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Biomedical subjects

G M Barrington

Publications and source records attributed to G M Barrington.

At least 19 recordsLinked to original sources

Escherichia coli isolates' serotypes, genotypes, and virulence genes and clinical coliform mastitis severity.

Dairy cattle with clinical mastitis caused by Escherichia coli exhibit a wide range of disease severity, from mild, with only local inflammatory changes of the mammary gland, to severe, with significant systemic derangement. The present study was designed to examine the relationship between serotype and virulence genes of E. coli mastitis isolates, different levels of systemic disease severity, and farm from which the E. coli strain was obtained. One hundred twenty-three E. coli milk isolates were obtained from cows with clinical mastitis of varying systemic disease severity from 6 different farms. No predominant serotype was identified by farm or by systemic disease severity; however, the most frequent serotype, O158:NM (n = 3), was isolated from cows in the moderate severity group. Virulence genes evaluated were identified infrequently and were not associated with systemic disease severity. Evaluation of genetic similarity showed no clustering assigned by farm or mastitis severity based on systemic disease signs. We concluded that a high degree of genotypic variability is characteristic of E. coli strains causing clinical mastitis within and between different farms and systemic severity groups, and that specific cow factors probably play a more important role in determining systemic disease severity.

Animals↗

The innate immune response in calves to Boophilus microplus tick transmitted Babesia bovis involves type-1 cytokine induction and NK-like cells in the spleen.

The innate immune response to Babesia bovis infection in cattle is age-related, spleen-dependent and, in stabilate inoculated calves, has type-1 characteristics, including the early induction of IL-12 and IFN-gamma. In this study with three calves, parameters of innate immunity were followed for 2 weeks after tick transmission of B. bovis. Each calf survived the acute disease episode without drug intervention, and responded with increased levels of plasma interferon-gamma and type-1 cytokine expression, monocyte/macrophage activation, and CD8+ cellular proliferation in the spleen. The proliferating CD8+ population consisted primarily of NK-like cells, and the expansion occurred in parallel with an increase in IL-15 mRNA expression in the spleen.

Animals↗

IL-4 and IL-10 inhibition of IFN-gamma- and TNF-alpha-dependent nitric oxide production from bovine mononuclear phagocytes exposed to Babesia bovis merozoites.

The requirement for IFN-gamma and/or TNF-alpha as co-stimulants with Babesia bovis merozoites for nitric oxide (NO) production was examined, as well as the regulatory role of IL-4 and IL-10. Purified B. bovis merozoites did not induce the production of NO in undifferentiated monocytes without addition of exogenous IFN-gamma and TNF-alpha unless the monocytes taken ex vivo were producing TNF-alpha endogenously. Under the latter condition, the NO production resulting from merozoite stimulation remained IFN-gamma-dependent. There was no evidence for endogenous synthesis of TNF-alpha in monocyte-derived macrophages (MDM), and merozoites alone were incapable of inducing TNF-alpha mRNA in MDM. However, while merozoites plus IFN-gamma induced TNF-alpha mRNA expression in MDM, NO was not produced. Both IL-4 and IL-10 inhibited expression of iNOS and production of NO in merozoite-stimulated monocytes.

Animals↗

Age-related innate immune response in calves to Babesia bovis involves IL-12 induction and IL-10 modulation.

There is a strong innate immunity in calves to infection with Babesia bovis. Interleukin (IL)-12 and IL-10 have been shown in vitro to be important immunoregulatory cytokines. Here we demonstrate in vivo that the protective innate response in young calves to infection with virulent B. bovis involves the early appearance of IL-12 and interferon-gamma (IFN-gamma) transcripts in the spleen. In contrast, IL-12 and IFN-gamma mRNA expression in the spleens of adult cattle that succumbed to the infection was delayed and depressed and occurred within the context of IL-10 expression. Also in contrast with calves, there was no detectable antibody response before death in adults. A vigorous CD8+ T-cell expansion occurred in the spleens of both calves and adults.

Age Factors↗

Bacteremia associated with naturally occuring acute coliform mastitis in dairy cows.

