epithelium glandular part of gastric mucosa crypt of gastric gland ✓ obvious dilatation of gastric glands ✓ epithelial hyperplasia and mucosal hypertrophy without inflammatory exudate Pathological changes in rodents with gastric cryptosporidiosis Cryptosporidum fragile • luminal surface of gastric epithelium and the crypts of gastric glands • from naturally infected black-spined toads Duttaphrynus melanostictus obtained via import of pet animals from the Malay Peninsula in 2006 • infection spontaneously disappeared 24–51 days after arrival https://folia.paru.cas.cz/artkey/fol-200802-0001_new_species_of_cryptosporidium_tyzzer_1907_apicomplexa_from_amphibian_host_morphology_biology_and_phyloge.php healthy gastric epithelium parasitised gastric epithelium Pathological changes in toads with gastric cryptosporidiosis • animal and human diseases • significance: cattle++++, sheep++, goat++, poultry++, reptiles++, human+, horse+, pig+, dog+, cat+ • no effective chemotherapy • emerging infectious diseases, zoonotic character, primary symptoms are acute, watery, and diarrhoea without blood • other symptoms may include anorexia, nausea, vomiting and abdominal cramping/pain, weight loss, electrolyte imbalance • self-limiting in immunocompetent people – parasite life-cycle ends when host immune response eliminates the reproductive cycle • in immunosuppressed patients (diarrhoea can reach 10–15 times per day), such as those with AIDS or those undergoing immunosuppressive therapy, the parasite overwhelms its host due to recycling of type I meronts and infection may lead to dehydration and even death - opportune parasites • extra-gastrointestinal sites include lung, liver and gall bladder, where it causes respiratory cryptosporidiosis, hepatitis, and cholecystitis, respectively Cryptosporidiosis Clinical signs of human cryptosporidiosis Diagnosis of cryptosporidiosis ✓ flotation methods ✓ stained smears ✓ immunoassays ✓ molecular methods – multipathogen molecular panels, molecular typing Water-borne cryptosporidiosis in Milwaukee (1993) The largest waterborne disease outbreak in documented US history (infection caused by C. hominis). It is suspected that The Howard Avenue Water Purification Plant was contaminated due to an ineffective filtration process. This abnormal condition lasted from March 23 through April 8, after which, the plant was shut down. Over the span of approximately two weeks, 403,000 of an estimated 1.61 million residents in the Milwaukee area (of which 880,000 were served by the malfunctioning treatment plant) became ill with the stomach cramps, fever, diarrhoea and dehydration caused by the pathogen. Over 100 deaths were attributed to this outbreak, mostly among the elderly and immunocompromised people, such as AIDS patients. The total cost of the outbreak, in productivity loss and medical expenses, was $96 million Cryptosporidium starring in the movie THIRST (1998) The town of San Paulo is suffering from a heatwave. Engineer Bob Miller turns up at work to learn that the new water filtration plant he has been building has been cancelled and that he is now unemployed. At the same time, Bob’s wife Susan has to deal with a series of mystery deaths at the hospital where she works as a nurse. Bob realizes that the cause might be the water supply. Dr Lawrence Carver confirms that the water is infected with a deadly bacteria Cryptosporidium. They discover that the Cryptosporidium is throughout the town’s drinking water. The only choice is to turn the water plant off and for everybody to drink only bottled or boiled water. As the death toll grows, they realize that they are dealing with a mutated form of the bacteria that is resistant to boiling. Meanwhile, the state governor’s office makes the decision to put a military quarantine around the town...... . Water-borne cryptosporidiosis in Ostersund (2010) https://doi.org/10.3201/eid2004.121415 Water-borne cryptosporidiosis Blastogregarinea Siedleckia nematoides • only two genera (Chattonaria, Siedleckia) known from polychaeta • blastogregarines possess both gregarine and coccidian features • permanent multinuclearity and gametogenesis (merogamont) • epicellular development - brush border of enterocytes • myzocytosis via mucron (conoid, rhoptries) + subpellicular microtubules = plesiomorphic features („hypersporozoite“) • wavy and bending motility https://doi.org/10.1016/j.protis.2018.04.006 