Genetic characterization, pathogenicity, and protection studies with an avian adenovirus isolate associated with inclusion body hepatitis

An avian adenovirus (AAV) was isolated from liver samples of two 2-wk-old broiler-breeder flocks obtained from grandparents vaccinated at 10 and 17 wks of age with an autogenous inactivated vaccine containing the European AAV 8 (8565 strain) and 11 (1047 strain) serotypes (AAV8/11 vaccine). Affected broiler-breeders exhibited clinical signs and macroscopic and microscopic lesions associated with inclusion body hepatitis (IBH). The isolated adenovirus, identified as Stanford, was molecularly characterized as European serotype 9. The pathogenicity of the Stanford strain was confirmed after inoculation of specific-pathogen-free (SPF) chickens at 1-7 days of age, causing 100% and 20% mortality, respectively. The level of protection against IBH was evaluated in two broiler-breeder progenies from AAV 8/11-vaccinated grandparent flocks and a commercial broiler flock by challenge at 1 or 7 days of age with the AAV 8 and 11 serotypes and/or the Stanford strain. The broiler-breeder progenies and the commercial broiler flock exhibited protection against IBH after challenge. No significant differences in mean body weights were observed at 3 wk of age in any of the evaluated groups. We conclude that broiler-breeder progenies from 30- to 50-wk-old grandparents vaccinated with the AAV 8/11 vaccine were adequately protected against challenge with the AAV 8 and 11 serotypes and the Stanford strain.     

Hagberg falling number and the nutritional value of wheat in broiler chicken diets

1. Three experiments were performed to study the effect of Hagberg falling number in wheat on performance, nutrient digestibility and AMEN in broilers. In two experiments, one hard and one soft wheat variety were used to study the interaction between falling number and hardness of wheat with regard to nutritional value. In these experiments, wheat batches with high falling number when harvested under dry conditions were used in broiler diets. 2. Wheat with reduced falling numbers (high, medium and low) was obtained by controlled germination. In the third experiment, wheat with reduced falling numbers were obtained by delayed harvesting times. 3. In each experiment, a total of 4 cereal batches with different falling numbers from each wheat variety were used to produce corresponding experimental diets with wheat as the major ingredient. Each diet was fed to broiler chickens ad libitum from d 1 to d 17 of age. 4. There was no consistent effect of falling number on performance. Low falling number did not improve feed utilisation or AMEN compared to the original wheat, despite a higher AMEN associated with higher starch digestibility. This phenomenon was not observed after reduction of falling number by delayed harvesting. Apparently, natural reduction of falling number resulted in enhanced degradation of arabinoxylans compared to controlled germination.     

Pharmacological characterisation of the electrically evoked release of monoamines from chicken brain in vitro

1. A study was conducted to develop an in vitro model for examining the basal and electrical-stimulation-induced release of [3H]monoamines from chicken hyperstriatal neurones in order to demonstrate the presence of presynaptic autoreceptors for the three main monoamine transmitters: noradrenaline, dopamine and 5-HT. 2. Two sets of experiments were carried out: the first was to evaluate the effect of calcium and tetrodotoxin (TTX, sodium channel conductance inhibitor) in order to demonstrate that evoked release of monoamines was a consequence of exocytotic processes; the second to investigate the effect of selective agonists and antagonists on neurotransmitter release. 3. Ross and Cobb broiler chickens of either sex (approximately 7 to 8 weeks old) were used. Slices of hyperstriatal tissue were preincubated with [3H]noradrenaline, [3H]dopamine or [3H]5-hydroxytryptamine (5-HT), washed, perfused and electrically stimulated at three time points (S1, S2 and S3) which released [3H]noradrenaline, [3H]dopamine and [3H]5-HT, as determined by scintillation spectrometry. 4. When calcium was removed from, or TTX added to, the superfusion medium prior to and including the second period of electrical stimulation (S2) the evoked releases of [3H]noradrenaline, [3H]dopamine and [3H]5-HT at S2 were abolished. 5. In the presence of the selective alpha2-adrenoceptor agonist UK 14304 during the S2 period, the S2/S1 ratio was lower than the control ratio due to a reduction in the stimulated release of [3H]noradrenaline. The selective alpha2-adrenoceptor antagonist RX 821002 blocked the UK 14304-induced reduction of evoked release and the S2/S1 ratio was similar to the control ratio. 6. The D2-like receptor agonist quinpirole reduced the S2/S1 ratio for [3H]dopamine release, an effect blocked by the antagonist AJ 76. The 5-HT1B receptor agonist CP 94253 during S2 reduced the S2/S1 ratio due to a reduction in evoked [3H]5-HT. This effect was blocked by the 5-HT1B receptor antagonist GR 55562. 7. The results demonstrate, for the first time, the functional presence of presynaptic alpha2-adrenoceptors, presynaptic 5-HT1B autoreceptors and presynaptic D2-like autoreceptors in broiler chicken hyperstriatal neurones in vitro.     

