Humoral and cellular responses to homologous extracts of Nematospiroides dubius and Nippostrongylus brasiliensis

and 1986. Humoral and cellular responses to homologous extracts of Nematospiroides dubius and Nippostrongylus brosiliensis. International Journal for Parasitology 16: 601–606. Humoral and cellular responses to homologous parasite extracts were studied in C57BL mice infected with Nematospiroides dubius or Nippostrongylus brasiliensis, to determine whether these parasites induced specific immunosuppression which might facilitate their survival. IgG and IgM titres to adult, excretory-secretory, larval and egg antigens from N. dubius and adult antigen from N. brasiliensis increased progressively for several weeks, irrespective of parasite rejection. Delayed-type hypersensitivity responses to the same antigens peaked after about 2 weeks and then remained constant in N. dubius -infected mice, but declined after the rejection of N. brasiliensis. The specific lymphoproliferative responses of spleen cells to these extracts reached peak values 1–3 weeks after infection and were anti-Thy 1.2-sensitive. They then declined sharply, but remained above the levels seen in uninfected mice. These results should be considered in the interpretation of any investigations into the stable host-parasite relationship which exists during a primary N. dubius infection.     

Immunosuppressive effects of extracts of helminthic parasites in C57BL mice

and 1986. Immunosuppressive effects of extracts of helminthic parasites in C57BL mice. International Journal for Parasitology 16: 607–615. Crude saline extracts of adult Nematospiroides dubius or Nippostrongylus brasiliensis worms administered with an i.p. immunization of ovalbumin (OA) in Al(OH)₃ depressed primary and secondary IgG responses, delayed-type hypersensitivity and in vitro splenic lymphoproliferative responses to the OA. The suppressive agent was trypsin-sensitive and was also present in larval extracts of the above parasites at levels proportional to their protein contents. Residual gut contents from infected mice caused no suppression. The extracts had little effect on the uptake of ¹²⁵I-PVP from the peritoneum or on peritoneal macrophage activation as assessed by cell numbers, lymphocystostatic potential and acid phosphatase activity. When present in cultures of normal spleen cells, the extracts impaired mitogen-induced lymphoproliferation if added with or before the mitogen. Possible roles for suppressor T cells, suppressive peritoneal or splenic macrophages and direct inhibition of clonal expansion in the suppression of the responses to OA are discussed.     

The response of the small intestine of the protein-deficient rat to infection with Nippostrongylus brasiliensis

The intestinal response of the protein-deficient Wistar rat was examined after primary infection with 1500 larvae of Nippostrongylus brasiliensis. Protein-deficient animals failed to expel N. brasiliensis after 15 days at a time when nutritionally normal animals had expelled more than 99% of the worm burden. Morphology of the small intestine of protein-deficient animals before infection showed small villi and crypt hypoplasia, followed after infection by sustained crypt hyperplasia and increased mitotic index of crypts. Protein deficiency was associated with fewer mucosal mast cells, goblet cells and intraepithelial lymphocytes. There was an impaired response of mucosal mast cells and goblet cells to infection. This could explain the deficiency of worm expulsion in these protein-deficient animals.     

The molecular size of nematode acetylcholinesterases and their separation from nematode allergens

The molecular size of nematode acetylcholinesterases, and their separation from nematode allergens. International Journal for Parasitology 3: 735–741. Acetylcholinesterases and allergens were derived from two parasitic nematodes, Nippostrongylus brasiliensis which parasitises rats and Trichostrongylus colubriformis which infects sheep and guinea pigs.

Chromatographic studies indicated that the mol. wt of nematode acetylcholinesterases lies between 65,000 and 75,000. The acetylcholinesterases of both species were separated from nematode allergens whose mol. wt is in the range of 10,000–50,000.

γG binding antigens of T. colubriformis were located in fractions with a mol. wt range of 30,000–150,000. γE binding activity was confined to allergenic material with a mol. wt of less than 70,000.     

Cross-reactivity between Necator americanus and Schistosoma mansoni in mice.

