Distribution of immunoglobulin-containing cells in calves inoculated orally with ruminal Bacteroides succinogenes and Selenomonas ruminantium

Distribution of immunoglobulin(Ig)-containing cells was investigated in calves inoculated orally with live organisms of both Bacteroides succinogenes and Selenomonas ruminantium. Pathological changes and many Ig-containing cells were observed in calves which inoculated three times at 2, 3 and 26 days of age. Follicular germinal center was increased in number and size of the lymph nodes associated with the forestomach, suggesting activation of lymph apparatus. In the associated lymph nodes, IgG-containing cells were predominant and were located in both cortex and medulla, mainly in the medullary cord, B lymphocyte areas. Only a few IgA- and IgM-containing cells were observed in the lymph nodes. Accordingly, the inoculated bacteria may stimulate IgG-containing B lymphocyte populations. A few IgG-containing cells were detected in the mucosa of the forestomach. Ig-containing cells, predominantly IgG, were observed in the mucosa of the abomasum and intestine, and in the mesenteric lymph nodes. However, number of the cells in the mesenteric lymph nodes was smaller than that of the forestomach associated lymph nodes. The results suggest that the intraorally inoculated bacteria may stimulate the maturation of IgG positive lymphocytes in the lymph nodes associated with the forestomach.     

Localization of bacterial antigens in calves inoculated orally with ruminal Bacteroides succinogenes and Selenomonas ruminantium

Localization of bacteria and bacterial materials was investigated in calves inoculated orally with live organisms of both Bacteroides succinogenes and Selenomonas ruminantium by a immunohistological method using rabbit antiserum against the outer membrane of those organisms and by a scanning electron microscope. The intact organisms of both inoculated bacterial species were detected on the rumen wall and in the lamina propria of the forestomach, and S. ruminantium also in the lymph nodes associated with the rumen. The bacterial materials were observed inside of macrophage-like cells in the lamina propria of the forestomach and of lymphoid cells in the lymph nodes. Number of the cells in the mesenteric lymph nodes was smaller than that of the forestomach associated lymph nodes. The results suggest that the orally inoculated bacteria may act as antigenical stimulant in the mucosa of the forestomach and associated lymph nodes of calves.     

Immune responses of the bovine fetus and neonate to Escherichia coli: plaque-forming and intestinal immune responses.

The immune responses of 26 Angus-Hereford fetuses and neonates to Escherichia coli O26:K60:NM were studied after bacterin or saline solution was injected (in utero) into the amniotic fluid. Calves were euthanatized at birth or were orally revaccinated; some were challenge exposed with live organisms. The hemolytic plaque assay was used to determine the presence of cells producing immunoglobulins M, G1, and G2 (IgM, IgG1, and IgG2) in 4 segments of the small intestine, mesenteric lymph nodes, and spleen. The passive hemagglutinin activity of intestinal washings was also determined. Anti-O26 passive hemagglutinin activity in the intestinal washings of principal calves was greater than in that of control calves, but in a given segment of the small intestine, usually this activity was relatively small and less consistent than the plaque-forming response. Greater numbers of plaque-forming cells were observed in the small intestine of 14 of the 15 principal calves when compared with the control calves tested.     

Induction of systemic and mucosal immune response in cattle by intranasal administration of pig serum albumin in alginate microparticles