Cell organisation in blastogregarines https://doi.org/10.1016/j.protis.2018.04.006 Protococcidiorida • incertae sedis Apicomplexa reported from polychaetes • life cycle of protococcidia not completely understood • epicellular within parasitophorous sac at the brush border of host enterocytes • apical complex detected only at sporozoite stage • trophozoites and gamonts appear nonmotile Eleutheroschizon duboscqi Epicellular niche of Eleutheroschizon duboscqi young trophozoite microgamont macrogamont F-actin Agamococcidiorida • agamococcidia • incertae sedis Apicomplexa • intracellular in polychaete enterocytes • apical complex not seen in trophozoites • trophozoites appear nonmotile • merogony and gametogony not observed • life cycle not known Rhytidocystis pertsovi https://doi.org/10.1038/s41598-020-72287-x Apicomplexa Coccidia • important parasites of humans and animals • monoxenous or heteroxenous • all endogenous stages are intracellular, no mucron or epimerite • merogony, gamogony, and sporogony normally present • mostly without syzygy - macrogamonts and microgamonts develop independently • microgametocytes usually produce many microgametes (with 1 – 3 flagella) • normally is zygote non-motile • sporozoites in oocyst • most coccidia develop resistant oocysts are passed in the faeces ▪ Adeleorina ▪ Eimeriorina Apicomplexa Adeleorina • monoxenous in arthropods (often referred to as adelines) x heteroxenous cycling between blood-feeding invertebrates (definitive hosts) and various vertebrates (intermediate hosts), usually referred to as haemogregarines • usually complex life cycle – 1 or more merogonies  gametogony  sporogony • meronts often of morphologically distinct types: 1) meront producing large merozoites that initiate another round of merogony, 2) meront producing smaller merozoites that represent the gamonts progenitors • microgamonts produce 1-4 microgametes that associate with macrogamete in syzygy • endodyogony absent, sporozoites in sporocysts and/or oocysts Apicomplexa Monoxenous adelines • e.g. Adelina and Adelea from insects, Klossia from kidney of molluscs, Klossiella from kidney of vertebrates, ... sporogony oocyst syzygy merogony Adelina tribolli Adelina melolonthae Apicomplexa Polysporocystic oocysts from kidney of Triodopsis hopetonensis Klossia helicina Klossia razorbacki Monoxenous adelines Klossiella equi K. equi multifocally expanding tubular epithelial cells (schizont - arrow; sporocysts - arrowheads) Sporocysts found on centrifugal flotation of horse urine Apicomplexa Heteroxenous haemogregarines • Hepatozoidae, Haemogregarinidae and Dactylosomatidae comprising several genera, including pathogens of vertebrates: ▪ Hepatozoon from carnivores and reptiles ▪ Haemogregarina from fish and turtles ▪ Dactylosoma from fish or amphibian (possibly also reptiles) • in all of them, invertebrate plays role of the definitive host with gamogony in its digestive tract, and parasite transmission occurs in 2 modes: 1) inoculation - infectious sporozoites enter the vertebrate host during blood feeding of vector (Haemogregarina, Dactylosoma) 2) ingestion of infected definitive (invertebrate) host by vertebrate host – may involve a paratenic host when the next definitive host is infected exclusively via blood feeding (Hepatozoon, Haemolivia, Karyolyssus) Apicomplexa genus Hepatozoon • more than 300 species • sporogony in invertebrate host  oocysts with large number of sporocysts with sporozoites • sporozoites in liver of the vertebrate host (asexual reproduction)  merozoites • merozoites in bloodstream  gametocytes = conspicuous organisms in erythrocytes / lymphocytes • sexual reproduction in haemocoel of invertebrate host • hepatozoonosis – when animal eats the invertebrate host (e.g. tick, disease is not spread by tick bites!), infection may cause polyostotic aggressive lesions Hepatozoon americarum Skeletal muscle containing a developing meront, mucopolysaccharide layers produced by host cell protect the developing organisms from the dog’s immune system Pyogranuloma in skeletal muscle of a infected dog, zoites displace the nucleus in several of the inflammatory cells Oocysts in hemocoel of Amblyomma maculatum tick, each oocyst with hundreds of small round sporocysts containing 10-6 sporozoites. Pelvic limb of infected dog showing smooth periosteal proliferation on the cranial aspect of the femur. Moderate to