Numbers Of Globule Leucocytes Research Articles

Differences in immune responses to Haemonchus contortus infection in the susceptible Ile de France and the resistant Santa Ines sheep under different anthelmintic treatments regimens

Understanding the immunological basis of resistance to gastrointestinal nematode infections in livestock is important in order to develop novel methods of parasite control such as vaccination or genetic selection for parasite resistance. The present study aimed to investigate differences in immune response between parasite resistant Santa Ines and susceptible Ile de France sheep breeds to natural Haemonchus contortus infection. Parasitological parameters, humoral immunity, local and circulating cellular immune responses were evaluated in 19 Santa Ines and 19 Ile de France lambs undergoing different anthelmintic treatments regimens: suppressive treatments (SUP) or targeted selective treatments (TST) over a 5-month grazing period. Santa Ines lambs had significantly lower Haemonchus faecal egg count and worm burden compared to Ile de France regardless of treatment regime. In addition, circulating blood eosinophils count and parasite-specific IgG levels were significantly higher and more rapidly induced in Santa Ines lambs. Abomasal immune responses were generally greater in the resistant breed, which had significantly higher levels of parasite-specific IgA in mucus, and elevated number of globule leukocytes and CD3+ T cells within the abomasal mucosal. Furthermore, numbers of POU2F3+ epithelial cells, a tuft-cell specific transcription factor, were also elevated in the Santa Ines breed, suggesting that this breed is better able to initiate T-helper type 2 immune responses within the abomasum. In conclusion, the differential immunological responses detailed here are relevant to understanding resistance to gastrointestinal nematodes in other host breeds, as well as to resistance breeding as a sustainable control approach for parasitic infections.

The relationship between the rapid rejection of Haemonchus contortus larvae with cells and mediators in abomasal tissues in immune sheep

Rapid rejection or immune exclusion of challenge larvae is a well recognised phenomenon in sheep hypersensitised by repeated infection with gastrointestinal nematodes. While mast cells and globule leukocytes (GLs) are typically associated with this rapid rejection response, the exact mechanisms and mediators involved are not known. This study has adapted a recently developed ex vivo tissue explant model to examine in more detail the cells and mediators involved in preventing establishment of Haemonchus contortus L3s in abomasal tissue of sensitised sheep. Hypersensitisation of sheep by repeated larval infection resulted in a significant inhibition of larval establishment in abomasal tissue cultures and the extent of inhibition was dependent on the sensitisation dose. Both mast cells and GLs, but not eosinophils, were increased in abomasal tissues of hypersensitised sheep. Globule leucocyte numbers decreased significantly after 3 h of culture, independent of the addition of L3s. In contrast, mast cell numbers only decreased after addition of L3s to the tissue cultures and this was associated with an increased release of histamine in tissue washes after incubation with L3s. Although, there was no significant difference in the number of tissue eosinophils between the groups, there was a marked increase in the eosinophil-specific protein, galectin-14, in tissue washes of the hypersensitised sheep after culture, suggesting eosinophils and their products may play a hitherto unrecognised role in the rapid rejection response. Further studies using specific inhibitors in this ex vivo tissue explant model may delineate the relative role of each cell population and mediator in the rapid rejection process.

The effect of corticosteroid treatment on local immune responses, intake and performance in lambs infected with Teladorsagia circumcincta

