Failed attempt of Cryptosporidium andersoni infection in lambs

Four lambs 4 months old were inoculated with Cryptosporidium andersoni oocysts (bovine isolate) in a dose of 5 x 10(6) per animal followed later by 1 x 10(7) oocysts per animal. No animal shed Cryptosporidium oocysts during 56 days post infection, no lamb showed clinical symptoms of cryptosporidiosis and no macroscopic changes were detected in the abomasum immediately after autopsy. Histological examinations did not demonstrate the occurrence of Cryptosporidium in the abomasum and other selected organs.     

The endogenous development, described by light and electron microscopy, of Eimeria jamescooki sp. n. (Apicomplexa: Eimeriidae) from the skink Cryptoblepharus virgatus

Eimeria jamescooki sp. n. was recovered from the skink Cryptoblepharus virgatus (Garman) found on the grounds of James Cook University, Townsville (type locality), North Queensland, Australia. Oocysts were 17.5-25.0 (22.1 +/- 1.9) x 15-22.5 (17.7 +/- 1.6) microm and sporocysts 6.25-10.0 (7.9 +/- 1.15) x 3.75-6.25 (5.3 +/- 1.0) microm in size. Endogenous stages are described from histological material examined by light microscope and by transmission electron microscope. Both merogony stages and gamonts were found to develop in the cytoplasm of the anterior gut mucosal epithelium. Meront progeny were comprised of 10 to 21 merozoites. Premature macrogamonts were elongate; some host cells contained two elongate macrogamonts. Unique to the presently described species were the Golgi "plaques" and an enclosure of tubuli. Mature macrogamonts and young oocysts ranged in size from 14 x 7 to 21 x 11 microm and contained two types of wall-forming bodies, canaliculi and amylopectin granules. Differentiating microgamonts conformed in fine structure with that observed in other eimerians. Their sizes increased from 15.4 x 4.2 to 28 x 8.4 microm while dividing to over 70 nuclei, which formed a corresponding yield of microgametes.     

Redescription of Eimeria dorcadis Mantovani, 1966 (Apicomplexa: Eimeriidae) from the dorcas gazelle (Gazella dorcas) in Saudi Arabia

Eimeria dorcadis Mantovani, 1966 is redescribed from dorcas gazelle (Gazella dorcas (L.)) from Saudi Arabia. Oocysts were detected in 7 out of 22 faecal samples (32%) using floatation method. The sporulated oocysts are cylindrical, slightly flattened at the micropylar pole, measure in average 32 x 19 microm (27-36 x 16-24 microm), length/width ratio being 1.7 (1.5-2.1). Oocyst wall is 1.2 microm thick, smooth, double-layered; outer layer is slightly thicker, light blue in colour; inner layer brownish, with micropyle in the inner layer and apparently continual outer one, measures 2.2 microm, but lacks a micropylar cap. The sporocyst elongate-ellipsoidal, measures 14 x 8 microm (12-17 x 6-9 microm), length/width ratio being 1.8, with sporocyst residuum as circular compact, coarse, refractile granules. Stieda body is present, while substieda body is absent. Sporozoites banana-shaped, measure 11 x 2.5 microm, each with a large spheroidal refractile body at the wider pole. Sporulation time is 2-3 days at 25 +/- 2 degrees C.     

Studies on the life cycle of Cryptosporidium coccidia in experimentally infected chickens

Experimental infections of 7-28-day-old chickens with Cryptosporidium oocysts isolated from spontaneously infected chickens demonstrated that the endogenous development of this parasite takes place simultaneously in the organs of digestive, respiratory and excretory systems and in bursa of Fabricius. It was demonstrated for the first time that the oocysts of Cryptosporidium are shed through the respiratory tract into the beak cavity. A novel rapid and simple method has been developed for the detection of oocysts. Its principle is the rinsing of the beak cavity. This method enabled to isolate the oocysts from experimentally infected chickens and immediately use them in the dose of 6 X 10(5) for peroral infection of a 37-day-old chicken. The prepatent period was 8 days, patent period 12 days. On days 4-7 after the first detection of Cryptosporidium oocysts in chicken excrements, the oocysts were detected also by the method of beak cavity rinsing. This indicates that the oocysts released from the respiratory tract are infective. This fact is important from the epizootological viewpoint in relation with possible spreading of Cryptosporidium infections in chicken farms.     

