Pathological changes associated with white striping in broiler breast muscles.

White striping is a condition in broiler chickens characterized grossly by the occurrence of white striations, seen parallel to the direction of muscle fibers, on broiler breast fillets and thighs. Based on visual evaluation of the intensity of white striping, breast fillets can be categorized into normal (NORM), moderate (MOD), and severe (SEV) categories. This study was undertaken to evaluate the details of changes in histology as well as proximate composition occurring in the fillets with respect to the 3 degrees of white striping. In experiment 1, representative breast fillets for each degree of white striping (n = 20) were collected from 45-d-old broilers, approximately 2 h postmortem. From each fillet, 2 skeletal muscle samples were obtained and fixed in 10% neutral buffered formalin. To identify and differentiate the histological changes, slides were prepared and stained using hematoxylin and eosin, Masson's Trichrome, and Oil Red O stains. In experiment 2, samples with 3 degrees of white striping were collected from 57-d-old birds for conducting proximate analysis. Major histopathological changes observed in the MOD and SEV samples consisted of loss of cross striations, variability in fiber size, floccular/vacuolar degeneration and lysis of fibers, mild mineralization, occasional regeneration (nuclear rowing and multinucleated cells), mononuclear cell infiltration, lipidosis, and interstitial inflammation and fibrosis. Microscopic lesions were visually scored for degeneration and necrosis, fibrosis, and lipidosis. The scale used to score the samples ranged from 0 (normal) to 3 (severe). There was an increase (P < 0.05) in mean scores for degenerative or necrotic lesions, fibrosis, and lipidosis as the degree of white striping increased from NORM to SEV. The results from the histopathological study were supported by the findings from proximate analysis confirming that the fat and protein contents of muscle increased (P < 0.05) and decreased (P < 0.05), respectively, as the degree of white striping increased. In conclusion, the histopathological changes occurring in white striping indicate a degenerative myopathy that could be associated with increased growth rate in birds.

Joined by geochemistry, divided by history: PCBs and PBDEs in Strait of Georgia sediments.

Polychlorinated biphenyls (PCBs) are relict contaminants, while polybrominated diphenyl ethers (PBDEs) are in increasing use. Using sediment cores collected in the Strait of Georgia, we demonstrate that the surface sediment concentration of PCBs is largely determined by environmental processes, such as sediment accumulation and mixing rates, while that of PBDEs is strongly influenced by proximity to source. The Iona Island wastewater outfall appears to be a primary pathway for PBDEs. As well, Vancouver Harbour is highly contaminated with both classes of chemical. BDE-209, the main component of deca-BDE, is the dominant PBDE congener. Environmental debromination is not evident. Currently, the ranges of the surface concentration of PCBs and PBDEs are similar to one another, but that will change in the future, as the concentration of PBDEs continues to rise. The experience with PCBs suggests that if PBDEs were banned today, it would take decades for inorganic sediment to bury them.

The efficacy of raw and concentrated bentonite clay in reducing the toxic effects of aflatoxin in broiler chicks.

The objective of this study was to evaluate the efficacy of two adsorbents, a raw bentonite clay (RC) and a concentrated bentonite clay (CC), in ameliorating the toxic effects of aflatoxin B1 (AFB1). Results of the in vitro study (pH 3.0) indicated the CC adsorbed more AFB1 than RC (93.39 mg/g vs. 79.30 mg/g) suggesting that CC may be more effective than RC in reducing the toxic effects of AFB1. One hundred and eighty day-old straight run broiler chicks were assigned to 6 replicate pens of 5 chicks each and assigned to 6 dietary treatments from hatch to day 21. Dietary treatments included: 1) basal diet (BD) containing no AFB1 or adsorbents; 2) BD plus 0.50% RC; 3) BD plus 0.50% CC; 4) BD plus 2.0 mg AFB1/kg; 5) BD plus 2.0 mg AFB1/kg plus 0.50% RC; and 6) BD plus 2.0 mg AFB1/kg plus 0.50% CC. Dietary AFB1 concentrations were confirmed by analysis and diets were screened for other mycotoxins prior to the start of the experiment. The addition of AFB1 to the feed reduced (P < 0.05) growth performance and increased (P < 0.05) relative liver weight (RLW) and kidney weight (RKW) of chicks fed AFB1 compared to control chicks on day 21. These changes were ameliorated (P < 0.05) by the addition of RC and CC to the AFB1 diet. Mild to moderate lesions of aflatoxicosis (2.25) were observed in chicks fed AFB1 alone on day 21. The addition of both RC and CC to the AFB1 diet decreased (P < 0.05) but did not prevent liver lesions (0.92 and 1.42, respectively). Results indicate that both RC and CC were effective in reducing the toxic effects of AFB1, however the cost of processing of CC would make the RC a more economical product for reducing the effects of AFB1 in young broiler chicks.

