Infections with Eimeria maxima and Eimeria acervulina in the fowl: effect of previous infection with the heterologous organism on oocyst production.

Judged by oocyst production, infections with Eimeria acervulina in birds immunized with E. maxima were consistently higher than in birds which had not been immunized. Oocyst production of E. maxima in birds previously infected with E. acervulina was greater, in three out of four experiments, than in control chickens, but some of the differences were slight. The findings are discussed but no satisfactory explanation can be offered. In general there was little or no difference between the oocyst production of the individual species when present as single or mixed infections.     

Modelling host cell availability and the crowding effect in Eimeria infections.

Within-host mathematical models of Eimeria maxima and Eimeria praecox infections of the chicken are presented and used to investigate the role of host cell availability as a possible determinant of the so-called 'crowding effect'; whereby the fecundity of the parasites decreases as infectious dose increases. Assumptions about the number of available host cells, the average lifespan of these cells and the age structure within the host-cell population were made and mathematical models were constructed and combined with experimental data to test whether these conditions could reproduce the crowding effect in the two species. Experimental data demonstrated that crowding during in vivo infections was apparent following very low infectious doses, but none of the models could adequately reproduce crowding at the same doses while maintaining realistic estimates of the dynamics of the enterocyte pool. However, both the size and lifespan of the enterocyte pool were demonstrated to have substantial effects on the fecundity of the infections, particularly at higher doses. These data indicate that host cell availability cannot be solely responsible for the crowding effect. Alternative factors such as the influence of the primary immune response to the parasite may also be explored using within-host models and other applications of these models are discussed.     

Eimeria stiedae: weight, oocyst output, and hepatic function of rabbits with graded infections

Groups of 5 (38–40 days old) Eimeria stiedae-naive rabbits were infected with 0 (mock infection), 10², 10³, 10⁴, and 10⁵ sporulated oocysts of Eimeria stiedae (groups A, B, C, D, and E, respectively) and body weight, oocyst output, serum glutamic pyruvic and serum oxalacetic transaminases, bilirubinemia. lipemia, glycemia, and proteinemia were measured on diverse occasions for 50 days. Mortality and carcass and liver weights at the end of the experiment were also recorded. Mortality was 80% in group E, 40% in group D, and 0% in the other groups. Reduction of weight gain was observed from the 8th day of infection and actual loss from the 15th day in infected animals. On Day 50, the average body and carcass weights of all infected rabbits were 71.2 and 63.2%, respectively, of group A. Only groups D and E had absolute hepatomegaly and group C had relative liver enlargement. Patency and rate of increase of oocyst output were not related to dose but peak and declination of oocysts production were delayed in proportion to the infective dose.Serum glutamic pyruvic and glutamic oxalacetic transaminases were increased from Day 8 to Day 36, bilirubinemia and lipemia augmented from Day 22, and glycemia and total serum proteins decreased from Days 22 and 29, respectively. Bilirubinemia tended to recover sooner (toward Day 50) in rabbits with lighter infection and lipemia recovered from Day 36 in proportion to the size of the infective dose. Modifications of glycemia and total proteinemia were consistent but reached statistical significance only occasionally. The asexual reproduction of the parasites caused transient damage to the hepatocytes during the second week of infection, and later sexual stages altered the ductal epithelium from the fourth week.     

Eimeria spp. of domestic fowl: the migration of sporozoites intra- and extra-enterically