Evaluation of the effectiveness of two infectious bronchitis virus vaccine programs for preventing disease caused by a California IBV field isolate

Infectious bronchitis virus CA99 serotype was isolated from several broiler flocks in Northern California. The virus caused late-onset respiratory disease and increased airsacculitis condemnation in affected flocks despite the use of an established infectious bronchitis virus vaccination program. An experimental study compared Holland/Arkansas and Massachusetts/Arkansas vaccination protocols to determine the efficacy of commercial infectious bronchitis virus vaccines in reducing respiratory disease and airsacculitis lesions found at processing that were associated with a CA99 field isolate. All vaccination groups were given Massachusetts/Connecticut strains of infectious bronchitis virus vaccines at age 1 day followed by vaccination with either Holland/ Arkansas or Massachusetts/Arkansas vaccine strains at 18 days of age. Birds were challenged at age 31 days with a CA99 field isolate. Gross pathology, histopathology, and virus isolation were evaluated. Chickens vaccinated with Holland/Arkansas had marginally better protection against CA99 challenge than chickens vaccinated with Massachusetts/Arkansas, although differences were not statistically significant.     

Epidemiologic investigation of an outbreak of goose parvovirus infection in Sweden

An outbreak of goose parvovirus (GPV) infection on a Swedish goose farm in the spring of 2004 increased the mortality rates from 2% in the early unaffected hatches to 90% and 99% respectively in the two hatches following virus introduction and 40% in goslings hatched later in the same breeding season. In this paper we describe the clinical observations, diagnostic procedures, and epidemiologic investigation carried out to elucidate the source of the infection. The diagnosis was confirmed by serology, virus isolation, and sequence analysis of a 493-bp-long fragment of the VP1 gene. Phylogenetically the causative virus was closely related to pathogenic GPV strains isolated in 2003 and 2004 from Poland and the United Kingdom, respectively. The Swedish isolate exhibited less homology with pathogenic strains from Hungary and Asia and with attenuated vaccine strains. The epidemiologic investigation showed that the virus was first introduced to a contract farm (farm A) and then was transferred with newly hatched goslings to the farm that had submitted the birds for necropsy (index farm). The exact time and source of the virus introduction to farm A could not be determined with absolute certainty. Possible sources of the infection included backyard goose eggs that had been delivered to farm A for subcontract incubation and hatching, wild geese that frequented the flock of breeding geese on pasture on farm A, and a clutch of Canada goose eggs (Branta canadensis) that had been produced by wild geese and was hatched in the same machine as the eggs produced by farm A.     

Rapid quantification of avian eggshell microstructure and crystallographic-texture using two-dimensional X-ray diffraction

1. It is important to quantify eggshell microstructure because it may influence eggshell mechanical properties and could be of importance for eggshell quality assessment. It also provides information about changes in eggshell associated with different hen physiological conditions. 2. This paper describes an alternative methodology which, based on 2D X-ray diffraction, allows eggshell microstructure quantification (for example, crystal size and orientation) much more efficiently than using traditional techniques (optical or scanning electron microscopy and conventional X-ray diffraction). 3. For such analyses, an X-ray diffractometer equipped with an area detector is used. Such equipment collects the diffraction pattern of the sample in one single exposure (taking a few tens of seconds), thus considerably reducing data acquisition time. 4. The two-dimensional diffraction patterns obtained contain detailed information about size and crystallographic orientation of crystals forming the eggshell. This information can be automatically extracted using specially designed software. 5. Access to the software is available on a website, the address of which is given in the Materials and Methods section. 6. In this paper, specific examples of how these analyses are applied are set out and recommendations for obtaining different types of microstructure information are given. In addition, the limitations of the technique are discussed.     