Ribavirin mitigates wart growth in rabbits at early stages of infection with cottontail rabbit papillomavirus.

The challenge to develop antiviral agents effective against DNA viruses such as human papillomavirus (HPV) has been dependent on finding an animal model which mimics the human forms of the disease. We have used an existing model system for the purpose of measuring the effect of antiviral drugs on the inhibition of growth of these lesions. This was based upon domestic rabbits which efficiently grow cutaneous papillomas (warts) when infected with cottontail rabbit papillomavirus (CRPV). One agent which had shown significant success in achieving these goals was ribavirin. Ribavirin was administered intradermally shortly prior to infection at multiple sites with CRPV. Following daily injections of this drug for eight weeks, we have shown a dose-dependent response which had markedly reduced the number of warts, the time of first appearance of warts and reduced the tumor mass as compared to placebo-treated control animals. At the highest dose of ribavirin tested, 30 mg/kg/day, compared to controls, the average reduction in the number of warts was 52%, the average time of first appearance of warts was 49% longer, and the average mass of the warts was reduced by 98%. No detectable antibodies to CRPV were observed in any of the animals. The only side effects which were observed was focal alopecia, and a decrease in body growth upon prolonged treatment, both of which were completely reversible. Pharmacokinetic studies established the metabolism of ribavirin over a 24-h period of time. Ribavirin administered beginning 12 or 30 days post-infection, while not reducing the number of warts, slightly retarded the growth of warts as determined by date of first appearance of warts and mass of warts.

Fiberoptic cardioscopy under cardiopulmonary bypass: potential for cardioscopic surgery?

Attempts at cardioscopy have been hampered by the inability to see through blood. We describe a new method of intracardiac fiberscopic visualization performed in sheep using cardiopulmonary bypass, asanguineous cardioplegic arrest, and replacement of intracardiac blood with a clear fluid. Right heart endoscopic visualization was achieved in 4 sheep using a method that could allow extrapolation to a percutaneous approach. Two of these animals were weaned successfully from cardiopulmonary bypass. Venting and flushing aspects of the procedure were of primary importance. Further development of this technique may open the way to percutaneous endoscopically guided intracardiac operations in humans.

The mechanism of blood flow in cardiopulmonary resuscitation--introducing the lung pump.

The mechanism of blood flow in cardiopulmonary resuscitation remains controversial. Inconsistencies in the cardiac pump model necessitated the development of the thoracic pump. Neither hypothesis alone can account for blood flow during external massage. Using the postulates that a cyclical pump has three components, an inlet valve, a compression chamber and an outlet valve, we propose the lung pump hypothesis where these components are represented by the pulmonary valve; the lung vasculature/left atrium/left ventricle; and the aortic valve. We explore the mechanism and effect of this hypothesis and match it to previous observations. The implications to cardiopulmonary resuscitation are addressed.

Immunohistochemical detection of avian pneumovirus in formalin-fixed tissues.