Chickens were dosed orally with sporulated oocysts of Eimeria acervulina, E. brunetti, E. maxima, or E. praecox and the subsequent presence, in various tissues, of parasites capable of inducing patent infections was detected by transferring the tissues to coccidia-free recipients. Similar results were obtained with each of the 4 species studied, irrespective of whether initial development occurs in the superficial (E. praecox, E. brunetti) or crypt (E. acervulina, E. maxima) epithelium. Infection was transferable by gut scrapings and liver homogenates at all time intervals (3, 6, 12, 18, 24, and 36 hr postinoculation) studied. Infection was also transferable with blood and with splenic homogenates but not consistently. Transfers made within a short time of the inoculation of donors were more successful in producing patent infections in the recipients. In all transfers the prepatent period was normal for the species. These findings suggest that sporozoites enter the mucosa very shortly after inoculation, and some of them pass to the liver and spleen and then leave these tissues at a somewhat slower rate, possibly to reenter the mucosa. Sporozoites in the lamina propria of the gut were found within host mononuclear cells in all 4 species studied. Most of the cells harbouring E. maxima and some of those with E. praecox were identified as intraepithelial lymphocytes while all others could only be identified as agranular mononuclear cells that were not characteristically macrophages.     

Eimeria stiedae: experimental infection in rabbits and the effect of treatment with toltrazuril and ivermectin

The aim of this study was to investigate the clinical, haematological, biochemical, lipid peroxidation, ultrasonographic and pathologic findings in hepatic coccidiosis induced by Eimeria stiedae in rabbits, and also to compare the treatment effects of both toltrazuril and ivermectin separately and in combination. In this study, 56 rabbits were divided into eight groups. The first group was designated as healthy control group. Rabbits were infected with 40.000 sporulated oocysts of E. stiedae. Groups 2, 3, 4, 5, 6, 7 and 8 were allocated as the infected control group, infected+toltrazuril-treated group, infected+ivermectin-treated group, infected+toltrazuril+ivermectin-treated group, non-infected+toltrazuril-treated group, non-infected+ivermectin-treated group, non-infected+toltrazuril+ivermectin-treated group, respectively. Haematocrit, Haemoglobin and MCV values as well as percentage of lymphocyte decreased in Groups 2 and 4 whereas leucocyte counts and percentage of granulocyte leucocyte increased. Serum GGT, ALT and AST activities increased but albumin value decreased. Plasma MDA concentrations increased whereas erythrocyte CAT, GSH-Px, and SOD activities decreased. Mean oocyst numbers in per gram faeces (epg values) increased in both groups during the study. Ultrasonographic examination revealed that the liver was enlarged and had hyperechogenic parenchyma. Bile ducts were dilated and hyperechogenic and the gall bladder was dilated. The livers of these animals were enlarged and typical macroscopic and microscopic findings of coccidiosis were present. Treatment with toltrazuril and toltrazuril+ivermectin combination were highly effective in reducing faecal oocyst output in infected rabbits. Haematological, biochemical and lipid peroxidation parameters and, ultrasonographic findings of the liver were close to control values for Groups 3 and 5. Necropsy of these animals showed no visible lesions related to hepatic coccidiosis although a few oocysts were detected in the bile duct epithelial cells.     

Genetic differences among brown trout, Salmo trutta, stocks and their importance for the conservation and management of the species

SUMMARY. 1. Review of published studies on genetic variation, as shown by electrophoretic studies of protein variation, in natural brown trout ( Salmo trutta L.) populations from Britain and Ireland, Finland, France, Greece, Iceland, Norway, Sweden, U.S.A. and U.S.S.R., revealed abundant geographical variation in gene frequency with individual populations containing only a limited part of the gene diversity of the species.
2. Thirty-eight (54%) of the seventy gene loci examined have been found to be polymorphic in the species with an average population showing polymorphism at 16% of its loci (range 0-34.8%).
3. The brown trout is naturally subdivided into a large number of reproductively isolated and genetically distinct populations within, as well as among, drainages.
4. Two independent post-glacial colonizations, by genetically distinct races, followed by independent evolution in separate drainages over the past 13,000 years is seen as responsible for the genetic diversity of brown trout in north-western Europe.
5. Many genetically unique populations have been lost in the past 100 years and there is an urgent need to identify and conserve the remaining genetic diversity. Genetically unique populations are an irreplaceable resource for rational management in relation to angling and future aquaculture potential.     

