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  • Author or Editor: Bradley W. Fenwick x
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Abstract

Objective

To determine whether bovine tear film contains the iron-binding glycoprotein, lactoferrin.

Animals

40 Adult Hereford, Angus, and Simmental cattle.

Procedure

Protein analysis: pooled bovine tears were used for protein analysis (size exclusion high-performance liquid chromatography [HPLC] fractionation). HPLC was used for tear analysis. A diode array detector was used (215 and 280 μm) for chromatogram analysis and comparisons.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE): protein electrophoresis was performed, using 7.5% running gels with 4% stacking gels. Molecular weight of proteins in the unknown samples was determined as recommended by the manufacturer of the standards.

Protein sequencing: amino acid sequencing, using automated Edman degradation of HPLC purified protein, was performed. The sequence obtained was compared with the known protein sequence of bovine lactoferrin.

Results

HPLC analysis of whole bovine tears resulted in a consistent chromatogram. Peak collection was performed to recover a protein from the bovine tear film with chromatogram characteristics nearly identical to purified bovine lactoferrin. Silver-stained SDS-PAGE of this peak revealed a band with molecular mass consistent with bovine lactoferrin (estimated mass of 78 kd). The first 13 amino acid residues of this protein were identical to the amino acid sequence of bovine lactoferrin.

Conclusion

Analysis of whole bovine tears, using size exclusion HPLC, SDS-PAGE, and amino acid sequencing, provided evidence that bovine tears contain lactoferrin.

Clinical Relevance

Lactoferrin probably exerts a bacteriostatic effect in bovine tear film. Locally produced lactoferrin may bathe the ocular surface and sequester iron from potential pathogens. (Am J Vet Res 1996;57:1369-1372)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To determine the relationship between ambient temperature and mean body surface temperature (MBST) measured by use of infrared thermography (IRT) and to evaluate the ability of IRT to detect febrile responses in pigs following inoculation with Actinobacillus pleuropneumoniae.

Animals—28 crossbred barrows.

Procedures—Pigs (n = 4) were subjected to ambient temperatures ranging from 10 to 32 C in an environmental chamber. Infrared thermographs were obtained, and regression analysis was used to determine the relationship between ambient temperature and MBST. The remaining pigs were assigned to groups in an unbalanced randomized complete block design (6 A pleuropneumoniae-inoculated febrile pigs [increase in rectal temperature ≥ 1.67 C], 6 A pleuropneumoniae-inoculated nonfebrile pigs [increase in rectal temperature < 1.67 C], and 12 noninoculated pigs). Infrared thermographs and rectal temperatures were obtained for the period from 2 hours before to 18 hours after inoculation, and results were analyzed by use of repeated-measures ANOVA.

Results—A significant linear relationship was observed between ambient temperature and MBST (slope, 0.40 C). For inoculated febrile pigs, a treatment X method interaction was evident for rectal temperature and MBST, whereas inoculated nonfebrile pigs only had increased rectal temperatures, compared with noninoculated pigs. A method X time interaction resulted from the longer interval after inoculation until detection of an increase in MBST by use of IRT.

Conclusions and Clinical Relevance—Infrared thermography can be adjusted to account for ambient temperature and used to detect changes in MBST and radiant heat production attributable to a febrile response in pigs. (Am J Vet Res 2001;62:676–681)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate lactoferrin and lysozyme content in various ocular glands of bison and cattle and in tears of bison.

Sample Population—Tissues of ocular glands obtained from 15 bison and 15 cattle and tears collected from 38 bison.

Procedure—Immunohistochemical analysis was used to detect lysozyme and lactoferrin in formalin-fixed, paraffin-embedded sections of the ocular glands. Protein gel electrophoresis was used to analyze ocular glands and pooled bison tears by use of a tris-glycine gel and SDS-PAGE. Western blotting was used to detect lactoferrin and lysozyme.

Results—Immunohistochemical staining for lactoferrin was evident in the lacrimal gland and gland of the third eyelid in cattle and bison and the deep gland of the third eyelid (Harder's gland) in cattle. Equivocal staining for lactoferrin was seen for the Harder's gland in bison. An 80-kd band (lactoferrin) was detected via electrophoresis and western blots in the lacrimal gland and gland of the third eyelid in cattle and bison, Harder's glands of cattle, and bison tears. An inconsistent band was seen in Harder's glands of bison. Lysozyme was not detected in the lacrimal gland of cattle or bison with the use of immunohistochemical analysis or western blots. Western blots of bison tears did not reveal lysozyme.

Conclusion and Clinical Relevance—Distribution of lactoferrin and a lack of lysozyme are similar in the lacrimal gland of cattle and bison. Differences in other tear components may be responsible for variability in the susceptibility to infectious corneal diseases that exists between bison and cattle. (Am J Vet Res 2003;64:104–108)

Full access
in American Journal of Veterinary Research