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Biomedical subjects

G T Pharr

Publications and source records attributed to G T Pharr.

At least 19 recordsLinked to original sources

The role of feline aminopeptidase N as a receptor for infectious bronchitis virus. Brief review.

Feline aminopeptidase N (fAPN) has been shown to serve as a receptor for feline, canine, porcine and human coronaviruses. Our objective was to determine if fAPN can serve as a receptor for infectious bronchitis virus (IBV). Feline kidney cells that express fAPN and hamster kidney fibroblasts that do not express fAPN were inoculated with IBV and monitored for replication by indirect fluorescent assay and confocal microscopy and in chicken embryonated eggs. The results showed that the feline cells were permissive to IBV but the hamster cells were not. The hamster cells became permissive to IBV after transfection with a fAPN cDNA suggesting that the feline APN molecule plays a role in IBV entry.

Animals↗

Molecular characterization of Cryptosporidium sp. isolated from northern Alaskan caribou (Rangifer tarandus).

Cryptosporidium sp. was found in 3 out of 49 caribou (Rangifer tarandus) from northern Alaska. Segments of both the 18S ribosomal RNA and the heat shock protein genes were amplified from the caribou isolate and compared with that obtained from an isolate from a wild white-tailed deer (Odocoileus virginianus) in Virginia as well as other species and isolates available from GenBank. Analyses showed the white-tailed deer isolate to be identical with the C. parvum cattle genotype; however, the caribou isolate represents a new genotype closely related to C. serpentis, C. muris, and C. andersoni. Giardia sp. was not detected in any of the caribou samples nor was Cryptosporidium sp. or Giardia sp. detected in any of the 42 moose (Alces alces) samples examined.

Alaska↗

Natural selection of the Pol gene of bovine immunodeficiency virus.

Genetic variability is a salient feature of lentiviruses, contributing to the pathogenesis of these viruses by enabling them to persist in the host and to resist anti-retroviral treatment. Bovine immunodeficiency virus (BIV), a lentivirus of unknown pathology, infects cattle in the United States and worldwide. Genetic diversity of BIV that is associated with naturally infected cattle is not well studied. We examined the genetic diversity and natural selection of a segment of the BIV pol gene amplified from the leukocyte DNA of naturally infected cattle. A portion of the reverse transcriptase domain (183 bp) of the pol region was targeted for amplification by PCR. PCR products were sequenced directly and aligned. When compared to the sequences of BIV R29-127, a molecular clone of the original BIV R29 isolate, all isolates were greater than 91% identical in nucleotide sequences and 77% identical in amino acid sequences. Pol genotypes were polymorphic at 14% of the nucleotide sites. The ratio of nonsynonymous to synonymous nucleotide substitutions (relative to the number of respective sites, Ka/Ks) was 0.16, indicating that this region of the BIV genome, like that of HIV-1, is subject to purifying selection. Based on the McDonald-Kreitman analysis, this region also was under positive Darwinian selection as HIV-1 and BIV diverged from a common progenitor. Phylogenetic analysis revealed that genotypes were geographically distinct, possibly indicating a common source of infection for animals within a herd.

Amino Acid Sequence↗

Class II MHC cDNAs in 15I5 B-congenic chickens.

cDNA was obtained from the bursae of Fabricius of chickens from six B-congenic lines developed at this laboratory and studied for expression of class II B-LB genes. Following cDNA amplification, cloning and sequencing, genes were assigned to B-LB loci based on characteristic DNA sequences, amino acid relatedness to characterized genes, and level of expression. Genes from the B-LBI, B-LBII, and B-LBVI loci were differentially expressed in chickens with the B2, B5, B13, B15, or B21 haplotypes. Chickens of all haplotypes expressed a B-LBII gene. Additional B-LB genes expressed included: B-LBI genes in the B5 and B19 haplotypes; a B-LBI/VI recombinant gene in the B2 haplotype; and a B-LBVI gene in the B13 haplotype. The B-congenic lines have demonstrable differences in resistance to Marek's disease (MD), and in responses to MD viral vaccines. This variability in disease resistance may be correlated with polymorphisms in the expressed B-LB genes, or with differential expression of genes at different loci.

