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[Antimicrobial activity of human colostrum against enteropathogens. Preliminary study].

The antimicrobial activity of the colostrum serum from ten women has been evaluated. The activity against Escherichia coli, Shigella sonnei and Klebsiella pneumonia at three different bacterial populations was determined (1 x 10(4), 1 x 10(5) and 1 x 10(6) UFC/ml). Antimicrobial activity against the three strains was found. Bactericidal activity was observed to inocula of 1 x 10(4) UFC/ml in 8/10 sera against E. coli, in 6/10 sera against S. sonnei, and in 3/10 sera against K. pneumonia; in the rest of the cases, sera were bacteriostatic. With inocula of 1 x 10(5) UFC/ml there was bactericidal activity in 4/10 sera against E. coli and S. sonnei and in 1/10 against K. pneumonia. Lastly, with inocula of 1 x 10(6) UFC/ml there was bactericidal activity in 4/10 sera against E. coli, 1/10 against S. sonnei and none against K. pneumoniae. The results suggest that because of the antimicrobial properties of human milk the risk of intestinal infections by enteropathogens is less.

Colony Count, Microbial↗

[Effect of colostrum polypeptide on structure and lipid peroxidation in liver nuclei of rats late after gamma-irradiation].

Rats were irradiated by 8Gr in a doze power of 2.33 R per second. A portion of animals were injected subcutaneously with a cow colostrum polypeptide (CP) in a doze of 1 mg per g of body weight before the irradiation and daily after the irradiation during 4 days. In irradiated rat liver nuclei injected with CP the lipid peroxide oxidation was restored. At 90th day after the irradiation the production of dienes and dieneketones in liver nuclei oscillated in normal limits. The structure and cytochrome-c-oxidase activity of the nuclei was restored to 60-90th day after the irradiation.

Animals↗

[Change in the proteins in the colostrum of a cow during the 1st hours after calving].

Studied were the changes taking place in the protein content of the colostrum of cows during the first six hours following calving. The protein fractions were identified by the method of disk polyacrylamide electrophoresis. It was established that the amount of the total protein dropped with the increase in the secretion time. Changes were found to take place in the gamma-globulin fractions, the casein, and the soluble proteins.

Animals↗

Enzymatic properties of beta-D-galactoside alpha2 leads to 6 sialytransferase from bovine colostrum.

The substrate specificity and kinetic properties of a pure sialyltransferase from bovine colostrum have been examined. The transferase appears to incorporate sialic acid into the sequence, NeuAcalpha2 leads to 6Galbeta1 leads to 4GlcNAc, which is commonly found in glycoproteins. It has a strict substrate specificity for CMP-NeuAc and forms only the alpha2 leads to 6 sialyl linkage with beta-D-galactosides. N-Acetyllactosamine (Galbeta1 leads to 4GlcNAc) and asialo-glycoproteins containing the N-acetyllactosaminyl linkage at the nonreducing ends of the oligosaccharides prosthetic groups are the best acceptor substrates. Isomers of N-acetyllactosamine with beta1 leads to 3 or beta1 leads to 6 glycosidic linkages are less than 1% as effective as acceptor substates as the beta1 leads to 4-linked isomer. Lactose (Galbeta1 leads to 4Glc) is also a poor acceptor, indicating the importance of the 2-acetamido group in the N-acetylglucosaminyl residues. The unnatural substrate beta-methyl-L-arabinopyrano-side, a five-carbon analog of beta-methyl-D-galactoside which contains no 6-hydroxyl, also acts as a poor acceptor of the transferase and the sialylated product has been partially characterized. Kinetic properties of the enzyme in the presence and absence of inhibitors suggest that the transferase has an equilibrium random order mechanism.

Animals↗

Infection and cross-protection studies of winter dysentery and calf diarrhea bovine coronavirus strains in colostrum-deprived and gnotobiotic calves.