An immunohistochemical staining technique (IHC) was developed to detect avian pneumovirus (APV) antigen in formalin-fixed, paraffin-embedded tissue sections using streptavidin-biotin immunoperoxidase staining. Samples of nasal turbinates and infraorbital sinuses were collected from 4-week-old poults experimentally inoculated with APV and from older turkeys infected during naturally occurring outbreaks of avian pneumovirus. Tissue was fixed in 10% buffered neutral formalin, embedded in paraffin, sectioned and stained. Inflammatory changes were observed microscopically in the mucosa and submucosa of the nasal turbinates and infraorbital sinuses of both experimentally inoculated poults and naturally infected birds. Viral antigen was detected by IHC in the ciliated epithelial cells of nasal turbinates and infraorbital sinuses.

Isolation of avian pneumovirus from an outbreak of respiratory illness in Minnesota turkeys.

Antibodies to avian pneumovirus (APV) were first detected in Minnesota turkeys in 1997. Virus isolation was attempted on 32 samples (28 tracheal swabs, 4 pools of trachea and turbinates) that were positive for APV by reverse transcriptase polymerase chain reaction (RT-PCR). The cell cultures used were chicken embryo fibroblast (CEF), Vero cells, and QT-35 cells. Five virus isolates were obtained from these samples, and the identity of the isolates was confirmed by RT-PCR. Four isolates were obtained by inoculation of CEF cells, and 1 isolate was obtained in QT-35 cells after 3-7 blind passages in cell cultures. Vero cells did not yield any isolate on primary isolation; however, all 5 isolates could be adapted to grow in Vero cells following primary isolation in CEF or QT-35 cells. This is the first report of isolation of APV in Minnesota and also the first report of primary isolation of APV in QT-35 cells.

Antimicrobial susceptibility and serotypes of Actinobacillus (Haemophilus) pleuropneumoniae recovered from Missouri swine.

The antimicrobial susceptibility of 73 Actinobacillus (Haemophilus) pleuropneumoniae isolates from swine in Missouri was determined with a microdilution minimal inhibitory concentration test system. Serotyping was accomplished by means of co-agglutination. Serotype 1 (39/73) and serotype 5 (30/73) were commonly found, whereas serotype 7 (4/73) was infrequently encountered. Most isolates (MIC90) were found susceptible to ampicillin (amoxicillin), cephalothin, penicillin, erythromycin, gentamicin, and kanamycin. Marked resistance was found with oxytetracycline, tylosin, and sulfadimethoxine. The data indicate that use of ampicillin (amoxicillin) or penicillin may correlate well with the favorable outcome of treatment.

Isolation of swine infertility and respiratory syndrome virus (isolate ATCC VR-2332) in North America and experimental reproduction of the disease in gnotobiotic pigs.

A recent epizootic of swine infertility and respiratory syndrome (SIRS) in a Minnesota swine herd was investigated. Examination of a sow, neonatal piglets, and stillborn fetuses obtained during the epizootic from the affected herd revealed interstitial pneumonitis, lymphomononuclear encephalitis, and lymphomononuclear myocarditis in the piglets and focal vasculitis in the brain of the sow. Fetuses did not have microscopic lesions. No cause for the infertility and respiratory syndrome was determined. Therefore, attempts were made to experimentally reproduce the disease. Eleven 3-day-old gnotobiotic piglets exposed intranasally to tissue homogenates of piglets from the epizootic became inappetent and febrile by 2-4 days postexposure and had interstitial pneumonitis and encephalitis similar to that seen in the field outbreak. After 2 blind passages in gnotobiotic piglets, tissue homogenates were cultured on continuous cell line CL2621, and a cytopathic virus (ATCC VR-2332), provisionally named SIRS virus, was isolated. Gnotobiotic piglets exposed intranasally to the SIRS virus developed clinical signs and microscopic lesions that were the same as those in piglets exposed to the tissue homogenates, and the virus was reisolated from their lungs. This is the first isolate of SIRS virus in the United States that fulfills Koch's postulates in producing the respiratory form of the disease in gnotobiotic piglets and the first report of isolation and propagation of the virus on a continuous cell line (CL2621). The virus is designated as American Type Culture Collection VR-2332.

Early chick mortality associated with rupture of the yolk sac.

