Establishment of a quantitative ELISA for the measurement of allergen-specific IgE in dogs using anti-IgE antibody cross-reactive to mouse and dog IgE

As IgE plays a pivotal role in type I hypersensitivity-mediated allergic diseases, it is valuable to measure absolute quantity of serum antigen-specific IgE for clinical and research purposes. Here we describe a novel ELISA system that enables quantification of antigen-specific IgE in ng/ml in dogs. A newly developed monoclonal antibody (CRE-DM) was shown to recognize canine and mouse IgE equally in a dose dependent manner, but it did not recognize canine IgG. The reactivity of CRE-DM to canine IgE was also confirmed by an inhibition ELISA using canine IgE as an inhibitor and the maximum inhibition rate was 91.3%. In order to know whether canine IgE specific to an allergen could be quantitatively measured with an ELISA using CRE-DM, we established a quantitative ELISA that could measure canine IgE recognizing Cry j 1, one of the major allergens of Japanese cedar pollen. In this ELISA, a standard curve was created by using concentration-predetermined Cry j 1-specific monoclonal mouse IgE. According to the standard curve, the concentration of Cry j 1-specific IgE in dogs that were experimentally sensitized to Japanese cedar pollen could be calculated and determined in ng/ml. The specificity of the Cry j 1-specific IgE ELISA using CRE-DM was also confirmed by inhibition ELISA using canine IgE as an inhibitor and the inhibition rate was 97.0%. Reproducibility of the ELISA in three independent assays was determined using groups of pooled canine sera whose Cry j 1-IgE titers ranged from 155.9 to 888.2 ng/ml. Intra- and inter-assay reproducibility was determined with coefficient of variation ranging between 3.1-5.2% and 2.2-8.0%, respectively. These results demonstrated that the ELISA utilizing CRE-DM was a specific, reliable and robust new laboratory test that could quantify absolute amount of antigen-specific IgE in canine serum. The ELISA will serve as a useful tool in the clinics to evaluate the change of serum IgE titers during anti-allergic treatments as well as during seasonal fluctuation of allergen exposure.     

Embryo collection from pigs post‐pseudopregnancy induced by estradiol dipropionate

We aimed to define whether embryo collection carried out after pseudopregnancy was of similar outcome and quality as after artificial abortion. To induce pseudopregnancy, 30 gilts or sows were given 20 mg intramuscular estradiol dipropionate (EDP) 10–11 days after the onset of estrus. Ten additional pigs were inseminated artificially at natural estrus as a control group. Prostaglandin F_2α (PGF_2α) was administered twice with a 24 hr interval beginning 15, 20, or 25 days after EDP‐treatment (n = 10 per group) or between 23 and 39 days after artificial insemination in control pigs. Following this, all pigs were given 1,000 IU equine chorionic gonadotropin and 500 IU human chorionic gonadotropin (hCG) and then inseminated. Embryos were recovered 6 or 7 days after hCG treatment and outcome was recorded. There was no significant difference in the number of normal embryos collected from the pigs with PGF_2α initiated at different time points or from the control group. Embryonic developmental stages 7 days after hCG treatment also did not differ among groups. These results indicate that the use of EDP to induce pseudopregnancy, followed by PGF_2α administration to synchronize estrus for subsequent embryo harvest, is a suitable alternative to the artificial abortion method.     

Effect of supplementation of mustard oil cake on intake,digestibility and microbial protein synthesis of cattle in a straw-based diet in Bangladesh

The objective of this study was to analyse the effects of different levels of rumen-degradable protein (RDP) on intake, digestibility and microbial protein synthesis by supplementing mustard oil cake (MOC) on rice straw-based diet of cattle (Bos indicus) in Bangladesh. A 4 × 4 Latin square design was applied. Four diets having constant energy (7.0 MJ/kg of dry matter (DM)) with varying levels of RDP (M ₀ = 4.1 g/MJ (control), M ₁ = 6.3 g/MJ, M ₂ = 8.3 g/MJ and M ₃ = 12.4 g/MJ of metabolizable energy (ME)) were received by each animal for a period of 28 days. A metabolism trial was conducted for 7 days. Results indicate that with increasing levels of RDP, crude protein (CP) and RDP intake increased significantly (P < 0.01). The significant (P < 0.01) increase in digestibility values are obtained for DM, organic matter, CP and digestible organic matter in the rumen. The digestibility of neutral detergent fibre and acid detergent fibre was also increased significantly (P < 0.05). The total nitrogen (N), ammonia-N and total volatile fatty acids increase significantly (P < 0.01) while the rumen pH increased from M ₀ to M ₂ and decreased thereafter. The efficiency microbial N intake increased significantly (P < 0.01) but showed a curvilinear response with higher RDP level (12.40 g/RDP/MJ ME). This study concludes that supplementation of RDP from MOC enhances the intake, digestibility and microbial protein synthesis which ultimately increases utilization of low-quality feed resources that can be used for developing cost-effective feeding systems on a straw-based diet in tropical regions.     