OBJECTIVE: To investigate in vitro antigenic relations, in vivo cross-protection, and isotype antibody responses to a winter dysentery (WD) and calf diarrhea strain of bovine coronavirus (BCV). DESIGN AND ANIMALS: Gnotobiotic and colostrum-deprived calves were inoculated oronasally with a WD (DBA) or a calf diarrhea (DB2) BCV, and were challenge exposed with the heterologous BCV. PROCEDURE: Nasal swab and feces specimens and blood samples were collected. Fecal and nasal specimens were assayed for BCV shedding by antigen-capture ELISA or immune electron microscopy. Bovine coronavirus antigens were detected in nasal epithelial cells by immunofluorescence. Antibody titers to BCV in serum were assayed by virus neutralization (VN), and BCV antibody isotype titers in feces and sera were quantitated by ELISA. RESULTS: All calves developed diarrhea and shed BCV nasally and in feces, then recovered and were protected from BCV-associated diarrhea after challenge exposure with the heterologous BCV. After challenge exposure with either strain, fecal shedding of DBA was detected in 1 of 4 calves and nasal shedding of DB2 was detected in 2 of 4 calves. Immunoglobulin M was the principal coproantibody to BCV early, followed predominantly by IgA. Immunoglobulin G1 coproantibody titers to BCV were low, but increased after challenge exposure. Immunoglobulin G1 antibodies were predominant in serum. After challenge exposure, all serum antibody isotype titers increased except IgG2. The VN antibody responses paralleled serum IgG1 antibody responses. CONCLUSIONS AND CLINICAL RELEVANCE: Immunoglobulin A coproantibodies at challenge exposure were associated with protection against diarrhea. Nasal shedding of BCV after challenge exposure confirmed field data documenting reinfection of the respiratory tract of cattle, suggesting that, in closed herds, respiratory tract infections constitute a source of BCV transmission to cows (WD) or young calves.

Animals↗

Colostrinine: a proline-rich polypeptide from ovine colostrum is a modest cytokine inducer in human leukocytes.

A proline-rich polypeptide (PRP), now named colostrinine, molecular weight 18,000, was isolated from ovine colostrum and characterized by Janusz, Lisowski et al. The nonapeptide (NP) which is an active fragment of PRP was obtained by chemical synthesis. In mice, PRP has many regulatory effects on the humoral and cellular immune response. The present paper describes PRP as a cytokine inducer. PRP at concentration of 1-100 micrograms/ml was found to induce production of interferon (IFN) and tumor necrosis factor (TNF) in human peripheral blood leukocytes and in whole blood cultures. The effects were dose related. The identified till now cytokines induced by PRP were IFN-gamma and TNF-alpha but many other cytokines may be stimulated also. NP was considerably less active as the cytokine inducer than the natural PRP. Two volunteers given orally once daily for two to three weeks 100 or 200 micrograms PRP in tablets were found to develop the tolerance of IFN induction and had the modified TNF response. Furthermore, the PRP-treated volunteers showed signs of psycho-stimulation. Taken together our observations suggest that ovine PRP is active in humans and may have therapeutic value as an immunostimulant and/or neurotropic cytokine.

Administration, Oral↗

Interaction of Haemophilus parasuis with nasal and tracheal mucosa following intranasal inoculation of cesarean derived colostrum deprived (CDCD) swine.

Twenty-three cesarean derived, colostrum deprived pigs were obtained at 5 wk of age and inoculated intranasally with either 1.4 x 10(8) colony forming units of Haemophilus parasuis or sterile phosphate buffered saline. Pigs were euthanized at 4, 8, 12, 18, 26, or 36 h post-inoculation and tissues from the oropharynx and respiratory tract were obtained for qualitative bacterial culture, immunohistochemistry for H. parasuis antigens, and light and transmission electron microscopy. Haemophilus parasuis was consistently isolated from the nasal cavity (17/17, 100%) and trachea (13/17, 76%) and rarely isolated from the lung (3/17, 18%) and blood stream (1/17, 6%) of infected pigs. Antigens of H. parasuis were sporadically detected on the nasal mucosa (6/17, 35%) and trachea (8/17, 47%). Light microscopic lesions included submucosal and intraepithelial infiltrates of neutrophils and infrequent, patchy loss of cilia. Ultrastructural changes in nasal mucosal epithelial cells included cell protrusion, loss of cilia, and dilation of the cytocavitary network. Bacteria were infrequently identified and were either within an amorphous material at the apical surface of the cilia or were between individual cilia. These results suggest H. parasuis associates with the nasal mucosa and can induce a suppurative rhinitis with nasal mucosal epithelial cell degeneration. This process may represent an initial event in the pathogenesis of H. parasuis infection of swine.

Administration, Intranasal↗

Antibody reactivity and fecal recovery of bovine immunoglobulins following oral administration of a colostrum concentrate from cows (Lactobin) to healthy volunteers.

