Experimental haemonchosis in white-tailed deer

White-tailed deer (Odocoileus virginianus) were successfully infected with Haemonchus contortus of sheep origin. Individual deer in each of three groups were inoculated with 0, 25,000, and 100,000 larvae respectively. Severity of infection was related to dose and signs of infection were most obvious in the heavily inoculated animals. Infected deer were weak, emaciated, and anemic, similar to the clinical response in sheep. Hemoglobin, packed cell volume, and total serum protein values for both infected groups were significantly lower than for the controls. Inhibition of larval growth was noted in both infected groups, but was most pronounced in the group which received 100,000 larvae. Inhibition of egg production was also noted in this group. The potential importance of H. cortortus in deer populations was discussed.     

Variation in resistance to haemonchosis: selection of female sheep resistant to Haemonchus contortus.

Seventy female lambs (6-7 months old) which were exposed to natural infections of Haemonchus contortus were designated as responders or non-responders on the basis of 10 weekly cumulative faecal egg counts. Selected responder and non-responder lambs were treated with ivermectin, housed separately and 6 weeks post-housing, seven lambs from each group were given a trickle infection of Haemonchus contortus at 1000 L3 daily for 5 days per week up to 2 weeks and examined weekly for 10 weeks after first infection. Analysis of data revealed significantly lower mean faecal egg counts and non-significantly less weight loss in responder than non-responder lambs. Mean values of haemoglobin, packed cell volume, total serum protein and peripheral eosinophil counts were significantly higher in responders than non-responders. In contrast, serum pepsinogen concentration was significantly less in responders than in non-responders. At 10 weeks post-infection, there were fewer pathological lesions and significantly lower worm burdens in responders than in non-responders. These results demonstrate a distinct resistance in responders to Haemonchus contortus infection.     

火炬松胚性细胞悬浮培养物的生长参数变化

以火炬松（PinustaedaL.）成熟合子胚来源的胚性愈伤组织为材料建立了胚性细胞悬浮系,测定了其培养物的鲜重、干重、细胞体积和胚数及培养液中的pH值、电导率和蔗糖浓度等生长参数在培养过程中的变化动态。结果表明,在培养周期内,培养液中的pH值、电导率和蔗糖浓度的逐步降低与培养物的鲜重、干重、细胞体积和胚胎数的逐步增加保持一致性。在培养至18—21d,pH值、电导率和蔗糖浓度均接近或降到最低点,而胚数及细胞体积的增长都达到最高点。     

The intramammary inflammatory response of genetically resistant Merino ewes infected with Haemonchus contortus.

The mammary glands of 103 pasture-reared non-lactating, non-pregnant Merino ewes were infused via the teat canal with antigens prepared from the nematode Haemonchus contortus, and the inflammatory response to infusion assessed by washing the gland of its contents after 24 h and 14 days. The ewes were of two genotypes: one with proven high levels of resistance to infection with the nematode H. contortus, the other random-bred animals with relative susceptibility to infection. On day 0 of a H. contortus infection, one gland of the subgroups of both genotypes was infused with the antigen preparation. At the same time, the other gland of the random-bred ewes was infused with sterile physiological saline. A third group of infected random-bred ewes was infused with only sterile physiological saline. Similar infusions were performed on other subgroups on days 12, 21 and 35 of infection, which was then terminated with anthelmintic. A fourth group of uninfected random-bred control ewes was given both infusions 35 days after the other groups were infected. Sheep of the resistant genotype had lower worm egg counts and smaller reductions in blood packed cell volumes from day 21 of infection. Infusion of antigen had no effect on the course of infection and no effect on the response of the other gland, which had been infused with saline alone. The dominant leukocyte response from the antigen-infused gland was eosinophilia. On all days of infusion, and after both 24 h and 14 days, eosinophil counts from the resistant genotype were higher than those from their random-bred counterparts. The sheep mammary gland provides a source of eosinophils whose number is related to host genotype and stage of infection and may provide a model for the investigation of cellular responses in mucosal immunity to nematode infections.     