Animals↗

Non-association between Rfp-Y major histocompatibility complex-like genes and susceptibility to Marek's disease virus-induced tumours in 6(3) x 7(2) F2 intercross chickens.

Marek's disease (MD) is a lymphoproliferative disease caused by a member of the herpesvirus family, and the best understood genetic resistance to MD involves the chicken major histocompatibility complex (MHC) B-complex. Preliminary observations have suggested that MHC-like Rfp-Y genes might also influence the incidence of MD. This study describes the differentiation and definition of unique Rfp-Y genes in inbred lines 6(3) and 7(2), lines that possess identical B-complex genes, but that are resistant or susceptible to MD, respectively. To assess if Rfp-Y genes affect susceptibility to MD, 265 6(3) x 7(2) F2 chickens were challenged with the JM strain of MD virus at 1 week of age and were evaluated for MD lesions at up to 10 weeks of age. Genotyping of the F2 chickens for Rfp-Y haplotypes was performed by restriction fragment length polymorphism analysis of genomic DNA using TaqI and a B-FIV probe. Analysis of variance and interval mapping procedures were used to determine association between the Rfp-Y haplotypes and the phenotypic MD values of the F2 chickens. The cosegregation analysis of 265 F2 chickens indicated that there was no association between Rfp-Y haplotypes and MD susceptibility. Furthermore, the fact that the Rfp-Y haplotypes fit the 1:2:1 segregation ratio and the Rfp-Y allele frequencies did not differ significantly from 0.5 in the full population or in selected subpopulations (of either 40 MD-resistant or 39 MD-susceptible chickens) also indicated that Rfp-Y haplotypes do not significantly influence MD susceptibility. We conclude that Rfp-Y haplotypes do not play a major role in determining the genetic susceptibility to MD in 6(3) x 7(2) F2 White Leghorn chickens.

Animals↗

Avian B cell development.

Development of B cells in chickens proceeds via a series of discrete developmental stages that includes the maturation of committed B cell progenitors in the specialized microenvironment of the bursa of Fabricius. The bursa has been shown to be required for the amplification of the B cell pool and selects for cells with productive immunoglobulin rearrangement events. Other events regulating chicken B cell development such as lymphocyte trafficking and apoptosis are just beginning to be elucidated. Within the bursa, the variable regions of immunoglobulin genes of B cell progenitors are diversified by a process of intrachromosomal gene conversion, where blocks of sequence information are transferred from pseudo-V regions to the recombined variable regions of the immunoglobulin genes. Recently gene conversion has been determined to play a role in the diversification of the immune repertoire in other species. In this review we focus on the current understanding and recent advances of B cell development in the chicken.

Animals↗

Histocompatibility antigen(s) linked to Rfp-Y (Mhc-like) genes in the chicken.

Major histocompatibility complex (Mhc) genes influencing transplantation rejections were first described in mice within the H2 complex and secondly in chickens within the B complex. In chickens, Rfp-Y haplotypes have recently been identified which contain class I and class II Mhc-like genes that assort independently of the B complex. Three Rfp-Y haplotypes have been defined in a closed breeding flock of line N chickens. In this study, progeny were obtained from line N Rfp-Y heterozygous matings to establish the role of Rfp-Y in transplantation immunity. Rfp-Y incompatibility did not induce significant one-way mixed lymphocyte responses. However, Rfp-Y-incompatible skin grafts were rejected more frequently and at a faster rate than Rfp-Y-compatible grafts by two-week-old chicks. The control Mhc B-incompatible grafts were rejected faster than the Rfp-Y-incompatible grafts; the latter were rejected at speeds that resemble rejection of minor histocompatibility antigens. We conclude that Rfp-Y class I and II Mhc-like genes are linked to the expression of minor histocompatibility antigens in chickens.

