Biomedical subjects

J J Giambrone

Publications and source records attributed to J J Giambrone.

At least 19 recordsLinked to original sources

PubMed2005

Yeast-derived sigma C protein-induced immunity against avian reovirus.

Avian reoviruses (ARVs) can result in disease and economic losses in the poultry industry. Vaccines against ARV may not provide full protection and can cause adverse reactions. The coding sequence of the sigma C protein from strain S1133 of avian reovirus was expressed in Schizasaccharomyces pombe. Sigma C protein expression was demonstrated by Western blotting, and the protein was evaluated for its ability to protect specific-pathogen-free (SPF) chickens against challenge with the virulent S1133 strain. Serologic and challenge-infection data showed the efficacy of the recombinant vaccine administered orally each week for 3 consecutive wk. Sigma C protein induced antibody, as determined by enzyme-linked immunosorbent assay. Percentage (%) protection induced by the low dose (125 microg purified yeast-expressed sigma C protein/chicken) or the high dose (250 microg purified yeast-expressed sigma C protein/chicken) was 64 and 91, respectively. The commercial vaccine administered once or twice provided 82% protection. Results supported the feasibility of a plant-derived vaccine for use in poultry immunization schemes.

Animals↗

PubMed2004

Expression of immunogenic VP2 protein of infectious bursal disease virus in Arabidopsis thaliana.

VP2 protein is the major host-protective immunogen of infectious bursal disease virus (IBDV) of chickens. Transgenic lines of Arabidopsis thaliana expressing recombinant VP2 were developed. The VP2 gene of an IBDV antigenic variant E strain was isolated, amplified by RT-PCR and introduced into a plant expression vector, pE1857, having a strong promoter for plant expression. A resulting construct with a Bar gene cassette for bialaphos selection in plant (rpE-VP2) was introduced into Agrobacterium tumefaciens by electroporation. Agrobacterium containing the rpE-VP2 construct was used to transform Ar. thaliana and transgenic plants were selected using bialaphos. The presence of VP2 transgene in plants was confirmed by PCR and Southern blot analysis and its expression was confirmed by RT-PCR. Western blot analysis and antigen-capture ELISA assay using monoclonal anti-VP2 were used to determine the expression of VP2 protein in transgenic plants. The level of VP2 protein in the leaf extracts of selected transgenic plants varied from 0.5% to 4.8% of the total soluble protein. Recombinant VP2 protein produced in plants induced antibody response against IBDV in orally-fed chickens.

Animals↗

PubMed2000

The use of monoclonal antibody probes for the detection of avian reovirus antigens.

Two monoclonal antibodies (MAb), E9 and H3, prepared against avian reovirus (ARV) S1133, were used in an immuno-dot assay to detect ARV antigens from cell culture and from tendon tissue samples of chickens. The limit of viral antigens detected was 8 ng using both MAb probes. The probes detected 10 ARV isolates representing at least two serotypes or pathotypes. The results indicated that these probes had broad specificity. The probes, however, did not cross-react with viral antigens prepared from six unrelated avian viruses. The ARV antigens in tendon tissue samples were detected by both probes, and it is possible, therefore, to use either of the two MAb probes for detection of ARV infections.

Animals↗

PubMed1997

Amplification, cloning and sequencing of the sigmaC-encoded gene of avian reovirus.

The sigmaC-encoding cDNA of avian reovirus (ARV) 1733 strain was amplified, cloned and sequenced using double nested polymerase chain reaction (PCR). The ARV sigmaC protein is a minor component of the outer capsid that induces type-specific neutralization antibodies. Four overlapping sigmaC-encoding cDNA fragments were obtained. Together, the four fragments represented the whole coding sequence. The nucleotide and deduced amino acid sequences of sigmaC-encoded gene of U.S. (S1133 and 1733) and Australian isolates (RAM-1 and SOM-4) were compared. The U.S. isolates were closely related, but different from Australian isolates. The degree of differences between the U.S. and Australian isolates was over 44.89% at both the nucleotide and deduced amino acid levels and suggested that the virus is evolving separately in different continents. The deduced amino acid sequences of ARV sigmaC indicated a heptapeptide repeat in the N-terminal region of ARV sigmaC existed in all ARVs. The results suggested that ARV sigmaC is structurally related to mammalian reovirus (MRV) sigma1.