Dielectric spectroscopy as a novel and convenient tool for the study of the shear sensitivity of plant cells in suspension culture.

Plant cell suspensions of different species and different age were subjected to hydrodynamic stress while following the decline in the volume fraction of intact cells by measuring the permittivity of the cell suspension at radio frequencies. Results were compared with the fresh weight, dry weight, packed cell volume and cell number of the suspensions. At first a rapid decline is seen as the most shear-sensitive cells are broken up, followed by a slower decline as less sensitive cells are broken up. The sensitivity of the cells to shear stress depended strongly on the cell line used but only slightly on their age, older cells being more sensitive. The dependence of the shear sensitivity on the cell line might be an effect of the species investigated, the culturing conditions of the cell line, or both. It was found that cells that grow in a finely dispersed suspension are much less prone to shear stress than is often assumed.     

Infection of immunosuppressed mice with the abomasal nematode parasite of ruminants, Haemonchus contortus

Infective larvae of Haemonchus contortus established in mice either immunosuppressed with the corticosteroid, dexamethasone, or the cytotoxic drug, cyclophosphamide, or treated with the histamine H2 inhibitor, cimetidine. Infections persisted for as long as the immunosuppressive treatment (7 days) and growth of larvae was similar to that seen in sheep. Virtually no larvae survived in untreated mice. Accordingly, it would appear that adaptive immunity is an important barrier against primary infection by H. contortus in mice and is a determinant of host-range for this parasite. Antibody raised in either sheep or mice against soluble extracts of adult H. contortus precipitated with different but overlapping sets of worm antigens. This suggests that the unique antigens recognized by the mouse compared with the sheep are crucial for the rapid protective responses which prevent primary infection.     

Experimental transmission of gastro-intestinal nematodes between sheep (Ovis aries) and Thomson's gazelles (Gazella thomsonii).

Experimental cross-transmission of gastro-intestinal nematodes between Merino sheep (Ovis aries) and Thomson's gazelles (Gazella thomsonii) from similar but separate grazing habitats in Kenya was studied. Cross-transmission did occur with some species but the faecal egg counts of sheep were higher than the gazelles' following infection with larvae isolated from either sheep or gazelles. Of the 11 gastro-intestinal nematodes which became established in gazelles following infection with larvae cultured from nematodes in gazelles, only Haemonchus contortus, Trichostrongylus probolurus and Cooperia hungi were infective for sheep. Following infection with larvae of either sheep or gazelle origin, the H. contortus recovered from the sheep at necropsy were more numerous and had greater average weights, lengths and spicule lengths than those recovered from the gazelles. This would suggest that H. contortus is primarily a sheep parasite. It is concluded that Thomson's gazelles probably would not contribute significantly to the problem of haemonchosis in sheep in areas of shared grazing, unless efforts were being made to eradicate the parasite from sheep, in which case the gazelles would act as a continuing reservoir of infection.     

A rapid method of determining growth characteristics of plant cell populations in batch suspension culture

A non-destructive, simple and accurate method of determining the relative growth rate (RGR) of the packed cell volume (PCV) of plant suspension cells in one Erlenmeyer flask at any time during the incubation period is described. The Erlenmeyer flask was tilted and the length of the chord formed by the surface of the packed cells across the bottom of the flask was measured. The chord length and the log PCV were correlated in a calibration line. The method enables the RGR during the exponential growth phase to be calculated by multiplying the slope of the linear part of the curve of the chord length in time with the slope of the calibration line. In order to investigate other growth parameters and to analyse the accuracy of the method statistically, a four-parameter function for the chord length and a computer program were used. The RGR during the exponential growth phase of cell suspensions of Solanum tuberosum and Haplopappus gracilis appeared to be independent of the PCV of the inoculum. The method appeared to be sufficiently accurate.     