OBJECTIVE: To determine the incidence of bacteremia in dairy cows with naturally occurring acute coliform mastitis (ACM) with a wide range of disease severity. DESIGN: Cohort study. ANIMALS: 144 dairy cows with ACM from 6 herds. PROCEDURE: Cows were examined at time of identification of ACM (time 0) and classified as having mild, moderate, or severe mastitis on the basis of rectal temperature, hydration status, rumen contraction rate, and attitude. Cows were reexamined at 24 or 48 hours. Bacteriologic culturing of milk and blood (30 ml), CBC, and serum biochemical analysis were performed at each time point. Appropriate samples were obtained at a single point from herdmates without mastitis (controls) that were closely matched for lactation number and days since parturition. Blood culture results were compared among severity groups and controls by use of chi2 tests, as was outcome of an ACM episode for cows grouped by blood bacterial isolates. RESULTS: Bacteria were isolated from 52 blood samples from 46 of 144 (32%) cows with ACM, which was significantly more than control cows (11/156; 7.1%). Group-1 isolates (Escherichia coli, Pasteurella multocida, Mannheimia haemolytica, Klebsiella pneumoniae, Enterobacter agglomerans, and Salmonella enterica serotype Typhimurium) were identified in 20 of 144 (14%) cows with ACM and 0 of 156 control cows. Group-1 isolates were identified in 4.3, 9.1, and 42% of cows classified as having mild, moderate, and severe ACM, respectively. Escherichia coli and K pneumoniae milk and blood isolates obtained from the same cow were of the same genotype. Bacillus spp were identified in 21 of 144 (15%) cows with ACM, which was significantly more than control cows (3/156; 1.9%). Thirty-five percent of cows with a group-1 isolate died during the mastitis episode. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that bacteremia develops in a substantial proportion of cows with ACM. Classification of severity of disease is important for establishment of effective treatment protocols; parenteral antimicrobial treatment may be indicated in cows with ACM.

Acute Disease↗

Use of systemic disease signs to assess disease severity in dairy cows with acute coliform mastitis.

OBJECTIVE: To evaluate the use of systemic disease signs for classifying severity of acute coliform mastitis in dairy cows. DESIGN: Prospective cohort study. ANIMALS: 144 dairy cows. PROCEDURE: Cows were examined at the time of initial identification of disease (time 0) and classified as having mild, moderate, or severe disease on the basis of rectal temperature, hydration status, rumen contraction rate, and attitude. A CBC and serum biochemical analyses were performed, and milk samples were submitted for bacterial culture at time 0 and 48 hours later. RESULTS: 69 cows were classified as having mild disease, 44 as having moderate disease, and 31 as having severe disease. Median WBC and neutrophil counts were significantly lower in cows with moderate or severe disease at time 0 than in cows with mild disease. Band neutrophil count was significantly higher at 48 hours and serum calcium concentration was significantly lower at time 0 and at 48 hours in cows with severe or moderate disease, compared with cows with mild disease. Twenty-eight, 51, and 77% of cows with mild, moderate, and severe disease, respectively, had > 100,000 colony-forming units/ml of milk at time 0. The odds that a cow with severe disease would die or be culled were 3.6 times the odds for a cow with moderate disease and 11.2 times the odds for a cow with mild disease. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a classification scheme based on readily observable systemic disease signs can be used to classify disease severity in cows with acute coliform mastitis.

Acute Disease↗

Bovine neonatal immunology.

The majority of early, in utero immune development occurs independent of antigen exposure. Only later during development can a fetus respond to antigens, and even then the response depends on the stage of fetal development and the nature of the antigen. At birth, the neonate is rapidly exposed to large numbers of potential pathogens. Although immunocompetent, the neonate is immunonaive and dependent on passively acquired maternal immunoglobulins, immune cells, and other substances from colostrum for protection. Neonates that suffer failure of passive transfer of maternal immunoglobulins may be at increased risk for disease; however, many other factors interact in conjunction with the level of passively acquired immunoglobulin to determine the occurrence of disease. These include, but are not limited to, management, environment, hygiene, infection pressure, virulence of organisms, and antibody specificity. In addition to immunoglobulins, colostrum contains large numbers of immune cells and cytokines. It is thought that the primary role for the cellular component of colostrum is to interact with the development of local immunity and to modulate active immunization of the neonatal intestine. In particular, T lymphocytes are thought to transfer immune functions and secrete cytokines. Although most of the major cytokines have been identified in colostrum and milk, their biologic effects on the neonate have yet to be determined.