A description of two new species of coccidia (Apicomplexa: Eimeriidae) from African reptiles with nomenclatural corrections for two Caryospora and one Eimeria species from snakes

Two new species of coccidian parasites are described from African reptiles. Oocysts of Eimeria foulshami sp. n. from the plated lizard Gerrhosaurus major bottegoi Del Prato of Sudan are ellipsoidal, 24.1 x 14.9 (23-26.5 x 14-17.8) microm with a bilayered, colourless oocyst wall and lack polar granules. The ellipsoidal sporocysts average 8.6 x 4.6 (7-10.6 x 4.4-7) microm and possess a prominent, globular, sporocyst residuum. Oocysts of Caryospora regentensis sp. n. from the Eastern green mamba Dendroaspis augusticeps Smith, 1849 [corrected] of Kenya are spherical to subspherical, 16.8 x 16.4 (16-17.6 x 15-17.2) microm with a bilayered oocyst wall and a single polar granule. The ellipsoidal sporocysts average 13.0 x 10.3 (10.2-14 x 9.2-11) microm and possess a Stieda and substieda body and a prominent globular sporocyst residuum. Oocysts of Caryospora legeri Hoare, 1933 are reported from a hissing sand snake, Psammophis sibilans sibilans L. from Nigeria, representing a new geographical record. The oocysts are slightly larger than the type, but otherwise identical. Caryospora psammophi Bray, 1960 and C. hermae Bray, 1960 from Psammophis sibilans phillipsi, oocysts of which are morphologically similar to and overlap in dimensions with C. legeri Hoare, 1933, are synonymised with the latter species. Eimeria samiae Iskander et Tadros, 1979 is emended to E. samyadeli to reflect the gender of the person the species was named after and because E. sani is preoccupied. In addition to these findings, Eimeria bohemi Modry, Slapeta et Koudela, 2000 and oocysts of an unidentified spherical Eimieria sp. are reported from Chamaeleo dilepis dilepis Leach from Cameroon.     

Two new species of isospora (Apicomplexa: Eimeriidae) from geckoes of the genus Rhacodactylus (Sauria: Gekkonidae) endemic to New Caledonia

Coprological examination of New Caledonian geckoes of the genus Rhacodactylus Fitzinger, 1843 revealed two new species of coccidia. Isospora leachiani sp. n. from R. leachianus (Cuvier, 1829) has oval, colourless oocysts, measuring 21-26 x 16-18.5 microm. Sporocysts are ellipsoidal, 11-12.5 x 6.5-8 microm, with distinct Stieda and substieda bodies. Oocysts of Isospora sykorai sp. n. from R. ciliatus (Guichenot, 1866) are elongately oval to cylindrical, 20-23.5 x 11-14 microm; sporocysts of this species are ellipsoidal, 10-11.5 x 7-8 microm, with a slightly pointed end and Stieda and substieda bodies. Infected geckoes did not exhibit any alteration of their health status.     

Ultrastructural characteristics of Nematopsis sp. oocysts (Apicomplexa: Porosporidae), a parasite of the clam Meretrix meretrix (Veneridae) from the Arabian Gulf, Saudi Arabia

This paper describes the fine structure of oocysts of Nematopsis sp. (Apicomplexa, Porosporidae) found in the abductor muscles of seawater clams, Meretrix meretrix (Linnaeus, 1758) (Veneridae), collected near the city of Dammam (6 degrees 17'0"N, 50 degrees 12'0"E) in the Arabian Gulf off the coast of Saudi Arabia. Oocysts of an ellipsoidal shape were found among myofibrils of the abductor muscles of infected clams. Each oocyst is composed of an oocyst wall surrounding a single uninucleate vermiform sporozoite located in the lumen of the oocyst wall. The thin oocyst wall (0.70-0.85 microm thick) is composed of homogenous electron-lucent material formed by three layers of equal-thickness. The oocyst wall contains a plano-convex opercular-like structure about 2.5 microm in diameter and 0.75-0.90 microm thick, composed of a homogenous material with moderate electron density. The oocyst is of an ellipsoidal shape and is 15.6 +/- 0.6 microm long and 11.1 +/- 0.7 microm wide. Externally, the oocyst wall is surrounded by a complex dense network of numerous anastomosed microfibrils, which are attached to the oocyst wall, forming 2-3 layers and extending towards the periphery, at some points penetrating amongst the host cells. The myofibrils in some cases show evident aspects of lysis as a consequence of the appearance of lysosome-like vesicles. Lacking knowledge of a complete life cycle and/or molecular data precluded the conclusive identification of this species.     