Spatial and temporal analysis of non-steady elongation of rice leaves

A precise knowledge of the temporal and spatial distributions of cell division and tissue expansion is essential for appropriate leaf sampling in omics studies and for analyses of plant–environment relations. Elongating leaves of rice were studied during their whole development for elongation rate, distribution of cell length, cell production rate and spatial distribution of growth in the leaf. In seven genotypes, the pattern of leaf elongation rate followed three phases: (1) an exponential increase before leaf appearance; (2) a short phase (2–4 d at 20 °C) with a stable leaf elongation rate around leaf appearance; and (3) a phase of 8–10 d with a progressive decrease in elongation rate. The profile of cell length along the leaf changed with time during the first and last phases, but was time invariant around appearance. We propose a method adapted to non-steady elongation based on anatomical measurements, which was successfully tested by comparing it with the pricking method. It allowed analysis of the change with time in the spatial distribution of growth from initiation to end of leaf growth. The length of leaf zones with cell division and tissue elongation varied with time, with maximums of 21 and 60 mm respectively around leaf appearance.     

Detection of anti-Haemonchus contortus antibodies in sheep by dot-ELISA with immunoaffinity purified fraction of ES antigen during prepatency

Haemonchosis is a very common disease in small ruminants caused by H. contortus, a blood sucking parasite causing anaemia that may be fatal particularly to young animals. Therefore, detection of the infection during prepatent period is important for early treatment. Excretory-Secretory (ES) protein of H. contortus was purified through immunoaffinity chromatography. Dot -ELISA was performed with crude ES antigen as well as immunoaffinity purified fraction (F-1) with experimental and natural sera of sheep infected with H. contortus. Solid dot formation took place with 4 day, 1, 2 and 3 weeks post infection sera. Dot formation did not take place with negative control serum and uninfected control animal serum. When crude ES antigens was reacted to natural sheep sera having H. contortus infection, 60% sera samples showed solid dot formation whereas in F-1 fraction 75% of the sera samples showed solid dot indicating purified fraction was a more potent antigen. Crude ES and F-1 were also fractionated through SDS-PAGE. ES antigen revealed polypeptides in the range of 10 to 200 kDa of which 26, 32, 60 and 120 kDa were found more prominent. F-1 fraction on SDS-PAGE analysis revealed only four polypeptides of 26, 32, 60, and 120 of which 60 and 120 kDa were found to be most prominent. Results indicate that the purified fraction of ES antigen may be utilized for early diagnosis of haemonchosis. Further studies on cross antigenicity of this fraction with other nematode and trematode needs to be conducted.     

Haemonchus contortus: in vitro and in vivo anthelmintic activity of aqueous and hydro-alcoholic extracts of Hedera helix

In vitro anthelmintic activity of crude extracts of the ripe fruits of Hedera helix was investigated on eggs and adult nematode parasites Haemonchus contortus. Aqueous extract of H. helix was also evaluated for in vivo anthelmintic activity at dose of 1.13 and 2.25 g/kg in sheep artificially infected with H. contortus. ED(50) for egg hatch inhibition was 0.12 and 0.17 mg/ml for aqueous and hydro-alcoholic extracts, respectively. There was no statistically significant difference in the activity of the two extract types (p>0.05). Hydro-alcoholic extract showed better in vitro activity against adult parasites compared to the aqueous extract. Significant faecal egg count reduction (FECR) was detected in groups treated with both doses of H. helix (p<0.05) on day 2 post-treatment. On day 7 post-treatment significant reduction was detected only for higher dose of H. helix (p<0.05) while on day 14 post-treatment there was no significant FECR in both groups treated with H. helix. The percentage of larvae recovered from culturing faeces obtained from groups of sheep treated with lower and higher doses of H. helix was 47.52% and 36.07%, respectively, which was significantly lower than (p<0.05) that recovered from the control group (60%). Significant (p<0.05), dose dependent total worm count reduction (WCR) was observed for groups of sheep treated with H. helix. Increasing the dose of H. helix improved the efficacy against the male than the female parasites. Treatment with both doses of H. helix helped the animals maintain their packed cell volume (PCV) unlike the untreated control group. The overall findings of the current study indicated that H. helix has a potential anthelmintic benefit and further in vitro and in vivo evaluation of the different parts and fractions is needed to make use of this plant for therapeutic purposes.     