Characterization of indigenous chicken production systems in Kenya

Indigenous chicken (IC) and their production systems were characterized to understand how the whole system operates for purposes of identifying threats and opportunities for holistic improvement. A survey involving 594 households was conducted in six counties with the highest population of IC in Kenya using structured questionnaires. Data on IC farmers’ management practices were collected and analysed and inbreeding levels calculated based on the effective population size. Indigenous chicken were ranked highest as a source of livestock income by households in medium- to high-potential agricultural areas, but trailed goats in arid and semi-arid areas. The production system practised was mainly low-input and small-scale free range, with mean flock size of 22.40 chickens per household. The mean effective population size was 16.02, translating to high levels of inbreeding (3.12%). Provision for food and cash income were the main reasons for raising IC, whilst high mortality due to diseases, poor nutrition, housing and marketing channels were the major constraints faced by farmers. Management strategies targeting improved healthcare, nutrition and housing require urgent mitigation measures, whilst rural access road network needs to be developed for ease of market accessibility. Sustainable genetic improvement programmes that account for farmers’ multiple objectives, market requirements and the production circumstances should be developed for a full realization of IC productivity.     

Prevalence and antimicrobial resistance profiles of <Emphasis Type="Italic">Salmonella enterica</Emphasis> serovars isolated from slaughtered cattle in Bahir Dar,Ethiopia

A study was undertaken from October 2006 to March 2007 to determine the prevalence and antimicrobial resistance patterns of Salmonella serovars. Liver, mesenteric lymph nodes, intestinal content, and carcass swab samples (each n?=?186) were collected from 186 apparently healthy slaughtered cattle at Bahir Dar abattoir. Bacteriological analysis was done according to the International Organization for Standardization (ISO 6579 2002). Isolates were serotyped at Agence Française de Securite Sanitaire des Aliments, Cedex, France. Twenty-eight isolates consisting of Salmonella Typhimurium, Salmonella Newport, Salmonella Haifa, Salmonella Heidelberg, Salmonella Infantis, and Salmonella Mishmarhaemek were identified. Salmonella Typhimurium and Salmonella Newport were most frequently isolated while Salmonella Heidelberg and Salmonella Mishmarhaemek were isolated least. Eleven of the 28 (39.3%) were resistant to one or more of the antimicrobials tested. Resistance was shown to ampicillin, chloramphenicol, gentamycin, norfloxacin, polymyxin-B, streptomycin, tetracycline, and trimethoprim. Four of 11 (36.4%) were multiple antimicrobial resistant. All the isolates tested were susceptible to the antimicrobial effects of gentamycin, norfloxacin, and trimethoprim. Eleven, four, and two isolates of the 28 were resistant to streptomycin, tetracycline, and ampicillin, respectively. All isolates of Salmonella Infantis, Salmonella Typhimurium (except one), and Salmonella Mishmarhaemek were susceptible to the tested antimicrobials. One Typhimurium isolate was resistant to chloramphenicol, streptomycin, and tetracycline. Salmonella Haifa was multiply antimicrobial resistant to ampicillin, tetracycline, and streptomycin. All isolates of Salmonella Heidelberg were resistant to streptomycin. Results of this study indicated high level of carcass contamination with antimicrobial-resistant Salmonella serovars which could pose public health risk; suggests need for hygienic slaughtering operations and proper cooking of meat before consumption. Further detailed studies involving different abattoirs, animal products, food items, and animals on different settings were recommended in the study area.     

Responses to graded replacement of urea by maize steep liquor in diets for intensively fed lambs for meat production

Urea is a common ingredient of the diets of intensively fed lambs, but is increasingly required for industrial processes. Maize steep liquor (MSL) is a by-product of maize grain degradation to produce starch that may be a suitable replacement. Fifty growing lambs were fed on equinitrogenous diets in which between 0% and 80% of the urea was replaced by MSL; their growth and metabolism were recorded over 70 days. Increasing replacement of urea by MSL increased feed intake and nutrient digestibilities, leading to increased growth rates, more efficient feed conversion, and increased nitrogen retention. Concentrations of triiodothyroxin, thyroxin, glucose, and methionine were increased by replacement of urea by liquor, and plasma urea was reduced. This study suggests that MSL is a suitable replacement for up to 80% of urea in the diet of rapidly growing lambs.     

