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PubMed1989

Flagellar movement in demembranated preparations of ejaculated fowl spermatozoa.

Triton X-100 at a concentration of 0.1% in the extraction medium was optimal for demembranating fowl spermatozoa. The most suitable conditions for reactivation were obtained when a medium composed of 0.5 mM-ATP, 25 mM-potassium glutamate, 10(-7) M-CaCl2, 20 mM-Tris-HCl(pH 7.9), 1 mM-MgSO4, 1 mM-dithiothreitol and 0.2 M-sucrose was used. More than 60% motile spermatozoa were obtained under these conditions. The addition of 1 or 10 microM-cAMP did not appreciably affect motility. Intact and demembranated spermatozoa were immotile at 40 degrees C, whilst at 30 degrees C motility was restored.

Animals↗
PubMed1990

Effects of high-density lipoproteins on storage at 4 degrees C of fowl spermatozoa.

Qualitative and quantitative characterization of lipoproteins found in seminal plasma from domestic cocks was performed after isolation by density gradient ultracentrifugation. Trigyceride-rich lipoproteins (very low, intermediate- and low density lipoproteins) were not detectable in seminal plasma. High-density lipoproteins (HDL), identified on the basis of size, chemical composition and protein moiety, were present at a concentration of 66 micrograms/ml. A fraction possibly corresponding to VHDL (very high density lipoproteins, 77% protein, 23% lipid) was also detected but appeared contaminated by a protein-rich opalescent material. Since HDL contains mostly phospholipid and cholesterol, the physiological role of these lipoproteins on the storage of fowl spermatozoa was studied. Replacing seminal plasma with a solution containing chicken HDL at physiological concentration (66 micrograms/ml) had no effect on fertilizing ability of spermatozoa stored at 4 degrees C for 24 h. However, higher concentrations of HDL (560 micrograms/ml) had deleterious effects on spermatozoa stored in vitro.

Animals↗
PubMed1991

Temperature-mediated regulation of calcium flux and motility in fowl spermatozoa.

The motility of fowl spermatozoa at various temperatures was shown to be a function of their intracellular calcium content, measured after hypotonic lysis of the cells. Retention of calcium by spermatozoa, with consequent enhancement of motility, increased as the temperature was lowered from 40 degrees to 30 degrees C. Raising the temperature within this range subsequently reduced calcium retention and motility again. The temperature-dependent retention of calcium was a function of the rate of calcium efflux rather then influx. The temperature-sensitive efflux mechanism appeared to involve a Ca2+ ATPase which was relatively inactive at 30 degrees C, but active at 40 degrees C.

Animals↗
PubMed1992

Induction of immunity to spermatozoa in male domestic fowl and effects on fertility.

Male fowl were immunized intravenously (i.v.) or intramuscularly (i.m.) with spermatozoa to assess the effects of immunity to spermatozoa on fertility. Histological and immunofluorescence evaluations of testis and ductus deferens tissues after 24 weeks of immunizations revealed immune cell infiltration and immunoglobulin associated with spermatozoa. The long-term immunization regimen resulted in significant antisperm antibody titres in the immunized groups. When semen from i.v.-immunized males was used to inseminate females, fertility over 7 days was reduced (P less than 0.05). A subsequent experiment using a 10-week i.v. immunization scheme also led to high antisperm titres. Spermatozoa from these males were characterized by lower fertility and duration of fertility than those of controls (P less than 0.05). As in mammals, a reduction in fertility may result from exposure of avian males to sperm antigens.

Animals↗
PubMed1992

Maintenance of motility of fowl spermatozoa in vitro is prolonged by a low molecular weight factor derived from cultured chick embryo cells.

Gel filtration of a conditioned medium composed of the supernatant fluid removed from a 5-day culture of skeletal muscle cells from 9-day-old chick embryos with Bio-Gel P-2 revealed one peak of motility-prolonging activity (about 0.3 kDa), which was not present in fresh medium. Spermatozoa incubated in this fraction of the conditioned medium maintained their motility for at least 36 h at 37 degrees C. Both the formation of lipid peroxide and the leakage of lactic dehydrogenase of spermatozoa incubated in the conditioned medium fraction were lower than those incubated in the corresponding fresh medium. Initial rate of oxygen consumption of the spermatozoa incubated in the conditioned medium fraction increased compared with that of the fresh medium fraction. These results suggest that a low molecular weight factor(s) supplied by cultured cells effectively prolongs the motility of fowl spermatozoa, and that the effect could result from inhibition of the structural damage to the sperm membrane.