, , and 1992. Cross-reactivity between Necator americanns and Schistosoma mansoni in mice. International Journal for Parasitology 22: 1143–1149. Poly-parasitism is common in endemic communities and reactivity of sera from hookworm-infected patients against schistosomular antigens has been reported. Protective cross-immunity between N. americanus and S. mansoni was investigated in NIH and BALB/c mice. Protective resistance to homologous challenge with both parasites was confirmed in this model, however, functional immunity to heterologous challenge was not demonstrated. Sera from animals which had received homologous challenge with N americanus and from hookworm-infected mice, which had previously been exposed to radiation-attenuated S. mansoni, exhibited an enhanced IgGAM response to infective stage N. americanus somatic antigens. The implications of these results with respect to serodiagnosis are discussed.     

Oocysts of Neospora caninum, Hammondia heydorni, Toxoplasma gondii and Hammondia hammondi in faeces collected from dogs in Germany

Faecal samples of 24,089 dogs were examined coproscopically in two veterinary laboratories in Germany between March 2001 and October 2004. In 47 dogs, oocysts of 9–14 μm size were found. Their morphology was similar to those of Hammondia heydorni and Neospora caninum. Samples of 28 of these dogs were further examined by inoculation into gerbils: seven isolates induced a specific antibody response against antigens of N. caninum NC-1 tachyzoites. This response suggests that the isolates contained N. caninum. In addition to H. heydorni (12 times isolated), Toxoplasma gondii occysts (twice) and Hammondia hammondi oocysts (twice) were observed in dog faeces. The latter findings suggest that coprophagia with a subsequent intestinal passage by dogs plays a role in the dissemination of coccidian parasites for which cats are definitive hosts. Five of the seven N. caninum (NC-GER2, NC-GER3, NC-GER4, NC-GER5, NC-GER6) and the two T. gondii isolates (TG-dgGER1, TG-dgGER2) were successfully passaged into cell culture and are now available for detailed characterization. In contrast to oocysts of other parasites, N. caninum oocysts were predominantly found between January and April (Fisher exact; P=0.038). In the sera of dogs shedding N. caninum, no reactions against the immunodominant antigens with apparent molecular weights of 19, 29, 30, 33 and 37 kDa of N. caninum tachyzoites were observed 3–5 weeks after shedding. However, the animals recognized a 152-kDa N. caninum antigen. Compared with those identified as H. heydorni, T. gondii or H. hammondi, N. caninum oocyst isolates were significantly smaller in length with the 75th percentiles ≤10.7 μm when measured in concentrated sucrose solution and smaller length–width ratios with the 75th percentiles ≤1.06. It may thus be possible to develop criteria for a preliminary identification of N. caninum in dog faeces based on the oocyst morphology.     

Lattice deformation and thermal stability of crystals in spider silk

The X-ray diffraction of dragline silks, produced by Nephila and Cyrtophora spiders, were measured by synchrotron radiation in their original states or in situ during stretching and heating. Nephila pilipes spiders construct a two-dimensional orb web that must be rebuilt in one or 2 days, but Cyrtophora spiders form a three-dimensional tent web that can exist for several weeks in a tropical forest. Diffraction patterns of N. pilipes and Cyrtophora draglines resemble each other. Crystals of two kinds are identified in these draglines; one is aligned parallel to the silk direction and another is less oriented. The less oriented crystal in Cyrtophora dragline is aligned better than that in N. pilipes dragline, which generates about three times stronger diffract intensity. Crystals in N. pilipes and C. moluccensis dragline silks have remarkable thermal stability. Equatorial reflections remain undiminished until 350 and 450 °C for N. pilipes and C. moluccensis, respectively. In contrast, the meridional reflections S and (0 0 2), which are parallel to the silk thread, disappear at a temperature less than 100 °C for C. moluccensis but remain for Nephila up to 100 °C. Meridional reflections S and (0 0 2) shift to a smaller angle during stretching, whereas equatorial reflections remain constant in a range 1.0–1.3 times the original length. The position of the S reflection shifts rapidly in the first 10% of elongation from the original length but remains constant during subsequent stretching, whereas the (0 0 2) reflection shifts rapidly during the first 5% elongation from the original length and continues to shift subsequently. In contrast, the features of N. pilipes dragline alter insignificantly during stretching. Examination of the composition of amino acids of the draglines of N. pilipes and C. moluccensis indicates that a dragline of N. pilipes contains more glycine, but much less alanine, than that of C. moluccensis.     