Biodegradable microparticles are an efficient mucosal delivery system that protect antigens from the harsh mucosal environment and facilitate their uptake by M cells at the epithelium of mucosal-associated lymphoid tissue. In this study, we determined the systemic and mucosal immune response in calves following intranasal and oral immunization with pig serum albumin (PSA) encapsulated in alginate microparticles. The size of the particles ranged from 1 to 50 microm in diameter, with 95% of the particles being smaller than 5 microm. High levels of anti-PSA IgG1 antibodies were found in the serum, nasal secretions, and to a less extent in saliva of calves vaccinated intranasally, but not orally, with PSA-microparticles. There was no significant increase of PSA-specific IgA. A weak lymphocyte proliferative immune response was observed in peripheral blood mononuclear cells (PBMCs), and few anti-PSA antibody-secreting cells (ASC) were detected in the blood of calves immunized intranasally. The combined systemic and mucosal response observed in intranasally immunized animals may be attributed to the wide variation in the size of the alginate microparticles, with smaller particles translocating to regional lymph nodes and inducing a systemic immune response, and larger particles being retained in the NALT and inducing a mucosal immune response. The procedure presented here may be useful as an intranasal vaccine against respiratory diseases in cattle.     

Direct inoculation of Mycobacterium avium subspecies Paratuberculosis into ileocecal Peyer's patches results in colonization of the intestine in a calf model

The objective of this study was to develop an intestinal model of Mycobacterium avium subspecies paratuberculosis (Map) infection in the calf for evaluation of mucosal pathology and local and systemic immunologic responses. Map was inoculated into Peyer's patches of young calves using a right flank surgical approach in standing calves to exteriorize the ileocecal junction. Inoculum doses ranging from 10(3) to 10(9) colony-forming units of strain K10 Map were injected through the serosal surface into Peyer's patches of the distal ileum near the ileocecal valve. Fecal samples were collected for culture from each calf weekly until termination of the study. Calves were necropsied at 7, 30, 60, and 90 days after infection, when inoculation sites, lymph nodes, spleen, and peripheral blood were collected for evaluation. Ileocecal lymph nodes were consistently colonized by Map in the 10(5) to 10(9) groups. The ileocecal valve was also colonized in 10(7) and 10(9) groups. This correlated with fecal culture results as infected calves intermittently shed Map in their feces throughout the study. Granulomatous lesions with giant cells and acid-fast bacilli at the ileocecal junction, ileocecal lymph nodes, and lamina propria of high-dose animals (10(7) and 10(9)) were identified from each time point. Flow cytometry was used to detect antigen-specific production of interferon-γ and interleukin-4 locally (ileocecal lymph node) and systemically (peripheral blood mononuclear cells), which defined distinct immunologic profiles in low-dose and high-dose calves. This study demonstrates intestinal Map infection via Peyer's patch inoculation, a novel model with many shared features of natural Map infection.     

Gut-associated lymphoid tissue in the large intestine of calves. I. Distribution and histology

Gut-associated lymphoid tissue (GALT) in the large intestine was characterized in 12 calves (10 to 84 days old) obtained at necropsy (7, group A) or healthy animals (5, group B). Patches of mucosal lymphoid follicles were in all calves at ileocecal entrances (ICE), 23-42 cm distal to the ICE in the proximal loop of the ascending colon (proximal colon [PC] patch), and in the terminal rectum. PC patches varied from 8 to 30 cm in length. Solitary lymphoid follicles were found in the cecum of three calves, between the ileocecal entrances and the PC patch in four calves, adjacent to the PC patch in all calves, and in the ampulla recti. GALT occupied 7.8% of the large intestinal wall in animals of group A: 0.6% at the ileocecal entrance, 4.8% in the proximal colon, and 2.4% in the rectum. There were two different types of mucosal lymphoid follicles in group B: propria nodules with lymphoid follicles predominantly in the lamina propria, and lymphoglandular complexes with lymphoid follicles in the submucosa. In three 3-, 6-, and 7-day-old, germfree calves, distinct follicle-associated epithelium covered propria nodules and covering folds in depths of the lymphoglandular complexes; it was characterized by numerous intraepithelial cells and lack of goblet cells.     

