Development and characterization of monoclonal antibodies to subgroup A avian leukosis virus

Avian leukosis virus subgroup A (ALV‐A) is a retrovirus which infects egg‐type chickens and is the main pathogen of lymphoid leukosis (LL) and myeloid leukosis (ML). In order to greatly enhance the diagnosis and treatment of clinical avian leukemia, two monoclonal antibodies (MAbs) to ALV‐A were developed by fusion between SP2/0 and spleen cells from mice immunized with expressed ALV‐A env‐gp85 protein. Using immunofluorescence assay (IFA), two MAbs reacted with ALV‐A, but not with subgroups B and J of ALV. Western blot tests showed that molecular weight of ALV‐A envelope glycoprotein recognized by MAbs was about 53 kD. Isotyping test revealed that two MAbs (A5C1 and A4C8) were IgG1 isotypes. These MAbs can be used for diagnosis and epidemiology of ALV‐A.     

Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.     

Long-term study of partial arytenoidectomy with primary mucosal closure in 76 Thoroughbred racehorses (1992-2006)

REASON FOR PERFORMING STUDY: The effectiveness and best method to perform a partial arytenoidectomy in racehorses is unclear. This study was performed to evaluate the success of and complications that can occur after a unilateral partial arytenoidectomy with primary mucosal closure in Thoroughbred racehorses. HYPOTHESIS: Partial arytenoidectomy is an effective surgical procedure to return Thoroughbred racehorses, afflicted by arytenoid chondropathy or a failed laryngoplasty, to preoperative levels of performance. METHODS: Seventy-six Thoroughbred racehorses admitted to the New Bolton Centre between 1992 and 2006 were assessed. Information was obtained from the medical records about the horse, laryngeal abnormalities, surgery and other findings during hospitalisation. Racing information was evaluated relative to those independent variables by an analysis of variance with a level of significance of P<0.05. RESULTS: Arytenoid chondropathy was the presenting complaint in 54 horses and failed laryngoplasty in 22 horses. Thirteen horses (17%) underwent a second surgery for laser excision of intralaryngeal granulation tissue at the arytenoidectomy site. Seventy-three horses were discharged from the hospital and racing outcome was evaluated. Sixty horses (82%) raced after surgery and 46 horses (63%) raced 5 or more times after surgery. The median time from surgery to the first start was 6 months. The average earnings/start was not significantly different before and after surgery. There was no association between earnings after surgery and age, gender, location of lesion, type of lesion, duration of tracheal intubation or undergoing a second surgery. CONCLUSIONS AND POTENTIAL RELEVANCE: A Thoroughbred racehorse is likely to race after a unilateral partial arytenoidectomy with primary mucosal closure and return to a preoperative level of performance.     

Multiple virus infections of lablab [<Emphasis Type="Italic">Lablab purpureus</Emphasis> (L.) Sweet] in Nigeria

Leaf samples of Lablab purpureus collected from two agroecological zones of Nigeria—the northern guinea savanna zone (NGSZ) and the derived savanna zone (DSZ)—were infected with viruses when serologically indexed against available antisera. Approximately 31.1 and 81.1% of the leaf samples collected from the NGSZ and DSZ, respectively, were infected. Seven viruses were found: Bean common mosaic virus (BCMV), Cowpea aphid-borne mosaic virus (CABMV), Cucumber mosaic virus (CMV), Cowpea mottle virus (CPMoV), Cowpea severe mosaic virus (CPSMV), Southern bean mosaic virus (SBMV) and Tobacco mosaic virus (TMV) were detected from samples collected from NGSZ, while CMV, CPMoV, Cowpea mosaic virus (CPMV) and CPSMV were detected from samples from DSZ.     

Surgical correction of gastro‐oesophageal intussusception with bilateral incisional gastropexy in three dogs

Three dogs presented for evaluation of acute onset tachypnoea and dyspnoea following episodes of vomiting and/or regurgitation. Thoracic radiographs were suggestive of a gastro‐oesophageal intussusception in all three dogs; one dog also showed evidence of aspiration pneumonia. All three dogs underwent surgical correction with a bilateral incisional gastropexy. All dogs recovered from anaesthesia uneventfully and were discharged from the hospital 3 days after presentation. Persistent megaoesophagus was evident in all three dogs, and they are being chronically managed with a strict feeding regime and pro‐motility agents.     

Effectiveness of strategic anthelmintic treatments in the control of gastrointestinal nematodes and <Emphasis Type="Italic">Fasciola gigantica</Emphasis> in cattle in Iringa region,Tanzania