The nutritional cost of, and the sequential cellular changes associated with the developing immune response to the abomasal parasite Teladorsagia circumcincta were investigated using corticosteroid-induced immune-suppression. Six-month-old lambs with minimal nematode experience were either infected with 4000 L3 T. circumcincta per day (group IF), similarly infected and concurrently immune-suppressed with methylprednisolone acetate (group ISIF), immune-suppressed only (group IS) or remained as controls (group C). Food intake, faecal egg count (FEC) and antibody titres in plasma were recorded weekly, worm burden at necropsy on day 63 p.i. and body composition by X-ray computed tomography on days −2 and 62 p.i. Furthermore, sequential immunological changes at the site of parasite infestation in the abomasal mucosa were measured from serial biopsy tissue samples taken from additional animals that were fitted with an abomasal cannula and either infected with the same regime as IF animals above (group CnIF) or concurrently infected and immune-suppressed as above (group CnISIF). Corticosteroid treatment resulted in greater FECs (P<0.01) and worm burdens (P<0.01) in both ISIF and CnISIF compared with IF and CnIF sheep, respectively. Infection reduced feed intake by 17% between 14 and 28 days p.i. (P<0.05) and efficiency of energy utilisation by 20% (P=0.07) in IF animals but not in ISIF animals. Mast cells, globule leukocytes and IgA in tissue biopsy samples were elevated in CnIF from 42 days p.i., all of which were abrogated by corticosteroid treatment. The ability to regulate the worm population appeared to be associated with a rise in tissue IgA concentration and numbers of globule leucocytes (GL). The results support the hypothesis that a majority of the production losses that occur during infection of lambs with T. circumcincta in lambs are a consequence of the host immune response. These findings may have implications for regimes that promote the development of a strong host immune reaction to gastrointestinal parasites in lambs.

Parasitological characteristics and tissue response in the abomasum of sheep infected with Haemonchus spp.

A histopathological study was conducted on the abomasal mucosa of 40 Corriedale sheep exposed to natural infection with Haemonchus spp. The sheep were allowed to graze on contaminated pastures for 14 days and, after being housed for 28 days, they were slaughtered. Fecal samples were collected for fecal egg counts (FEC) and abomasum samples were obtained for histopathological examination and eosinophil, mast cell and globule leucocyte counts. The number of Haemonchus spp. present in the abomasum was estimated from a 10% aliquot of its content. The number of eggs per female was estimated for 10 Haemonchus spp. females collected from each animal. The following significant correlation coefficients were obtained between the characteristics analyzed: number of Haemonchus and FEC (r = 0.86), female length and number of eggs per female (r = 0.60), female length and FEC (r = 0.53), number of eosinophils and number of Haemonchus (r = 0.48), number of eosinophils and number of globule leucocytes (r = 0.54), number of mast cells and female length (r = -0.39), and number of mast cells and number of globule leucocyte (r = 0.34). The characteristics that showed the highest correlation with animal load was FEC.

The effect of truncated infections with Ostertagia ostertagi on the development of acquired resistance in calves

The relative contribution of the third (L 3), fourth (L 4) and adult stages of Ostertagia ostertagi to the development of immunity was assessed in calved which were either continuously infected during 21 weeks or subjected to infections truncated by anthelmintic treatment at the L 3 or L 4 stage. A fourth group remained uninfected (control group). Faecal samples and blood samples were collected weekly for faecal egg counts and determination of pepsinogen and antibody levels. Only the continuously infected animals showed positive egg counts, which fell towards the end of the primary infection period. Pepsinogen and antibody levels remained high in the continuously infected group until the end of the primary infection period. At that time, they were significantly higher compared to the control calves, with intermediate values in the truncated infection groups. After the 21 weeks primary infection period all animals were dewormed. To evaluate the protection provided by the different immunisation protocols, all animals were challenged 1 week later with 156 000 Ostertagia L 3, spread over 12 consecutive days. The marked reduction in egg counts following challenge infection indicated a certain degree of immunity in the continuously infected calves, which was confirmed at necropsy by the reduced worm burdens, the high percentage of inhibited early L 4 larvae, the reduced size of the adult worms and the higher numbers of mucosal mast cells in this group. Numbers of globule leucocytes and eosinophils were not significantly different from the control group. Infections truncated by anthelmintic treatment elicited poor development of immunity as shown by the egg output after the challenge infection and the percentages of arrested larvae and the lengths of adult worms which were intermediate to those of the continuously infected calves and control animals.