Pathogenic Shiga toxin-producing Escherichia coli in the intestine of calves

The purpose of this study was to compare the pathological effects of Shiga toxin-producing Escherichia coli (STEC) that vary in their association with bovine and human disease. Shiga toxin-producing E. coli of serotypes associated with both dysentery in calves and hemolytic uremic syndrome (HUS) in humans (O5:H-, O26:H11, O111:H-, O113:H21) were compared with O157:H7 STEC, which are associated with HUS in humans but not with disease in calves. The STEC were administered orally to 80 day-old chicks and into ligated loops in the ileum and colon of four 2- to 6-day-old calves. Examination of the ceca of the chickens 10 d postchallenge showed no adherence or tissue abnormality for any isolate. The calves were euthanized 8 to 10 h postinoculation, and sections of the intestinal loops were examined by light microscopy, transmission and scanning electron microscopy, and immunohistochemistry. All strains showed consistent focal adherence associated with mild lesions in the colon. Attaching and effacing lesions were observed with the eae-positive strains. Ileal lesions were similar to the colonic ones but were sometimes severe, with marked polymorphonuclear leukocyte proliferation in the lamina propria. It is concluded that chickens were unsuitable for studying interaction of STEC with the intestine and that there was no difference in the interaction of the ligated calf intestine with STEC of serotypes associated with disease in calves compared with O157:H7 STEC.     

Mast cells in the bovine lower respiratory tract: morphology, density and distribution

The lower respiratory tract of six 8-day-old calves and six adult cows was fixed in either isotonic formal-acetic-acid or neutral buffered formalin in order to study the morphology, density and distribution of mast cells. They were found at all levels of the tract with the highest density in the major bronchi. Tissues from cows had significantly more mast cells than those from calves. There were significantly more mast cells detected in calf airways fixed with isotonic formal-acetic-acid compared with those seen in the same tissues fixed with neutral buffered formalin. Regardless of the age and fixation, mast cells were located predominantly in the alveolar septa and in the lamina propria of airways. They were also commonly encountered within the mucosal epithelium of the trachea. Ultrastructurally, mast cells in bovine airways and lung contained two types of intracytoplasmic granules as described in other species.     

Neospora caninum in persistently infected, pregnant cows: spontaneous transplacental infection is associated with an acute increase in maternal antibody.

Nine cows which were naturally and persistently infected with Neospora caninum were housed and observed intensively throughout pregnancy. No recrudescence of a latent infection was detected by PCR tests on maternal blood but fetal infection, implying a recrudescence of maternal parasitosis, was associated with a marked increase in maternal antibody. The increase occurred in the second half of pregnancy in five cows which infected their calves, and before mid-pregnancy in one cow which aborted. There was no change in the avidity of the antibody, which remained high and characteristic of long-term infection. In three infected cows that gave birth to uninfected calves there was no marked increase in maternal antibody. Antigen-specific interferon gamma responses of peripheral blood mononuclear cells were observed in all the infected cattle but they did not vary significantly either during pregnancy, or whether the cows did or did not infect their calves, although the responses were consistently higher in the latter. There was no change in the plasma concentrations of cortisol or acute phase proteins associated with the recrudescence of the parasite. Three uninfected cows housed with the infected cows remained uninfected throughout the experiment. No immunosuppressive event was detected which might have provoked parasite recrudescence but the acute antibody rise associated with transplacental infection provides a valuable, non-invasive marker for further studies to investigate the cause and consequences of parasite recrudescence in N caninum infection in cattle.     

In vitro lymphocyte proliferative responses and gamma-interferon production as measures of cell-mediated immunity of cattle exposed to Pasteurella haemolytica.