Ultrastructural studies on experimental primary amoebic meningo-encephalitis (PAME) of mouse due to Naegler1a aerobia and Hartmannella culbertsoni

Primary amoebic meningo-encephalitis (PAME) in mouse can be easily produced by intranasal inoculation of Naegleria aerobia Singh & Das, 1970 and Hartmannella culbertsoni Singh & Das, 1970. The ultrastructural characteristics of N. aerobia and H. culbertsoni from infected mouse brain tissues showed many similar characteristics. The distinctive features between the two were the retention of phagocytic activity by N. aerobia as well as pinocytic activity in the ingestion of food as compared to H. culbertsoni where only pinocytic activity was observed. Staining characteristics of mitochondria suggested that N. aerobia was more active in the brain tissues than H. culbertsoni. Membrane-bound darkly stained bodies of unknown function were found in the brains infected with both N. aerobia and H. culbertsoni. They may have some bearing on the pathogenicity of these amoebae. Human cases of PAME so far reported are mostly due to N. aerobia and rarely with H. culbertsoni. This may be due to certain inherent characteristics present in N. aerobia which help them to survive on human brain tissue. On the other hand, the same may be lacking in H. culbertsoni. These ultrastructural differences in both N. aerobia and H. culbertsoni, as revealed by electron microscope studies of the infected brain tissues of mice, are presented in this paper.     

A genetic approach to species criteria in the amoeba genus Naegleria using allozyme electrophoresis

The present study employs allozyme electrophoresis to characterize and inter-relate 61 isolates of Naegleria. Diploidy was confirmed, with heterozygotes observed at 29 of the 33 loci established and in all but two isolates. With a single exception, isolates clustered at two levels of similarity, either below 21% or above 52%. It is argued that such a major discontinuity provides a sound biological basis for a species concept in Naegleria. On this basis the present species-level taxonomy does not reflect the genetic diversity of the genus. The study recognized 18 genetic groups of species rank. The subspecies N. australiensis italica deserves specific rank; additional thermophilic species not closely related to N. fowleri and N. lovaniensis are recognized; and N. gruberi as currently conceived is a complex of 10 species, at least five of which are represented in the formal culture collections. Most species are genetically too different for relationships to be elucidated by allozyme electrophoresis, supporting the view that some of the times of divergence within the genus are extremely ancient.     

Nicotiana benthamiana LRR‐RLP NbEIX2 mediates the perception of an EIX‐like protein from Verticillium dahliae

Verticillium wilt diseases caused by the soil‐borne fungus Verticillium dahliae result in devastating yield losses in many economically important crops annually. Here, we identified a novel ethylene‐inducing xylanase (EIX)‐like protein, VdEIX3, from V. dahliae, which exhibits immunity‐inducing activity in Nicotiana benthamiana. In vitro‐purified VdEIX3 can induce strong oxidative burst, activate the expression of defense‐related genes, and increase resistance against oomycete and fungal pathogens in N. benthamiana. VdEIX3 orthologs of other Verticillium pathogens also induce cell death in N. benthamiana, which form a new type of EIX protein family that is distinct from the known EIX proteins. A leucine‐rich repeat receptor‐like protein, NbEIX2, regulates the perception of VdEIX3 in N. benthamiana. Our results demonstrate that VdEIX3 is a novel EIX‐like protein that can be recognized by N. benthamiana NbEIX2, and also suggest that NbEIX2 is a promising receptor‐like protein that is potentially applicable to transgenic breeding for improving resistance to Verticillium wilt diseases.     

锁阳及其寄主白刺内生真菌的生态分布及遗传关系

首次研究了寄生植物锁阳及其寄主白刺内生真菌的分布特征及遗传关系.采用组织块法分离天然白刺、寄生体中锁阳和白刺的内生真菌,利用ITS-rDNA分子序列并结合形态学方法鉴定菌种,研究内生真菌的分离率、定殖率、分离频率、多样性指数、均匀度指数及相似性系数等的差异,以及寄生关系中内生真菌的多样性、遗传关系及分布特征等.结果表明: 本次获得的49株内生真菌隶属于18个分类单元,95.9%为子囊菌,4.1%为担子菌;内生真菌总分离率为15.3%,总定殖率为25.0%;天然白刺中内生真菌Shannon多样性指数最大,为2.13;锁阳花序与锁阳茎的内生真菌相似性系数最大,为0.50;镰孢菌属为白刺的优势菌属,青霉属为锁阳的优势菌属.锁阳与白刺寄生体中真菌类群的差异性分布表明寄生关系对内生真菌群落存在一定影响.     

