Research Note: Bovine lactoferrin in chickens: an investigation into its viability as an antibiotic alternative.

Finding effective antibiotic alternatives is crucial to managing the re-emerging health risk of Clostridium perfringens (CP) type A/G-induced avian necrotic enteritis (NE), a disease that has regained prominence in the wake of governmental restrictions on antibiotic use in poultry. Known for its antimicrobial and immunomodulatory effects, the use of bovine lactoferrin (bLF) in chickens is yet to be fully explored. In this study, we hypothesized that bLF can accumulate in the small intestines of healthy chickens through gavage and intramuscular supplementation and serves as a potential antibiotic alternative. Immunohistochemistry located bLF in various layers of the small intestines and ELISA testing confirmed its accumulation. Surprisingly, sham-treated chickens also showed the presence of bLF, prompting a western blotting analysis that dismissed the notion of cross-reactivity between bLF and the avian protein ovotransferrin. Although the significance of the route of administration remains inconclusive, this study supports the hypothesis that bLF is a promising and safe antibiotic alternative with demonstrated resistance to the degradative environment of the chicken intestines. Further studies are needed to determine its beneficial pharmacological effects in CP-infected chickens.

Dramatic differences in the response of macrophages from B2 and B19 MHC-defined haplotypes to interferon gamma and polyinosinic:polycytidylic acid stimulation.

The chicken MHC has been associated with disease resistance, though the mechanisms are not understood. The functions of macrophages, critical to both innate and acquired immunity, were compared between the more infectious bronchitis virus-resistant B2 and the more infectious bronchitis virus-susceptible B19 lines. In vivo peripheral blood concentrations of monocytes were similar in B2 or B19 homozygous haplotypes. Peripheral blood-derived macrophages were stimulated with poly I:C, simulating an RNA virus, or IFNγ, a cytokine at the interface of innate and adaptive immunity. Not only did B2-derived peripheral monocytes differentiate into macrophages more readily than the B19 monocytes, but as determined by NO production, macrophages from B2 and B2 on B19 genetic background chicks were also significantly more responsive to either stimulant. In conclusion, the correlation with resistance to illness following viral infection may be directly linked to a more vigorous innate immune response.

In vitro effects of lactoferrin on lipopolysaccharide-induced proliferation, gene expression, and prostanoid production by bovine peripheral blood mononuclear cells.

OBJECTIVE: To evaluate the effect of lactoferrin on lipopolysaccharide (LPS)-induced proliferation of bovine peripheral blood mononuclear cells (PBMCs), gene expression of inflammatory mediators, and production of prostanoids in vitro. SAMPLE POPULATION: PBMCs isolated from 15 Holstein bull calves. PROCEDURES: Mixed populations of PBMCs were isolated by differential centrifugation. Proliferation assays were conducted in 96-well plates designed to allow addition of lactoferrin (200 ng/mL) with and without LPS (1 microg/mL) in a checkerboard design. Incorporation of 3H-thymidine was used to determine proliferation of PBMCs. Prostaglandin E2 production was determined in culture-conditioned medium by use of enzyme immunoassay. Effects of lactoferrin on LPS-induced gene expression of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 were monitored by use of PCR assays. RESULTS: Lactoferrin supplementation significantly reduced LPS-induced incorporation of 3H-thymidine and production of prostaglandin E2 by PBMCs. Lactoferrin reduced LPS-induced expression of COX-2 and MMP-9 mRNA. CONCLUSIONS AND CLINICAL RELEVANCE: Lactoferrin reduced LPS-induced cellular proliferation, inflammatory mediator gene expression, and prostaglandin E2 production by bovine PBMCs in vitro. These effects may be beneficial in reducing the impact of endotoxemia in neonates.

Evaluation of a cow-side immunoassay kit for assessing IgG concentration in colostrum.

OBJECTIVE: To determine sensitivity and specificity of a cow-side immunoassay kit for assessing IgG concentration in colostrum. DESIGN: Prospective study. ANIMALS: 76 dairy and 11 beef cows of various parities. PROCEDURE: Colostrum from first, second, and third milkings and milk samples were collected, and IgG concentration was determined by means of radial immunodiffusion. The immunoassay was performed according to the manufacturer's instructions, and sensitivity and specificity were calculated by comparing results of the immunoassay (positive vs negative) with results of immunodiffusion (< 50 g/L vs > or = 50 g/L). RESULTS: 135 colostrum or milk samples were collected. Mean +/- SD colostral IgG concentrations, determined by means of radial immunodiffusion for dairy and beef cows were 65.4 +/- 51.4 g/L and 114.8 +/- 42.7 g/L, respectively. Mean IgG concentrations for first-, second-, and third-milking colostrum samples and for milk samples were 92 +/- 49.0 g/L, 74.6 +/- 45.1 g/L, 47.5 +/- 32 g/L, and 6.8 +/- 3.8 g/L, respectively. Sensitivity of the immunoassay (ie, percentage of samples with IgG concentration < 50 g/L with a positive immunoassay result) was 93%, and specificity (ie, percentage of samples with IgG concentration > or = 50 g/L with a negative immunoassay result) was 76%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the immunoassay kit was an acceptable cow-side test to identify colostrum samples with IgG concentrations < 50 g/L. The immunoassay kit should be useful in screening colostrum for adequate IgG concentration before feeding to calves or storage.

Effect of delayed colostrum collection on colostral IgG concentration in dairy cows.

OBJECTIVE: To determine the effect of timing of first-milking colostrum collection on colostral IgG concentration. DESIGN: Prospective study. ANIMALS: 13 healthy Holstein cows. PROCEDURES: All calvings were observed. After parturition, calves were not allowed to suckle and were separated from the dam. Colostrum was collected from a single randomly selected quarter at 2, 6, 10, and 14 hours after parturition until all 4 quarters were sampled. Colostral IgG concentration was determined via radial immunodiffusion. RESULTS: Mean colostral IgG concentration was 113, 94, 82, and 76 g/L at 2, 6, 10, and 14 hours after calving, respectively. Colostrum collected 6, 10, and 14 hours after calving had significantly lower IgG concentrations than did colostrum collected 2 hours after calving. Mean colostral IgG concentration at 14 hours after calving was significantly lower than that at 6 hours after calving. Cows in their third or greater lactation had mean colostral IgG concentrations 2 hours after calving (132 g/L) that were greater than the first and second lactation cows (mean, 95 and 100 g/L, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that early or immediate colostrum collection from dairy cows will maximize colostral IgG concentration. Adjustment of routine dairy farm management procedures may be required to maximize colostrum quality and minimize prevalence of failure of passive transfer in dairy calves.

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.

Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves.

OBJECTIVE: To evaluate diagnostic utility of a commercially available immunoassay for assessing adequacy of passive transfer of immunity in neonatal calves. DESIGN: Prospective study. ANIMALS: 123 calves. PROCEDURE: Blood and serum samples were obtained from the calves prior to 2 weeks of age. The immunoassay was performed, along with refractometry and an 18% sodium sulfite turbidity test. Serum IgG concentration was determined with a radial immunodiffusion assay. Sensitivity and specificity of the immunoassay, refractometry, and the sodium sulfite test were calculated by comparing results with results of the radial immunodiffusion assay. RESULTS: Sensitivity and specificity of the blood IgG immunoassay were 0.93 and 0.88, respectively, compared with 1.00 and 0.53 for the sodium sulfite test. For refractometry, sensitivity and specificity were 0.71 and 0.83, respectively, when a serum total solids concentration of 5.2 g/dl was used as the cutoff between positive and negative test results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the immunoassay performs well in detecting calves with inadequate passive transfer of immunity.