Using immunoblot techniques, we investigated the immunoglobulin G (IG) reactivity present in Lactobin, an immunoglobulin concentrate (prepared from colostrum pools from non-immunized cows) against potential pathogenicity factors from Yersinia enterocolitica and Campylobacter jejuni. A strong reactivity against Yersinia outer proteins (Yops), against the Yersinia adhesin A (Yad A) as well as a high reactivity against flagellin and the outer membrane proteins (OMP) of C. jejuni was demonstrated. The IgG antibody reactivity against these antigens was also assessed in vitro after incubation of IG with stools from healthy adults for different time intervals. Minimal loss occurred within 2 hours of incubation at 37 degrees C and complete loss after 24 hours. In a clinical study stool specimens from 8 healthy volunteers were analyzed 1-4 days after oral administration of the drug for the presence of bovine IgG and its antibody reactivity against Yersinia antigens. Small amounts of the bovine immunoglobulins were detected in stools from 3 of the 8 subjects, however, without antibody reactivity. Additional pharmacokinetic investigations in patients with gastrointestinal diseases are necessary to determine the optimal therapeutic regimen for these patients.

Adhesins, Bacterial↗

The concentration of cotinine in urine, colostrum and amniotic fluids within the system mother-baby.

The findings obtained by the authors of the thesis submit the new cognitive values to the diagnosis of pathology of pregnancy i.e. the influence of nicotine on the organisms of a mother and a new-born child, estimated by the assay of cotinine, the most important metabolite of nicotine. The authors lay a particular stress on the "colostrum-milk way" in the mother-child relationship and this is that needs to be emphasized in this thesis.

Adult↗

Effect of colostrinin, an immunomodulatory proline-rich polypeptide from ovine colostrum, on sialidase and beta-galactosidase activities in murine thymocytes.

Colostrinin: a proline-rich polypeptide (PRP) from ovine colostrum and its nonapeptide active fragment (NP) induce maturation and differentiation of murine thymocytes, formation of helper cells from PNAhigh thymocytes and cytotoxic T cells from PNAlow thymocytes. These processes are accompanied by changes in expression of receptors for peanut agglutinin (PNA), PNAhigh thymocytes were transformed into PNAlow cells, and vice versa. It was shown, in various laboratories, that sialyltransferases are involved in the transformation of PNAhigh thymocytes into PNAlow cells. To find out whether the expression of receptors for PNA on murine thymocytes might also be influenced by other enzymes, we decided to study the effect of PRP and NP on sialidase and beta-galactosidase activities in these cells. The results obtained showed that the most of sialidase activity of murine thymocytes is present in the plasma membrane compartments. Both thymocyte subpopulations PNAhigh and PNAlow, showed similar sialidase activity, which was not affected either by PRP or NP. In contrast to sialidases, most of beta-galactosidase activity was present in the cytosol. PNAhigh, thymocytes showed a higher beta-galactosidase activity than PNAlow cells. Incubation of immature, PNAhigh, thymocytes with PRP or NP enhanced the beta-galactosidase activity in these cells. The presented results suggest that sialidases seem not to be involved in modulation of surface sialic acid content during murine thymocyte maturation. On the other hand, stimulation of activity of beta-galactosidase in PNAhigh, immature thymocytes by PRP and NP suggests that beta-galactosidase in murine thymocytes might be involved in transformation of PNAhigh into PNAlow cells.

Adjuvants, Immunologic↗

Evaluation of killed and modified live porcine rotavirus vaccines in cesarean derived colostrum deprived pigs.

Twenty-eight cesarean derived, colostrum deprived (CDCD) piglets were used to evaluate the efficacy of killed and modified live rotavirus (MLV) vaccines against challenge with virulent A-1 and A-2 rotaviruses. Two killed rotavirus vaccines were evaluated: an experimental vaccine and a commercially available vaccine. Efficacy parameters included: average daily weight gains, rotavirus shedding in feces, morbidity incidence and duration, and rotavirus serum antibody conversion post-vaccination and post-challenge. Piglets vaccinated orally/intramuscularly with the modified live vaccine were completely protected from A-1 and A-2 virulent rotavirus challenge. Nonvaccinated control piglets and piglets receiving killed rotavirus vaccines developed diarrhea, shed virus and exhibited reduced weight gains post-challenge. Only the MLV rotavirus vaccine was able to prevent virus shedding in feces after virulent challenge. Both controls and pigs which received killed vaccines intraperitoneally, orally or intramuscularly shed virus in the feces for 7 days post-challenge and virus peak titers approached 10(7) fluorescent antibody infectious dose (FAID)50/g feces. These studies clearly reflected the inability of killed rotavirus vaccines to induce active local immunity to rotaviral diarrhea in piglets.

