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Biomedical subjects

P Villegas

Publications and source records attributed to P Villegas.

At least 19 recordsLinked to original sources

Detection of Massachusetts and Arkansas serotypes of infectious bronchitis virus in broilers.

The objective of this study was to compare the presence of the Arkansas (Ark) and Massachusetts (Mass) serotypes of infectious bronchitis virus (IBV) in the tracheas and cecal tonsils of commercial broilers after vaccination at 1 day of age by coarse spray. When given as a single serotype vaccine, the Mass strain was detected by reverse transcriptase-polymerase chain reaction (RT-PCR)-restriction fragment length polymorphism (RFLP) only in the tracheas, whereas the Ark strain was detected in both the tracheas and cecal tonsils. By in situ hybridization, the Mass and Ark nucleocapsid (Nc) genes were detected only at 7 days in the tracheas. When both strains were given in the mixed vaccine, the Mass strain was more consistently detected by RT-PCR-RFLP in the tracheas and cecal tonsils at early stages of infection (up to 14 days) and the Arkansas strain was more consistently detected at late stages of infection (21 and 28 days). By in situ hybridization, the IBV Nc gene was more consistently detected in the trachea at early stages of infection (7, 14, and 21 days) and in the cecal tonsils at late stages of infection (21, 28, and 35 days). In general, the Mass strain was more frequently recovered from the tracheal and cecal tonsil tissues at earlier stages of infection and the Ark strain was recovered at later stages of infection.

Animals↗

Molecular characterization of avian infectious bronchitis virus strains isolated in Colombia during 2003.

Sixteen infectious bronchitis virus (IBV) isolates were recovered from broilers and layers from five geographic poultry regions in Colombia. The viruses were isolated from tracheas, lungs, and cecal tonsils of birds, previously vaccinated with the Massachusetts strain, that were showing respiratory signs. Further analysis of the IBV isolates was achieved by phylogenetic analysis comparing their deduced amino acid sequences in the hypervariable region 1 of the S1 gene with reference strains. Four unique genotype clusters containing isolates with indigenous genotypes were observed. One isolate was found to be the Connecticut genotype and three isolates were found to be the Massachusetts genotype.

Animals↗

Sequence analysis, viral rescue from infectious clones and generation of recombinant virions of the avian adeno-associated virus.

Aiming at the generation of a viral-vectored system for gene delivery and vaccination in poultry, the entire genomes of the VR-865 and DA-1 strains of the avian adeno-associated virus have been cloned and sequenced. Sequence analysis of the clones showed that the genomic distribution of the structural and non-structural protein-coding genes of these viruses is conserved and in agreement with what has been previously described for the primate adeno-associated viruses. Amino acid differences between the avian adeno-associated viruses and the primate adeno-associated viruses are more evident in the genes that code for the non-structural (Rep) proteins of the virus, while the Cap region amino acid sequence was found to be more conserved. Since all the regulatory and coding sequences of the virus were present in the plasmids obtained, complete infectious viral particles were rescued from these clones, and these rescued viral populations were amplified by co-infecting primary embryo liver cells with the rescued virus and the CELO strain of the avian adenovirus type 1. As a proof of concept of the validity of this system for the purpose of gene delivery, recombinant viruses encoding for the LacZ gene as a reporter system were also generated. These recombinant viruses were used to express beta galactosidase activity in primary chicken embryo cell cultures.

Animals↗

Viral diseases of the respiratory system.

Infectious bronchitis, Newcastle disease, infectious laryngotracheitis, avian influenza, and pneumovirus are the viruses that more frequently affect the respiratory tract of chickens. Because of the tendency to change its antigenic properties, infectious bronchitis is currently the viral disease present in most poultry producing areas of the world. New serotypes and variant strains are reported in several countries. Current commercially available vaccines do not always provide protection against new field isolates. Vaccination programs are constantly adjusted in an attempt to improve protection against this disease. Infectious laryngotracheitis has appeared in the broiler industry as a serious disease. Improved vaccines are needed to control the disease in broilers. In the U.S., the control of the highly pathogenic forms of avian influenza and the velogenic forms of Newcastle disease have been achieved by eradication. In other countries, effective vaccines have been used to control Newcastle and avian influenza. Avian pneumovirus infection is also an emerging disease of chickens and turkeys.