Animals↗
PubMed1992

The embryo pancreas is a source of increased yolk amylase in the fertilized eggs of domestic fowls.

The amylases were studied in the yolk of fertilized eggs and in the pancreases of the embryos of domestic fowls. The amylase activity in the yolk increased markedly from 13 days of incubation until hatching, but the activity decreased when the embryos were taken out of the eggs. The isoamylases in the yolk and in the pancreas of the embryo were identical electrophoretically. The amylase occurs mainly in the pancreas of the embryo. We think that the increase in amylase activity in the yolk of fertilized eggs during incubation depends upon the accumulation of pancreatic amylase synthesized by the developing embryo in the egg.

Amylases↗
PubMed1988

Light and dark cells in the lumbar sensory ganglia of pre and post-hatching domestic fowl.

Clear and dark satellite cell classes were identified by electron microscopy in the lumbar sensory ganglia of domestic fowl in 8 pre and 4 post-hatching stages of development. Some cytologic differences found between the two classes relating to the rough-endoplasmic reticulum, ribosomes, Golgi apparatus and junctional complexes suggest the existence of distinct functional features for both types of satellite cells.

Animals↗
PubMed2000

Presence-absence sequential sampling plan for northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae), on caged-layer hens.

Caged-layer hens were scored as infested or uninfested by visual examination of the vent region, and the number of northern fowl mite, Ornithonyssus sylviarum (Canestrini & Fanzago), per hen was estimated. The proportion infested and average number of mites per hen were shown to have a highly significant, positive relationship (r = 0.936). Sampling among houses within a flock, and rows and sections within houses were analyzed to determine the reliability of sampling a representative portion of a flock. Low- and moderate-tolerance treatment thresholds, based on percentage of hens infested with mites, were developed from sampling 1 wk before and 1 wk after acaricide treatments determined necessary by the producer. These thresholds were used to compare a fixed (single) sampling plan, a curtailed procedure of the fixed sampling plan, and a sequential sampling plan based on a sequential probability ratio test, by sampling 174 hens (the maximum number needed for the single sampling plan). The sequential sampling plan required fewer hen examinations on average to reach a treatment decision than did the other plans, depending on the infestation tolerance limits. Using a low tolerance approach in which infestations below 15% are considered noneconomic (safe threshold) and infestations above 25% are considered economically important (action threshold), as few as 5 hens required examination to reach a treatment decision. Sequential sampling plan graphs are presented for 2 tolerance threshold scenarios (a 15% safe-threshold paired with a 25% action threshold and a 35% safe-threshold paired with a 45% action threshold). These sequential sampling plans using presence absence assessments should greatly facilitate monitoring and treatment decisions for this important pest.

Animals↗
PubMed2000

Monitoring northern fowl mites (Acari: Macronyssidae) in caged laying hens: feasibility of an egg-based sampling system.

Northern fowl mites were monitored on a caged-layer operation in southern California for 22 mo. Three experienced observers underestimated actual numbers of mites in the vent region approximately 80% of the time. Errors were higher for heavy infestations. Observer estimates were highly correlated with each other (r > 0.89, P < 0.01) and with mite numbers estimated by vent feather removal (r > 0.82, P < 0.01). Mites on hens varied between houses and over time. Molting consistently reduced mite numbers, but did not eliminate then in a flock. Long-term monitoring of individual sentinel hens demonstrated that some hens would support high numbers of mites for several months or more. Use of a new sequential hen sampling plan required approximately 1 min per hen, if mite numbers were estimated. At this site, treatment decisions often could be reached in < 20 min per house. Mite scores (index of estimated mites per hen) were well correlated with percentage of hens infested in both test houses. In a chronically infested house, prevalence of mites on eggs averaged 8.5%, with a range of 0-55%. Applications of tetrachlorvinphos-dichlorvos by the producer appeared to be based on mites on > about 20% of eggs. The chemical was marginal for controlling mites on hens (25% reduction in percentage of hens infested), but effectively reduced mites on eggs (95% fewer mites on eggs at 1 wk and 90% at 2 wk). When data were grouped by mite index score on hens, there was a strong relationship (r2 = 0.83, P < 0.01) between mite prevalence on eggs and the scores of the hens which laid them. Sampling 100 eggs evenly spaced in a house required < 7 min, and adult mites were easily seen. Sampling mites on eggs appears to be useful to localize at least high-level infestations, and egg-based sampling for mites merits further investigation.

Animals↗
PubMed1982

Delayed neurotoxicity resulting from administration of leptophos to the comb of domestic fowl.