Cell-mediated immune mechanisms may play a role in the pathogenesis and prevention of pneumonia in cattle caused by Pasteurella haemolytica serotype A1. To determine the circumstances required to stimulate and identify cell-mediated immune responses, calves were vaccinated with a commercial P. haemolytica bacterin or a live commercial P. haemolytica vaccine, or were infected intratracheally with virulent P. haemolytica. All calves were challenge-exposed intratracheally with P. haemolytica 31 d after vaccination or prior infection. Peripheral blood mononuclear cells and mediastinal and superficial cervical lymph node cells were stimulated with antigens prepared from P. haemolytica to evaluate in vitro proliferative responses and gamma-interferon production as measures of cell-mediated immunity. Strong proliferative responses and gamma-interferon production were detected in lymph node cells from calves vaccinated with the live vaccine and from infected calves, especially in response to stimulation with an outer membrane protein preparation from P. haemolytica. Greater proliferative responses and gamma-interferon production were associated with the lymph node nearer the site of bacterin administration (superficial cervical lymph node) or the site of infection (mediastinal lymph node), whereas greater proliferative responses and gamma-interferon production were associated with the more distant lymph node (mediastinal lymph node) in calves vaccinated with the live vaccine. Neither proliferative responses nor gamma-interferon production were detected in peripheral blood mononuclear cells from calves that were vaccinated for or infected with P. haemolytica. Antileukotoxin antibody titers were determined by a serum neutralization assay, and protection against pneumonic lesions was more closely correlated with antileukotoxin antibody responses than with lymphocyte proliferation or gamma-interferon responses.     

Isotype-specific antibody-secreting cells in systemic and mucosal associated lymphoid tissues and antibody responses in serum of conventional pigs inoculated with PEDV

An enzyme-linked immunospot (ELISPOT) has been developed to detect porcine epidemic diarrhea virus (PEDV)-specific antibody secreting cells (ASC) in gut associated lymphoid tissues (duodenum and ileum lamina propria and mesenteric lymph nodes) and systemic locations (spleen and blood) of conventional pigs so as to characterise the mucosal and systemic antibody response generated by the infection with PEDV. A total number of 28 eleven-day-old conventional pigs were orally inoculated with the field isolate of the PEDV strain CV-777. Diarrhea was observed in 32% of the pigs and virus shedding was demonstrated in 100% between postinoculation day (PID) 1 and 8. Serum IgG and IgA antibodies to PEDV were detected by isotype ELISA from PID 12 and 15, respectively, reaching maximum values at PID 32 (IgG) and 21 (IgA). PEDV specific IgM ASC occurred in all the tissues between PID 4 and 7, with the strongest response in the intestinal lamina propria. IgA and IgG ASC responses were evident in the intestinal lymphoid tissues from PID 21, the highest number of specific ASC corresponded to the duodenum lamina propria. In the systemic lymphoid tissues the number of IgG and IgA ASC detected were lower than in the mucosal tissues, however, in the blood, presence of IgA ASC was constantly detected from PID 14 until the end of the experiment. Memory antibody response to the PEDV was also studied by secondary in vitro stimulation of the mononuclear cells (MNC) isolated from mesenteric lymph nodes, spleen and blood. The memory B cell response was prominent at PID 21 and 25 and consisted in IgG and IgA ASC. To our knowledge, this is the first report to research into the presence and distribution of specific ASC in different locations of the systemic and the gut associated lymphoid tissues after a PEDV infection as well as the presence of memory B cells.     

Mucosal immune response in cattle with subclinical Johne's disease

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease, a chronic granulomatous enteritis of wild and domestic ruminants. During a long subclinical period, the organism persists in the intestine despite systemic cellular and humoral immune responses. To explore the mucosal immune response in Johne's disease, we isolated mononuclear leukocytes from the ileum of cows naturally infected with M. avium subsp. paratuberculosis and from cows that were not infected. We evaluated the immunophenotype of these cells and the proliferative responses after the addition of M. avium subsp. paratuberculosis sonicate or B-cell or T-cell mitogens. Although the percentage of T cells was increased in infected cows, these cells consisted mostly of memory (CD2+CD62L-) and regulatory (CD4+CD25+) T cells. Further evidence of immune hyporesponsiveness included a decrease in the percentage of T cells with an activated phenotype and a decrease in cells expressing major histocompatibility factor class II (MHC class II). Unlike the spleen, ileal lymphocytes from infected cows failed to proliferate in response to M. avium subsp. paratuberculosis sonicate. Additionally, ileal lymphocytes from infected cows proliferated poorly in response to concanavalin A and pokeweed mitogen, suggesting generalized T cell and B cell hyporesponsiveness. These results indicate that a state of tolerance may exist in the intestine of cows subclinically infected with M. avium subsp. paratuberculosis organisms in subclinically infected cows. This effect may be induced, at least in part, by proliferation of regulatory T cells that nonspecifically suppress mucosal immune responsiveness.     