Animals↗

Serologic responses of young colostrum fed dairy calves to antigens of Pasteurella haemolytica A1.

The potential and limitations of early calfhood vaccination to induce active immunity to Pasteurella haemolytica A1 in conventional colostrum fed calves were investigated. Holstein dairy calves (n = 29) were vaccinated at 2 and 4 weeks of age, or at 6 and 8 weeks of age with a commercial culture supernatant vaccine (Presponse, Langford Inc., Guelph, Ont., Canada), or remained unvaccinated as controls. Serum antibody titres were measured using an indirect bacterial agglutination assay, a leukotoxin neutralization assay, and enzyme immunoassays for antibodies of the IgM, IgG1, and IgG2 isotypes binding purified capsular polysaccharide of P. haemolytica A1. Seroconversion (fourfold or greater increase in serum antibody titre) rates were compared using Fisher's exact test. The effects of passive antibody titres and age on response to vaccination were assessed by linear modelling. Vaccination at 2 and 4 weeks of age was associated with 40%, and 0% of calves seroconverting on the basis of agglutinating antibody titres, and leukotoxin neutralizing titres respectively, and 50%, 0%, and 0% seroconverting on the basis of IgM, IgG1 and IgG2 antibodies to capsular polysaccharide, respectively. Agglutinating antibody responses were not related to prevaccination antibody titres, or to age at vaccination. Higher responses (p = 0.08) to leukotoxin were observed in older calves (after taking differences in prevaccination titres into account). Statistical analyses of responses to capsular polysaccharide among calves with comparable prevaccination IgG1 antibody titres revealed significantly higher IgM, IgG1 and IgG2 responses in older calves. Rising titres of IgM antibodies in nonvaccinated calves after 5 weeks of age suggest natural exposure to P. haemolytica A1 or antigens which result in serologic cross-reactions as a means of priming immune responses.

Animals↗

Clinical effects of experimental dual infections with porcine reproductive and respiratory syndrome virus followed by swine influenza virus in conventional and colostrum-deprived pigs.

Previous studies demonstrated that experimental dual infections of pigs with porcine reproductive and respiratory syndrome virus (PRRSV) followed by H1N1 influenza virus cause more severe disease and growth retardation than the respective single virus infections. Here three experiments were undertaken to better define the clinical impact of combined PRRSV-H1N1 infections in conventional and caesarean-derived colostrum-deprived (CDCD) pigs. Groups of pigs were inoculated by aerosol with PRRSV followed by H1N1 at 3-, 7- or 14-day intervals. During the post-H1N1 period, mean body temperatures, respiratory signs and mean weight gains in the PRRSV-H1N1 inoculated groups were recorded and compared with those in uninoculated controls (experiments 1 and 2) or in singly virus-inoculated pigs (experiment 3). In a first experiment with conventional pigs, the PRRSV-3d-H1N1 and PRRSV-7d-H1N1 infections induced mean body temperatures > 40.5 degrees C during 8 days (peaks 41.1 and 41.6 degrees C, respectively) and mean growth reductions of 3.4 and 4.8 kg, respectively, during the 2 weeks after H1N1, along with marked depression and respiratory disease. The PRRSV-14d-H1N1 infection, on the contrary, was largely subclinical. In a second experiment with conventional pigs, PRRSV-3d-H1N1 and PRRSV-7d-H1N1 infections were clinically milder, with smaller increases in mean body temperatures (peak 40.5 degrees C in both groups) and growth reductions (1.4 and 1.6 kg, respectively). In both groups, only one pig showed prominent general and respiratory signs. In a final experiment with CDCD pigs, PRRSV-7d-H1N1 infection had minimal effects on mean clinical performances and growth and, except for one pig that was severely affected, differences with the single virus inoculations were negligible. Thus, both the time interval between infections and the sanitary status of pigs can affect the clinical outcome of dual PRRSV-H1N1 infections. However, factors so far unknown seem to cause large variations in the clinical response between individual pigs.

Animals↗

Impact of Yersinia enterocolitica enteritis on disaccharidase activity and small intestinal morphology in colostrum-deprived newborn piglets.