Animals↗

The age-related immunity in cattle to Babesia bovis infection involves the rapid induction of interleukin-12, interferon-gamma and inducible nitric oxide synthase mRNA expression in the spleen.

Young calves possess a strong innate immunity against Babesia bovis infection that lasts for approximately 6 months after birth and is abrogated with the removal of the spleen. This immunity is characterized as cellular involving a soluble mediator. Nitric oxide has been implicated by virtue of its babesiacidal affects in vitro, but questioned to be as effective in vivo, due to its ability to downregulate type-1 immunity. Spleen cells were obtained from 4-month-old calves and adult steers and processed for monitoring cytokine and inducible nitric oxide synthase (iNOS) mRNA expression during the response to initial B. bovis infection. The data provided evidence of a transient role for nitric oxide in innate immunity, characterized by brief iNOS induction in the spleen of calves that was not detectable in the spleens of adults. The iNOS message followed the early induction of interleukin (IL)-12 and interferon (IFN)-gamma message in calves. The induction of IL-12 and IFN-gamma message in adults was delayed until IL-10 message was induced. Transformation growth factor-beta mRNA expression levels were greater in spleen cells from adults early in infection and then declined, whereas expression levels increased in spleen cells from calves later in the infection process. Together, the data support the concept of 'first come, first serve' cytokine influence over cellular activities, the importance of a type-1 response in the control of an initial infection and the need for tight regulation in order to prevent pathology associated with over production of nitric oxide and inflammatory cytokines.

Age Factors↗

Cutaneous metastases of a mammary carcinoma in a llama.

An 8-year-old, female llama was evaluated for nonhealing, ulcerative, cutaneous lesions, which also involved the mammary gland. Biopsies of the lesions distant from and within the mammary gland area revealed an aggressive carcinoma. The tumor was confirmed at necropsy to be a mammary gland adenocarcinoma with cutaneous metastasis.

Adenocarcinoma↗

Flow cytometric analysis of an immunodeficiency disorder affecting juvenile llamas.

The present study was undertaken to characterize the immune system of llamas and alpacas and establish the basis for an immunodeficiency disorder affecting juvenile llamas. Flow cytometric (FC) analysis of the immune system with a panel of monoclonal antibodies (mAbs) revealed the immune system of llamas and alpacas is similar in leukocyte subset composition to that in ruminants. Peripheral blood mononuclear cells in adults are comprised of surface immunoglobulin (sIg(+)) B-cells (31%+/-8 S.D.), alphabeta T-cells (27%+/-12 S.D.), WC1(+) gammadelta T-cells (16%+/-11 S.D.), and 5-16% monocytes. In contrast to cattle, goats, and sheep, however, the frequency of WC1(+) gammadelta T-cells is not high in juveniles but similar to the frequency in adults. Also, sIg(+) B-cells are present in high concentration in juveniles (43%+/-11 S.D. ). Expression of major histocompatibility class II molecules on resting T-cells was low or absent. Comparative analysis of peripheral blood lymphocyte composition in normal juvenile llamas and llamas presenting with the signs of the juvenile llama immunodeficiency syndrome (JLIDS) revealed the concentration of B-cells is extremely low (1-5%) in affected animals. The findings suggest JLIDS is attributable to an autosomal recessive genetic defect in the development of B-cells.

Animals↗

Septic orchitis in an alpaca.

An adult, intact male alpaca was presented with an acute onset of unilateral scrotal swelling. Following complete physical and ultrasonographic examination, the most likely differential diagnoses were orchitis, hematoma, and testicular torsion. The animal was castrated and histopathologic evaluation revealed unilateral orchitis. Streptococcus equi subsp. zooepidemicus was cultured.

Animals↗

Regulation of the immunoglobulin G1 receptor: effect of prolactin on in vivo expression of the bovine mammary immunoglobulin G1 receptor.

