Biomedical subjects

B Amorena

Publications and source records attributed to B Amorena.

36 records · Page 2Linked to original sources

PubMed1994

Staphylococcus aureus capsule and slime as virulence factors in ruminant mastitis. A review.

Staphylococcus aureus is one of the most prevalent causes of ruminant mastitis. The interaction of this microorganism with the host is strongly dependent on its cell surface properties, specially concerning the presence of the exopolysaccharide-containing outer layers (glycocalyx), which appear to play an important role in virulence. In this article, the definition and recognition of the types of exopolysaccharide layers are described, together with their likely role in the pathogenesis of mastitis.

Animals↗

PubMed1993

Adherence of ruminant mastitis Staphylococcus aureus strains to epithelial cells from ovine mammary gland primary cultures and from a rat intestinal cell line.

Staphylococcus aureus isolated from mastitis (14 bovine and 11 ovine strains) exhibited an ability to adhere to epithelial primary cultures from ovine mammary gland and to a rat epithelial cell line, RIE-1. Strain differences in the degree of adherence were observed in both cases. These differences were maintained when comparing different epithelial sources (rat vs. ovine). RIE-1 cells can thus be used as a model for studying staphylococcal adherence to epithelial cells. Changes in bacterial adherence were observed according to the bacterial growth phase. The magnitude of these changes differed among strains. Bacterial cell surface hydrophobicity was not related to the degree of adherence to mammalian epithelial cells.

Animals↗

PubMed1993

Phase variation of slime production in Staphylococcus aureus: implications in colonization and virulence.

Two methods commonly used for slime detection in coagulase-negative staphylococci (tube biofilm formation and colony morphology in Congo red agar) were used to study 144 ruminant mastitis Staphylococcus aureus strains. Slime production was detected in 21 strains. A majority of cells (85%) in slime-producing (SP) strains and a minority of cells (5%) in non-slime-producing (NSP) strains showed a condensed exopolysaccharide matrix (slime) surrounding the bacterial cell wall, as revealed by electron microscopy and immunofluorescence. In vivo slime production was also detected immunohistochemically after experimental infection of the mammary gland in sheep. Upon repeated subcultures in Congo red agar, NSP variants were obtained from four ovine and four bovine SP strains at a frequency ranging from 0.5 x 10(-4) to 10(-4). Because SP variants could not be obtained from NSP strains within this range or at a higher frequency, they were obtained by the tube biofilm formation (requiring repeated subculturing of NSP strains in tryptic soy broth containing 2% glucose for subsequent recovery of colonies adherent to the walls of the culture tubes). In experimental challenge, the SP variant showed a significantly higher colonization capacity than did the NSP variant of the same strain used (P < 0.001). However, the NSP variant had a higher virulence than did the SP variant (P < 0.001). These results may help to explain the different roles of S. aureus slime production cell types (SP and NSP) coexisting in disease.

Animals↗

PubMed1993

Role of an intramammary device in protection against experimentally induced staphylococcal mastitis in ewes.

An intramammary device (IMD) was adapted for use in ewes; this device was made of abraded polyethylene material (1.7 mm in diameter, 47 mm long) and formed a 15-mm-diameter loop in the gland cistern. The IMD was inserted in 1 gland in each of 43 ewes. A significant (P < 0.0001) increase in milk somatic cell count (SCC) was observed in glands provided with an IMD. This increase was attributable to an increase in neutrophil numbers and was observed during the first 12 weeks after insertion. The IMD had a protective effect against experimentally induced staphylococcal mastitis (Staphylococcus aureus and S epidermidis), although different milk SCC were required for protection from each bacterial species in most ewes (10(6) and 2 x 10(5) cells/ml, respectively). Histologic studies revealed that the IMD induced local squamous metaplasia in the glandular part of the lactiferous sinus. Erythrocytes were found in milk from glands provided with an IMD throughout the studied period (35 days of the 45-day lactation) and, in some cases, blood clots were observed during the first 2 weeks of lactation. Glands with IMD also had lower milk production and quality at 30 and 32 days of lactation. Eight ewes with IMD were studied throughout a subsequent lactation. Milk from the IMD-containing glands had an increase in SCC, as in the previous lactation period; did not contain blood clots or erythrocytes; and had normal composition (similar to that in glands without the IMD).

Animals↗

PubMed1992

An efficient microtest to study adherence of bacteria to mammalian cells.

A simple, fast and highly reproducible microtest was developed for in vitro adherence studies. A rat epithelial cell line was investigated for the adherence of clinical and subclinical ovine and bovine Staphylococcus aureus strains isolated from mastitis. Staphylococcus aureus strains differed in their ability to adhere to epithelial cells, the degree of adherence being dependent on the concentration of bacteria used in the test.