severe illness: fever, lethargy,  appetite, weight loss, muscle pain/weakness, reluctance to move, discharge from the eyes/nose, enlarged lymph nodes. Periods of illness and apparent resolution; without treatment  damage to the blood vessels and kidneys often with death within 1 year. Hepatozoon canis Meront in splenic tissue of a dog demonstrating the typical “wheel spoke” pattern Gamonts in neutrophils on a blood smear • most dogs infected show no or only mild signs of illness: fever, weight loss, lethargy, anaemia, enlarged spleen • in some dogs with high parasite numbers, the disease can become more severe and may be potentially life-threatening • immunosuppression or concurrent disease is an important factor in expression of illness in H. canis (but not in H. americanum) Hepatozoon ayorgbor • African Python regius • merogony in liver and spleen, gametocytes in erythrocytes • transmitted by Culex quinquefasciatus • rodents as a reservoir https://www.parazitologie.cz/protozoologie/Personal%20homepages/Votypka/Hepatozoon%20ayrbor.pdf Apicomplexa genus Haemogregarina • infected leech feeding on a reptile or amphibian injects sporozoites into the bloodstream where they enter erythrocytes  trophozoites • erythrocytes with trophozoites lodge in the capillaries of the bone marrow  macromeronts  merozoites infect new erythrocytes  micro-/macrogametocytes • fusions of gametes  oocyst in the gut of leech where they hatch and sporozoites migrate through the intestinal wall into the circulatory system, sporozoites ending up in the proboscis are injected into the next host Haemogregarina bigemina https://doi.org/10.1016/j.ijpara.2012.11.012; https://doi.org/10.1016/j.ijppaw.2019.01.006 • ectoparasitic gnathiid isopods likely serve as vectors Haemogregarina stepanowi https://doi.org/10.1017/S0031182013001820 • turtle Emys orbicularis (not only) • trasmitted by leeches genus Dactylosma • amphibians, fish • transmitted by leeches and possibly mosquitos Blood smear from Pelophylax frog with merogony (primary meront) Blood smear from Pelophylax frog with gamont Apicomplexa Eimeriorina • homoxenous or heteroxenous life cycles • syzygy is absent - macrogametes and microgametes develop independently • anisogamous microgamonts produce a large number of flagellated microgametes • zygote is nonmotile, sporozoites enclosed in a sporocyst https://www.youtube.com/watch?v=euHBghFhJRc Sporulation in coccidia Excystation of coccidian oocyst Factors affecting the significance of individual coccidian species Factors affecting the host ✓ "technology" of animal breeding ✓ host immunity (age category) ✓ nutrition, other diseases Factors of individual species of coccidia ✓ pre-patent period ✓ patent period ✓ number of merogonies ✓ size of developmental stages ✓ sporulation time ✓ extraintestinal development Chicken coccidiosis (vaccination available) https://www.youtube.com/watch?v=NFh2aPmuYQE https://www.youtube.com/watch?v=u1iIZQrB_B4 https://www.youtube.com/watch?v=HnVCmZUoork Taxonomy of coccidia Sporocysts: 2 Sporozoites: 8 Eimeriorina Eimeriidae - monoxenous Sarcocystidae - heteroxenous Eimeria (4-2) Isospora (2-4) Caryospora (1-8) Tyzzeria (0-8) Wenyonella (4-4)… x Morphology of coccidian oocyst Oocyst of Eimeria Oocyst of Isospora Oocyst of Caryospora Coccidia in poultry 9(7) Eimeria species - „pathogenicity top-ten“ 1. Eimeria necatrix - small intestine 2. Eimeria tenella - ceca 3. Eimeria maxima - small intestine 4. Eimeria brunetti - small intestine, ceca, rectum 5. Eimeria mitis - small intestine, ceca, rectum 6. Eimeria acervulina (E.mivati ??) - duodenum 7. Eimeria praecox (E. hagani ??) - duodenum 632 Localisation of developmental cycle and pathological changes in the intestine E. tenella E. necatrix E. acervulina E. maxima Clinical signs of poultry coccidiosis Pathology of poultry coccidiosis Coccidia in rabbits 11 Eimeria species - „pathogenicity top-ten“ 1. Eimeria intestinalis - jejunum, cecum, colon 2. Eimeria flavescens - jejunum, cecum, colon 3. Eimeria stiedai - liver 4. Eimeria magna - jejunum, cecum, colon 5. Eimeria irresidua - jejunum, ileum 6. Eimeria media - jejunum, ileum 7. Eimeria piriformis - jejunum, ileum 8. Eimeria vejdovski - jejunum, ileum 9. Eimeria coecicola - cecum 10. Eimeria exigua - ileum 11. Eimeria perforans - duodenum Coccidia in rabbits Coccidia in pigs 7 Eimeria species - „pathogenicity top-ten“ 1. Eimeria scabra - small intestine 2. Eimeria polita - small intestine 3. Eimeria debliecki - small intestine 4. Eimeria spinosa - small intestine 5. Eimeria neodebliecki - small intestine 6. Eimeria porci - ? 