We investigated the occurrence of delayed neurotoxicity in domestic fowl following percutaneous application of leptophos. Five groups of 5 adult hens received daily percutaneous doses of 1.0 ml/hen of leptophos emulsion (leptophos; 340 mg/hen/day) for 2, 5, 10, 15 or 20 days. There was no abnormal gait in the 2-day group. Two out of 5 hens in the 5-day group showed mild ataxia from about 2 weeks after the final administration, but did not develop severe neuropathy. On the contrary, 4 out of 5 birds in the 10-day group and all hens in the 15- and 20-day groups were affected by various stages of neurotoxicity. Some of them died from neurotoxicity. Ten of young male chickens were given the same dermal dose for 5 or 10 days. Although no abnormal chicken was observed in the 5-day group, all chickens in the 10-day group showed severe paralysis and two of them died. We studied the incidence rates of delayed neurotoxicity resulting from respective applications of the emulsion and the acetone solution of leptophos. No significant difference was observed between them. These results suggest that the daily dermal application of the relatively high dose of leptophos, even if for the short term, can cause the same delayed neurotoxic effects as by the oral administration in hens or chickens.

Animals↗
PubMedDate not supplied

Growth and infectivity assays of the Israeli vaccine strain of fowl poxvirus in chicken embryo fibroblasts.

The Israeli vaccine strain of fowl poxvirus grows efficiently in chicken embryo fibroblasts but not in cell lines derived from monkey kidney or human fibroblasts. We developed two assays for the titration of the infectivity of this virus in secondary cultures of chicken embryo fibroblasts. The first is a focus assay, in which minimum essential medium and SeaKem ME agarose were used for the overlay media. Under these conditions, clear virus foci appeared after 5 days of incubation at 37 C. The second assay is a semiautomatic colorimetric test based on the ability of live cells in culture to reduce the yellow tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT; thiazolyl blue) to its formazan derivative. The reagent was added to infected chicken embryo fibroblasts in 96-well plates 10 days after infection. The formazan formed during 2 hr was extracted with dimethyl sulfoxide, and its absorbance was read by an automatic microplate spectrophotometer. A good correlation of the infectivity titers of the virus was obtained by the two methods.

Animals↗
PubMedDate not supplied

Fowl adenovirus serotype 4 associated with outbreaks of infectious hydropericardium in Haryana, India.

During 1998, hydropericardium syndrome was observed among 3-to-6-wk-old broilers in 45 different flocks of Haryana, India, with mortality ranging between 10% and 30%. Fowl adenovirus (FAV) was isolated from one of the affected flocks by chicken embryo liver cell culture. Serum neutralization test and polymerase chain reaction assay coupled with restriction enzyme analysis confirmed that the isolated virus belonged to FAV serotype 4. The disease was reproduced in 28-day-old broilers by subcutaneous and oral inoculation of isolated FAV4 alone. Typical hydropericardium and basophilic intranuclear inclusions in hepatocytes were observed in experimental birds by day 4 postinoculation.

Adenoviridae Infections↗
PubMedDate not supplied

Prevention of inclusion body hepatitis/hydropericardium syndrome in progeny chickens by vaccination of breeders with fowl adenovirus and chicken anemia virus.

The hypothesis that an effective protection of progeny chickens against inclusion body hepatitis/hydropericardium syndrome (IBH/HP) can be achieved by dual vaccination of breeders with fowl adenovirus (FAV) serotype 4 and chicken anemia virus (CAV) was tested. Thus, 17-wk-old brown leghorn pullet groups were vaccinated by different schemes including single FAV (inactivated), single CAV (attenuated), FAV and CAV dually, or were not vaccinated (controls). Subsequent progenies of these breeders were challenged with the virulent strains FAV-341 and CAV-10343 following three strategies: 1) FAV-341 intramuscularly (i.m.) at day 10 of age (only FAV-vaccinated and control progenies); 2) FAV + CAV i.m. simultaneously at day 10 of age (all progenies); 3) CAV i.m. at day 1 and FAV orally at day 10 of age (all progenies). The induction of IBH/HP in these progenies was evaluated throughout a 10-day period. Both breeder groups vaccinated against FAV and those vaccinated against CAV increased virus neutralizing specific antibodies. Challenge strategy 1 showed 26.6% mortality in control progeny chickens and 13.3% in the progeny of FAV-vaccinated breeders. Presence of lesions in the liver of these groups showed no significant differences (P > 0.05), suggesting a discreet protective effect of the vaccine. Challenge strategy 2 showed 29.4% mortality in controls and 94% of chickens showed hepatic inclusion bodies (HIB). Single CAV vaccination of breeders did not demonstrate a beneficial effect, with both mortality and liver lesions resembling the nonvaccinated controls. FAV vaccination of breeders significantly reduced both mortality (7.4%) and liver lesions (26% HIB) (P < 0.05), providing protection against this challenge strategy. Dual vaccination of breeders with FAV and CAV proved to be necessary to achieve maximum protection of the progeny (no mortality and 7% HIB). Challenge strategy 3 produced no mortality but consistent liver damage in controls (96% HIB). In this case, both CAV and FAV + CAV-vaccinated breeders showed best protection results in terms of liver histopathology (8% and 0% HIB, respectively). FAV vaccination alone produced 24% HIB, similar to challenge strategy 2, demonstrating a lower protective effect.