The immune response of sheep surgically modified with intestinal loops to challenge with Trichostrongylus colubriformis

Aspects of the local immune response to nematode challenge were investigated in vivo in isolated loops of the upper small intestine of mature sheep that were immunised by repeated infections with Trichostrongylus colubriformis infective larvae (L3). Groups of 3 sheep were challenged either through the loop (Group 1) or orally (Group 2) with T. colubriformis L3, the third group served as unchallenged controls (Group 3). Nematode specific antibody levels, mast cell proteinase levels (SMCP) and larval migration inhibition (LMI) activity were determined in loop secretions for 4 weeks after challenge. The intestinal loops remained functional throughout the experiment. Groups 1 and 2 were re-challenged 2 weeks after the first challenge, and all 3 Groups were slaughtered 2 weeks later. Histopathological examination showed elevated numbers of globule leukocytes (GL) in both the nematode-challenged loop and unchallenged small intestine of Group 1 and small intestine of Group 2 indicating that nematode infections induce the local appearance of large numbers of GL. Oral, but not loop challenge caused increased antibody levels in loop secretions when compared to unchallenged controls. Only loop-challenged sheep showed a peak in loop fluid SMCP levels 10–13 days after the first challenge which coincided with a peak in numbers of mucosal GL. The isolated loops of all 3 groups showed highly elevated numbers of eosinophils when compared to the intact small intestine. Loop fluid of all 3 groups showed a high level of LMI activity reflecting the high level of nematode-resistance induced by the immunisation procedures. Sheep in Groups 1 and 2 were both able to expel challenge infections, and when compared to Group 3, showed higher blastogenic activity of unstimulated cells derived from a mesenteric lymph node in the region of the challenged part of the intestine. The present experiment showed that surgically constructed intestinal loops provided a model system by which the substantial changes associated with the local intestinal immune response to challenge with T. colubriformis could be investigated.

Studies on caprine responsiveness to nematodiasis: Segregation of male goats into responders and non-responders

Eighty-three 2–4-year-old intact male goats exposed to a combination of artificial and natural challenge were segregated into responders and non-responders by ranking of weekly faecal egg counts (FECs). Retrospective analysis of samples over a 15-week-period showed responders had a statistically lower mean FEC than non-responders. Estimates of repeatability between consecutive egg counts were significant in both groups. The 6 top responders and bottom non-responders were subsequently given an artificial trickle challenge with Teladorsagia circumcincta and Trichostrongylus vitrinus. Mean faecal egg output was significantly lower in responders than non-responders. Peripheral eosinophil numbers following challenge were significantly greater in responder than non-responder goats. Abomasal and intestinal worm burdens were considerably lower in responders, with evidence of retardation of worm development compared to non-responders. Both abomasal and jejunal tissue eosinophil numbers were significantly higher in responders, although there was no difference in mucosal mast cell or globule leucocyte numbers. These results suggest that under temperate climatic conditions, it is possible to segregate male goats into responders and non-responders on the basis of simple parasitological criteria.

Regulation of egg production, worm burden, worm length and worm fecundity by host responses in sheep infected with Ostertagia circumcincta.

Following infection with Ostertagia circumcincta there was considerable variation in worm burdens, worm size and number of inhibited larvae even among sheep matched for age, sex, breed, farm of origin and history of parasite exposure. There was also substantial variation among sheep in the concentration of mast cells, globule leucocytes, eosinophils, IgA-positive plasma cells and parasite-specific IgA in the abomasal mucosa. With the exception of faecal egg counts over time, the parasitological and immunological traits were all continually distributed among animals and sheep did not fall into discrete high and low-responder categories. The responses were correlated. Sheep with more mast cells also had more globule leucocytes, more eosinophils, more IgA plasma cells and greater amounts of parasite-specific IgA in the abomasal mucosa. Female worm length was strongly and positively correlated with the number of eggs in utero. Faecal egg counts were associated with variation in worm number and with variation in the number of eggs in utero. The worm burden was negatively correlated with the number of globule leucocytes in the abomasal mucosa, suggesting that worm numbers are regulated by immediate hypersensitivity reactions. Decreased female worm length was associated with an increased local IgA response to fourth stage larvae. The number of inhibited larvae was positively associated with the size of the local IgA response and positively associated with the size of the worm burden. The results suggest that variation among mature sheep in faecal egg counts is due, at least in part, to variation in local IgA responses which regulate worm fecundity and to variation in local immediate hypersensitivity reactions which regulate worm burdens.