New species of Choleoeimeria (Apicomplexa: Eimeriidae) from the veiled chameleon, Chamaeleo calyptratus (Sauria: Chamaeleonidae), with taxonomic revision of eimerian coccidia from chameleons

Coprological examination of 71 samples from a breeding colony of veiled chameleons, Chamaeleo calyptratus Duméril et Duméril, 1851, revealed a presence of two species of coccidia. In 100% of the samples examined, oocysts of Isospora jaracimrmani Modry et Koudela, 1995 were detected. A new coccidian species, Choleoeimeria hirbayah sp. n., was discovered in 32.4% of samples from the colony. Its oocysts are tetrasporocystic, cylindrical, 28.3 (25-30) x 14.8 (13.5-17.5) microm, with smooth, bilayered, -1 microm thick wall. Sporocysts are dizoic, ovoidal to ellipsoidal, 10.1 (9-11) x 6.9 (6-7.5) microm, sporocyst wall is composed of two plates joined by a meridional suture. Endogenous development is confined to the epithelium of the gall bladder, with infected cells being typically displaced from the epithelium layer towards lumen. A taxonomic revision of tetrasporocystic coccidia in the Chamaeleonidae is provided.     

Phylogenetic characterization of a microsporidium (Nosema sp.) isolated from the mulberry pest, Hemerophila atrilineata

Microsporidia are a group of obligate intracellular unicellular eukaryotes that can parasitize a wide variety of other eukaryotes ranging from protists to invertebrates and vertebrates. In this study, we examined the microsporidium Nosema sp. isolated from the mulberry pest, Hemerophila atrilineata Butler, 1881, named herein "Nosema sp. HA". The fresh spores were long oval in shape, 3.8 +/- 0.4 microm in length and 1.9 +/- 0.3 microm in width. Analysis of tissue infection of silkworm, Bombyx mori Linnaeus, 1758, indicated that the midgut, Malpighian tubules, muscle, fat body, silk glands, hemocytes, nerve tissue and gonads of silkworm were infected with Nosema sp. HA. The complete rRNA gene sequence of this microsporidium contained 4 305 base pairs (GenBank Accession JN882299), including the large subunit rRNA (2492 bp), the internal transcribed spacer (187 bp), the small subunit rRNA (1232 bp), the intergenic spacer (279 bp) and the 5S region (115 bp). The organization of the rRNA gene is 5'-LSU-ITS-SSU-IGS-5S-3'. Phylogenetic analysis, comparison of sequence identities and the arrangement in the rRNA gene subunits suggested that this isolate is separate from other Nosema species.     

Light and scanning electron microscopy of Myxobolus porofilus sp. n. (Myxosporea: Myxobolidae) infecting the visceral cavity of Prochilodus lineatus (Pisces: Characiformes: Prochilodontidae) cultivated in Brazil

Myxobolus porofilus sp. n. is described infecting the visceral cavity of Prochilodus lineatus (Valenciennes, 1836) cultivated in S?o Paulo State, Brazil. The plasmodial form of the parasite is 3-5 mm in length and appeared compressed between the wall of the visceral cavity and the pyloric caecum, reposing on this organ. The spores are small (length 5.7 +/- 0.3 microm, width 4.8 +/- 0.2 microm; mean +/- SD) and round to elliptical in frontal view. The valve surfaces are smooth and have sutural folds. The polar capsules are ovoid, small (length 1.6 +/- 0.1 microm, width 1.1 +/- 0.1 microm) and equal in size. The polar filaments have three turns aligned perpendicularly to the longitudinal axis of the capsule. A conspicuous polar filament pore is arranged at the anterior end of the spore. The only reaction observed upon histological analysis was the presence of a capsule of connective tissue surrounding the plasmodia. This is the first report of a myxosporean parasite in the Prochilodontidae.     

Eimeria burdai sp. n. (Apicomplexa: Eimeriidae), a new parasite species from subterranean African silvery mole-rat, Heliophobius argenteocinereus

A new coccidian parasite of the genus Eimeria Schneider, 1875 is described from the subterranean African silvery mole-rat Heliophobius argenteocinereus Peters, 1846. Oocysts of Eimeria burdai sp. n. were subspherical to broadly ellipsoidal 17.8 (16-19) x 14.1 (12-15), with a shape index 1.2 (1.1-1.4). Oocyst wall was bilayered, smooth and colourless, approximately 1.0 thick. Outer layer was significantly thicker than inner one. A micropyle and oocyst residuum were absent. One or two ellipsoidal or spherical polar granules were present. Sporocysts were ellipsoidal, 10.8 (9-12) x 6.2 (5-8) with a shape index 1.7 (1.5-1.9). Sporocyst wall was single-layered, thin, smooth and colourless, with small Stieda body at the pointed end. In freshly sporulated oocysts, spherical sporocyst residuum was composed of small granules enclosed by a thin membrane. Sporozoites were elongate, lying length-wise in the long axis of the sporocyst, partially curled around each other, with single large refractile body located posteriorly. Faintly distinguishable nucleus was in the central part of the sporozoite. This eimerian represents the first coccidian species described from subterranean African silvery mole-rat (Rodentia: Bathyergidae).     