Immunity of sheep to homologous challenge with dog-borne Sarcocystis species following varying levels of prior exposure

Ford G. E. 1985. Immunity of sheep to homologous challenge with dog-borne Sarcocystis species following varying levels of prior exposure. International Journal for Parasitology15: 629–634. Grazing sheep in which sarcocysts develop (sarcosporidiosis) survive the pre-cyst stages of the parasite infection which cause pathophysiological changes (sarcocystosis). A peak temperature rise occurs at 3–4 weeks post-infection in experimental animals. A maximum fall in packed cell volume occurs at 5–6 weeks post-infection. Groups of sheep maintained specific pathogen free were experimentally infected with sporocysts of dog-origin to determine the dose level required for protection or resistance, and 13 weeks later challenged with a pathogenic dose of 50,000 sporocysts. Prior exposure levels (PEL) were used of nil, 5, 50, 500 and 5000 sporocysts. All prior infections elicited an antibody response to a complement fixation test in all sheep. A peak antibody level was detected within 2 weeks of challenge, with a higher titre for all previously exposed groups than for the nil PEL controls. Resistance of the sheep to the challenge dose, based on reduction of counts of parasites developing (sarcosporidiosis), was shown by significantly lower counts for the 50 PEL group (85%), as well as for the 500 (92%) and 5000 PEL (98%) groups. The 5 PEL group was not resistant. The protection of sheep, based on reduction of the above pathophysiological changes (sarcocystosis), was highly correlated with the level of prior exposure (r = ?0.80, P < 0.001). The changes in body temperature and packed cell volume were significantly less in groups with 500 and 5000 PEL than with nil and 5 PEL. The 50 PEL group was intermediate.     

The mechanism of cell elongation during lens fiber cell differentiation

Lens fiber formation is characterized by extensive cell elongation. Earlier studies have shown that lens cell elongation in vitro can occur in the absence of microtubules and is associated with a proportional increase in cell volume. We have previously suggested that lens fiber cell elongation is directly caused by an increase in cell volume. In this report, lenses from 3- and 6-day-old chicken embryos were three-dimensionally reconstructed from serial sections to provide a measure of cell volume and length during various stages of primary and secondary lens fiber formation. In both cases, cell volume was highly correlated with cell length during lens cell elongation. In addition, during primary lens fiber formation, large intercellular spaces between lens vesicle cells disappeared as these cells began to elongate to form lens fibers. Loss of intercellular spaces would be expected if increasing cell volume were responsible for cell elongation. Finally, results of experiments in which the lens capsule was cut with a fine tungsten needle suggested that the capsule was elastic and normally under tension. These findings were used to formulate a model which accounts for the major events in lens morphogenesis based on (1) the regulation of cell volume, (2) the junctions present between lens cells, and (3) the constraint provided by the elasticity of the lens capsule.     

The Effect of Nutrient Medium Composition on the Growth Cycle of Catharanthus roseus G. Don Cells Grown in Batch Culture

Growth of C. roseus cell suspension culture was defined in termsof dry weight, cell number, mitotic index, and packed cell volume.Removal of major nutrients from the medium was monitored asa function of culture growth. Phosphate and sucrose were theonly macronutrients completely exhausted. Utilization of thesetwo nutrients occurred parallel with increments in dry weightand cell number. Increasing the nutrient medium levels of sucroseand phosphate prolonged growth of this culture; lag and exponentialphases were extended; cell number and dry weight yield weredoubled. Dry weight assimilation was enhanced by increasingthe nutrient medium level of sucrose, whereas increments incell number were related to phosphate level. Two alkaloid fractions(fractions 1 and 2) were identified in this cell line. Fraction2 alkaloid level declined as the nutrient medium supply of nitrogenwas depleted.     