Modulation by colostrum-acquired maternal antibodies of systemic and mucosal antibody responses to rotavirus in calves experimentally challenged with bovine rotavirus

The effect of colostral maternal antibodies (Abs), acquired via colostrum, on passive protection and development of systemic and mucosal immune responses against rotavirus was evaluated in neonatal calves. Colostrum-deprived (CD) calves, or calves receiving one dose of pooled control colostrum (CC) or immune colostrum (IC), containing an IgG1 titer to bovine rotavirus (BRV) of 1:16,384 or 1:262,144, respectively, were orally inoculated with 105.5 FFU of IND (P[5]G6) BRV at 2 days of age. Calves were monitored daily for diarrhea, virus shedding and anti-BRV Abs in feces by ELISA. Anti-rotavirus Ab titers in serum were evaluated weekly by isotype-specific ELISA and virus neutralization (VN). At 21 days post-inoculation (dpi), all animals were euthanized and the number of anti-BRV antibody secreting cells (ASC) in intestinal and systemic lymphoid tissues were evaluated by ELISPOT. After colostrum intake, IC calves had significantly higher IgG1 serum titers (GMT=28,526) than CC (GMT=1195) or CD calves (GMT<4). After BRV inoculation, all animals became infected with a mean duration of virus shedding between 6 and 10 days. However, IC calves had significantly fewer days of diarrhea (0.8 days) compared to CD and CC calves (11 and 7 days, respectively). In both groups receiving colostrum there was a delay in the onset of diarrhea and virus shedding associated with IgG1 in feces. In serum and feces, CD and CC calves had peak anti-BRV IgM titers at 7 dpi, but IgA and IgG1 responses were significantly lower in CC calves. Antibody titers detected in serum and feces were associated with circulation of ASC of the same isotype in blood. The IC calves had only an IgM response in feces. At 21 dpi, anti-BRV ASC responses were observed in all analyzed tissues of the three groups, except bone marrow. The intestine was the main site of ASC response against BRV and highest IgA ASC numbers. There was an inverse relationship between passive IgG1 titers and magnitude of ASC responses, with fewer IgG1 ASC in CC calves and significantly lower ASC numbers of all isotypes in IC calves. Thus, passive anti-BRV IgG1 negatively affects active immune responses in a dose-dependent manner. In ileal Peyer's patches, IgM ASC predominated in calves receiving colostrum; IgG1 ASC predominated in CD calves. The presence in IC calves of IgG1 in feces in the absence of an IgG1 ASC response is consistent with the transfer of serum IgG1 back into the gut contributing to the protection of the intestinal mucosa.     

Variation in the immuno-pathological responses of lambs after experimental infection with different strains of Mycobacterium avium subsp. paratuberculosis