Interactive effect of ractopamine and dietary fat source on quality characteristics of fresh pork bellies

Crossbred pigs (n = 216) were used to test the interaction, if any, of ractopamine (RAC) and dietary fat source on the characteristics of fresh pork bellies. Pigs were blocked by BW (77.6 +/- 6.5 kg) and allotted randomly to pens (6 pigs/pen). After receiving a common diet devoid of RAC for 2 wk, pens within blocks were assigned randomly to 1 of 4 treatments arranged in a 2 x 2 factorial design, with 5% fat (beef tallow vs. soybean oil) and RAC (0 vs. 10 mg/kg). At the conclusion of the 35-d feeding period, pigs were slaughtered at a commercial pork packing plant (average BW of 108.8 +/- 0.6 kg), and fresh bellies were captured during carcass fabrication. Neither RAC (P = 0.362) nor fat source (P = 0.247) affected belly thickness. Subjective (bar-suspension) or objective (compression test) measures of belly firmness were not (P > or = 0.148) affected by the inclusion of RAC in the diet; however, bellies from pigs fed soybean oil (SBO) were softer than those from pigs fed beef tallow (BT), as indicated by perpendicular (P < or = 0.005) and parallel (P < 0.001) suspensions. Moreover, bellies from BT-fed pigs required more (P = 0.096) force to compress 50% of their thickness than bellies from SBO-fed pigs (52.29 vs. 43.51 kg). Color (L*, a*, and b* values) of the belly lean and fat was not (P > or = 0.131) affected by RAC, and lean color was similar (P > or = 0.262) between fat sources; however, belly fat from BT-fed pigs was lighter (P = 0.030) and redder (P = 0.013) in color than belly fat from SBO-fed pigs. Bellies of SBO-fed pigs had greater (P < 0.001) proportions of PUFA and lower (P < 0.001) proportions of SFA and MUFA than belly fat from pigs fed BT. Regardless of the RAC inclusion level, PUFA:SFA and iodine values were lower in belly fat from pigs fed BT than SBO; however, within SBO-fed pigs, PUFA:SFA and iodine values were further increased by feeding RAC (RAC x fat source, P < 0.001). As expected, dietary fat source altered the fatty acid composition of fresh pork bellies, which subsequently impacted fresh belly firmness. Interestingly, including RAC in swine finishing diets exacerbated the effect of feeding SBO on pork fat polyunsaturation.     

The effect of induced forelimb lameness on thoracolumbar kinematics during treadmill locomotion

REASONS FOR PERFORMING STUDY: Lameness has often been suggested to result in altered movement of the back, but there are no detailed studies describing such a relationship in quantitative terms. OBJECTIVES: To quantify the effect of induced subtle forelimb lameness on thoracolumbar kinematics in the horse. METHODS: Kinematics of 6 riding horses was measured at walk and at trot on a treadmill before and after the induction of reversible forelimb lameness grade 2 (AAEP scale 1-5). Ground reaction forces (GRF) for individual limbs were calculated from kinematics. RESULTS: The horses significantly unloaded the painful limb by 11.5% at trot, while unloading at walk was not significant. The overall flexion-extension range of back motion decreased on average by 0.2 degrees at walk and increased by 3.3 degrees at trot (P<0.05). Changes in angular motion patterns of vertebral joints were noted only at trot, with an increase in flexion of 0.9 degrees at T10 (i.e. angle between T6, T10 and T13) during the stance phase of the sound diagonal and an increase in extension of the thoracolumbar area during stance of the lame diagonal (0.7degrees at T13, 0.8 degres at T17, 0.5 degres at L1, 0.4 degrees at L3 and 0.3 degrees at L5) (P<0.05). Lameness further caused a lateral bending of the cranial thoracic vertebral column towards the lame side (1.3 degrees at T10 and 0.9 degrees at T13) (P<0.05) during stance of the lame diagonal. CONCLUSIONS: Both range of motion and vertebral angular motion patterns are affected by subtle forelimb lameness. At walk, the effect is minimal, at trot the horses increased the vertebral range of motion and changed the pattern of thoracolumbar motion in the sagittal and horizontal planes, presumably in an attempt to move the centre of gravity away from the lame side and reduce the force on the affected limb. POTENTIAL RELEVANCE: Subtle forelimb lameness affects thoracolumbar kinematics. Future studies should aim at elucidating whether the altered movement patterns lead to back and/or neck dysfunction in the case of chronic lameness.     