Diarrhoea in Merino ewes during winter: association with trichostrongylid larvae

A study on 3 farms, each having a preventive strategy for the control of nematode infections, determined the risk factors associated with diarrhoea among Merino ewes in winter. Comparisons were made among groups of mated and unmated ewes with and without treatment with controlled-release capsules containing albendazole. Diarrhoea was assessed from the accumulations of faeces around the breech (dag score) and related to worm egg counts, total worm counts and histopathological findings. The overall prevalence of severe dag in ewes not treated with a capsule was 26% (221/844). In comparison, only 2.7% of treated ewes had severe dag. The adjusted odds ratio on each farm indicated that untreated ewes were between 12 and 16 times more likely to be affected with severe dag than ewes treated with a capsule. The effect of lactation was significant on only one farm whereas initial body weight had no significant effect. There was no significant association between worm egg counts and the occurrence of severe dag. There were significantly more eosinophils in the small intestine of ewes affected with severe dag compared with unaffected ewes. There were no significant differences in mast cell and globule leucocyte numbers between affected and unaffected ewes. This study provides strong evidence that the main cause of diarrhoea among Merino ewes grazing winter and early spring pastures is the ingestion of trichostrongylid larvae even by sheep that have a well-developed protective immune response to these parasites. New strategies for the control of nematode infections are needed to prevent diarrhoea and dag formation in adult sheep.

Resistance of St. Croix lambs to Haemonchuscontortus in experimentally and naturally acquired infections

Parasitological and immunological parameters of experimental or naturally acquired infections with Haemonchus contortus were compared in St. Croix and Dorset lambs. In experimental infections, St. Croix lambs developed significantly greater levels of resistance to H. contortus, following primary exposure, as compared with Dorset lambs. This resistance was influenced both by age and by prior exposure to parasites. In grazing experiments on H. contortus-infected pasture, St. Croix lambs shed significantly fewer eggs as early as 5 weeks following initial exposure. Further, St. Croix lambs had more than 99% fewer worms in the abomasum at necropsy compared with age-matched Dorset lambs. Lymphoproliferative assays using peripheral blood mononuclear cells and antigen-specific serological tests demonstrated only minor differences in immune responsiveness between the two breeds despite the dramatic parasitological differences. Similarly, abomasal mucus from both breeds had elevated levels of parasitic-specific antibodies and contained substances mediating larval paralysis. In contrast, St. Croix lambs which had become resistant to nematode infection had dramatically higher numbers of globule leukocytes in the abomasal mucosa compared with Dorset lambs.

An unusually rich occurence of globule leucocytes in the bileducts of a roedeer (Capreolus capreolus)

In the epithelium of affected bileducts was found a considerable number of globule leukocytes. The identification of the globule leucocytes was confirmed by the use of staining methods according to Rahko and Nikander and by electron microscopy. The globules of each individual cell were numerous and of varying size, electrondensity and shape.

Changes in epithelial secretory cells and potentiation of neurogenic inflammation in the trachea of rats with respiratory tract infections

SummaryIn rats respiratory tract infections due to Sendai virus and coronavirus usually are transient, but they can have long-lasting consequences when accompanied by Mycoplasma pulmonis infections. Morphological alterations in the tracheal epithelium and a potentiation of the inflammatory response evoked by sensory nerve stimulation (“neurogenic inflammation”) are evident nine weeks after the infections begin, but the extent to which these changes are present at earlier times is not known. In the present study we characterized these abnormalities in the epithelium and determined the extent to which they are present 3 and 6 weeks after the infections begin. We also determined the magnitude of the potentiation of neurogenic inflammation at these times, whether the potentiation can be reversed by glucocorticoids, and whether a proliferation of blood vessels contributes to the abnormally large amount of plasma extravasation associated with this potentiation. To this end, we studied Long-Evans rats that acquired these viral and mycoplasmal infections from other rats. We found that the tracheal epithelium of the infected rats had ten times as many Alcian blue-PAS positive mucous cells as did that of pathogen-free rats; but it contained none of the serous cells typical of pathogen-free rats, so the total number of secretory cells was not increased. In addition, the epithelium of the infected rats had three times the number of ciliated cells and had only a third of the number of globule leukocytes. In response to an injection of capsaicin (150 μg/kg i.v.), the tracheas of the infected rats developed an abnormally large amount of extravasation of two tracers Evans blue dye and Monastral blue pigment, and had an abnormally large number of Monastral blue-labeled venules, particularly in regions of mucosa overlying the cartilaginous rings. This abnormally large amount of extravasation was blocked by dexamethasone (1 mg/day i.p. for 5 days). We conclude that M. pulmonis infections, exacerbated at the outset by viral infections, result within three weeks in the transformation of epithelial serous cells into mucous cells, the proliferation of ciliated cells, and the depletion of globule leukocytes. They also cause a proliferation of mediator-sensitive blood vessels in the airway mucosa, which is likely to contribute to the potentiation of neurogenic inflammation that accompanies these infections.