Animals↗

PubMed1991

Infection of rabbit mammary glands with ovine mastitis bacterial strains.

An experimental model was developed in rabbits to study ovine mastitis. A total of 19 ovine mastitis bacterial strains (seven Staphylococcus aureus, four Staph. chromogenes, four Staph. hyicus and four Escherichia coli) were used for mammary gland infections. The histopathological results showed that the ovine mastitis types corresponded to experimental infections produced in the rabbit with the ovine strains. These results helped the grading of the bacterial species tested according to the severity of their effects on the mammary gland. The most pathogenic species was Staph. aureus, followed by E. coli, Staph. hyicus and Staph. chromogenes, in that order. There was, however, variation among strains within a given species (e.g. one out of seven Staph. aureus strains gave rise to a mild infection in sheep and rabbits). The procedure was simple and consisted of introducing bacterial suspensions through alternate teat ducts of does with the help of a cannula. It helped minimize the number of animals required in the experiments.

Animals↗

PubMed1990

Factors influencing the degree of in vitro bacterial adhesion to ovine mammary gland epithelial cells.

Bacterial adhesion to mammary gland epithelial cells (EC) may play a role in the pathogenesis of mastitis. In vitro adherence systems have been developed to study mastitis in cattle but little has been done in sheep. In this work, a method is described for obtaining mammary gland cell preparations containing greater than or equal to 65% EC from live or dead ewes, using a Ficoll-Hypaque flotation method (cell viability = 70-90%). An in vitro adhesion assay procedure was also developed to study the interaction between EC and ovine mastitis bacterial strains. It was observed that, under the test conditions, adherence increased as the incubation time was prolonged from 30 to 120 min (P less than 0.05). Adhesion was greater at incubation temperature of 37 degrees C than at 22 degrees C (P less than 0.001). An acidic pH (5.9) was associated with an increase in adhesion, when compared with a higher pH (7.2; P less than 0.05). Tween 20, Tween 80 and bovine serum albumin helped to eliminate a background of unbound bacteria from the test slides, but they also inhibited adhesion to some strains. Strain differences in adhesion and in ability to form a background were also observed. Some of these findings may have in vivo implications.

Animals↗

PubMed1990

Milk whey induction of agglutination in ovine and bovine mastitis Staphylococcus aureus.

A total of 59 mastitis staphylococcic strains were tested for growth agglutination upon supplementation of growth media with ovine and bovine milk whey and mammary secretions from dry cows. Differences were observed when comparing bacterial species or origins (ovine vs. bovine) of bacteria and whey. All of the ovine and bovine S. aureus strains tested, but only 4 among 22 other ovine mastitis staphylococcic strains, showed growth agglutination in Todd Hewitt broth (THB) supplemented with greater than or equal to 30% (v/v) ovine milk whey. None of the strains agglutinated during growth in regular THB medium. Ovine whey had an agglutination induction capacity higher than bovine whey (P less than 0.005), concerning the number of responsive ovine and bovine S. aureus strains. There were no differences between whey samples from different ewes with regard to their capacity to induce agglutination. Ovine S. aureus strains were more responsive than bovine strains of this bacterial species, concerning the number of responsive strains (P less than 0.001) to bovine whey (greater than or equal to 30% in THB), the proportion of responsive strains at low (10%) ovine whey concentration (P less than 0.001), and the strength of reaction (precipitation timing and clump size). Secretions from dry cows systematically induced agglutination in all of the bovine and ovine S. aureus strains tested.

Agglutination↗

PubMed1989

Blood biochemical polymorphisms as markers for genetic characteristics of wild Spanish and domestic rabbits.

Seventeen blood proteins were studied in a sample of 412 Spanish wild rabbits and in 598 domestic rabbits belonging to various breeds. The wild rabbit populations showed a high level of genetic polymorphism. Six loci were monomorphic, while the remaining ten loci were segregating for at least two alleles. Two of the loci that were polymorphic in the wild rabbits were monomorphic in the domestic ones. Wright's inbreeding coefficient in the total Spanish wild rabbit population was F = 5.66, indicating subdivision of the total population. Inbreeding coefficients, estimated by Kidd et al.'s method (Anim. Blood Grps, Biochem. Genet. 11: 21-38), differed significantly from zero, being 15.62%, in wild rabbits and 6-12% in domestic breeds, indicating consanguinity. Genetic distances between wild rabbit populations showed that factors other than geographic distance (e.g., bottlenecks, barriers such as rivers, mountains, etc.) may explain the result that a northern population forms a cluster with two central populations whereas the northeastern populations form a different cluster with another central population. Populations of the first cluster are more closely related to the captive populations than others. There are three population clusters of domestic rabbits, namely (1) New Zealand White and a hybrid combination; (2) Spanish Common, Butterfly, Burgundy, and Californian; and (3) Spanish Giant.