Amino Acid Sequence↗

PubMed1997

Molecular characterization of avian reoviruses using nested PCR and nucleotide sequence analysis.

A nested polymerase chain reaction (PCR) with subsequent nucleotide sequence analysis identified and differentiated avian reoviruses (ARVs). PCR products amplified from the S1 gene segment of ARV of USA isolates were 738 and 342 bp, respectively. PCR products were conformed by Southern and dot blot hybridizations. The amplified cDNA fragments were cloned into the pUC18 vector and subjected to DNA sequencing. The nucleotide and deduced amino acid sequences of four USA (S1133, 1733, 2408, and CO8) and two Australian isolates (RAM-1 and SOM-4) were compared. Results of paired difference analysis and a predicted dendrogram revealed that USA isolates were closely related, but different from, Australian isolates. The deduced amino acid sequences of the N-terminal region of ARV sigma C showed a heptapeptide repeat of hydrophobic residues in all ARV isolates.

Amino Acid Sequence↗

PubMed1996

Promoter and transcription of type X collagen gene in broiler chickens with tibial dyschondroplasia.

Type X collagen is produced exclusively in hypertrophic chondrocytes of the growth plate of the proximal tibiotarsus and is believed to play an important role during normal development from chondrogenesis to osteogenesis. Chondrocytes of chickens with tibial dyschondroplasia (TD) fail to attain full hypertrophy and the amount of type X collagen, being a marker of hypertrophy, is likely to be reduced. It is not clear whether transcriptional regulation is functional for expression of the type X collagen gene in TD birds. Nucleotide sequence of the type X collagen gene promoter was determined by sequencing PCR-based DNA clones. Nucleotide identity of this fragment between the normal and TD carriers was 97.6%. Both normal and TD birds were similar in a putative transcription start site, the site of TATAA box, and neither had a CCAAT box. However, there were two gaps in TD carriers, four gaps in normals, and five nucleotide substitution sites. By rapid amplification of cDNA ends by PCR (RACE-PCR), transcription of the gene was assessed using total RNA and mRNA from both normal chondrocytes and TD lesions at 3 and 4 wk of age. The RACE-PCR product for type X collagen mRNA was detectable in both normal and TD birds at two stages. No difference was found between them. This result does not support the hypothesis that transcriptional regulation of type X collagen gene is important in TD development of chickens. Variations in the promoter region did not affect transcription of type X collagen gene in TD carrier chickens.

Animals↗

PubMed1995

Random amplified polymorphic DNA comparisons among broiler lines selected for incidence of tibial dyschondroplasia.

Lines selected for high (H) and low (L) incidence of tibial dyschondroplasia (TD) for eight generations and a randombred control (C) line of broiler chickens were fingerprinted by random amplification of genomic DNA mixed from 20 individuals of each line with 20 oligonucleotide primers. Among these 20 primers, 15 could distinguish the H from the L line, 14 the H from the C line, and 13 the L from the C line. Band sharing (BS), on the average over 20 primers, was .7 for the H vs L comparison and .8 for both H vs C and L vs C comparisons. The levels of BS calculated from individuals was .6 between the H and L line, .7 between the H and C line, and .7 between the L and C line. The ranking of BS values obtained from individual DNA samples was consistent with that obtained from the mixed DNA samples. Genomic distance between divergently selected lines (H vs L) was larger than that between the divergently selected lines and randombred line (H vs C and L vs C). Individual variation within lines was detected in spite of eight generations of selection. Results showed that eight generations of divergent selection for TD incidence in broiler chickens had resulted in genetic variation among lines. The procedure of random amplified polymorphic DNA assay using mixed DNA samples could be used to evaluate genetic distance among lines of chickens.

Animals↗

PubMed1994

Detection of genetic variations in serotype I isolates of infectious bursal disease virus using polymerase chain reaction and restriction endonuclease analysis.

Reverse transcription with polymerase chain reaction (PCR) followed by restriction endonuclease analysis detected genetic variations among serotype I isolates of infectious bursal disease virus (IBDV). Using a set of synthetic primers derived from the large genome segment of APHIS-IBDV, the hypervariable region (AccI-SpeI fragment) located in the VP2 gene was amplified. With all strains, a cDNA fragment of approximately 643 bp was amplified, indicating that there were no apparent deletions or insertions in this region among isolates. Fragments amplified from 9 isolates were digested with 14 restriction enzymes. Restriction fragment profiles generated by restriction enzymes NaeI, StuI, TaqI, and SacI, showed genetic variations among isolates. This study provided a simple and sensitive method for detection of genetic variations among isolates that are closely related serologically and could not be differentiated using current serologic methods.