Cross-transmission studies with Eimeria arizonensis, E. arizonensis-like oocysts and Eimeria langebarteli: host specificity at the genus and species level within the Muridae

Cross-transmission experiments were done using sporulated oocysts of Eimeria arizonensis from Peromyscus truei and Peromyscus maniculatus, and oocysts of 2 putative species that resemble E. arizonensis, i.e., Eimeria albigulae from Neotoma albigula, and Eimeria onychomysis from Onychomys leucogaster. Oocysts of each species were inoculated into representatives of P. maniculatus and the latter 2 rodent species. Other experiments were conducted wherein oocysts of Eimeria langebarteli from Peromyscus leucopus were given to P. truei and P. maniculatus. Oocysts of E. arizonensis from P. truei and P. maniculatus could be transmitted only to P. maniculatus; likewise, oocysts of E. albigulae and E. onychomysis produced patent infections only in N. albigula and O. leucogaster, respectively. Oocysts of E. langebarteli from P. leucopus could be transmitted to P. truei, but not P. maniculatus. These results indicate that E. arizonensis, and the morphologically similar E. albigulae and E. onychomysis, are distinct species that are not transmissible between the genera of their respective hosts (Peromyscus, Neotoma, Onychomys), and that some isolates of E. langebarteli, reported from 6 species of Peromyscus and Reithrodontomys megalotis, may not always be infective to P. maniculatus.     

A pathogenic new species of Eimeria from the pygmy rabbit, Brachylagus idahoensis, in Washington and Oregon, with description of the sporulated oocyst and intestinal endogenous stages

In January 2003, fecal samples from 13 live pygmy rabbits, Brachylagus idahoensis (Merriam, 1891), were collected at the Oregon Zoo in Portland, Oregon, and sent to the University of New Mexico (UNM), Albuquerque, New Mexico, to be examined for coccidia. In July 2004, 14 more fecal samples were collected and sent to UNM, 6 from some of the same rabbits and 8 from 16 other rabbits (4 were pooled samples from siblings). In addition, tissue sections from 3 dead rabbits (2 from the Oregon Zoo, 1 from Washington State University) also were examined. Two of 4 (50%) pooled fecal samples and 8 of 17 (47%) 1-rabbit samples were positive for a single species of Eimeria, which we describe here as a new species. Sporulated oocysts were subspheroidal, 25.6 x 23.8 (22-28 x 21-27) microm, with a length:width (L:W) ratio of 1.1 (1.0-1.2). A micropyle (approximately 2 microm wide) and 0-1 polar granules were present, but an oocyst residuum was absent. Sporocysts were ellipsoidal, 13.4 x 8.1 (11-16.5 x 7.5-9) microm, with a L:W ratio of 1.7 (1.3-2.2), and they had a Stieda body and sporocyst residuum. Tissue sections showed a heavy infection of the villous epithelial cells of the proximal and mid-small intestine with coccidial endogenous stages, but no stages were found in liver hepatocytes. Meronts with approximately 46 (26-70) merozoites per infected cell appeared to be fully developed and were subspheroidal, 14.8 x 13.9 (13-18 x 10.5-16.5) microm. Developing macro- and microgamonts were indistinguishable from each other and were spheroidal to subspheroidal, 10.4 x 9.5 (9-11 x 7.5-10.5) microm. Mature macrogamonts were spheroidal to subspheroidal, 14.2 x 13.7 (12-17 x 11-16) microm, and mature microgamonts were smaller and subspheroidal, 11.9 x 10.8 (10.5-13 x 9-12) microm. This eimerian seems to be extremely pathogenic to young pygmy rabbits, and given the precarious nature of this unique genetic population, it appears to be an emerging pathogen that deserves immediate further study.     