Ruminant infection by Mycobacterium avium subsp. paratuberculosis (MAP) causes a granulomatous inflammatory response in the intestine and associated lymph nodes. Differences either in the affected organs or in the inflammatory infiltrate were observed between species and individuals. Such differences are usually attributed to variations in host immune responses or to inconsistent effects among different MAP strains. To evaluate if different MAP strains induce different immuno-pathological responses in lambs, 28 one-month-old individuals were divided into six groups and inoculated with different MAP strains. Groups 1 and 2 were inoculated with two bovine strains isolated in Argentina that showed different genetic patterns after BstEII-IS900-RFLP (hereafter strains E and A respectively). Group 3 was inoculated with a bovine strain isolated in Spain obtained after a previous step of culture (patterns C1). Group 4 was inoculated with a homogenate of intestinal mucosa of a clinical case affected by the same bovine strain as that of group 3. Group 5 was inoculated with an ovine strain that was directly purified from the intestinal mucosa of a clinical case, and group 6 was kept as control (i.e. no inoculation). Peripheral immune responses were assessed until 150 days post-infection (dpi), when lambs were humanely killed. Pathological studies were performed in tissues from the intestine and lymph nodes. Lesion types and inflammatory infiltrates were examined as indicators of pathogenicity. All the lambs infected with bovine MAP strains showed a common lesion pattern regardless of the strain type. Such pattern was characterized by focal lesions mainly in the mesenteric lymph nodes, the presence of fibrous tissue, and, occasionally, necrosis in the granulomas as well as the presence of numerous giant cells. Differences in lesion severity were observed among groups: lambs from groups 1 and 2 had the highest number of granulomas and the largest lymph node area affected. Lesions in animals from group 5 (infected with an ovine strain) were more severe and occurred mostly in the intestinal lymphoid tissue; necrosis, fibrosis or giant cells were never detected in this group. These results indicate that the MAP strain type induces different pathological responses in lambs.     

Local and systemic immune responses to Echinococcus granulosus in experimentally infected dogs

Local and systemic immune responses were studied in six dogs experimentally infected with the dog/sheep tapeworm Echinococcus granulosus. All dogs developed similar IgG antibody response to parasite antigens. In contrast, IgE and IgA responses differed widely. No relationship between IgA responses and parasite burden at the end of the infection were observed. Further, clear differences in the anti-parasite IgA response in serum as compared with specific IgA forming cells in mesenteric lymph nodes were observed within the same dog. An inverse association of anti-parasite IgE and parasite load seemed to be present, with the strongest IgE response in the one dog that had no worms in the intestine at the end of the experiment. No differences were observed in the numbers of intestinal mast cells and goblet cells among all infected dogs. However, the dog with no detectable parasite load had a marked reduction of detected mast cells in the submuscular and muscular layer of the mucosa. Our data give new insight into the immune response of dogs during E. granulosus infection and provide information that may be useful for the rational design of vaccines for the control of hydatid disease.     

Experimentally induced Cooperia oncophora infection in calves: lymphocyte blastogenic and delayed hypersensitivity responses

Lymphocytic responses in peripheral blood and visceral lymph to Cooperia oncophora antigen and skin tests were determined in 35 Holstein male calves that were inoculated orally with single or multiple doses of C oncophora infective larvae. Several calves were vaccinated or given immune serum before larvae were inoculated. Antigen-specific in vitro blastogenesis of blood and lymph lymphocytes and delayed-type hypersensitivity reactions were observed in several inoculated, vaccinated, and/or passively immunized calves. Most calves that had delayed skin reactions also had in vitro lymphocyte responses to C oncophora antigen. The lymphocyte and skin responses were inconsistent and variable in time of onset--the earliest lymphocyte response occurring 7 days after calves were inoculated. A cellular immune response was induced by both dermal vaccination and oral inoculation; however, passive immunization by IV administration of immune serum simultaneously with inoculation did not have an apparent effect on the cellular response, as measured by the lymphocyte blastogenesis test or dermal testing. Although cellular immune responses were observed in several calves infected with C oncophora, there was no apparent relationship between the specific responses and number of nematodes establishing infection in calves after either single- or multiple-dose oral inoculations.     

Enteritis associated with Yersinia pseudotuberculosis infection in a buffalo

SUMMARY: A week after transport a 4 month old buffalo calf developed diarrhoea. Its condition gradually deteriorated and it died. Necropsy revealed acute haemorrhagic enteritis and enlarged mesenteric lymph nodes. Haemorrhages and numerous microabscesses were detected in the lamina propria of the small intestine associated with colonies of Gram negative bacteria. Yersinia pseudotuberculosis was isolated from the small intestine and from mesenteric lymph node. Enteritis caused by Y pseudotuberculosis does not appear to have been reported previously in buffalo in Australia.     