Animals↗

PubMed1989

Efficacy of simultaneous administration of Marek's disease and viral tenosynovitis vaccines to day-old broiler chickens.

Experiments were conducted to determine the efficacy of simultaneous administration at 1 day of age of the herpes virus of turkeys vaccine (HVT) and viral tenosynovitis (VT) vaccine. Day-old broilers from commercial breeders, which were immunized against both MD and VT, were used. The vaccines were injected at full dosage or diluted 1 to 4. Challenge with either Marek's disease virus (MDV; intraperitoneal) or VT virus (footpad) was at 7 days of age. At 7 wk of age, birds were weighed, killed, and examined for gross lesions. Either vaccine given at full dosage alone, or in combination, rendered birds resistant to homologous viral challenge. However, when either vaccine was diluted and administered alone, the efficacy of the VT but not the HVT vaccine was reduced, resulting in increased lesions and mortality and decreased weight gain after VT challenge. When the HVT vaccine was diluted and combined with undiluted VT vaccine, interference occurred, resulting in reduced efficacy of the HVT vaccine (increased tumors and mortality and reduced body weight). In contrast, combining the HVT at full dosage and the diluted VT vaccine given alone. The data indicate that caution should be maintained when mixing HVT and VT vaccines. For maximum efficacy, both vaccines should be mixed undiluted when administered simultaneously.

Analysis of Variance↗

PubMed1988

Effect of broiler chicken age on susceptibility to experimentally induced Cryptosporidium baileyi infection.

Clinical signs of respiratory tract disease were observed in chickens that were inoculated intratracheally with 1 x 10(6) oocysts of Cryptosporidium baileyi at 2 or 14 days of age (10 chickens/group), but not in chickens inoculated at 28 or 42 days of age (10 chickens/group). Orally inoculated chickens in all age groups (10 chickens/group) did not develop clinical signs of disease. Orally and intratracheally inoculated chickens in all age groups were infected, as determined by the finding of cryptosporidia in tissue sections of the trachea, bursa of Fabricius, and cloaca, and by the recovery of oocysts from their feces. Chickens inoculated at 2 and 14 days of age excreted oocysts for a longer period and had greater numbers of cryptosporidia in their tissues, compared with chickens inoculated at 28 and 42 days of age.

Aging↗

PubMed1987

Experimental cryptosporidiosis in broiler chickens.

Seven-day-old conventional broilers were inoculated either orally or intratracheally (IT) with 2.5 X 10(5), 5.0 X 10(5), or 2.0 X 10(6) oocysts of Cryptosporidium baileyi (32 birds for each dosage level per group; 192 birds total). Thirty-two birds served as unninoculated controls. Mean weekly weight gain and feed conversion were determined during a 5-week period. Carcass pigment was graded using a Roche Color Fan. Fecal oocysts were calculated from random cage samples 6, 8, 11, 13, 15, 18, 20, 22, and 25 days after inoculation (DAI). Effects of C. baileyi on immune responses were examined for Newcastle disease virus-hemagglutination inhibition (NDV-HI) antibody, infectious bursal disease virus-enzyme-linked immunosorbent assay (IBDV-ELISA) antibody titers and delayed hypersensitivity (DH) in half of the birds in each group. Disease or death from cryptosporidiosis did not result from oral inoculation of C. baileyi. Signs of respiratory disease, consisting of rales, sneezing, and dyspnea were observed in all IT-inoculated birds 7 to 21 DAI. Seven deaths occurred in the IT-inoculated groups 14 to 21 DAI. At necropsy, lung parenchyma was gray, firm, and wet in the ventral region. Air sacs contained a foamy, white to gray, mucoid fluid. Histologic lesions in the air sacs and bronchi were epithelial hyperplasia, discharge of mucocellular exudate to the mucosal surface, thickening of the mucosa by cellular infiltrates, loss of cilia, and dilation of mucous glands. Weight gains for IT-inoculated birds were lower (P less than .05) than controls from 14 to 21 DAI, although weight gains for the 5-week period were not significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

Evaluation of broiler breeder pullet vaccination programs for preventing clinical reovirus infections in the progeny.

