Modulation of host immune responses by protozoal DNA

The pathology caused by acute Babesia bovis infection is similar to that seen in severe human malaria caused by Plasmodium falciparum infection, which is related to dysregulated production of inflammatory cytokines and nitric oxide (NO). We have observed induction of NO, inducible nitric oxide synthase (iNOS) and inflammatory cytokines in macrophages by B. bovis. Furthermore, proliferation of lymphocytes from individuals never exposed to certain protozoal pathogens can be induced by crude protozoal parasite extracts. We have repeatedly observed stimulation of naive PBMC from cattle to antigenic extracts of Babesia bovis. Based on recent studies demonstrating the mitogenicity of bacterial and other non-vertebrate DNAs for murine B cells and macrophages, the mitogenic properties of B. bovis DNA were examined. B. bovis and E. coli DNAs induced proliferation of PBMC and purified B cells from non-exposed cattle. Stimulatory activity was reduced by DNase treatment and methylation with CpG methylase, indicating the presence of stimulatory non-methylated CpG motifs in the B. bovis genome. B. bovis and E. coli DNAs enhanced IgG secretion by cultured B cells, stimulating IgG1 and more strongly, IgG2. Several hexameric CpG immunostimulatory sequences (ISS) active for murine B cells were identified in an 11 kb fragment of B. bovis DNA. An oligodeoxyribonucleotide containing one of these (AACGTT), located in the rhoptry associated protein-1 (rap-1) open reading frame, stimulated B cell proliferation. These studies identify a potential mechanism by which protozoal parasites may modulate host immune responses, leading to consequences such as hypergammaglobulinemia and splenomegaly. These results also support the use of ISS as vaccine adjuvants to enhance Type 1 immune responses in cattle.     

Experimental infection of normal and immunosuppressed pigs with Pseudomonas pseudomallei

A single dose of 5 x 10(8) bacilli of Pseudomonas pseudomallei by intratracheal injection resulted in acute (21 cases) or chronic (19 cases) melioidosis in 40 of 48 pigs. Fifteen (10 acute and 5 chronic) had been immunosuppressed by cyclophosphamide before inoculation. The major clinical signs were initial fever, marked neutrophilia and, in the acute cases, respiratory distress. There were no signs of the nasal and ocular discharge, paresis or diarrhoea seen in acute cases in south-east Asia. The cyclophosphamide treatment caused a significant decrease in the neutrophil count by 7 d after inoculation in all 15 immunosuppressed pigs, and all were culture positive at necropsy. Eight of the 33 non-treated pigs were culture negative at necropsy. Pigs overcoming the initial phase of infection had more abscess-like nodules that were bacteriologically sterile at necropsy than the pigs with acute cases of melioidosis. P. pseudomallei was isolated predominantly from the spleen, lungs and the injection site. Although only one strain was used in this study, it is likely that Australian strains of P. pseudomallei are not as virulent as the south-east Asian isolates.     

Effects of supplemental lactoferrin on serum lactoferrin and IgG concentrations and neutrophil oxidative metabolism in Holstein calves

Lactoferrin (LF) is an iron-binding protein present in both colostrum and secondary granules of polymorphonuclear neutrophils (PMNs). We hypothesized that supplemental LF enhances neutrophil function in neonatal calves. Newborn calves were assigned to receive colostrum (C), colostrum + LF (CLF, 1 g/kg), or milk replacer + LF (MRLF, 1 g/kg). Serum (LF and IgG) and whole blood (neutrophil isolation) samples were obtained prior to treatment (day 0) and at 24 hours and 9 days of age. Serum IgG concentrations (mean +/- SD) in C, CLF, and MRLF calves at 24 hours were 1,911 +/- 994 mg/dL, 2,181 +/- 625 mg/dL, and 0 mg/ dL, respectively. Serum LF concentrations in C, CLF, and MRLF calves on day 0 were 324 +/- 334 ng/mL (range 0-863 ng/mL), 135 +/- 158 ng/mL (range 0-429 ng/mL), and 318 +/- 337 ng/mL (range 0-964 ng/mL), respectively. LF concentrations in C, CLF, and MRLF calves at 24 hours were significantly higher (P < .05), at 1,564 +/- 1,114 ng/mL (range 335-3,628 ng/mL, 2,237 +/- 936 ng/mL (range 31-3,287 ng/mL), and 3,189 +/- 926 ng/mL (range 1,736-4,120 ng/mL), respectively. Cytochrome c reduction in opsonized zymosan-treated or phorbol ester-treated cells was not significantly affected by supplemental LF provided at birth. Oral LF is absorbed in calves but does not alter PMN superoxide production and does not alter IgG absorption.     

Tracking the long-term decline and recovery of an isolated population

Effects of small population size and reduced genetic variation on the viability of wild animal populations remain controversial. During a 35-year study of a remnant population of greater prairie chickens, population size decreased from 2000 individuals in 1962 to fewer than 50 by 1994. Concurrently, both fitness, as measured by fertility and hatching rates of eggs, and genetic diversity declined significantly. Conservation measures initiated in 1992 with translocations of birds from large, genetically diverse populations restored egg viability. Thus, sufficient genetic resources appear to be critical for maintaining populations of greater prairie chickens.