The postnatal development of the mucosal immune system and mucosal tolerance in domestic animals

The mucosal immune system is exposed to a range of antigens associated with pathogens, to which it must mount active immune responses. However, it is also exposed to a large number of harmless antigens associated with food and with commensal microbial flora, to which expression of active, inflammatory immune responses to these antigens is undesirable. The mucosal immune system must contain machinery capable of evaluating the antigens to which it is exposed and mounting appropriate effector or regulatory responses. Since the immune system is likely to have evolved initially in mucosal tissues, the requirement to prevent damaging allergic responses must be at least as old as the adaptive immune system, and studies of the mechanisms should include a range of non-mammalian species. Despite the importance for rational design of vaccines and for control of allergic reactions, the mechanisms involved are still largely unclear. It is not clear that the classical experimental protocol of "oral tolerance" is, in fact, measuring a biologically important phenomenon, nor is it clear whether tolerance is regulated in the evolutionarily recent organised lymphoid tissue (the lymph nodes) or the more ancient, diffuse architecture in the intestine. The capacity of the immune system to discriminate between "dangerous" and "harmless" antigens appears to develop with age and exposure to microbial flora. Thus, the ability of an individual or a group of animals to correctly regulate mucosal immune responses will depend on age, genetics and on their microbial environment and history. Attempts to manipulate the mucosal immune system towards active immune responses by oral vaccines, or towards oral tolerance, are likely to be confounded by environmentally-induced variability between individuals and between groups of animals.     

Is Lamina Muscularis Mucosae Present in the Ruminal Mucosa of Cattle?

The cells in the condensed fibrous layer in the lamina propria of the ruminal mucosa of the calves and cows were studied at the light microscopic level. Abundant actin-immunoreactive cells were detected both in the calves and cows, while desmin-immunoreactive cells were moderate in frequency in the cows and low or absent in the calves. Vimentin-immunoreactive cells were abundant in the cows but moderate to low in frequency in the calves. These cells were distributed to form a condensed layer being more abundant in the ruminal papillae than the interpapillar mucosae in the cows and equal in the papillae and interpapillar mucosae in the calves. At the ultrastructural level, smooth muscle cells and fibroblasts were abundant in the deep core of the ruminal papilla of the cows. The present results suggest that the condensed fibrous layer revealed in the ruminal mucosae contains smooth muscle cell population which could be regarded as the specialized lamina muscularis mucosae.     

Antibody responses in protein-energy restricted beef cows and their cold stressed progeny.

Antibody titers were measured in serum and colostral whey of pregnant beef cows immunized with tetanus toxoid and chicken red blood cells while being fed diets either restricted or nonrestricted in protein and/or metabolizable energy during the last 150 days of gestation. Serum antibody titers were also measured in the colostrum-fed, cold and noncold stressed progeny that were actively immunized with dinitrophenol conjugated to keyhole limpet hemocyanin. In general, there were no major or sustained differences in humoral immune responses to injection of tetanus toxoid or chicken red blood cells between cows fed diets that were adequate or restricted in protein or metabolizable energy. In the few cases where serum antibody titers to tetanus toxoid or chicken red blood cells differed (P less than 0.05) between adequately fed or restricted cows, the differences were no greater than twofold. Anti-chicken red blood cell titers were uniformly low (P less than 0.05) by a magnitude of two to threefold in colostral whey of cows restricted in protein and/or metabolizable energy when compared to titers in cows fed adequate amounts of protein and metabolizable energy. With one exception, neither maternal dietary restriction nor cold exposure had a major effect on the ability of the calves to absorb antitetanus toxoid and chicken red blood cell antibodies from colostrum. The humoral immune responses of all calves to injection of keyhole limpet hemocyanin and dinitrophenol were similar in magnitude.