Induction of colostrogenesis in non-pregnant cows was used to evaluate the relationship between prolactin (PRL) and mammary immunoglobulin G1 (IgG1) receptor expression. Six of eleven non-pregnant, non-lactating Holstein cattle responded to a standard lactation induction protocol by development of elevated IgG1 concentrations in mammary secretions. In order to increase the diversity in PRL concentrations, two of the six cattle were treated with bromocriptine, and two others were treated with recombinant bovine PRL. Serum alpha-lactalbumin, serum PRL and mammary secretion IgG1 concentrations were measured throughout the experiment. Biopsies of mammary tissue were collected after induction of lactation, and after treatments to alter serum PRL. Immunohistochemistry was used to evaluate IgG1 receptor expression. Administration of recombinant bovine (rbPRL) was associated with increased lactogenic activity, decreased secretion IgG1 concentrations, and decreased IgG1 receptor expression. Decreased serum PRL, due to bromocriptine, was associated with decreased lactogenic activity and maintenance of IgG1 receptor expression. Results of this experiment are consistent with an effect of PRL in decreasing the expression of the bovine mammary IgG1 receptor at the onset of lactogenesis.

Animals↗

Expression of immunoglobulin G1 receptors by bovine mammary epithelial cells and mammary leukocytes.

The objective of this study was to identify and evaluate expression of IgG1 receptors by different cell types in mammary tissue sections and digest-dispersed cells from the bovine mammary gland. An immunohistochemical system utilizing avidin-biotin-peroxidase complex demonstrated epithelial expression of IgG1 receptors in mammary tissue sections from cows producing colostrum but not from cows in lactation. Fluorescence flow cytometry demonstrated that cells dispersed in digests from both tissues producing colostrum and lactating tissues selectively bound IgG1. Fluorescence flow cytometry, using monoclonal antibodies to cell surface molecules, cytokeratin, and IgG1, revealed that leukocytes constituted the largest percentage of cells and were the predominant cell type binding IgG1 in mammary tissue digests. Although IgG1 binding to epithelial cells predominated in the gland during colostrum production in situ, digestion and filtration to produce single cell suspensions resulted in the loss of large numbers of epithelial cells. Studies of Ig binding of cells produced by enzymatic digestion must account for the types of cells surviving the digestion process.

Animals↗

Effect of prolactin on in vitro expression of the bovine mammary immunoglobulin G1 receptor.

Explants of mammary tissue from cows in late pregnancy were incubated for 72 h in serum-free, hormonally defined media to investigate the regulation of the bovine mammary IgG1 receptor. Treatments included incubation in basal medium alone, basal medium plus estradiol-17 beta, basal medium plus prolactin, or basal medium plus estradiol-17 beta and prolactin. alpha-Lactalbumin production was measured by radioimmunoassay in culture supernatants collected at 24, 48, and 72 h. Explants were examined immunohistochemically for expression of the IgG1 receptor at 24, 48, and 72 h. alpha-Lactalbumin concentrations increased, and IgG1 receptor expression decreased, by 72 h with explants cultured in the medium containing prolactin. Results suggest that, in addition to its positive lactogenic effect, prolactin decreases expression of the bovine mammary IgG1 receptor.

Animals↗

Squamous cell carcinoma originating from a cutaneous scar in a llama.

A nonhealing wound associated with a laceration in a 12-year-old llama was evaluated. Initial attempts at closure were unsuccessful and biopsy revealed scar tissue. Subsequent biopsies, 18 mo later, revealed squamous cell carcinoma with regional metastasis. This report describes squamous cell carcinoma, secondary to a traumatic wound in a llama.

Animals↗

Tick paralysis in two llamas.

An 18-month-old sexually intact male llama and a 7-month-old female llama were examined because of weakness, lethargy, and recumbency. Both had signs of ascending motor paralysis with minimal or no afferent sensory loss. Tick paralysis was diagnosed on the basis of complete clinical recovery following removal of a single, attached and engorged female tick (Dermacentor andersoni) from each llama. The male llama recovered within a few hours after removal of the tick, but recovery in the female llama required several days. Serum creatine kinase activity was high in both llamas, possibly as a result of direct effects of toxin on muscles or nerves.

Animals↗

Ruptured urinary bladder after dystocia in a cow.

Rupture of the urinary bladder of a cow occurred secondary to prolonged dystocia. Primary surgical closure of the tear was performed after drainage of the uroperitoneum, and the cow recovered without complications.

Animals↗