Globule leukocytes and mast cells in the rat trachea: their number, distribution, and response to compound 48/80 and dexamethasone.

SummaryGlobule leukocytes in the epithelium of the rat trachea may be counterparts of mucosal mast cells that are located in the gastrointestinal tract. If they are indeed similar to mucosal mast cells, globule leukocytes would be expected to decrease in number in rats treated with dexamethasone but not in rats treated with compound 48/80, an agent which causes non-antigenic degranulation of connective tissue mast cells. In this study, we determined the number and compared the distribution of globule leukocytes and connective tissue mast cells in the tracheas of pathogen-free rats. We then determined whether the number of these two types of cells changes in rats treated for 5 days with compound 48/80, dexamethasone, a combination of compound 48/80 and dexamethasone, or saline. We identified globule leukocytes and mast cells in whole mounts and histological sections of rat tracheas by using a histochemical reaction that demonstrates the chymotrypsin-like protease (chloroacetate esterase) present in mast cell granules. Using this method, we found that aproximately 225000 globule leukocytes were present in the epithelium of the trachea. These cells were most abundant in the rostral trachea. Rats treated with dexamethasone had a 91% reduction in the number of globule leukocytes with protease-containing granules, but rats treated with compound 48/80 had a normal number of these cells. We found some 55000 connective tissue mast cells in the same tracheas. Mast cells were most abundant in the posterior membrane of the caudal trachea and in the lamina propria between cartilaginous rings. Rats treated with compound 48/80 had a 96% reduction in mast cells with protease-containing granules, but rats treated with dexamethasone had a normal complement of mast cells. We conclude that globule leukocytes are abundant in the tracheas of healthy rats, are similar in morphology and pharmacological responses to mucosal mast cells located in other organs of rats, and are more numerous than and have a different distribution than connective tissue mast cells. Globule leukocytes in the tracheal epithelium may have a role in respiratory defenses similar to that of mucosal mast cells in other organs.

Relationship of nematode cholinesterase activity and nematode burdens to the development of resistance to trichostrongyle infections in sheep

The changes in nematode cholinesterase (ChE) activities were examined in relation to the development of resistance in (1) a flock of young grazing sheep, (2) grazing and penned sheep treated with dexamethasone and (3) penned sheep receiving a single mixed infection. Nematodes from grazing sheep with high faecal egg counts (FECs) had higher ChE activities than those from sheep with low FECs. Female nematodes tended to have higher ChE activities than males, and ChE activities in both tended to decline with increasing age of the sheep. The decline in female Trichostrongylus colubriformis ChE activity was associated with a decline in both worm length and in utero egg count. No decline in nematode ChE activity was observed when grazing sheep were treated with dexamethasone. ChE activity of T. colubriformis established in immunosuppressed penned sheep declined 10–20 fold 8 weeks after cessation of treatment. Nematode burdens in the small intestine and abomasum of grazing sheep were significantly correlated, and in individual species they were also correlated with ChE activities. The development of resistance in sheep and the elimination of adult nematode burdens is discussed in relation to gastrointestinal mucosal globule leucocyte numbers, mucus antiparasite activity and the impairment of nematode metabolic function.

