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.     

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.     

Effect of orally administered Lactobacillus casei on porcine reproductive and respiratory syndrome (PRRS) virus vaccination in pigs

The purpose of this study was to determine whether intranasal/oral administration of probiotics can assist vaccination efficacy against an important swine pathogen, porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV). A controlled challenge trial was performed employing: (a) pigs vaccinated against PRRS and treated with a Lactobacillus casei, (b) pigs vaccinated against PRRS only, (c) pigs treated with L. casei only, and (d) pigs neither vaccinated against PRRS nor treated with L. casei. All pigs were challenged intranasally with a wild PRRSV strain. There was no difference in clinical signs or rectal temperature among the four groups. However, pigs that received L. casei gained significantly more weight than pigs that did not. Vaccinated pigs did not gain more weight than nonvaccinated pigs. Vaccinated groups had significantly fewer viraemic pigs on days post-challenge 4, 11 and 17 than nonvaccinated groups of pigs. There was no effect of probiotic on prevalence or duration of viraemia. Among viraemic pigs, there was no significant difference in mean log base(10) titer of PRRS virus among groups. These results suggest that orally administered L. casei does not affect immune response in such a way as to affect PRRS viraemia or nasal shedding. However, it still appears to provide significant benefit when administered during vaccination as indicated by the higher bodyweight gain following PRRS virus infection.     

Molecular cloning of a Mycoplasma meleagridis-specific antigenic domain endowed with a serodiagnostic potential

A recombinant phage library harbouring Mycoplasma meleagridis (MM) genomic DNA fragments was generated in the bacteriophage lambda gt11 expression vector. The library was screened for expression of MM specific antigens with a polyclonal antiserum that had been preadsorbed with antigens of the most common unrelated avian mycoplasma species. A 49-amino acid antigenic domain unique to MM was isolated, expressed in Escherichia coli, and its serodiagnostic potential was demonstrated. An antiserum raised against this MM-specific antigenic domain recognized a cluster of seven membrane-associated MM proteins with molecular masses ranging from 34 to 75 kDa. Overall, this study resulted in the identification of a potent serodiagnostic tool and revealed the complex antigenic nature of MM.     

Onset of immunity following in ovo delivery of avian metapneumovirus vaccines

Avian metapneumovirus (aMPV) is an important cause of disease in chickens and turkeys. As infection can occur early in life and spread of the virus throughout a flock is rapid, an early onset of immunity post-vaccination would be advantageous. We have studied the serological immune response and the onset of protective immunity of an aMPV vaccine delivered to chickens via the in ovo route compared to oculonasal delivery at day old. A 1000-fold lower dose delivered in ovo to chicken specific pathogen free (SPF) embryos, than vaccination at day old, provided a significantly higher antibody response. In the presence of maternally derived antibody (MDA), there was no significant difference in antibody response between the vaccination routes. However, the onset of immunity (OOI) for the vaccine delivered to MDA positive chicken embryos was 5 days post-hatch in comparison to 8 days post-hatch for the same dose of vaccine given at day old indicating that chicks would be protected against disease earlier in the field if vaccinated by the in ovo route. In further experiments the OOI for a turkey vaccine delivered to MDA positive turkey embryos was shown to be 8 days post-hatch.     

Evaluation of radiometric faecal culture and direct PCR on pooled faeces for detection of Mycobacterium avium subsp. paratuberculosis in cattle

Dilution rates for pooled faecal culture (PFC) and direct IS900 polymerase chain reaction (D-PCR) tests were evaluated on faecal samples from infected cows mixed with uninfected faeces in dilutions from 1 in 5 to 1 in 50. PFC was performed by radiometric culture, with confirmation by IS900 PCR and restriction endonuclease analysis (PCR/REA) on growth, and by mycobactin dependency testing on solid medium. Using 37 culture positive faecal samples from 12 subclinical cows, 83.8% and 94.6% of samples were confirmed positive in the PFC assay at dilutions of 1 in 50 and 1 in 30, respectively. Lower dilutions (1 in 5 to 1 in 20) provided only marginally better sensitivity, and confirmation of PFC growth by PCR/REA was significantly more sensitive than mycobactin dependency. D-PCR had significantly lower sensitivity than PFC confirmed by PCR/REA, with pools of 1 in 50, 30, 10 and 5 yielding positive results in 64.9%, 70.3%, 78.4% and 83.8% of samples, respectively. Cattle considered to be shedding 1.5 x 10(6) viable M. avium subsp. paratuberculosis (Map)/g faeces (on the basis of estimated losses in processing and growth rates in radiometric broth) were positive at dilutions up to 1 in 50 in the PFC and D-PCR. Those shedding 5 x 10(5) viable Map/g were positive in the PFC at dilutions up to 1 in 40, but required a 1 in 10 dilution or less for D-PCR. The results suggest that for cattle shedding relatively high concentrations of Map in faeces (>2 x 10(5) viable Map/g), maximal dilutions of 1 in 30 for PFC and 1 in 10 for D-PCR would be applicable.