Adenoviridae Infections↗
PubMedDate not supplied

Field trial in commercial broilers with a multivalent in ovo vaccine comprising a mixture of live viral vaccines against Marek's disease, infectious bursal disease, Newcastle disease, and fowl pox.

A multivalent in ovo vaccine (MIV) was tested for safety and efficacy in a commercial broiler complex. The MIV comprised five replicating live viruses including serotypes 1, 2, and 3 of Marek's disease virus (MDV), an intermediate infectious bursal disease virus (IBDV) and a recombinant fowl poxvirus (FPV) vector vaccine containing HN and F genes of Newcastle disease virus (NDV). The performance of MIV-vaccinated broilers was compared with that of hatchmates that received turkey herpesvirus (HVT) alone (routinely used in ovo vaccine in the broiler complex). The chickens that hatched from the MIV-injected and HVT-injected eggs were raised under commercial conditions in six barns. Barn 1 housed 17,853 MIV-vaccinated chickens and each of the barns 2-6 housed 18,472-22,798 HVT-vaccinated chickens. The HVT-vaccinated chickens were given infectious bronchitis virus (IBV) and NDV vaccines at hatch and at 2 wk of age. The MIV-vaccinated chickens received IBV vaccine at hatch and IBV + NDV at 2 wk of age. The relative values of hatchability of eggs, livability and weight gain of chickens, and condemnation rates at processing were comparable between the MIV and the HVT groups (P > 0.05). Chickens from the MIV- and the HVT-vaccinated groups were challenged with virulent viruses under laboratory conditions. The resistance of vaccinated chickens against Marek's disease could not be assessed because of high natural resistance of unvaccinated commercial broilers to virulent MDV. The relative resistances of the MIV- and the HVT-vaccinated groups, respectively, against other virulent viruses were as follows: IBDV, 100% for both groups; NDV, 81% vs. 19%; FPV, 86% vs. 0%. The successful use of MIV under field conditions expands the usefulness of the in ovo technology for poultry.

Animals↗
PubMedDate not supplied

Origins of coccidiosis research in the fowl--the first fifty years.

In 1910, H. B. Fantham described the life cycle of a coccidian parasite in birds. Fantham was a parasitologist at Cambridge University in the United Kingdom working for an enquiry into diseases affecting the red grouse. Despite the growing importance of the poultry industry and the realization that coccidiosis was an important disease of the fowl, little further work was carried out in the United Kingdom until coccidiosis research was initiated at the Veterinary Laboratory, Weybridge almost 30 yr later. Further progress depended upon research carried out at academic and agricultural institutions in the United States. E. E. Tyzzer at Harvard University provided the solid foundation upon which our present knowledge of coccidiosis, and the species of Eimeria involved in the disease, is based. Agricultural experiment stations (AESs) throughout the nation played an important role in communicating advances to the agricultural community. W. T. Johnson at Western Washington and, subsequently, Oregon AES made significant contributions to our understanding of the disease, as did C. A. Herrick and coworkers at Wisconsin AES, J. P. Delaplane and coworkers at Rhode Island AES, and P. P. Levine at Cornell University.

Animals↗
PubMedDate not supplied

A consideration of previously uncharacterized fowl poxvirus unidirectional and bidirectional late promoters for inclusion in homologous recombinant vaccines.