Animals↗

PubMed1987

Relationship between rabbit transferrin electrophoretic patterns and plasma iron concentrations.

Rabbit transferrin (Tf) was studied electrophoretically using 1141 blood samples from individuals belonging to seven populations (Spanish Common, Spanish Giant, Butterfly, Lyoné de Bourgogne, New Zealand White, Californian and New Zealand White X Californian hybrids). No Tf polymorphism was found by starch gel electrophoresis, but six patterns, differing in the presence and/or intensity of three bands ('a', anodic; 'b', intermediate; and 'c', cathodic) were observed by polyacrylamide gel electrophoresis. No genetic model could explain these patterns, since they reflect differences in plasma Tf iron content. The electrophoretic test allowed a direct observation of the relative in vivo levels of the different Tf molecular species; saturated (band 'a', Fe2Tf); semi-saturated (band 'b', Fe1Tf); and without iron (band 'c' Fe0Tf, apotransferrin). The degree of iron saturation of Tf varied among individuals and throughout the individual's life. Specifically, in pregnant females, Fe2Tf and Fe1Tf are generally observed, except in late pregnancy (from day 25 to parturition), when mainly apotransferrin is observed. Significantly, within 24 h post-partum, high levels of Fe2Tf are reached in the female's serum.

Animals↗

PubMed1987

Evidence for transferrin polymorphism in Spanish wild rabbits.

Serum samples from 412 Spanish wild rabbits were analysed by starch and polyacrylamide gel electrophoresis. Three different transferrin (Tf) phenotypes (A, AB and B) were observed by both methods. The occurrence of two codominant alleles (TfA and TfB with frequencies of 0.89 and 0.11 respectively) at an autosomal locus (Tf) was supported by the population data on genetic equilibrium. Electrophoretic mobility differences between the Tf variants A and B could not be explained by differences in sialic acid or iron contents. Each of the two Tf variants were shown to have two sialic acid residues by neuraminidase treatment. These variants had similar affinities for iron, and iron binding did not lead to the conversion of one variant into the other.

Alleles↗

PubMed1987

Linkage studies in blood biochemical polymorphic markers of rabbits.

Five erythrocyte proteins (Adenosine deaminase, Ada; 6-Phosphogluconate dehydrogenase, Pgd; Esterase 1, Es-1; Esterase 3, Es-3; NADH-Diaphorase 2, Dia-2) and a serum beta-globulin protein (Esterase 7, Est-7) were studied in rabbits using starch gel electrophoresis. Samples were obtained from 317 Spanish Common individuals (38 families, 317 individuals). For the analyses of linkage, Morton's sequential probability ratio test was applied. Clear evidence for linkage between Es-1 and Est-7 (theta 0.2) was obtained, and no evidence of linkage was obtained for the remaining pairwise combinations of loci studied.

Alleles↗

PubMed1983

Computer methodology for simulation and prediction of alloimmune responses: expected antibody specificities.

The methodology presented here was developed for simulating planned alloimmunization results, with regard to the type and number of expected antibody specificities. The computer program designed for this purpose was adapted to an immunogenetic model using Boolean algebra. It was written to help immunogeneticists avoid handling routine data preceding selection of donor-recipient pairs, specially concerning blood type alloimmunizations in animals: all of the donor-recipient combinations and the expected antibody specificities (their limit number being specified in each particular immunization program) are provided in the print-out.

Animals↗

PubMed1982

Sources of bovine lymphocyte antigen (BoLA) typing reagents.

Sera from about 1000 cows were tested for cytotoxicity against a panel of up to 100 lymphocyte samples. Cytotoxic antibodies presumably resulting from transplacental immunization of the cow by her calf were found in about 45% of these sera. The antibody titers of sera from parous cows rarely exceed 4(2), some persisted for over one year, but decreased notably at calving. Thirty-five immune sera were also produced by alloimmunization with lymphocytes. They usually reached peak titers of up to 4(4) at 2 or 3 weeks after the initial immunization. Subsequent immunizations produced sera with very high titers but they were much more polyspecific. High-titered antibodies were also produced by skin graft recipients. Useful cytotoxic antibodies were found in 19 of 111 colostrum whey samples. Studies on 13 dam-calf pairs showed that the newborn calf may acquire cytotoxic antibodies from its mother's colostrum, but the only cytotoxic antibodies detectable in this calf's serum are those not directed against its own lymphocyte antigens. It is concluded that efficient lymphocyte typing requires antibodies from a variety of sources.

