Analyses of monoclonal antibodies reacting with porcine CD3: results from the Second International Swine CD Workshop

Among the 57 monoclonal antibodies (mAb) analyzed within the T-cell group from the Second Swine CD Workshop, six mAb fell within clusters T10 and T11 (No. 088, STH164; No. 148, FY1A3; No. 149, FY2C1; No. 150, FY1H2; No. 151, FY2A11; No. 169, BB23-8E6). The mAb within these two groups gave a similar appearance on flow cytometry and stained all peripheral blood T-cells as defined by CD4 and wCD8 staining. All six mAb precipitated a 24 kDa protein. On the basis of inhibition analyses performed as part of the workshop and from published data, the mAb define at least three epitopes. There is only minimal stimulation of resting peripheral lymphocytes, but four of the mAb produce strong stimulation in the presence of PMA. With the exception of STH164, all have been shown to react with CD3-transfected COS cells. The new mAb, therefore, react with three epitopes on porcine CD3 designated CD3a (BB23-8E6, FY2A11), CD3b (FY1A3, FY2C1), and CD3c (FY1H2). mAb STH164 appears to be reactive with another epitope, however, since its reactivity with CD3 has not been confirmed it is designated as wCD3.     

Intercostal thoracotomy in 20 dogs: muscle-sparing versus traditional techniques

The levels of pain, duration of approaching and closure, and surgical exposure associated with intercostal thoracotomy were compared between muscle-sparing and traditional techniques in 20 dogs. Postoperative pain was assessed based on numerical pain scores using behavioral observation, heart rate, respiratory rate, and wound palpation. Time for approaching and closure were measured, and the extent of intrathoracic organ exposure for the surgical procedures was described for each technique. There were significant differences in numerical pain scores at 2 h as well as 1, 2, 3, 4, 5, 6, and 7 days after surgery between the two groups (p < 0.0001). There was no significant (p = 0.725) difference in times for approaching and closure between the two groups. Compared to the traditional method, the muscle-sparing technique also achieved the desired exposure without compromising exposure of the target organs. Our results suggest that the muscle-sparing technique is more effective than the traditional method for providing a less painful recovery during the first 7 days after intercostal thoracotomy. Additionally, the muscle-sparing technique is as effective as the traditional modality for providing an appropriate time for approaching and closure during intercostal thoracotomy as well as adequate organ exposure for the surgical procedures.     

Computed tomographic bronchioarterial ratio for brachycephalic dogs without pulmonary disease

The bronchoarterial (BA) ratio measured with computed tomography is widely used in human medicine to diagnose bronchial dilation or collapse. Although use of the BA ratio in veterinary medicine has been recently studied, this has not been evaluated in brachycephalic dogs predisposed to bronchial diseases including bronchial collapse. The purpose of this study was to establish BA ratios for brachycephalic dogs and compare the values with those of non-brachycephalic dogs. Twenty-three brachycephalic dogs and 15 non-brachycephalic dogs without clinical pulmonary disease were evaluated. The BA ratio of the lobar bronchi in the left and right cranial as well as the right middle, left, and right caudal lung lobes was measured. No significant difference in mean BA ratio was observed between lung lobes or the individual animals (p = 0.148). The mean BA ratio was 1.08 ± 0.10 (99% CI = 0.98~1.18) for brachycephalic dogs and 1.51 ± 0.05 (99% CI = 1.46~1.56) for the non-brachycephalic group. There was a significant difference between the mean BA ratios of the brachycephalic and non-brachycephalic groups (p = 0.00). Defining the normal limit of the BA ratio for brachycephalic breeds may be helpful for diagnosing bronchial disease in brachycephalic dogs.     

Study on the porcinophilic foot-and-mouth disease virus I. production and characterization of monoclonal antibodies against VP1

Monoclonal antibodies (MAbs) reported here were produced against the porcinophilic foot-and-mouth disease virus (FMDV) that caused the devastating swine disease on 1997 in Taiwan. A panel (25) of MAbs were found to react with VP1 of O/Taiwan/97 (O/97) by ELISA with various potencies. The biological identities of these VP1 reacting MAbs, such as neutralization activity, isotype and capability to distinguish between two serotype O FMDVs, O/97 and O/Taiwan/KM1/99 (O/99), were further analyzed. Eleven out of the total eighteen O/97 neutralizing MAbs were able to neutralize heterologous O/99. Eight O/97 neutralizing and five non-neutralizing MAbs could differentiate two serotype O FMDVs by immunofluorescence assay (IFA) implied that these thirteen MAbs recognized O/97 specific epitope(s). Furthermore, reactivities of the VP1 reacting MAbs with a 29 amino acids synthetic peptide (P29) representing the betaG-betaH loop of VP1 were analyzed by ELISA and fourteen were found positive. MAb clone Q10E-3 reacting strongest with VP1 and P29, neutralizing both but not differentiating two serotype O viruses suggested that the antibody binding site might involve the RGD motif and its C terminal conserved region on betaG-betaH loop. MAbs with diverse characters presented in this study were the first raised against porcinophilic FMDV. The complete set of MAbs may be used for further studies of vaccine, diagnostic methods, prophylaxis, etiological and immunological researches on FMDV.     