Veterinary aspects of ecological monitoring: the natural history of emerging infectious diseases of humans,domestic animals and wildlife

Proliferation of disease pathogens capable of affecting humans, domestic livestock and wildlife increasingly threatens environmental security and biodiversity. Livestock and wild animals in proximity to human beings are often in the chain of transmission and infection. Globalization of industrial livestock production (especially poultry upon which so much of the burgeoning human population depends) often permits transcontinental disease spread. Rapidly expanding (and often illegal) international trade in wild and domestic animals and their products are increasingly involved in the emergence of new diseases that may have the ability to transmit among humans, livestock and wildlife. Rapidly increasing urbanization has led in many places to overcrowded townships that rely on “bushmeat” for sustenance and has contributed to the emergence of virulent zoonotic pathogens. The emergence and proliferation of pathogens are exacerbated by anthropogenic transformation of natural landscapes in order to increase agricultural and livestock production. This paper posits that data gathered by veterinary ecologists should be interpreted and used by other disciplines. The importance of a thorough knowledge of the “natural history” (ecology) of the disease agent and its human, domestic and wild hosts is stressed.     

Prognostic significance of surgical margin, Ki-67 and cyclin D1 protein expression in grade II canine cutaneous mast cell tumor

The prognosis for canine cutaneous mast cell tumor (CCMT) is thought to be correlated with histopathological grading. However, the wide variety of histopathologic types of grade II is one of the most troublesome and difficult points for prognosis. The objective of this study is to determine the prognostic value of surgical margin, ki-67 and cyclin D1 protein expression in grade II tumor. Surgically resected specimens of solitary grade II CCMT from 48 dogs with follow-up periods over 360 days (median was 1080 days) were used in this study. The expression of cyclin D1 and ki-67 proteins was determined by morphometrically using slides stained immunocytochemically, and the correlations among the results, survival rate, and recurrence and/or metastasis rate of each dog were analyzed statistically. The recurrence and/or metastasis and mortality rate in the incomplete surgical excision group within 30 months postoperatively were higher than that of the complete surgical excision group. In the incomplete surgical excision group, dogs with low positive staining of ki-67 had a significantly better survival, but the recurrence and metastasis rate and ki-67 positivity failed to show a significant correlation. Only a small number of cases showed cyclin D1-positive tumor cells, but most of them had a poor outcome with a high recurrence rate. In grade II CCMT, incomplete excision induced a relatively high metastasis rate and poor prognosis. Ki-67 positivity is a marker for the estimation of overall survival in incomplete surgical excision cases. Cyclin D1 positivity was low and may not have a prognostic role.     

Cell cycle analysis of bovine cultured somatic cells by flow cytometry

This study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells (fetal fibroblasts, adult skin and muscle cells, and cumulus cells) after culture under a variety of conditions; 1) growth to 60-70% confluency (cycling), 2) serum starvation, 3) culture to confluency. Cell -cycle phases were determined by flow cytometry with propidium iodide staining enabling the calculation of percentages of cells in G0 /G1, S and G2 /M. The majority was in G0/G1 regardless of cell type and treatment. Serum-starved or confluent cultures contained higher percentages of cells in G0/G1 (89.5-95.4%; P < 0.05). Percentages of cells in G0/G1 increased as cell size decreased regardless of the cell type and treatment. In the serum-starved and confluent cultures, about 98% of small cells were in G0/G1 . Serum-starved cultures contained higher percentages of small cells (38.5-66.9%) than cycling and confluent cultures regardless of cell type (P < 0.05) . After trypsinization of fetal fibroblasts and adult skin cells that were serum-starved and cultured to confluency, the percentages of cells in G0/G1 increased (P < 0.05) on incubation for 1.5 (95.7-99.5%) or 3 hr (95.9-98.6%). These results verify that serum starvation and culture to confluency are efficient means of synchronizing bovine somatic cells in G0/G1, and indicate that a more efficient synchronization of the cells in G0/G1 can be established by incubation for a limited time period after trypsinization of serum-starved or confluent cells.     

Socio-economic factors influencing control of vector-borne diseases in the pastoralist system of south western Uganda

Research in control of tick-borne diseases and trypanosomosis, and their vectors, namely, ticks and tsetse flies respectively, has been on going for decades. However, very little attention has been paid to the socio-economic factors that are likely to influence the outcome of the interventions in the control of these diseases. Thus, this study was designed to investigate these factors, mainly the intra-household factors influencing decision-making in the control of Vector-borne diseases in the pastoralist areas of Uganda. These factors included: indigenous technical knowledge, household economic factors, and gender. Both qualitative and quantitative methods were used in the collection and analysis of data. The tools used for data collection included among others, participatory learning and action (PLA), and Case studies. The findings included the following: In pastoralist households, a big proportion of the household budget was allocated to vector-borne diseases control. In the male-headed households, men dominated decision-making on vector-borne diseases control, although the goals and priorities of men and women in these households were not the same. Also, vector-borne disease control was predominantly by use of modern veterinary drugs, and pastoralists treated sick cattle by themselves even in situations where there were veterinary personnel.