A longitudinal field trial was conducted to determine the effectiveness of strategic anthelmintic treatments in the control of gastrointestinal (GI) nematodes and Fasciola gigantica in cattle. A total of 167 cattle (6–18 months) from three large-scale dairy farms, four traditional farms and nine small-scale dairy farms were randomly selected. The selected animals on each farm were ear tagged and allocated into three groups based on live weight and treated as follows: Group T4 was treated with albendazole 10% drench at 10 mg/kg four times a year (mid rainy, end of the rain, mid dry and late dry/early rainy season). Group T2 was treated with albendazole 10% drench at 10 mg/kg two times a year (mid rainy and late dry/early rainy season). Group UT remained as untreated control. Faecal, blood and pasture samples were taken every month for 13 months. In addition, individual body weight (BWT) was measured on every sampling date. Results showed that two and four strategic treatments significantly reduced faecal egg counts (FEC) by 49.5% and 62.3% respectively compared to untreated control animals (P < 0.001). Two and four strategic treatments per year significantly reduced the proportion of animals passing Fasciola eggs in faeces by 30.6% and 51.7% (P < 0.001), respectively. Animals treated two and four times a year significantly outgained untreated animals by 14.8 kg and 17.7 kg respectively at the end of the trial (P < 0.05). The management system had a significant effect on packed cell volume and the proportion of animals passing Fasciola eggs in faeces (P < 0.05). The programme of two strategic treatments per year was only effective in controlling GI nematodes. It is concluded that a programme of four strategic treatments per year was effective in controlling GI nematodes and F. gigantica and improved weight gain.     

Simulating the cumulative effects of multiple forest management strategies on landscape measures of forest sustainability

While the cumulative effects of the actions of multiple owners have long been recognized as critically relevant to efforts to maintain sustainable forests at the landscape scale, few studies have addressed these effects. We used the HARVEST timber harvest simulator to predict the cumulative effects of four owner groups (two paper companies, a state forest and non-industrial private owners) with different management objectives on landscape pattern in an upper Michigan landscape managed primarily for timber production. We quantified trends in landscape pattern metrics that were linked to Montreal Process indicators of forest sustainability, and used a simple wildlife habitat model to project habitat trends. Our results showed that most trends were considered favorable for forest sustainability, but that some were not. The proportion of all age classes and some forest types moved closer to presettlement conditions. The trend for the size of uneven-aged patches was essentially flat while the average size of patches of the oldest and youngest age classes increased and the size of patches of the remaining age classes decreased. Forest fragmentation generally declined, but edge density of age classes increased. Late seral forest habitat increased while early successional habitat declined. The owners use different management systems that cumulatively produce a diversity of habitats. Our approach provides a tool to evaluate such cumulative effects on other landscapes owned by multiple owners. The approach holds promise for helping landowner groups develop and evaluate cooperative strategies to improve landscape patterns for forest sustainability.     

Growth,feed efficiency and blood profile of buffalo calves consuming high levels of fluoride

Twelve male buffalo calves of 10 to 12 months of age were divided into 3 groups of four each. They were fed wheat straw+concentrate mixture +3 Kg greens. The chemical composition of the diet was same in all the three groups except fluoride which was added (as NaF) in concentrate mixture of group B and C to make the final fluoride concentration 30 ppm and 60 ppm respectively. The animals were kept on scheduled diet for a period of 90 days. Body weights were recorded at the start of the experiment and at fortnightly interval thereafter. Analysis of data revealed that the dry matter intake decreased non significantly in group B and C as compared to control group. A significant decrease in serum calcium and a significant increase in phosphorus concentration were observed in group C animals. A significant increase was observed in alkaline phosphatase activity in group C animals. A non significant decrease was observed in T₄ values in group C animals. On the basis of these results it could be concluded that fluoride in the diet of buffalo calves @ 30 ppm is a safe level whereas 60 ppm has affected the blood metabolites.     

Evaluation of Methods to Determine Sperm Density for the European eel,Anguilla anguilla

European eel, Anguilla anguilla, is a target species for future captive breeding, yet best methodology to estimate sperm density for application in in vitro fertilization is not established. Thus, our objectives were to evaluate methods to estimate European eel sperm density including spermatocrit, computer‐assisted sperm analysis (CASA) and flow cytometry (FCM), using Neubauer Improved haemocytometer as benchmark. Initially, relationships between spermatocrit, haemocytometer counts and sperm motility were analysed, as well as the effect of sperm dilution on haemocytometer counts. Furthermore, accuracy and precision of spermatocrit, applying a range of G‐forces, were tested and the best G‐force used in method comparisons. We found no effect of dilution on haemocytometer sperm density estimates, whereas motility associated positively with haemocytometer counts, but not with spermatocrit. Results from all techniques, spermatocrit, CASA and FCM, showed significant positive correlations with haemocytometer counts. The best correlation between spermatocrit and haemocytometer counts was obtained at 6000 ×  g (r = 0.68). Of two CASA variants, one or three photographic fields (CASA‐1 and CASA‐2), CASA‐2 showed a very high accuracy to haemocytometer counts (r  =  0.93), but low precision (CV: CASA‐2 = 28.4%). FCM was tested with and without microfluorospheres (FCM‐1 and FCM‐2), and relationships to haemocytometer counts were highly accurate (FCM‐1: r  =  0.94; FCM‐2: r  =  0.88) and precise (CV: FCM‐1 = 2.5; FCM‐2 = 2.7%). Overall, CASA‐2 and FCM‐1 feature reliable methods for quantification of European eel sperm, but FCM‐1 has a clear advantage featuring highest precision and accuracy. Together, these results provide a useful basis for gamete management in fertilization protocols.