Relationship of gastrointestinal histology and mucus antiparasite activity with the development of resistance to trichostrongyle infections in sheep

Changes in the numbers of globule leucocytes, mast cells, eosinophils and goblet cells in the gastrointestinal mucosa were examined in relation to the development of resistance and elimination of nematodes in grazing sheep in their firsty year of life. Sheep immunised against Trichostrongylus colubriformis, and sheep treated with dexamethansone were also examined. A strong association between resistance to infection and the presence of globule leucocytes was found. In contrast, the numbers of mast cells or goblet cells were not correlated with resistance. Globule leucocyte and eosinophil numbers were also correlated with antiparasite activity in mucus. Immunising infections of T. colubriformis given to 10-month-old sheep, their duration limited by thiabendazole treatment, gave rise to considerable immunity to homologous challenge infections. Larvae that developed to the 4th stage were as effective at stimulating immunity as those that developed to the 5th stage. Dexamethasone treatment abrogated resistance to trickle challenge infection with T. colubriformis and reduced mucosal globule leucocyte and mast cell numbers. After cessation of drug treatment, the re-establishment of resistance and adult worm elimination were associated with repopulation of the mucosa with large numbers of globule leucocytes and high antiparasite activities in mucus.

Experimental cholecystitis in goats caused by mature Fasciola gigantica

Mature Fasciola gigantica obtained from naturally infected cattle were surgically transferred into the gallbladders of six fluke-free goats. Two of the goats died at two and 19 weeks and the remaining four were slaughtered at 12, 48, 48 and 80 weeks after the transfer. Nine of 20 transferred F gigantica were recovered from the gallbladder and common hepatic duct of the experimental goats. The walls of gallbladders and bile ducts were thickened by epithelial proliferation, fibrosis and mononuclear cell infiltration. Large numbers of globule leucocytes were present in the epithelium of gallbladders and common hepatic ducts as well as in minute intrahepatic bile ducts. Very few mast cells were observed in these locations. Globule leucocytes in two goats were positive for argentaffin reaction with Gomori hexamine silver stain. The proliferative cholecystitis in experimental goats was similar to adenomatous cholecystitis in cattle naturally infected with fascioliasis. Globule leucocytes had no obvious deleterious effects on the parasites.

Relationship of Globule Leucocytes to Infections with Fasciola hepatica in Sheep

The presence of globule leucocytes (GL) within tissues of animal organs has been associated with various parasitic helminth infections. The GL is a nonepithelial cell found in and under epithelia. It is distinct from the Russell body cell, and evidence supports the thymus-dependent mast cell as its precursor (Gregory 1979, Vet. Bull. 49: 821-827). Sommerville (1965, Aus. Vet. J. 32: 237-240) found GL in abomasums of sheep with mixed nematode infections. Dobson (1965, Aus. J. Sci. 28: 434; 1966, Aus. J. Agric. Res. 17: 955-966; 1966, Nature 211: 875) reported GL in the gastrointestinal tracts of sheep with infections of Oesophagostomum columbianum and concluded that their presence reflected an immunological response. Lawrence (1977, Res. Vet. Sci. 23: 239-240) studied GL in lungs, livers, bladders, and kidneys, in addition to the gastrointestinal tracts, of cattle infected with Schistosoma mattheei. He concluded that they were most numerous and in more organs in animals subjected to repeated infections. Campbell et al. (1978, Vet. Parasitol. 4: 143-152) found GL in the bile ducts of sheep infected one time with Fasciola hepatica. In conjunction with another study, the opportunity arose to examine sheep infected singly and repeatedly with F. hepatica to determine whether GL might be present in tissues other than liver, and whether repeated infections would increase their numbers. Five groups of 10, 8-mo-old, helminth-free lambs were used. Groups 1 through 4 were inoculated with F. hepatica (Knight, 1980, Proc. Helminthol. Soc. Wash. 47: 186-191). Group 5 comprised uninfected lambs (Table I). At necropsy, a tissue sample was taken from liver, abomasum, kidney, cecum, lung, and small intestine and placed in buffered formolsaline. The tissues were imbedded in paraffin, sectioned, and stained with hematoxylin and eosin by conventional methods. One section of tissue from the excised sample was studied, and the area with the greatest number of GL was located under the low power of a compound microscope. Because the GL were not uniformly dispersed throughout the s ction of tissue, a coded value (score) from 0 to 10 was given to the tissue sample; this value was based on the number of GL present in a field of view under high dry (x40) power in that portion containing the densest concentration of GL (Table II). The coded values for each group were subjected to analysis of variance after square root transformation. Group means were separated by Kramer's modification (1956, Biometrics 12: 307-310) of Duncan's new multiple range test.