Animals↗

Chicken interferon-mediated induction of major histocompatibility complex class II antigens on peripheral blood monocytes.

Conditioned medium containing immune interferon (IFN) activity was prepared by stimulating spleen lymphocytes obtained from inbred SC chickens with 10 micrograms concanavalin A (Con A) for 48 h. Pretreatment of spleen cells with monoclonal antibody against CD4, but not CD8, abrogated IFN production suggesting that CD4+ lymphocytes are responsible for immune IFN production. Immune IFN was purified 25-fold from Con A conditioned medium using controlled-pore glass column chromatography resulting in an increase in specific antiviral activity from 7 to 3290 units mg-1. Partially purified immune IFN retained antiviral and macrophage-activating factor (MAF)-like activities. Normal peripheral blood macrophages, when cultured in the presence of partially purified immune IFN, showed a dose-dependent increase in cell surface major histocompatibility complex Class II antigen expression by flow cytometry. Northern blot analysis of mRNA obtained from IFN-treated macrophages showed a concomitant increase in Class II gene expression. This effect was more obvious in cells induced for 48 h than in those induced for 24 h. These results strongly suggest that existence of an avian homologue of the MAF-like activity.

Animals↗

A class I cDNA from SPAFAS line-11 chickens.

A chicken MHC class I (B-F) cDNA from SPAFAS line 11 embryonic liver tissue was isolated and characterized by nucleotide sequencing. Comparing this sequence with previously described B-F cDNAs highlights clustered nucleotide substitutions in exon 3, encoding amino acids located on the alpha-helical region of the alpha 2 domain.

Amino Acid Sequence↗

Identification of class II major histocompatibility complex polymorphisms predicted to be important in peptide antigen presentation.

Chickens of the B2, B5, B15, B19, and B21 B-congenic haplotypes differ in disease resistance. Complementary DNA from B-congenic chicken strains have been analyzed for allelic diversity of expressed Class II MHC genes. The predicted amino acid sequences of eight genes from five haplotypes were subjected to Wu-Kabat variability analysis. The B-L gene polymorphic regions and conserved regions are highly similar to the human leukocyte antigen Class II genes. Therefore, the present analysis reveals candidate polymorphisms important in determining the spectrum of antigenic peptides presented to T helper cells, and allelic differences possibly important in resistance to avian disease.

Animals↗

Avian thyroid isthmus.

An avian equivalent of the mammalian thyroid isthmus appeared in thiouracil-treated (.1% in feed nd hatching to 6 and 8 weeks of age) chickens. In two experiments in which chickens were fed a diet containing thiouracil, thyroid tissue was found connecting the thyroid glands. Histological examination of the connecting tissue revealed that it was of the same cellular composition as the thyroid glands.

Animals↗

Avian thyroid isthmus.

An avian equivalent of the mammalian thyroid isthmus appeared in thiouracil-treated (.1% in feed from hatching to 6 and 8 weeks of age) chickens. In two experiments in which chickens were fed a diet containing thiouracil, thyroid tissue was found connecting the thyroid glands. Histological examination of the connecting tissue revealed that it was of the same cellular composition as the thyroid glands.

Animals↗

Identification of Rfp-Y (Mhc-like) haplotypes in chickens of Cornell lines N and P.