Some ecological aspects of the copepod parasites of the common hake, Merluccius hubbsi, from the Argentine-Uruguayan coasts

A total of 2164 common hake Merluccius hubbsi captured in the Argentine-Uruguayan Common Fishing Zone was examined for parasitic copepods. The fish were infested with Chondracanlhus palpifer and Neobrachietia insidiosa f. lageniformis, the former parasitizing the buccal cavity and the latter the branchial arches. C. palpifer showed an increase in both prevalence and intensity in relation to the host size; infestation with N. insidiosa f. lageniformis decreased with increasing length of hake. Attachment site preferences and variations in the parasite distribution patterns as related to the host biology were observed. Evidence of negative association between copepod species and seasonal changes in the parasite composition was also found.     

Biochemical systematics and evolutionary genetics of the common shiner species group

Electrophoretic analyses were performed on blood proteins of the five members of the Notropiscornutus species group. The protein systems included two plasma esterases, transferrin, a pre-albumin and hemoglobin. Plasma protein polymorphism within and between taxa was common. Hemoglobin appeared to be a more consistent and conservative character for assessing phylogenetic relationships. As deduced by both biochemical and morphological evidence, N. cerasinus is the most primitive member of the species group. Uniqueness for several biochemical characters suggests that the striped shiner should be afforded full species recognition as N. isolepis but additional study is needed concerning its relationship with N. cornutus chrysocephalus. The closest biochemical similarity was between the forms interpreted here as subspecies N. cornutus cornutus and N. c. chrysocephalus. N. albeolus clearly evolved from N. c. cornutus but early hybridization with N. cerasinus resulted in limited introgression from that species.     

Metabolism of CO2 by leaves of different photosynthetic types of Neurachne species

Following a series of continuous exposures to ¹⁴CO₂ for different lengths of time, leaves from Neurachne munroi (C₄), N. minor (C₃-C₄) and N. tenuifolia (C₃| were estimated to assimilate 100%, 9% and 2–4%, respectively, of atmospheric CO₂ by the C₄ pathway. The percentage of ¹⁴C-label appearing in malate and aspartate in leaves of N. minor progressively increased with longer exposure times indicating that a significant proportion of its C₄ acids are formed as secondary products. In ¹⁴CO₂/¹²CO₂ pulse/chase experiments, the ¹⁴C-label in leaves of N. munroi was rapidly transferred from C₄ acids to sugar monophosphates plus sugar diphosphates, and finally to sucrose. In leaves of N. minor, the ¹⁴C-label was slowly metabolized from the C-4 carboxyl of malate and asparate (apparent half-time = 250 s), and the formation of C₄ acids as secondary products was again evident. ¹⁴C-label in serine/glycine accumulated to comparable magnitudes in both N. minor and in N. tenuifolia, but there was an initial lag phase in the accumulation of label in N. minor. C₄ photosynthesis is apparently of minimal importance in reducing photorespiration in N. minor, but leaf anatomical specializations and a possible compartmentation of photorespiratory metabolism may be of considerable importance.     

The physiology of the ovary: Maturation of ovarian granulosa cells and a novel role for antioxidants in the corpus luteum