The influence of thyroid and testicular hormones on globule leucocytes in the rat duodenal crypt epithelium.

The globule leucocyte is a cell with a wide distribution in the digestive, respiratory, biliary, urinary and genital tract epithelia of homeotherms. It occurs in the crypts of the rat small intestine and within the epithelium on the villous bases but not on the remainder of the villi. A characteristic feature of this cell is the presence of acidophilic cytoplasmic granules, 0.5 to a few micrometers in diameter. The nucleus resembles that of intraepithelial intestinal lymphocytes. In this investigation, a quantitative study was made of the effect of thyroidectomy and/or castration on the number of globule leucocytes in the duodenal crypts of immature and adult rats. In sham-operated rats, globule leucocytes were rare, occurring with a frequency of 1 or 2 per 1,000 epithelial cells. After thyroidectomy, they increased to 14--20 per 1,000 epithelial cells. Castration did not influence the number of globule leucocytes but resulted in an increase in the number, size and acidophilia of their cytoplasmic granules. In immature rats, but not in adults, castration combined with thyroidectomy enhanced the effect of thyroidectomy, increasing globule leucocyte number to 32 per 1,000 epithelial cells. Treatment of thyroidectomized-castrates with thyroxine, initiated 38 days after operation, reduced the crypt globule leucocyte population to normal, whereas treatment with testosterone did not. Growth hormone failed to influence the elevated number of globule leucocytes in thyroidectomized rats, suggesting that this action of thyroxine was not mediated via an influence on growth hormone release. Because of their similar nuclear morphology, intraepithelial crypt lymphocytes were also counted and there was no obvious relationship in the fluctuations of these two cell populations. It is suggested that the greater number of crypt globule leucocytes in thyroid deficiency may reflect changes in the intestinal secretory immunoglobulin system and this is being investigated, beginning with a study of the distribution of IgE.

Resistance to Haemonchus contortus in sheep given repeated daily doses of 10 000 infective larvae

Two experiments on the effects on sheep of prolonged repeated (5 days in every week) infection with 10 000 infective larvae of Haemonchus contortus are reported. In the first experiment 7 sheep were infected for 6 months. Regular observations were made of haematocrit and faecal egg count. Two sheep required anthelmintic treatment once, 2 sheep required 3 treatments, the remaining 3 sheep required no anthelmintic support. At the end of the 6-month period all 7 sheep had acquired a high level of resistance. In the second experiment, this acquisition of resistance while subject to a weekly intake of 5 × 10 000 L 3 was studied in 7 sheep which were fitted with abomasal cannulae. These sheep were infected repeatedly for 8 to 22 weeks. The acquired resistance was not associated with an abnormally high gastric pH. Large numbers of globule leucocytes were present in the fundic mucosa of all 14 sheep at the conclusion of the experiments.

Responses to infection with Haemonchus contortus and Trichostrongylus colubriformis in ewes of different reproductive status

Groups of ewes which were either lactating, pregnant, non-lactating (lambs removed at birth) or unmated were subjected to twice weekly infection with constant numbers of H. contortus and T. colubriformis infective larvae for 8 weeks. From the size, structure and fecundity of their worm burdens, lactating ewes, and those in late pregnancy did not differ from each other and showed evidence of diminished immunological responses to infection compared with unmated ewes. The worm burdens of non-lactating ewes, slaughtered less than 30 days after parturition, resembled those of lactating and pregnant animals, but substantially fewer T. colubriformis were found in two ewes which had been deprived of their lambs for 33 and 40 days respectively. Increased numbers of mast cells, eosinophils and globule leucocytes were found in the gut mucosa of animals showing parasitological evidence of an immune response, and in particular, very few globule leucocytes were present in lactating and pregnant animals. Within animals over all groups there was a pronounced negative correlation between numbers of T. colubriformis and numbers of globule leucocytes in the small intestine.