Expression of porcine endogenous retroviruses (PERVs) in melanomas of Munich miniature swine (MMS) Troll

Porcine endogenous retroviruses (PERVs) are integrated in the genome of all pig breeds. Since some of them are able to infect human cells, they might represent a risk for xenotransplantation using pig cells or organs. However, the expression and biological role of PERVs in healthy pigs as well as in porcine tumours is largely unknown. Since we and others have recently shown overexpression of a human endogenous retrovirus, HERV-K, in human melanomas, we studied the expression of PERVs in melanomas of selectively bred Munich miniature swine (MMS) Troll. This breeding herd of MMS Troll is characterised by a high prevalence of melanomas, which histologically resemble various types of cutaneous melanomas in humans. Several genetic factors have been defined when studying inheritance of melanomas and melanocytic nevi in MMS Troll. Here we show that the polytropic PERV-A and PERV-B as well as the ecotropic PERV-C are present in the genome of all melanoma bearing MMS Troll investigated. Most interestingly, in the spleen, but not in other organs, recombinant PERV-A/C proviruses were found. PERV expression was found elevated in melanomas when compared to normal skin and viral proteins were expressed in melanomas and pulmonary metastasis-derived melanoma cell cultures. During passaging of these cells in vitro the expression of PERV mRNA and protein increased and virus particles were released as shown by RT activity in the supernatant and by electron microscopy. Genomic RNA of PERV-A, -B and -C were found in pelleted virus particles. Although PERV expression was elevated in melanomas and pulmonary metastasis-derived cell cultures, the function of the virus in tumour development is still unclear.     

Characterization of multidrug resistant Salmonella recovered from diseased animals

Three hundred and eighty Salmonella isolates recovered from animal diagnostic samples obtained from four state veterinary diagnostic laboratories (AZ, NC, MO, and TN) between 2002 and 2003 were tested for antimicrobial susceptibilities and further characterized for bla(CMY) beta-lactamase genes, class 1 integrons and genetic relatedness using PFGE. Forty-seven serovars were identified, the most common being S. Typhimurium (26%), S. Heidelberg (9%), S, Dublin (8%), S. Newport (8%), S. Derby (7%), and S. Choleraesuis (7%). Three hundred and thirteen (82%) isolates were resistant to at least one antimicrobial, and 265 (70%) to three or more antimicrobials. Resistance was most often observed to tetracycline (78%), followed by streptomycin (73%), sulfamethoxazole (68%), and ampicillin (54%), and to a lesser extent chloramphenicol (37%), kanamycin (37%), amoxicillin-clavulanic acid (20%), and ceftiofur (17%). With regards to animal of origin, swine Salmonella isolates displayed the highest rate of resistance, being resistant to at least one antimicrobial (92%), followed by those recovered from turkey (91%), cattle (77%), chicken (68%), and equine (20%). Serovars commonly showing multidrug resistance (MDR) to > or =9 antimicrobials were S. Uganda (100%), S. Agona (79%), and S. Newport (62%), compared to S. Heidelberg (11%) and S. Typhimurium (7%). Class-1 integrons were detected in 43% of all isolates, and were found to contain aadA, aadB, dhfr, cmlA and sat1 gene cassettes alone or in various combinations. All ceftiofur resistant isolates (n=66) carried the bla(CMY) beta-lactamase gene. A total of 230 PFGE patterns were generated among the 380 isolates tested using XbaI, indicating extensive genetic diversity across recovered Salmonella serovars, however, several MDR clones were repeatedly recovered from different diseased animals.