7. Eimeria suis - ? Cystoisospora suis • non-haemorrhagic transient diarrhoea, resulting in poor weight gain • it appears that primary immune responses against C. suis cannot readily be mounted by neonates, contributing to parasite establishment and rapid, while in older pigs, age-related resistance prevents disease development (morbidity ++++, mortality +) C. suis in pigletsoocysts Cystoisospora suis Cystoisospora suis https://doi.org/10.3389/fvets.2015.00068 Coccidia in ruminants Cattle - 12 Eimeria species Eimeria zuernii - pathogenic Eimeria bovis - pathogenic Both species responsible for severe clinical disease characterised by haemorrhagic diarrhoea with sometimes fatal outcome. Eimeria alabamensis - also can cause clinical disease Prevalence of Eimeria infection in cattle is generally high and can reach 100% in calves. Eimeria ellipsoidalis - bloody diarrhoea in calves and young cattle on pasture, „red dysentery of cattle„, "neurological coccidiosis" Pathological changes in the small and large intestine - hyperaemia, haemorrhages, pseudomembranes. Coccidia in ruminants Sheep - 11 Eimeria species Eimeria bakuensis - pathogenic Eimeria ovinoidalis (E. ninakohlyakimovae) - middle pathogenicity Eimeria crandalis - pathogenic Eimeria ashata - pathogenic Goat -13 Eimeria species • similar morphology of oocysts to sheep, different species Eimeria ninakohlyakimovae - slightly pathogenic, high prevalence Eimeria arloingi - most pathogenic Eimeria arloingi gamonts macromeronts Coccidia in carnivores Dog - 3 (5) Isospora species Isospora canis Isospora ohioensis Isospora burrowsi • cause of diarrhoea in young animals • dormozoites (hypnozoites) in rodents • Neospora / Hammondia Cat - 3 (5) Isospora species Isospora cati Isospora rivolta • cause of diarrhoea in young animals • dormozoites (hypnozoites) in rodents • Toxoplasma / Hammondia genus Caryospora • mostly monoxenous • oocyst with 1 sporocyst with 8 sporozoites inside • parasites in snakes a raptors • about 70 described species Caryospora bigenetica C. simplex • facultatively heteroxenous • primary and secondary hosts • caryocyst (dormant stage) in secondary host • dermal coccidiosis in secondary host • potentially zoonotic Life cycle of Caryospora bigenetica A) First type of final host (rattlesnake) 1) The sporulated oocyst contains a single sporocyst with 8) sporozoites. 2–4) After oral infection with oocysts 2 generations of meronts are formed inside the intestinal epithelium. 5-6) Male and female gamonts are formed and later gametes occur. The male gamont (G) forms numerous gametes. 7-8) After fertilisation a thick-walled oocyst is formed inside the host cell and becomes free within the faeces. While sporulating on the ground a single sporocyst with 8 sporozoites is formed inside each oocyst. B) Second type of final host (mice, cotton rats, dogs) 2.1–2.4) Repetition of the merogonic and gamogonic development (described in 1–8). The sporulation of oocysts may occur in skin regions (2.4). 2.5) Sporozoites that were set free from their sporocysts while still inside the skin of their hosts enter... Caryospora simplex Caryospora bigenetica genus Lankesterella • parasites of amphibia reptiles, birds; transmission by a leeches • heteroxenous with entire replicative process occurring in vertebrate host (connective tissue, visceral organs - liver, spleen, intestine, lung, kidney) • absence of environmentally resistant oocysts • thin-walled oocysts harbour variable numbers (usually more than 32) of naked sporozoites (no sporocysts) which after exiting the oocyst in situ enter blood cells and become dormant until they are ingested by leech vectors during a blood meal Blood smear from Pelophylax frog with Lankesterella sporozoites Thank you for your attention ☺ Lectures ✓ Introduction: BPP 2022 I ✓ Euglenozoa (Excavata): BPP 2022 II ✓ Fornicata / Preaxostyla / Parabasala (Excavata): BPP 2022 III ✓ Apicomplexa I (SAR): BPP 2022 IV  Apicomplexa II (SAR): BPP 2022 V • Amoebae (Excavata, Amoebozoa): BPP 2022 VI • Ciliophora, Opalinata (SAR): BPP 2022 VII • Pneumocystis (Opisthokonta, Fungi): BPP 2022 VIII • Microsporidia (Opisthokonta, Fungi): BPP 2022 IX • Myxozoa (Opisthokonta, Animalia): BPP 2022 X