The Effects of Nutrient Deficiency and Rust Infection on the Relationship Between Root Dry Weight and Length in Groundsel (Senecio vulgaris L.)

Groundsel (Senecio vulgaris L.) was grown in sand culture ata range of nutrient concentrations. Except when nutrient deficiencywas severe, infection by the rust fungus Puccinia lagenophoraeCooke substantially reduced root dry weight but had little effecton root length. Thus, specific root length (SRL, cm root mg^–1d. wt) was significantly increased in rust-infected plants.The inhibition of root dry weight caused by rust infection wasmost pronounced late in development, especially after floweringwhen, in control plants, root elongation but not dry weightaccumulation ceased. In rusted plants, and in all plants subjectedto severe nutrient deficiency, dry weight accumulation in theroots ceased concurrently with root elongation. Late in developmentat high nutrient concentration adventitious roots with low SRLswere produced. However, infection did not modify the productionof such roots and increases in SRL could not be attributed tochanges in any single type of root. There was an inverse relationship between SRL and root diameter.This relationship was unaffected by rust infection whilst nutrientdeficiency changed only its intercept: at a given SRL rootsof nutrient stressed plants were thinner than those of plantswith adequate nutrient supply. Thus, the smaller diameter ofroots of nutrient-stressed plants occurred independently ofmeasured changes in SRL but, in the absence of nutrient stress,the decrease in root diameter caused by rust was closely relatedto increases in SRL. Changes in the root: length relationships in rusted plants mayhave important implications for root activity in the field.In view of the reported changes in SRL, inhibition of root growthin terms of dry weight may be a poor indicator of potentialchanges in activity. Senecio vulgaris, rust infection, nutrient deficiency, root weight: length ratio, root diameter     

Comparison of transfusion with DCLHb or pRBCs for treatment of intraoperative anemia in sheep.

Isoflurane-anesthetized sheep were transfused with packed red blood cells (pRBCs) or diaspirin cross-linked hemoglobin (DCLHb) for treatment of intraoperative hemorrhage. A rapid 15-min hemorrhage with lactated Ringer (LR) infusion maintained filling pressure at baseline and reduced blood hemoglobin (Hb) to ~5 g/dl. Sheep received 2 g/kg Hb, DCLHb (n = 6), or pRBCs (n = 7); control group received LR alone (n = 6). After 2 h, anesthesia was discontinued; sheep were monitored in the animal intensive care unit for 48 h. DCLHb expanded blood volume more, but increased total blood Hb less, than pRBCs. Lower Hb and increased methemoglobin resulted in lower arterial oxygen content compared with the pRBCs. DCLHb caused pulmonary hypertension (from 13 to 30 mmHg) and elevated filling pressure (from 6 to 15 mmHg). Cardiac outputs (CO) were similar for all groups during anesthesia; however, during recovery CO increased only in the LR and packed pRBCs groups. DCLHb may limit the reflex ability to increase CO after volume expansion. Hemodynamic effects of DCLHb may be exaggerated when infused after large-volume LR.     

Chlorophyll synthesis in chlorella I. Occurrence of a lag phase on initiation of a dilute culture

A culture of Chlorella established by 30-fold dilution of a culture already grown to a level of 15 ml packed cell volume per liter produces little chlorophyll for approximately 12 hours. Investigation of other characteristics such as nitrogen incorporation, increase in packed cell volume and dry weight as well as RNA level show all of these to increase without any significant lag. α-Linolenate, which can be considered as a chloroplast marker, increased markedly. Photosynthetic oxygen evolution and respiration as well as the heme enzyme, catalase, increase also, indicating that the lag in chlorophyll synthesis is not due to a general inability to produce the porphyrin moiety.     