Two strains of chickens selected for differential Marek's disease (MD) resistance or susceptibility were studied for the presence of the recently described Rfp-Y major histocompatibility complex (Mhc-like) haplotypes. MD resistant chickens from line N were fixed for the classical Mhc B21 haplotype, whereas MD susceptible line P chickens were fixed for the B19 haplotype. The Rfp-Y haplotypes were identified by restriction fragment polymorphism (RFP) analysis using enzymes and Mhc probes for B-G, B-L beta II, and B-FIV. In addition an Rfp-L beta III clone was developed that differentiated Rfp-L beta from B-L beta genes. Three Rfp-Y haplotypes, defined for both class I and class II Mhc-like loci, were identified in line N (Rfp-Y5, Rfp-Y7, and Rfp-Y8) and in line P (Rfp-Y5, Rfp-Y8, and Rfp-Y9), respectively. The Rfp-Y7, Rfp-Y8, and Rfp-Y9 haplotypes have not been described previously. The Rfp-Y5 haplotype was most frequent (0.70) in line N, but existed in low frequency (approximately 0.04) in line P; the Rfp-Y9 haplotype was most frequent in line P (0.63), but was absent in line N. The Rfp-Y haplotypes-segregated in a Mendelian fashion in each line based on analysis of progeny from Rfp-Y heterozygous matings. The frequency of recombination between the Rfp-Y F and L loci was estimated to be less than 0.25%.

Animals↗

The effects of 6/85 live Mycoplasma gallisepticum vaccine in commercial layer hens over a 43-week laying cycle on egg production, selected egg quality parameters, and egg size distribution when challenged before beginning of lay.

In each of two trials, 80 commercial leghorn-type pullets were separated into two treatments with four replicates of 10 chickens in each treatment. Forty pullets were designated as controls and received no inoculation, whereas the remaining 40 pullets received the 6/85 vaccine strain of Mycoplasma gallisepticum (MG) at 10 wk of age. Hen-day egg production, egg weight, eggshell strength, Haugh unit score, pimpling incidence, and blood/meat spot incidence were monitored and recorded weekly in each trial through an entire laying cycle of 43 wk. Further, eggs from all treatments were collected daily, Monday-Thursday, and individually weighed. No significant difference was observed between the treatments for 43-wk means for hen-day egg production, for any of the monitored egg or eggshell quality parameters, or for the number of extra large, large, medium, small, pee wee, or undergrade egg sizes. A significant (P < or = 0.05) difference was observed for the number of jumbo-sized eggs between the two treatments. Results of this study suggest that vaccination of commercial layer chickens at 10 wk of age with 6/85 strain MG does not detrimentally impact egg production, egg size distribution, or ovary/oviduct function as evidenced by selected egg parameters monitored in this study.

Animals↗

Effects of an S6 strain of Mycoplasma gallisepticum challenge at onset of lay on digestive and reproductive tract characteristics in commercial layers.

Mycoplasma gallisepticum (MG), a reproductive/respiratory pathogen in poultry, has been implicated in suboptimum egg production and decreased hatchability. Commercial layer hens raised in a controlled environment were inoculated with the S6 strain of MG at 20 wk of age. The S6 inoculation had no effect on bird weight, egg production, digestive tract weight and length, or histopathologic lesion scores, although significant differences were noted in the lengths and weights of various portions of the reproductive tract. This study shows that S6MG inoculation does not detrimentally affect layer hen performance when in the absence of environmental stressors customary to a caged layer facility.

Aging↗

Mycoplasma gallinarum infection in commercial layers and onset of fatty liver hemorrhagic syndrome.

Fatty liver hemorrhagic syndrome (FLHS) was observed in each of three trials in which commercial layers were utilized to determine the effect of Mycoplasma gallinarum (MGn) on egg and eggshell quality parameters and egg production. In each of three trials, FLHS occurred 31-54 days later in MGn-inoculated hens as compared with the Mycoplasma-clean (control) hens. In trials 1 and 2, no therapeutic intervention was initiated to ameliorate FLHS. In trial 3, therapeutic intervention was instituted and consisted of the addition of 1 pound of choline chloride/ton of feed. Total mortality recorded throughout the duration of each trial and attributable to FLHS was not significantly different between the control and the MGn-inoculated treatment. However, FLHS-associated mortality in each of the three trials was numerically greater for the control treatment.

Animals↗