The efficacy of various reovirus vaccination programs in commercial broiler breeder pullets for preventing clinical reovirus infections in broiler progeny was studied in three experiments. In each, 70 1-day-old broilers from each of four different commercial poultry companies that used various combinations of live and killed reovirus vaccines in breeder pullets were used. Broiler progeny from each company were bled and serum analyzed for neutralization antibody to the S1133 reovirus. Twenty chicks from each company were challenged by foot pad with a virulent reovirus (S1133) and 20 chicks challenged with another reovirus (81-B) isolate to determine their susceptibility. An additional 20 chicks from each of the four groups were maintained as unchallenged controls. Chicks were examined for morbidity, mortality, and weight gain for a 3-week period. Three weeks after challenge, all chickens were necropsied for gross lesions. There was a direct correlation between the number of doses of live inactivated reovirus vaccines that pullets received to antibody titer and resistance to challenge in the day-old progeny. Day-old progeny derived from breeder pullets that received one dose of live and two doses of inactivated reovirus vaccines had the highest numerical maternal antibody titer and best resistance to clinical infection following challenge.

Animals↗

PubMed1986

Efficacy of coarse spray administration of commercial intermediate infectious bursal disease vaccines.

The efficacy of two intermediate infectious bursal disease (IBD) vaccines (Clone Vac D-78 and S-706) for immunizing specific pathogen-free (SPF) White Leghorn chickens by coarse spray (CS) against subclinical IBD was compared. Both intermediate IBD vaccines were equally capable of immunizing day-old SPF chickens by CS and were safe as evidenced by the absence of morbidity, mortality, or severe gross and microscopic bursal pathology at 28 days of age.

Animals↗

PubMed1986

Evaluation of the immunogenicity, stability, pathogenicity, and immunodepressive potential of four commercial live infectious bursal disease vaccines.

The study was divided into three experiments. In each, day-old specific pathogen free (SPF) White Leghorns were injected subcutaneously with one of the following commercial live infectious bursal disease (IBD) vaccines: Burcell, Bursine, Clone Vac D-78, and S-706. In the first experiment, the immunogenicity of each vaccine was determined by challenge with virulent IBDV at 25 days of age. All four vaccines were equally efficacious in preventing clinical IBDV infection. The second experiment determined the stability and pathogenicity of the four vaccines by successive back passage of each virus in SPF chicks for 4 consecutive weeks. Although all vaccines were capable of spreading to contact controls, only the two intermediate vaccines (D-78 and S-706) produced slightly atrophied bursae and moderate microscopic bursal lesions. However, both vaccines were stable, because neither reverted back to increase virulence nor resulted in morbidity or mortality associated with virulent IBDV after successive passage. In the third experiment, the immunodepressive potential of each vaccine virus was determined by examining the ability of IBD-vaccinated birds to respond to Newcastle disease (ND) virus vaccination. None of the four vaccines was found to be immunodepressive, as all IBD vaccinated birds responded to NDV vaccination.

Animals↗

PubMed1986

Effects of aflatoxin on the growth performance and immune responses of weanling swine.

Aflatoxin (AF)-contaminated ground corn was mixed with a commercial swine ration to yield 2 concentrations (500 mg of AFB1/kg of feed [A] and 300 mg of AFB1/kg [B]) and was fed to 2 groups of pigs. Groups A and B were fed the AF-containing ration, whereas control group C was fed the same commercial ration mixed with ground corn devoid of AF. A comparative analysis of the average weight gain per pig in each of the treatment groups, compared with that in the control group, indicated a significantly (P less than 0.01) greater weight gain in the control group. The average feed conversion rate was also significantly (P less than 0.01) lower in group A pigs, compared with that in the control group. The humoral immune response to Erysipelothrix rhusiopathiae, measured by enzyme-linked immunosorbent assay, did not reveal a significant difference among groups; there were no consistent differences observed in the proliferative responses of lymphocytes to mitogens. In contrast, a significant (P less than 0.05) reduction in complement titers was observed, whereas an increase in serum immunoglobulin G and M values occurred in the AF-treated group A, compared with that in group C. Gross enlargement of the liver, substantiated by histologic evidence of toxic damage to the hepatic parenchyma, revealed that AF at concentrations of 500 mg/kg of feed was toxigenic and produced an adverse effect on the growth rate, feed efficiency, and general well-being of young pigs.