Animals↗

Growth and tissue lipid deposition of broilers fed alpha-ketoisocaproic acid.

Dietary alpha-ketoisocaproic acid (KIC) was studied for effects on growth, feed efficiency, tissue cholesterol (TC) content, and immune response to a Newcastle disease virus (NDV) vaccination in broilers. In two experiments, broiler chicks were fed diets supplemented with graded levels of KIC to .54% of the diet. Feed efficiency, fat deposition, BW gain, and plasma cholesterol levels were determined at 3 wk. In a third experiment, broilers were fed diets supplemented with 0, .1, .2, and .4% KIC throughout the starter (0 to 3 wk), grower (3 to 6 wk), and finisher (6 to 7 wk) periods. Abdominal fat and plasma and breast TC were measured at 7 wk. In Experiment 4, levels of KIC to .6% were fed to broilers to determine TC content and to measure the immune response to NDV vaccination. Feed efficiency, BW gain, and percentage abdominal fat were not affected by KIC in any of the broiler experiments. The addition of KIC to broiler diets resulted in a significant reduction in breast (Pectoralis major) TC. Biceps (Biceps femoralis) TC, adductor (Pubo-ishio-femoralis) TC, and plasma cholesterol content was not affected by KIC. Geometric mean titers to NDV were not improved by KIC. The data indicate that KIC has no effect on growth or abdominal fat deposition, but may have a role in TC homeostasis.

Adipose Tissue↗

Early region 4 sequence and biological comparison of two isolates of mouse adenovirus type 1.

The DNA sequence of 88-100 map units of mouse adenovirus type 1 (MAV-1) was determined. One translational open reading frame showed 48% sequence similarity to a human adenovirus type 2 early region 4 protein. Based on the protein similarity, genome location, and transcriptional polarity, we concluded that this region of MAV-1 corresponds to early region 4. A 241-bp sequence consisting of 10 imperfect direct repeats with sequence similarity to minisatellite DNA was found in this region. Two virus isolates with different passage histories were examined and were found to have a sequence polymorphism within this region. The two viruses were compared for growth in cell culture and mice and small quantitative differences were observed only in vivo.

Adenoviridae↗

Antibody response against newcastle disease in commercial broilers fed different dietary protein levels.

A factorial experiment was conducted with two levels of dietary protein (17.5 and 26%) and three vaccination regimens (inactivated oil-emulsion, live LaSota strain, and unvaccinated) against Newcastle disease at 10 days of age. There were two trials with 3 pens of 15 male broiler chicks each per cell. Birds fed the high protein diet grew significantly better (P less than .05) in the second trial (1731 g at 6 weeks vs. 1622 g) but not in the first (1304 vs. 1244 g); no growth differences were noted due to vaccine. At 3 weeks of age, the hemagglutination-inhibition antibody titers from the live vaccine were significantly (P less than .01) higher than from the inactivated vaccine. At 3 weeks of age, antibody titers from the inactivated vaccine were elevated (P less than .05) in birds fed the low protein diet in the second trial. At 6 weeks of age, antibody titers were affected by vaccine (P less than .05) but not by dietary protein. Antibody titer levels were higher from the inactivated vaccine than from the live vaccine in the first trial; these results were reversed in the second trial. Postchallenge results with the Texas GB strain of Newcastle did not reveal any significant differences due to dietary protein content, trial, or type of vaccine.

Animals↗

Further studies with an inactivated oil emulsion Newcastle disease vaccine in broiler breeders.