Wardium paucispinosum sp. n. (Eucestoda: Hymenolepididae), parasite of Larus maculipennis (Aves: Laridae) in Mar del Plata, Argentina; with comments on Wardium semiductilis (Szidat, 1964) comb. n

A new species Wardium paucispinosum (Eucestoda: Hymenolepididae) parasite from the intestine of Larus maculipennis (Lichtenstein) from Mar del Plata, Argentina is described. The distinctive features of the new species are: strobilar length 52.8 mm; 10 aploparaksoid rostellar hooks, 14 (12-17) microm long; ratio between cirrus pouch length and mature proglottid width (CPL/MPW) 0.38 (0.27-0.50); regular cylindrical evaginated cirrus, 90 x 10 microm, with distal end without spines and proximal and medium thirds covered with spines 7 microm long; simple tubular membranous vagina, 110 x 10 microm, without sclerotised portions and sphincters; eggs fusiform, 77 x 44 microm. Besides, Hymenolepis semidutctilis Szidat, 1964, from the intestine of Larus dominicanus and L. maculipennis from Santa Fe, Argentina is transferred to the genus Wardium Mayhew, 1925, based on the presence and shape of the rostellar hooks.     

Genetic resistance to bacterial blight disease in Persian walnut

Bacterial blight disease of Persian walnut (Juglans regia, L.), caused by Xanthomonas arboricola pv. juglandis (Xaj), leads to significant nut losses in northern, central and western areas of Iran. To identify the natural sources of resistance to disease in the endemic walnut genotypes of Iran, sixteen walnut genotypes, collected from different areas of Hamedan province, were inoculated with Xaj in a randomized complete block design with five replicates for each genotype. Two-year old genotypes were gently sprayed with a suspension of bacteria adjusted to approximately 2 × 10⁹ cfu ml⁻¹ of distilled water in May. Infected leaves were rated for disease 28 and 42 days after inoculation, using a 0 to 5 severity scale, based on the number, size and distribution of lesions on the leaves. Data analyses showed that there were variations among genotypes in response to pathogen. Upon inoculation by bacterial suspension genotype 94 showed the highest resistance to both disease incidence and its progress after 4–6 weeks of infection. Genotype 65 showed high susceptibility to disease and genotype 69 showed high susceptibilities both to disease incidence and its progress after 4–6 weeks of infection.     

Caryospora varaniornati sp. n. (Apicomplexa: Eimeriidae) in the Nile monitor, Varanus (Polydaedalus) niloticus species complex

Parasitological examination of two ornate Nile monitors Varanus ornatus (Daudin, 1803) imported from Benin revealed the presence of a new species of Caryospora. Oocysts of Caryospora varaniornati sp. n. are spherical to slightly subspherical, 12.0 (11-12.5) x 11.5 (11-12) microm, without amicropyle and oocyst residuum, and occasionally possessing one small polar granule. Sporocysts are broadly ellipsoidal, 8.8 (8.5-9.5) x 6.7 (6.5-7) microm; a lentil-like Stieda body is present, ca. 0.5 x 1 microm; substieda body not visible. Experimental infection of a closely related host, Varanus niloticus (L.), did not lead to the oocyst excretion despite the fact that one of the experimentally inoculated monitors was immunosuppressed by dexamethasone. Histological examination did not reveal stages of coccidian development. Therefore, it is possible that C. varaniornati is strictly host specific.     