THE ROLES OF PLANT HORMONES IN STYLE AND STIGMA GROWTH IN GAILLARDIA GRANDIFLORA (ASTERACEAE)

Style and stigma elongation and stigma unfolding, and the roles of plant hormones in these processes in Gaillardia grandiflora Van Houtte were investigated. Style and stigma elongation in vivo began just after anthesis, and style elongation was accompanied by epidermal cell elongation (greatest near the stigma) and a fresh weight increase, but not by cell division or a dry weight increase. The stigma unfolded after the style and stigma elongated. Style-stigma units excised from young disc flowers of this composite were measured as they responded to plant growth regulators applied singly, as well as in sequential and simultaneous combinations, in vitro. Style elongation was promoted by auxin, was inhibited by gibberellins and ethylene, and was unaffected by other growth regulators. Stigma elongation followed a similar pattern of response. Endogenous auxin levels and ethylene production showed parallel variation and endogenous gibberellin levels showed inverse variation with style and stigma elongation. Stigma unfolding was more sensitive to auxin applications and was promoted by applied ethylene. Ethylene production showed parallel variation and endogenous auxin levels showed inverse variation with stigma unfolding. AVG and Co²⁺ applications decreased auxin-induced style elongation and fusicoccin promoted all of the growth responses of style-stigma units in vitro. A gibberellin-auxin-ethylene-acid growth interaction mode of control is proposed for these three growth processes.     

Haemonchus contortus and Trichostrongylus colubriformis did not adapt to long-term exposure to sheep that were genetically resistant or susceptible to nematode infections

We tested the hypothesis that Haemonchus contortus and Trichostrongylus colubriformis would adapt to long-term exposure to sheep that were either genetically resistant or susceptible to H. contortus. Sheep genotypes were from lines with 10 years prior selection for low (resistant, R) or high (susceptible, S) faecal worm egg count (WEC) following H. contortus infection. Long-term exposure of H. contortus and T.colubriformis to R or S genotypes was achieved using serial passage for up to 30 nematode generations. Thus, we generated four nematode strains; one strain of each species solely exposed to R sheep and one strain of each species solely exposed to S sheep. Considerable host genotype differences in mean WEC during serial passage confirmed adequate nematode selection pressure for both H. contortus (R 4900 eggs per gram (epg), S 19,900 epg) and T. colubriformis (R 5300 epg, S 13,500 epg). Adaptation of nematode strain to host genotype was tested using seven cross-classified tests for H. contortus, and two cross-classified and one outbred genotype test for T. colubriformis. In the cross-classified design, where each strain infects groups of R, S or randomly bred control sheep, parasite adaptation would be indicated by a significant host genotype by nematode strain interaction for traits indicating parasite reproductive success; specifically WEC and, for H. contortus strains, packed cell volume. We found no significant evidence of parasite adaptation to host genotype (P > 0.05) for either the H. contortus or T. colubriformis strains. Therefore, we argue that nematodes will not adapt quickly to sheep bred for nematode resistance, where selection is based on low WEC, although selecting sheep using a subset of immune functions may increase adaptation risk. Our results support the hypothesis that nematode resistance is determined by many genes each with relatively small effect. In conclusion, selection of sheep for nematode resistance using WEC should be sustainable in the medium to long-term.     

Aspects of Growth and Metabolism in a Suspension Culture of Acer pseudoplatanus (L. ) Grown on a Glycerol Carbon Source

A suspension culture of Acer pseudoplatanus cells was transferredfrom medium containing 2% (w/v) glucose to an identical onecontaining glycerol at 2% (w/v) as the sole carbon source. Thepatterns of cell number increase, dry weight increase, and changesin packed cell volume showed marked differences as a resultof this transfer. The glucose-grown cultures contained a small proportion of cellsof exceptionally large diameter, and transfer to the glycerolcarbon source appeared to bring about a considerable increasein their number. These larger cells, in both glucose- and glycerol-containingcultures, exhibited considerable differences in cell wall architecturewhen compared with their smaller counterparts. They appearedmore irregular, and had much looser cellulose microfibril arrangementin their outer layers