Chickens vaccinated with a live Newcastle disease (ND) vaccine and subsequently revaccinated with an inactivated oil emulsion ND vaccine had higher and more persistent hemagglutination-inhibition (HI) antibody titers than birds vaccinated with live ND vaccine. The geometric mean HI antibody titers of flocks vaccinated with the inactivated ND vaccine ranged from 121 to 172, whereas the titers of flocks vaccinated every 90 days with a live ND vaccine ranged from 18 to 54. Breeder flocks revaccinated with live LaSota ND vaccine had lower egg production than the flocks vaccinated with inactivated vaccine. The average egg production per hen for the 40-week laying cycle was 174 and 170 eggs per hen for hens in the two houses vaccinated with the inactivated vaccine. Hens vaccinated with the live virus vaccine averaged 167 and 164 eggs per hen. The increase in egg production would more than offset the additional cost of the oil emulsion ND vaccine as well as the cost of injecting each individual bird at 18 weeks of age. Between 8 and 32 weeks after onset of egg production, geometric mean maternal antibody titers in 1-day-old chickens derived from breeders vaccinated with live ND vaccine ranged from 7 to 24, whereas, chicks from breeders vaccinated with the inactivated oil emulsion vaccine had maternal antibody titers that ranged from 64 to 89.

Animals↗

Vaccination of broiler chicks from breeder flocks immunized with a live or inactivated oil emulsion Newcastle disease vaccine.

One-day-old chicks used in this study were from breeder flocks vaccinated with live (B1 and LaSota) or inactivated oil emulsion Newcastle disease (ND) vaccine. Chicks were vaccinated against ND by various procedures. The vaccination response was evaluated by hemagglutination-inhibition antibody titers and by challenge. Chicks from breeder flocks vaccinated with live virus vaccine had a geometric mean hemagglutination-inhibition antibody titer (GMT) for Newcastle of 7 (low maternal antibody titer) at 1 day of age, whereas those chickens derived from breeder flocks vaccinated with inactivated oil emulsion ND vaccine had a GMT of 84 (high maternal antibody titer). One-day-old chicks injected with live B1 vaccine were not immunized against ND regardless of breeder flock source. However, chickens with low maternal antibody titers were effectively immunized against ND when injected at 1 day of age with an inactivated or inactivated plus live ND vaccine. Chicks with high maternal antibody titers were not effectively protected when vaccinated with inactivated vaccine at 1 day of age; however, these chicks were protected when injected with a combined live and inactivated ND vaccine. Chicks from both breeder flocks were effectively immunized against ND when injected at 1 day of age with a live or inactivated ND vaccine and revaccinated by aerosol at 21 days of age with live B1 ND vaccine. Even though they were protected against ND, there appears to have been an interference phenomenon in chicks derived from breeder flocks vaccinated with the live ND vaccine. Beak-O-Vac vaccinated chickens were not effectively protected against ND when compared with chicks vaccinated by aerosol at 1 day of age. Water vaccination at 7, 14, or 21 days of age was as effective as aerosol vaccination when administered to chicks with low maternal antibody titers. However, water vaccination was not as effective as aerosol vaccination when administered to chicks with high maternal antibody titers.

Administration, Oral↗

Field trials with an oil emulsion Newcastle disease vaccine in broiler breeders.

Chickens vaccinated with live Newcastle disease (ND) vaccine and subsequently revaccinated with an inactivated oil emulsion vaccine had high and persistent hemagglutination-inhibition (HI) antibody titers for at least 40 weeks. The geometric mean HI antibody titers of flocks vaccinated with the inactivated ND vaccine ranged from 48.8 to 91.9, whereas the titers of flocks vaccinated every 90 days with a live ND vaccine ranged from 8.6 to 43.5. Breeder flocks revaccinated with a liver LaSota ND vaccine had lower egg production than the flocks vaccinated with the inactivated vaccine. The average egg production per hen for the 40 week laying cycle was 177.8 and 174.8 eggs per hen for hens vaccinated with the inactivated vaccine, whereas those hens vaccinated with the live virus vaccine averaged 163.0 and 155.6 eggs per hen. The increase in egg production would more than offset the additional cost of the oil emulsion NDV vaccine as well as the cost of injecting each individual bird at point of lay. Maternal antibody titers in one-day-old chicks were related to the titers in the dams. Maternal antibody titers in chicks originating from breeder flocks vaccinated with the oil emulsion vaccine were higher than the maternal antibody titers of chickens from breeders vaccinated with the live NDV vaccine.

Animals↗

Comparison of inactivated and live infectious bursal disease virus vaccines in White Leghorn breeder flock.