The effect of enteritis on the development of the small intestine was examined in newborn, colostrum-deprived piglets infected with a human isolate of Y. enterocolitica (serotype 0:3, biotype 4) soon after birth. The piglets were killed 3 days (n = 6) or 5 days (n = 8) after infection, or antibiotic therapy was commenced on day 5 and the animals killed on day 14 (n = 5). Compared with the non-infected controls, infected animals had reduced mucosal lactase and sucrase, but not maltase activity, while after antibiotic therapy, previously infected piglets had a lower lactase and a higher maltase and sucrase activity. Lactase activity was significantly reduced in the duodenum and jejunum, and mean values were lower in the ileum, but the difference did not reach significance; maltase activity was greater at all ages from the distal jejunum to the mid-ileum; sucrase activity was reduced in all segments up to day 5 but after antibiotic therapy was increased in the jejunum and appeared early in the ileum. Enzyme profiles were more mature along the crypt-villus axis in some segments of the intestine in previously infected piglets. Sodium-potassium-ATPase activity was unchanged. There was a reduced villus height:crypt depth ratio, crypt hyperplasia and increased crypt cell proliferation. Morphological maturation, indicated by loss of vacuoles and location of the nucleus at the base of the enterocyte, proceeded distally from the duodenum to ileum from 3 to 14 days of age when only the ileum remained immature. In infected piglets, there was reduced vacuolation and earlier location of the nucleus at the base of the cell in the distal intestine. Accelerated maturity of specific disaccharidases and enterocyte morphology in infected piglets appears to be due to physical damage to the mucosa resulting in faster proliferation of crypt cells and migration of enterocytes. It is suggested that this may reduce macromolecular internalisation and impair the ability to utilise dietary carbohydrate and may have long-term effects on growth and immunological responses of the gut.

Journal Article↗

Protection studies in colostrum-deprived piglets of a bovine rotavirus vaccine candidate using human rotavirus strains for challenge.

Studies were performed to evaluate the potential use of the bovine RIT 4237 rotavirus strain as a vaccine candidate against infection with human rotaviruses. Initial experiments revealed that colostrum-deprived piglets were susceptible to infection with several human strains, except for those belonging to subgroup 1. Subsequently, different immunization procedures with RIT 4237 were studied in this animal model. It was found that a two-dose administration, either given intramuscularly (twice) or once intramuscularly and once intragastrically, was necessary to induce a significant serum antibody response. Finally, the protective effect of the latter vaccination schedules against subgroup 2 and 3 rotavirus strains of human origin was evaluated by artificial challenge. In both cases, prior administration of live RIT 4237 significantly decreased fecal shedding of the challenge virus when compared with control animals.

Animals↗

Isotype-specific antibody responses to bovine coronavirus structural proteins in serum, feces, and mucosal secretions from experimentally challenge-exposed colostrum-deprived calves.

Five newborn isolation-reared colostrum-deprived calves were inoculated orally and intranasally when they were 20 to 30 hours old and challenge exposed when they were 21 days old with a suspension of virulent bovine coronavirus (BCV). Blood, feces, nasal swabs, tears, saliva, and bronchoalveolar lavage (BAL) fluids were collected from each calf prior to inoculation and then weekly for 5 post-inoculation weeks. An ELISA was used to quantitate the immunoglobulin isotype titers of BCV antibodies in all samples. An immunoblot assay was used to determine the antibody isotype responses to BCV structural proteins in all the samples, except saliva. At postinoculation days 2 to 3, all calves had severe watery diarrhea, shed BCV in their feces, and had evidence of BCV replication in their upper respiratory tract. After challenge exposure, no calves became ill and no evidence of BCV replication in the respiratory or intestinal tracts was detected. At postinoculation week 1, IgM responses to the N protein were seen in mucosal secretions (except nasal fluid) and feces. At postinoculation weeks 2 and 3, IgA was predominant in mucosal secretions and feces directed toward all the BCV proteins (except the E2 protein in BAL fluid). After challenge exposure, an increase (or failure to decrease) in most IgA and some IgG1 titers to BCV proteins was seen. The increases in IgA titers were to all viral proteins in all mucosal secretions and feces, except to the N and E1 viral proteins in feces. The IgG1 titer increases were to the E2 proteins in tears and BAL fluid and to the E3 protein in BAL fluid.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