Screening for blast resistance in rice using AMMI models to understand G x E interaction in Guyana

In rice, totally one hundred and three genotypes were evaluated for resistance against blast disease in three hot spot locations of Guyana. The experiments were conducted in spring and autumn seasons of 2015, 2016 and 2017. The modified upland blast nursery (UBN) method was adopted for screening blast resistance. The Additive Main effects and Multiplicative Interaction (AMMI) analysis was applied to understand the interaction between genotype (G) and environment (E). The results of all seasons at three locations showed the consistent expression of high resistance by genotype FL 127 followed by FG12–08 and FG12–273. The AMMI analysis of variance (ANOVA) revealed that, resistance was dominated by the genotype main effect. The magnitude of genotypes by environment (G x E) sum of square (SS) was 3.64 times smaller than genotype SS, and 6.74 times larger than environment SS. The analysis has indicated that the differences of the genotypes across the environment were not substantial. All of the seven environment vectors appeared to be positively correlated. The screening at Onverwagt Back, autumn 2016 (E7) and Black Bush Polder, spring 2015 (E5) were observed to be the most representative environment. This was clearly revealed by smaller average - environment axis (AEA) angle. Likewise, E4 (Canje, autumn 2015) was observed closer to the “ideal test environment”, thus suggested as an ideal environment for blast resistant screening. Also, genotypes clustered near the origin have showed stable blast disease resistant status and contributed little to both G and G x E. These genotypes proposed as useful ones for breeding disease resistant varieties and hybrids in rice.     

Phytophthora infestans Populations from Tomato and Potato in North Carolina Differ in Genetic Diversity and Structure

ABSTRACT Phytophthora infestans causes a destructive disease on tomato and potato. In North Carolina (NC) potatoes are mostly grown in the east, whereas tomatoes are grown in the mountainous areas in the western part of the state. Five genotypes of P. infestans were identified from 93 and 157 isolates collected from tomato and potato over a 5 year period between 1993 and 1998. All isolates collected from potato in eastern NC were the US-8 genotype, whereas only a single isolate was the US-1 genotype. Tuber blight was found on immature daughter tubers in a single field in 1997, however infection on mature tubers was not observed. Within potato fields, a range of sensitivity to metalaxyl was observed among isolates but all were either intermediate or highly resistant to the fungicide. In contrast, isolates from tomatoes included previously reported US-7 and US-8 genotypes and two new genotypes called US-18 and US-19 (A2 mating type, allozyme genotype Gpi 100/100 and Pep 92/100). These genotypes had unique restriction fragment length polymorphism banding patterns, were sensitive to metalaxyl, and have not been reported elsewhere. All genotypes, with the exception of the US-1, were the Ia mitochondrial haplotype. Thus, isolates of P. infestans from tomato were more genetically diverse over time in NC than those from potato and include two new genotypes that are sensitive to metalaxyl.     

Slow wilting components in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis>(L.) Millsp.)

Partial resistance to Fusarium wilt was characterized based on root inoculation on seven pigeonpea genotypes with a virulent isolate of Fusarium udum. Resistance to wilt seems to be mainly due to localization of the pathogen in roots and vascular systems in the stem. Three components of resistance (wilting rate or incubation period, weighted wilt index and number of colonized plants) were determined and compared to those of susceptible genotypes Bahar and TTB7. There were significant differences between the genotypes for the resistance components. The mechanisms of resistance in the genotypes appeared to be different, with genotype ICP8863, having a longer incubation period, minimum wilt index and minimum pathogen colonization as compared to other resistant genotypes (ICP9174, ICP87119 and ICP8858). Wilting rate or incubation period and number of colonized plant were significantly correlated with resistance in adult plants in the field (AUDPC). Wilt index was useful in discriminating between genotypes that had a similar incubation period and number of colonized plants. The partial character of resistance is probably based on quantitative differences in localization capacity among the genotypes. A quantitative relationship between components, incubation period and number of colonized plants and the AUDPC, if verified for a large number of genotypes, may be used to obtain an index of resistance that may predict resistance levels in the field.     

Isospora carliae sp. n. (Apicomplexa: Eimeriidae) from the skink Carlia rhomboidalis (Peters) from Daintree Forest, North Queensland, Australia: description and fine-structural account of endogenous development

Isospora carliae sp. n. is described from the blue-throated rainbow skink Carlia rhomboidalis (Peters), from Daintree Forest, North Queensland, Australia. Oocysts are ellipsoidal, 16.8-21.0 x 12.6-15.4 microm in size, with their two sporocysts, 9.0-14.0 x 7.0-9.24 microm in size, positioned along the wide axis. Sporozoites contain a distinct refractile body and are accompanied by a residuum. All endogenous development occurs within the host-cell nucleus. Nuclei are sometimes invaded by several merozoites, but only infections by a single parasite persist. Nuclei lodging meronts, mature microgamonts and premature macrogamonts have an elongate shape. Some meronts exhibit a membrane-bound cytoplasmic inclusion that contains many micronemes.