Leghorn breeder chickens, which had received live infectious bursal disease (IBDV) virus vaccine at 12 weeks of age and had been injected subcutaneously at 20 weeks of age with an inactivated IBDV oil emulsion vaccine, produced very high antibody titers which persisted for at least 9 months and were more uniform than those obtained from breeders receiving only the live IBDV vaccine. Breeders vaccinated with the inactivated IBDV oil emulsion vaccine at 12 and 20 weeks of age had antibody titers higher than breeders which received only the live IBDV vaccine at 12 weeks of age; titers were lower than in the group which received both the live and inactivated oil emulsion IBDV vaccines. Maternal antibody levels in chickens derived from parent flocks vaccinated with the inactivated oil emulsion IBDV vaccine were higher and more uniform than in chickens derived from parent flocks vaccinated with the live IBDV vaccine. Maternal antibody persisted longer in chickens derived from parent flocks vaccinated at 20 weeks of age with the inactivated oil emulsion IBDV vaccine than in chickens vaccinated by the other procedures. The titers remained quite high up to 16 days of age.

Animals↗

Efficacy of intratracheal administration of Newcastle disease vaccine in day-old chicks.

Groups of day-old chicks with varying levels of parental antibody were vaccinated against Newcastle disease (B1 strain) with a commercially available device which simultaneously debeaks the chick and emits a fine spray of vaccine into its trachea. Some groups were also vaccinated (B1 or Lasota strain) with a commercially available vaccine sprayer at 9 days, 14 days, or 9 and 25 days of age. Response to vaccine was evaluated once each week during the experimental period of approximately 8 weeks HI titers were determined and 10 chicks were challenged with the Texas GB strain of Newcastle disease virus. In chicks with low to moderate levels of maternal antibody a satisfactory antibody response was attained by vaccination at 1 day of age, and in most cases resistance to challenge was evident by 3 weeks of age. Intratracheal vaccination of chicks with extremely high levels of maternal antibody had a minimal antibody response. All groups of chicks spray vaccinated at 9, 14, or 9 and 25 days of age showed a marked increase in antibody titer regardless of whether they had been vaccinated at 1 day of age.

Aerosols↗

Aerosol administration of Newcastle disease vaccines at one day of age.

Newcastle disease vaccines were administered at one day of age by 3 general procedures: 1) discharge of a vaccine aerosol over chicks confined in an enclosed space, 2) aerosol adminstration in a poultry house, 3) a commercial device which emits a fine spray into the nasopharynx while the chick is being debeaked. From this work the following conclusions have been drawn: 1) All 3 methods are of some value with respect to the development of antibody titers and resistance to challenge. 2) The efficacy is universely proportional to the maternal antibody titer. 3) Aerosol vaccination into an enclosed air space is superior to the other methods tested. 4) Day-old vaccination is useful primarily in conferring some early resistance until a subsequent aerosol vaccination at 9-14 days of age confers additional protection. Other important factors include strain of vaccine, vaccine titer, the nature of the diluent, and site of the aerolized particles.

Administration, Oral↗

Efficacy of lyophilized turkey herpesvirus vaccine against Marek's disease in broilers.

Field trials involving approximately 1,250,000 chickens vaccinated with cell-free turkey herpesvirus (HVT) vaccine and 635,000 chickens vaccinated with cell-associated HVT vaccine howed both vaccines equally effective in reducing condemnations (attributable to Marek's disease) at the poultry processing plant. Vaccinated chickens were progeny of hens vaccinated at 1 day of age with cell-associated HVT vaccine. The maternal antibody did not interfere with the ability of either the cell-free or cell-associated HVT vaccine to provide protection for chickens against challenge from the virulent Marek's disease virus.

Animals↗

Efficacy of turkey herpesvirus vaccine when administered simultaneously with fowl pox vaccine.

The efficacy of the turkey herpesvirus (HVT) vaccine in protecting chickens challenged with virulent Marek's disease (MD) virus was unaffected by the presence of either the chick embryo fowl pox vaccine or fowl pox vaccine derived from cell culture. Conversely the HVT vaccine did not affect the efficacy of the fowl pox vaccine in chickens challenged with pathogenic fowl pox virus. A combination of spectinomycin dihydrochloride pentahydrate and lincomycin hydrochloride monohydrate as well as spectinomycin sulfate tetrahydrate were found to be compatible with the HVT and fowl pox vaccines as demonstrated by resistance after challenge with virulent MD virus or fowl pox virus.