Because of the limited analysis of fowl poxvirus (FPV) promoters, expression of foreign proteins by recombinant FPV has usually been directed by heterologous vaccinia virus or synthetic poxvirus promoters. Thus, the impact of completely homologous recombinant virus vaccines has yet to be realized by the poultry industry. In an effort to increase the availability of such transcriptional regulatory elements, the modulation of gene expression by six previously uncharacterized FPV late promoters was examined. To simplify this comparison, each promoter region was separately coupled to the same reporter gene (lacZ) in individual plasmid constructs, and their activities in transfected, virus-infected cells were monitored. In each of the four selected unidirectional transcriptional regulatory elements as well as a 30-base pair representative of the bidirectional promoter region, the predicted temporal specificity of expressing at late stages of virus replicative cycle was verified. Stable lacZ gene transcripts arising from each plasmid varied less than threefold in quantity, whereas the amounts of beta-galactosidase product ranged within a 130-fold interval. Only the promoter that naturally regulates expression of the A type inclusion body protein gene directed production of beta-galactosidase at a level comparable with that associated with the strong vaccinia virus P11 promoter. Because one of the remaining unidirectional transcriptional regulatory elements, P174, was only 2.4-fold less efficient, both of these promoters, P174 and P190, should be satisfactory for directing the expression of poultry pathogen genes inserted into the genomes of FPV recombinant vaccines.

Animals↗
PubMedDate not supplied

Molecular characterization of reticuloendotheliosis virus insertions in the genome of field and vaccine strains of fowl poxvirus.

Evidence of the widespread occurrence of reticuloendotheliosis virus (REV) sequence insertions in fowl poxvirus (FPV) genome of field isolates and vaccine strains has increased in recent years. However, only those strains carrying a near intact REV provirus are more likely to cause problems in the field. Detection of the intact provirus or REV protein expression from FPV stocks has proven to be technically difficult. The objective of the present study was to evaluate current and newly developed REV and FPV polymerase chain reaction (PCR) assays to detect the presence of REV provirus in FPV samples. The second objective was to characterize REV insertions among recent "variant" FPV field isolates and vaccine strains. With REV, FPV, and heterologous REV-FPV primers, five FPV field isolates and four commercial vaccines were analyzed by PCR and nucleotide sequence analysis. Intact and truncated REV 5' long terminal repeat (LTR) sequences were detected in all FPV field isolates and vaccine strains, indicating heterogeneous REV genome populations. However only truncated 3' LTR and envelope sequences were detected among field isolates and in one vaccine strain. Amplifications of the REV envelope and 3' LTR provided strong evidence to indicate that these isolates carry a near intact REV genome. Three of the four FPV vaccine strains analyzed carried a solo complete or truncated 5' LTR sequence, indicating that intact REV provirus was not present. Comparison of PCR assays indicated that assays amplifying REV envelope and REV 3' LTR sequences provided a more accurate assessment of REV provirus than PCR assays that amplify the REV 5' LTR region. Therefore, to differentiate FPV strains that carry intact REV provirus from those that carry solo 5' LTR sequences, positive PCR results with primers that amplify the 5' LTR should be confirmed with more specific PCR assays, such as the envelope, or the REV 3' LTR PCR.

Animals↗
PubMedDate not supplied

Protection and synergism by recombinant fowl pox vaccines expressing multiple genes from Marek's disease virus.

Recombinant fowl poxviruses (rFPVs) were constructed to express genes from serotype 1 Marek's disease virus (MDV) coding for glycoproteins B, E, I, H, and UL32 (gB1, gE, gI, gH, and UL32). An additional rFPV was constructed to contain four MDV genes (gB1, gE, gI, and UL32). These rFPVs were evaluated for their ability to protect maternal antibody-positive chickens against challenge with highly virulent MDV isolates. The protection induced by a single rFPV/gB1 (42%) confirmed our previous finding. The protection induced by rFPV/gI (43%), rFPV/gB1UL32 (46%), rFPV/gB1gEgI (72%), and rFPV/gB1gEgIUL32 (70%) contributed to additional knowledge on MDV genes involved in protective immunity. In contrast, the rFPV containing gE, gH, or UL32 did not induce significant protection compared with turkey herpesvirus (HVT). Levels of protection by rFPV/gB1 and rFPV/gl were comparable with that of HVT. Only gB1 and gI conferred synergism in rFPV containing these two genes. Protection by both rFPV/gB1gEgI (72%) and rFPV/gB1gEgIUL32(70%) against Marek's disease was significantly enhanced compared with a single gB1 or gI gene (40%). This protective synergism between gB1 and gI in rFPVs may be the basis for better protection when bivalent vaccines between serotypes 2 and 3 were used. When rFPV/gB1gIgEUL32 + HVT were used as vaccine against Md5 challenge, the protection was significantly enhanced (94%). This synergism between rFPV/gB1gIgEUL32 and HVT indicates additional genes yet to be discovered in HVT may be responsible for the enhancement.

Animals↗