The effect of propolis and its components on eicosanoid production during the inflammatory response

To investigate the possible mechanism of the therapeutic action of propolis, we studied: (a) the effect of propolis, its components, caffeic acid phenethyl ester (CAPE), caffeic acid (CA), quercetin and naringenin, as well as the synthetic compounds indomethacin (IM) and nordihydroguaiaretic acid (NDGA), and a novel lipoxygenase inhibitor N,N′-dicyclohexyl-O-(3,4-dihydroxycinnamoyl)isourea (DCHCU) on eicosanoid production by mouse peritoneal macrophages in vitro; (b) the effect of IM, NDGA, CA, CAPE, DCHCU and propolis on eicosanoid production during acute inflammation in vivo; and (c) the ex vivo and in vivo effect of dietary propolis on arachidonic acid metabolism. The ethanol extract of propolis suppressed prostaglandin and leukotriene generation by murine peritoneal macrophages in vitro and during zymosan-induced acute peritoneal inflammation in vivo. Dietary propolis significantly suppressed the lipoxygenase pathway of arachidonic acid metabolism during inflammation in vivo. CAPE was the most potent modulator of the arachidonic acid cascade among the propolis components examined.     

RFLP- and RAPD-based genetic relationships of seven diploid species of Avena with the A genome.

Relatively few molecular analyses are available for diploid oat species, which constitute the majority of the wild species of Avena and, therefore, the principal natural reservoir of variability. The present work reports an RAPD-(random amplified polymorphic DNA) and RFLP-(restriction fragment length polymorphism) based study of the intra- and interspecific variability of seven diploid A-genome oat species. Both types of markers resulted in valid tools for identifying polymorphisms both within and between species. The two statistical analyses, UPGMA (unweighted pair group method, arithmetic mean) and PCoA (principal coordinate analysis), computed on the basis of genetic similarities estimated from RAPDs and RFLPs, showed that the different accessions grouped according to species, but the similarity coefficients were consistently higher in the RFLP analysis. Furthermore, slight differences were observed in the intra- and interspecific relationships found with the two types of markers. This may support the hypothesis that the polymorphisms revealed by the two types of markers may associate with regions of the genome having different evolutionary rates. The relationships among species are not identical to those deduced from previous karyotypic and morphological studies, thus suggesting a partially different evolutionary pathway in oat speciation.     

Eimeria tenella: experimental studies on the development of resistance to amprolium, clopidol and methyl benzoquate.

The development of resistance by the Houghton strain of Eimeria tenella to the anticoccidial drugs amprolium, clopidol and methyl benzoquate has been studied. Resistance to amprolium and clopidol developed more readily in experiments where a large number of coccidia were exposed to the drug, either by increasing the number of oocysts in the inoculum or by increasing the number of birds in the group. When 45 birds were given 2.0 X 10(6) oocysts, resistance to amprolium and clopidol appeared after 6 and 7 passages respectively. In previous experiments, under similar conditions, resistance to robenidine developed after 6 passages, suggesting little difference between these three drugs. Resistance to amprolium and clopidol arose gradually as the concentration of drug was increased, but resistance to methyl benzoquate appeared in a single step from sensitivity to high-level resistance. Both amprolium and clopidol-resistant lines showed an 8-fold reduction in drug sensitivity. Attempts to measure the degree of resistance by calculation of the ED50 were unsuccessful.     

Host bromeliads and their associated frog species: Further considerations on the importance of species interactions for conservation

Bromeliads constitute a good example of symbiosis with organisms that spend their entire life cycle inside the plants, and often depend on them to breed. The bromeliads benefit from this interaction by increasing their nutrients intake. Conservation efforts tend to focus on a single endangered species, but in symbiotic associations, the viability of one species depends on that of the other. Based on IUCN criteria, any species that depends on another to complete its life cycle should be assigned a conservation status equivalent to that of the host taxon, where appropriate. We gathered published plus fieldwork data on the frog-bromeliad mutualism and compiled a checklist of 99 bromeligenous frogs species associated to 69 bromeliad hosts, and found threatened bromeliads hosting non-threatened frogs. We found that 62% bromeligenous frogs inhabit unspecified bromeliads. Finally, we propose strategies for improving understanding and conservation of the frog-bromeliad mutualism.     