Animals↗

Molecular characterization of Spanish infectious bursal disease virus field isolates.

Nine Spanish isolates of infectious bursal disease virus (IBDV) were characterized and classified after reverse transcriptase-polymerase chain reaction of a 248-bp fragment of the VP2 gene hypervariable region and restriction fragment length polymorphism (RFLP). The restriction endonucleases (REs) used were BstNI, Sad, SspI, TaqI, DraI, and StyI. Sequencing of the amplified product and further comparison of these sequences with published sequence data from other IBDV strains were also performed. Very virulent and classic strains were identified. None of the strains identified had molecular characteristics similar to that of the American variant strains. Four very virulent strains (VG-248, 5939, 6145, and 7333) were digested by the TaqI, SspI, and StyI enzymes. The sequences of these strains were closely related to other European and Japanese very virulent IBDV (vvIBDV) strains. Strains VG-311, VG-262, and VG-208 were digested by the BstNI and Sad REs and were classified as classic strains. Strains VG-276 and VG-313 had unique RFLP patterns. VG-276 exhibited the SspI RE site, which has been reported as a characteristic of vvIBDV strains, whereas the VG-313 strain exhibited a Sad and StyI RE site indicative of the classic IBDV Edgar and 52-70 strains. However, nucleotide sequence analysis of the amplified hypervariable region strain VG-276 revealed a higher identity with the classic strains STC, 52/70, and 9109 IBDV strains, whereas strain VG-313 exhibited a higher identity with the vvIBDV strains.

Animals↗

Lack of interaction between avian leukosis virus subgroup J and fowl adenovirus (FAV) in FAV-antibody-positive chickens.

Unfounded field speculation has suggested that avian leukosis virus subgroup J (ALV-J) predisposes young meat-type chickens to inclusion body hepatitis caused by fowl adenovirus (FAV). To address this hypothesis, we infected 1-day-old grandparent meat-type chickens carrying maternal antibodies against FAV with a field isolate of FAV associated with inclusion body hepatitis in broilers, ALV-J, or both FAV and ALV-J. We examined the effects of FAV alone or in combination with ALV-J on the basis of clinical signs, overall mortality, growth rate, and gross and microscopic lesions. With such criteria for evaluating possible interactions, we found no significant differences in the dually infected birds in comparison with chickens that received a monovalent challenge with either FAV or ALV-J.

Adenoviridae Infections↗

Molecular characterization of infectious bursal disease virus from commercial poultry in the United States and Latin America.

From June 1999 to September 2001, 216 bursal samples from broiler farms in the United States and from countries of Latin America were submitted to the Poultry Diagnostic and Research Center at the University of Georgia for the purpose of genotyping field infectious bursal disease viruses (IBDVs). The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify a 248-bp product, encompassing the hypervariable region of VP2 gene. The genotyping was conducted by restriction fragment length polymorphism (RFLP) analysis with six restriction endonucleases, DraI, SacI, TaqI, Sty, BstNI, and SspI. For the 150 samples received from the United States, 125 samples (83.3%) were RT-PCR positive for the presence of IBDV. One hundred positive samples (80%) had RFLP identical to the variant Delaware E strain, whereas 10 samples (8.0%) exhibited a RFLP pattern similar to this antigenic variant. Other IBDV strains such as Grayson Laboratory strain (GLS), Lukert, PBG-98, Delaware A, and the vaccine strains Sal-1 and D-78 were also detected. Two samples exhibited a pattern similar to the standard challenge (STC) strain, and seven strains (5.6%) were not classified by RFLP. Sixty-six bursal samples previously inactivated with phenol were received from Latin American countries. IBDV strains with analyzed genotypes similar to the Lukert strain were predominantly detected in Mexico. IBDV strains similar to variant E were detected in Colombia and Ecuador. Peru and Venezuela exhibited a higher heterogeneity of IBDV strains due to the detection of classic Delaware type as well as GLS variant strains. IBDV strains detected from Brazil and Dominican Republic exhibited RFLP patterns identical to very virulent IBDV strains prevalent in several countries in Europe, Asia, and Africa.

Animals↗