Rupture of the yolk sac was identified in leghorn chicks from six placements of a commercial hatchery that had high mortality (up to 3%) during the first 3 days. The abdomens of these chicks were filled with cloudy yellowish fluid; kidneys were swollen, and many chicks also had severe visceral urate deposition (visceral gout). No significant bacterial growth was detected. The hatchery had recently hired new sexors. After the sexors were cautioned to handle the chicks more carefully, the problem abruptly subsided. In an experimental attempt to reproduce the condition, a graded series of doses (0, 0.5, 1.0, 2.0, and 4.0 ml) of egg yolk were injected intraperitoneally into five groups of twenty 1-day-old chicks. Twelve of the 20 chicks that received the 4-ml dose died. They developed swollen, pale kidneys and had cloudy yellow fluid in the abdominal cavity, but no visceral urate deposition. Only six of the remaining 80 chicks that received lesser doses or no yolk died.

Susceptibility of an avian pneumovirus isolated from Minnesota turkeys to physical and chemical agents.

Survival characteristics of a Minnesota avian pneumovirus (APV) isolated from a turkey nasoturbinate, propagated in tissue culture, and exposed to various physiochemical treatments were determined. These characteristics included survivability under various conditions. Specifically, APV was viable at temperatures of -70 C and -20 C for over 26 wk, 4 C for less than 12 wk, 20 C for less than 4 wk, 37 C for 48 hr, and 50 C for less than 6 hr. In addition, APV survived 12 freeze/thaw cycles with no loss of activity. With a variable pH for 1 hr, the titer was unaffected between the levels of pH 5 and 9. Several disinfectants, including quaternary ammonia, ethanol, iodophor, a phenol derivative, a biguanide, and bleach, were all effective in reducing the viability of the virus. After 7 days of drying at room temperature, the APV remained viable and was recovered on cell culture.

Ornithobacterium rhinotracheale infection in turkeys: immunoprophylaxis studies.

Ornithobacterium rhinotracheale has been shown to cause serious clinical illness and is a significant concern to the turkey industry because of its potential economic impact. In this study, 6-wk-old turkeys were vaccinated intranasally with a live or subcutaneously with a killed O. rhinotracheale vaccine. At 14 or 21 wk of age, the birds were challenged intratracheally with live O. rhinotracheale. Airsacculitis and pneumonia occurred less frequently in vaccinated birds than in unvaccinated birds after challenge with O. rhinotracheale. Ornithobacterium rhinotracheale was recovered from unvaccinated, challenged birds but not from vaccinated, challenged or from unchallenged birds. Thus, turkeys inoculated with live or killed O. rhinotracheale vaccine were protected from pathologic changes.

Susceptibility of broiler chicks to infection by avian pneumovirus of turkey origin.

In this paper we present the results of studies on the infectivity of an isolate of avian pneumovirus (APV) from turkeys to broiler chickens. Two-week-old broiler chicks free of antibodies to APV were exposed either by oculonasal or oral route with a cell cultured APV of turkey origin. Chickens from both APV-inoculated groups exhibited clinical signs that included coughing, sneezing, nasal discharge, and watery eyes during 2-8 days postinoculation. Tissue samples from birds in the APV-inoculated group were positive for APV by polymerase chain reaction (PCR) up to 9 days postinoculation. Samples of blood from both oculonasally and orally infected chickens were positive for APV. Intestinal samples from chickens infected with APV orally were positive for the presence of APV on PCR up to 9 days postinoculation. APV was reisolated from samples taken from chickens in both groups inoculated orally and oculonasally. Sera from birds exposed by the oculonasal or by the oral route showed the presence of APV-specific antibodies.

Outer membrane proteins for serologic detection of Ornithobacterium rhinotracheale infection in turkeys.