CANALOGRAPHIC EVALUATION OF THE EXTERNAL EAR CANAL IN DOGS

Canalography was performed to measure the diameter of the horizontal ear canal and to assess the clinical relevance of the measurements in identifying abnormalities of the ear canal. The diameter of the proximal and distal end of the annular cartilage in the horizontal ear canal was measured using canalography in 222 ears. The tympanic membrane could not be visualized with otoscopic examination in 70 ears even after ear cleansing. These canals were classified as being stenotic. The diameter of the proximal annular cartilage in the stenotic canals ranged from 0.8 to 3.1 mm (mean = 2.6 +/- 0.8 mm). The proximal annular cartilage was consistently smaller in diameter than the distal annular cartilage. The ratio between the diameter of the proximal and distal annular cartilage varied between dogs. In stenotic ear canals (70 ears) the ratio was less than 0.65. Total ear canal ablations and histopathological analysis were performed in 70 ear canals classified as stenotic after canalography. Hyperplasia of the epidermal layer was found in 56 stenotic canals. Stenosis of the canal due to otitis externa was found in the other 14 canals. Three canals with severe otitis externa were also diagnosed with canalography as having a ruptured tympanic membrane. The results suggest that canalography can be used to measure the diameter of the horizontal ear canal and to assess whether or not the tympanic membrane is ruptured prior to conservative or surgical therapy.     

Comparison of morphology, viability, and function between blood and milk neutrophils from peak lactating goats

The morphological features of blood and milk neutrophils from peak lactating goats were compared using light microscopy, scanning electron microscopy and flow cytometry in order to investigate the cytological changes of neutrophils after migration into the mammary gland. The kinetics of reactive oxygen intermediates (ROI) generation and gelatinase release of blood and milk neutrophils, with or without stimulation of phorbol 12-myristate, 13-acetate ester (PMA), were used to characterize their responses to inflammatory stimuli. Neutrophils isolated from goat milk were highly segmented and contained multi-lobed nuclei. Ultrastructurally, milk neutrophils were more ruffled on the surface compared to blood neutrophils. Approximately 30% of milk neutrophils were undergoing cell death, either necrosis or apoptosis, in contrast to 8% of blood neutrophils. The ROI production of activated milk neutrophils peaked earlier than blood neutrophils, but the duration and the intensity were much less. Neutrophils from both sources augmented the release of gelatinase in response to PMA (1 ng/mL). However, the amount of gelatinase released from milk neutrophils was lower (P < 0.05) than that of blood neutrophils. In summary, more neutrophils become apoptotic and necrotic in the mammary gland, presumably due to spontaneous aging, the process of diapedesis, and the interaction with milk components. Milk neutrophils have impaired functionalities in comparison with blood neutrophils. The information is relevant when studying mammary gland immunity and related diseases, such as mastitis.     

A longitudinal study of unweaned piglets raised by smallholder farmers in the Philippines

We conducted a longitudinal study to obtain detailed baseline information about the health and productivity of unweaned piglets raised by smallholder pig raisers in the Philippines, and to identify constraints and opportunities to smallholder-piglet production. At two sites in the Philippines, we collected data about 242 litters from 173 sows in 102 herds during on-farm visits over 12-18 months. Preweaning mortality at the northern and southern sites was 17 and 9%, respectively. Of the piglet deaths at the northern site, 49% occurred within 24h of birth. Pig raisers used few of the recognised interventions known to reduce neonatal piglet mortality, although >50% of litters were given iron injections at the northern site and kept in a farrowing pen with a barrier at the southern site. Creep feeding was more common at the northern site but the nutritive value of the feeds given to piglets was low. We used repeated-measures and time-to-event analyses. Our results suggest that targeted extension (addressing, in particular, management issues relating to neonatal piglets) can contribute to increased productivity of unweaned piglets.     

A PCR-based assay for discriminating Cervus and Rangifer (Cervidae) antlers with mitochondrial DNA polymorphisms

This study describes a method for discriminating Rangifer antlers from true Cervus antlers using agarose gel electrophoresis, capillary electrophoresis, quantitative real-time PCR, and allelic discrimination. Specific primers labeled with fluorescent tags were designed to amplify fragments from the mitochondrial D-loop genes for various Cervus subspecies and Rangifer tarandus differentially. A 466-bp fragment that was observed for both Cervus and Rangifer antlers served as a positive control, while a 270-bp fragment was specifically amplified only from Rangifer antlers. Allelic discrimination was used to differentiate between Cervus and Rangifer antlers, based on the amplification of specific alleles for both types of antlers. These PCR-based assays can be used for forensic and quantitative analyses of Cervus and Rangifer antlers in a single step, without having to obtain any sequence information. In addition, multiple PCR-based assays are more accurate and reproducible than a single assay for species-specific analysis and are especially useful in this study for the identification of original Cervus deer products from fraudulent Rangifer antlers.