Animals↗

PubMed1980

Bovine lymphocyte antigens (BoLA): a serologic, genetic and histocompatibility analysis.

Eleven lymphocyte antigens have been defined in cattle using 81 lymphocytotoxic sera. These sera (typing reagents) were selected from over 1,000 normal sera, 35 alloimmune sera, and 111 samples of colostrum whey. Absorptions revealed that nine of the 11 antigens detected by these reagents are serologically independent. Segregation of the genes controlling these antigens was observed among 470 calves from mating of 56 heterozygous sires and 360 dams, indicating that each of the 11 antigens is controlled by one of 11 alleles at a single autosomal co-dominant locus BoLA-A (Bovine Lymphocyte Antigen, locus A). To determine the relation between BoLA antigens and histocompatibility, skin allografts were exchanged among four unrelated adult cows. Incompatibilities for the SD antigens, but not MLR or blood group antigens, were predictive of early rejection of allografts. Furthermore, graft recipients produced antibodies against the SD antigens of incompatible donors. These results indicate that the BoLA-A locus is part of the major histocompatibility complex (MHC) of cattle.

Absorption↗

PubMed1979

Analysis of alloantisera against bovine lymphocytes. Joint report of the 1st International Bovine Lymphocyte Antigen (BoLA) workshop.

The results and agreements of the 1 international BoLA workshop, held in Edinburgh, Scotland in August 1978, are reported. Most of these concern the results from a comparison test of 249 alloantisera to bovine lymphocytes, the antisera being contributed by 9 laboratories. These sera were compared directly in Edinburgh on a panel of lymphocytes from 130 cattle of 21 breeds. In the microlymphocytotoxicity test used 75% of the sera reacted. Sixty eight of these sera were grouped into clusters according to their reaction patterns against the lymphocyte panel. Eleven of these clusters were clearly defined and were given workshop BoLA designations. In addition 22 sera were assigned to subgroups of the agreed clusters. There was no evidence that the method of production of the sera had any effect on their specificity. Although genetic data was not available, the phenotypes of the test panel of lymphocytes are consistent with the clusters detecting antigens controlled by multiple alleles at a single autosomal locus. It was agreed to name the genetic region where this putative locus is located BoLA (bovine lymphocyte antigen).

Animals↗

PubMed1978

Serologically defined (SD) locus in cattle.

Using cytotoxic serums obtained from multiparous cows or by alloimmunization, we have detected 11 lymphocyte antigens controlled by codominant alleles at a serologically defined locus called BoLA-A (bovine lymphocyte antigens). This locus, along with the lymphocyte defined loci previously reported, establishes the existence of a major histocompatibility system of cattle.

Alleles↗

PubMedDate not supplied

In vitro development of Staphylococcus aureus biofilms using slime-producing variants and ATP-bioluminescence for automated bacterial quantification.

In this work, a method was developed to establish Staphylococcus aureus biofilms on 96-well plates and automatically quantify viable cells within these biofilms by ATP-bioluminescence. Different strains were compared for biofilm formation. Cells from slime producing (SP) strain variants were more adherent (p < 0.001) and therefore more suitable for biofilm formation than non-slime producing original isolates. To compare biofilm support surfaces, SP biofilms were formed for 6, 24 and 48 h on 96-well polystyrene plates, containing wells coated with gelatin, poly-L-lysine or pre-treated for tissue culture and uncoated wells. Tissue culture-treated wells enhanced biofilm formation, allowing the highest growth (p < 0.001) in well-established biofilms (24 or 48 h old). For ATP quantification, the efficacy of different ATP extractants was compared: dimethyl sulphoxide (DMSO), trichloroacetic acid (TCA), a commercially available releasing reagent(R) (RR) and lysostaphin. A greater inhibitory effect on the ATP detection (p < 0.01), a more variable light emission (variation coefficient >/=50% vs. <19%, respectively) and a lower extraction efficiency (p < 0.05) were found in the case of TCA or lysostaphin in relation to RR or DMSO. DMSO was found preferable in relation to RR (upper detection limits 2.3 x 10(9) and 2 x 10(8) CFU/mL respectively) for bacterial ATP extraction from biofilms with high bacterial density. DMSO extracted ATP within seconds, light emission being stable for 6 h. The method developed allows automated viability determination of biofilm cells using bioluminescence and simultaneous study of factors affecting this viability (culture media, antibiotic types, antimicrobial concentrations, support surfaces and biofilm ages). It may be of use in bacteriological and antimicrobial research.

Adenosine Triphosphate↗