Ornithobacterium rhinotracheale (ORT) is a bacterium responsible for a respiratory disease in turkeys and chickens and has been identified as one of the emerging respiratory bacterial pathogens. The clinical signs and lesions caused by ORT are very similar to those caused by other respiratory infectious agents; therefore, an accurate diagnostic test is necessary to identify the infection. In this study, we have investigated the use of outer membrane proteins of ORT in an indirect enzyme-linked immunosorbent assay (ELISA) to detect the exposure to ORT infection. Outer membrane proteins of ORT were extracted and used as an antigen in ELISA to detect infection in turkeys exposed to different serotypes of ORT. The ELISA results were compared with the conventional serum plate agglutination test. The agglutination test detected specific antibodies for ORT in 65% of experimentally infected turkeys during the first 2 wk of infection. The ELISA detected up to 100% of infected birds for 8 wk postinfection. The results suggest that ELISA is able to detect the exposure to ORT in later stages of the infection and this assay can be used in serologic surveillance of ORT infection for poultry in the field.

A highly fatal vascular disease of broiler breeder chickens.

A broiler breeder operation experienced a sudden outbreak of high mortality in nine different houses of breeders at eight different locations. In the first house to be affected, the losses were mainly in the females, but later flocks had losses up to 6.4%, mainly in the males. Affected chickens ranged in age from 7 to 22.5 weeks. Multifocal areas of necrosis and vasculitis were found in skin, muscle, and internal organs. No involvement of the brain was identified, but many of the birds had uveitis. The outbreak lasted 3 weeks and ended without treatment. Neither infectious agents nor mycotoxins were identified.

Virulence factors of Escherichia coli associated with colisepticemia in chickens and turkeys.

Four hundred twenty turkey and 80 chicken Escherichia coli isolates from colisepticemic birds were examined for the following properties: heat-labile toxin (LT), heat-stable enterotoxin, verotoxin, colicinogenicity, hemolysin, and hydroxamate/aerobactin production. Twenty-four (5.7%) of the 420 turkey isolates and six (7.5%) of the 80 chicken isolates produced an LT that was cytotoxic for both Vero and Y-1 cells. In contrast, 48 (11.4%) of the turkey isolates and 18 (22.5%) of the chicken isolates produced a distinct LT that was cytotoxic only for Vero cells. In addition, 64 (80.0%) of the chicken isolates and 309 (74.0%) of the turkey isolates produced aerobactin. Colicinogenicity occurred in 51 (64.0%) of the chicken isolates, with 41 (51.0%) producing colicin V. By contrast, 254 (61.0%) of the turkey isolates produced a colicin, of which 176 (42.0%) produced colicin V. None of the chicken and turkey isolates produced hemolysin or heat-stable enterotoxin.

Pasteurella haemolytica as a co-pathogen in pullets and laying hens.

Pasteurella haemolytica was isolated from internal organs of laying hens and pullets of four flocks in which five incidents of increased mortality had occurred. There was inflammation of the trachea in all cases, and infectious laryngotracheitis virus was identified in three of the incidents. Rapid response to antimicrobial therapy was seen in two out of three outbreaks in the pullets. P. haemolytica was regarded as a significant secondary invader in these cases.

Seroprevalence of Ornithobacterium rhinotracheale infection in commercial laying hens in the north central region of the United States.

This study was the first to examine the seroprevalence of Ornithobacterium rhinotracheale (ORT) within a commercial egg layer population. Serum samples collected from egg production companies were examined by serum plate agglutination test (SPAT) and outer membrane protein-based enzyme-linked immunosorbent assay (ELISA). Results show that 90% of layer flocks were positive by SPAT and 100% by ELISA. Of the pullet flocks examined, 43% and 52% were positive by SPAT and ELISA, respectively. Our study indicates that the prevalence of ORT antibody is high in the commercial layer population, suggesting that this respiratory pathogen can easily spread through multiple-age layer farms from older flocks to newly housed pullet flocks.

Ornithobacterium rhinotracheale infection in commercial laying-type chickens.

Ornithobacterium rhinotracheale is a gram-negative, rod-shaped, pleomorphic bacterium that has been isolated from flocks of turkeys and broilers from around the world. Infections cause respiratory disease, mortality, and growth suppression, or clinical signs of infection may be absent. In layers, there have been few reports of disease caused by O. rhinotracheale. This is the first report of O. rhinotracheale infection in United States layer flocks.