During folliculogenesis the granulosa cells divide whilst in contact with each other, and so exhibit some of the characteristics of stem cells. In vitro we have shown that bovine granulosa cells from 3–7 mm follicles, like stem cells, divide without the need for a substratum, and produce colonies of cells. Growth factors, bFGF and IGF's, stimulate their division. These cells secrete and assemble a basal lamina, suggesting that the follicular basal lamina is produced by the granulosa cells. They have the morphological characteristics of follicular granulosa cells. Thus this system is ideal for studying the functions of immature granulosa cells because the cells do not spontaneously differentiate or luteinize into luteal cells, as occurs in culture on a substratum. On differentiation into luteal cells in vivo the cells express the steroidogenic enzymes for progesterone production and accumulate β-carotene. During culture of bovine luteal cells we observed that a proportion of the steroidogenic enzyme cholesterol side-chain cleavage cytochrome P450 enzyme became chemically cross-linked to its electron donor, adrenodoxin. P450 enzymes produce oxygen free radicals and oxygen free radicals can cause cross-linking between proteins in close proximity. Cell protect against this damage by the use of antioxidant vitamins. Repleting the cultured luteal cells with β-carotene reduced the amount of cross-linking. We conclude that the high levels of β-carotene in corpora lutea are to protect against damage due to oxygen free radicals generated in the course of progesterone synthesis.     

Differences in the lipid peroxidative status, cytochrome P-450 content and microsomal oxygen consumption between right and left lobes of the liver in fetal sheep. Comparison with maternal liver

The existence of the ductus venosus in mammalian fetal liver determines a different blood flow in the right and left lobes. This asymmetry is associated to a decrease in oxygen delivery to the right lobe. Some stages in oxygen metabolism are related to the formation of free radicals that can induce cellular damage dependent on the tissue oxygen concentration and on the activity of the enzymatic and non enzymatic systems involved in the tissue protection. In the present study, the differences in the lipid peroxidative status and some parameters related to the protection against oxygen toxicity between both lobes in the sheep fetal liver were analyzed and compared to the same parameters in the dam liver.     

The effect of light on oxygen consumption in two amphipod crustaceans-the hypogean Niphargus stygius and the epigean Gammarus fossarum

The effects of light on the metabolic rates of the hypogean amphipod Niphargus stygius and the epigean amphipod Gammarus fossarum were compared by measuring oxygen consumption and respiratory electron transport system (ETS) activity. They were exposed to light intensities of 720 and 4700 lx at 10°C. Oxygen consumption increased significantly in N. stygius exposed to both low and high intensities of light, but no significant increase was observed in G. fossarum at either intensity. The increase of oxygen consumption in N. stygius was significantly greater at the higher light intensity. This indicates a stress response in which exploitation of half the metabolic potential for energy production in N. stygius during exposure to high light intensity constitutes an adverse effect on its metabolism, since this species usually uses less than 25% of its total metabolic potential for standard metabolic demands.     

Cytokines, free radicals and resistance to Eimeria

The cytokine, gamma-interferon (IFN-gamma), which is produced by CD4(+) T cells, plays a crucial role in host resistance to Eimeria infections. Karen Ovington and Nick Smith propose that free oxygen radical generation by leukocytes in response to infection with Eimeria is the result of activation by IFN-gamma. The functional role of free oxygen radicals is unclear but these highly reactive radicals are produced by the leukocytes that infiltrate the intestine in large numbers during infection, and the parasites,enterocytes and cells of the immune system may all be vulnerable to oxidative damage. Gamma-interferon also appears to induce the enterocytes inhabited by Eimeria to turn against the parasite. The authors draw from literature documenting similar effects on other protozoa, especially Leishmania and Plasmodium, and speculate that reactive nitrogen intermediates produced by enterocytes have a functional role in resistance to Eimeria.     

Bile and bile salts and exsheathment of the intestinal nematodes Trichostrongylus colubriformis and Nematodirus battus

1972. Bile and bile salts and exsheathment of the intestinal nematodes Trichostrongylus colubriformis and Nematodirus battus. International Journal for Parasitology, 2: 433–438. Exsheathment of T. colubriformis was potentiated at physiological levels of CO₂ by bile and bile salts. Lamb bile and crude sodium taurocholate preparations produced the greatest increase in exsheathment while rabbit bile, sodium glycocholate and deoxycholate had less pronounced effects. Exsheathment in bile and taurocholate occurred at a pH above 4 and it is therefore suggested that infective larvae which fail to exsheath in the abomasum could well do so in the proximal part of the small intestine. Pure sodium taurocholate was found to greatly potentiate exsheathment of N. battus in vitro but this occurred at a pH below 3 and thus the action of this bile salt could not affect exsheathment in the host.