The importance of cuticular permeability, osmolyte production and body size for the desiccation resistance of nine species of Collembola

Euedaphic collembolans have recently been shown to actively regulate internal osmotic pressure by means of sugars and polyols in response to desiccation. In contrast, studies of cuticular permeability have shown that some, especially epedaphic, species of collembolans may primarily rely on a low cuticular permeability to survive desiccation. To elucidate to what extent these strategies are important for desiccation resistance, the survival of 7-day acute desiccation stress (LRH(50)), the cuticular water conductance constant and osmolyte production were investigated in nine species of collembolans, covering euedaphic, hemiedaphic and epedaphic species. The LRH(50) values ranging from 98.8% to 95.2% RH showed no correlation with the vertical distribution of species, since both the highest and lowest values were found in epedaphic species. The water conductance varied from 698+/-141 to 41+/-13 microg h(-1) cm(-2) mmHg(-1) and showed good agreement with the vertical distribution of species in their natural habitats. Modelling the drying curves showed that, in addition to cuticular permeability and osmolyte production, body size also plays an important role in the survival of short-term severe desiccation stress. Furthermore, the model pointed to the need for behavioural responses to desiccation, particularly in epedaphic species. Thus, in keeping with expected humidity regimes in their respective microhabitats, euedaphic species rely on small body size, high cuticular permeability and the ability to actively regulate the osmotic pressure of their body fluids, hemiedaphic species have similar strategies but with reduced cuticular permeability, whereas in epedaphic species, active regulation of osmotic pressure is replaced by combinations of greatly reduced cuticular permeability or greatly reduced surface area to volume ratio combined with behavioural responses to desiccation.     

Neural cost functions and search strategies for the generation of block designs: an experimental evaluation

A constraint satisfaction problem, namely the generation of Balanced Incomplete Block Designs (v, b, r, kappa, lambda)-BIBDs, is cast in terms of function optimization. A family of cost functions that both suit the problem and admit a neural implementation is defined. An experimental comparison spanning this repertoire of cost functions and three neural relaxation strategies (Down-Hill search, Simulated Annealing and a new Parallel Mean Search procedure), as applied to all BIBDs of up to 1000 entries, has been undertaken. The experiments were performed on a Connection Machine CM-200 and their analysis required a careful study of performance measures. The simplest cost function stood out as the best one for the three strategies. Parallel Mean Search, with several processors searching cooperatively in parallel, could solve a larger number of problems than the same number of processors working independently, but Simulated Annealing yielded overall the best results. Other conclusions, as detailed in the paper, could be drawn from the comparison, BIBDs remaining a challenging problem for neural optimization algorithms.     

The use of anti-Pfs 25 monoclonal antibody for early determination of Plasmodium falciparum oocyst infections in Anopheles gambiae: comparison with the current technique of direct microscopic diagnosis.

Experimental infections of laboratory-reared anopheline mosquitoes were carried out with 57 Plasmodium falciparum gametocyte carriers from Cameroon. Prevalence of infected mosquitoes and oocyst intensity were determined by two independent methods. Young P. falciparum oocysts were detected on day 2 after feeding using an immunofluorescent assay, and the results were compared with direct microscopic examination of midgut oocysts on day 7 postinfection. The immunofluorescent assay was based on a FITC-labeled anti-25-kDa monoclonal antibody, while the direct microscopy was performed on midguts stained with 2% mercurochrome. Young oocysts were easily detected by their typical and bright green-fluorescing Pfs25 positive coat and their characteristic pattern of pigment granules under transmitted white light examination. The agreement between the results of the two methods was assessed using the Kappa coefficient on prevalences of positive infections and the interclass correlation coefficient on arithmetic mean oocyst load per infected midgut. The results indicated a low agreement between the two methods for the comparison of prevalences of infected mosquitoes. However, this agreement was near perfect for the comparison of mean oocyst intensities. Prevalences of positive infections and the overall number of parasites per positive gut were significantly correlated for both methods. Thus, the immunofluorescent test could be an appropriate tool for early determination of malaria infection in mosquitoes, particularly under laboratory conditions. The possible applications of this immuno-fluorescent technique are discussed.