Aflatoxins↗

PubMed1985

Effects of purified aflatoxin on broiler chickens.

Purified aflatoxin B1 (AFB1) or AFB1 plus aflatoxin B2 (AFB2) was given daily for 5 weeks in gelatin capsules to 2-week-old feather-sexed broilers. In Experiment 1, pure AFB1 was given in doses equivalent to the quantity of toxin received, if diets containing either 0, 200, 500 or 1000 ppb of AFB1 were consumed. In Experiment 2, pure AFB1 or AFB1 plus B2 was administered in capsules in doses equivalent to the quantity of toxin received, if diets containing either 0, 100, 200, or 400 ppb of AFB1 were consumed. In Experiment 1, pure AFB1 greater than or equal to 500 ppb was only mildly toxic. These levels produced a significant decrease in the 5-week weight gain and microscopic lesions indicative of alfatoxicosis. No morbidity, mortality, or effects on feed conversion or immune responses, however, were noted in birds given pure AFB1 at these levels. Gross liver lesions indicative of aflatoxin toxicity occurred at the 1000 ppb only. Results of Experiment 2 were similar to the first. Weight gain and feed conversion were not affected for broilers receiving pure AFB1 as low as 200 ppb. No morbidity, mortality, or gross lesions were evident in birds given either pure AFB1 or AFB1 plus AFB2 as high as 400 ppb. However, cell-mediated immunity as measured by a delayed hypersensitive skin test was significantly affected in birds receiving 400 ppb AFB1 plus AFB2. No effects on humoral immunity or the development of acquired immunity to Newcastle disease or fowl cholera vaccination were noted.

Administration, Oral↗

PubMed1985

Effect of purified aflatoxin on turkeys.

Purified aflatoxin B1 (AFB1) or AFB1 plus aflatoxin B2 (AFB2) was given daily for 5 weeks in gelatin capsules to 2-week-old Nicholas X Nicholas turkeys. In Experiment 1, pure AFB1 was given in dosages equivalent to the quantity of toxin received if diets containing either 0, 200, 500, or 1000 ppb of AFB1 were consumed. In Experiment 2, either pure AFB1 or AFB1 plus AFB2 was given in capsules equal to the quantity of toxin received if a diet containing either 0, 100, 150, or 200 ppb of AFB1 was consumed. In Experiment 1, pure AFB1 administered by capsule equal to 500 and 1000 ppb was highly toxic and by the second week, it resulted in 100% morbidity, mortality, and gross and microscopic lesions. Aflatoxin B1 at 200 ppb caused none of these changes. However, it did cause a significant depression in feed conversion, but not weight gain, during all weeks and also a reduction in cell-mediated immunity (CMI) as measured by a delayed hypersensitive skin test to mycobacterium. Results in Experiment 2 were similar to the first. Aflatoxin B1 and AFB1 plus AFB2 as high as 200 ppb resulted in no morbidity, mortality, or gross or microscopic lesions. Also, no significant reductions in either weight gain or feed conversion were evident in any level of either AFB1 or AFB1 plus AFB2. However, numerical, but not statistically significant, dose-related reductions in all CMI tests were noted in birds receiving either AFB1 or AFB1 plus B2 at as low as 100 ppb.

Administration, Oral↗

PubMed1985

Effect of levamisole on the response of broilers to coccidiosis vaccination.

The effect of levamisole on the response of commercial broilers to coccidiosis vaccination was assessed. The concentration, frequency, and timing of levamisole were varied in three separate experiments to determine optimum conditions for potentiating the response of broilers to a single oral dose of commercial coccidiosis vaccine at 7 days of age. In all instances, levamisole was found to improve development of immunity to coccidiosis. Birds that received levamisole by intraperitoneal (IP) injection had better weight gains for 3 weeks following coccidiosis vaccination and improved resistance to challenge with Eimeria tenella when compared with vaccinated chickens that did not receive levamisole. However, levamisole given at .25 mg/kg body weight by IP injection 3 days prior to the coccidiosis vaccine resulted in the greatest potentiation of the immune response. Performance parameters following challenge included weight gain, oocyst production, fecal droppings, and livability. Findings confirm and extend observations regarding the ability of levamisole to potentiate vaccination responses in chickens.

Animals↗