Challenge studies of European stocks of redfin perch, Perca fluviatilis L., and rainbow trout, Oncorhynchus mykiss (Walbaum), with epizootic haematopoietic necrosis virus

A challenge model for comparison of the virulence of epizootic haematopoietic necrosis virus (EHNV) to European stocks of redfin perch, Perca fluviatilis L., and rainbow trout, Oncorhynchus mykiss (Walbaum), was tested. The model investigated intraperitoneal (IP), bath and cohabitation routes at 10, 15 and 20 °C for 5–6 g fish and 15 °C for 20 g perch. In the IP challenges of perch, significant mortality occurred at 15 °C and 20 °C. In challenge trials for rainbow trout, significant mortalities were observed in IP and bath challenges at 20 °C. The mortality observed in IP challenged 20 g perch was not significantly different from that recorded for 6 g fish challenged IP. No significant mortality was observed in any other treatment groups. Re-isolation of ranavirus was confirmed by IFAT and was consistently associated with dead or moribund fish in the trial groups challenged with EHNV. The findings indicate that EHNV does not pose a high risk for wild perch and trout populations in Europe by natural exposure. Mortality appears to be primarily a function of environmental factors, with temperature playing an important role, and not just the presence of the virus in the fish.     

Prevalence of and intrinsic risk factors for appendicular osteosarcoma in dogs: 179 cases (1996-2005)

OBJECTIVE: To determine the prevalence of appendicular osteosarcoma (OSA) in Greyhounds compared with other breeds and identify potential intrinsic risk factors associated with development of OSA. DESIGN: Retrospective case series. ANIMALS: 179 dogs with primary appendicular OSA. PROCEDURES: Medical records of dogs in which primary appendicular OSA had been diagnosed between 1996 and 2005 were reviewed. Prevalence and crude odds ratios for OSA were calculated for various breeds by comparison with a reference population of mixed-breed dogs. Age and sex were examined as potential risk factors for the 3 breeds with highest prevalence. RESULTS: Breed period prevalence of OSA was highest for Greyhounds (21/339 [6.2%]), followed by Rottweilers (51/969 [5.3%]) and Great Danes (13/297 [4.4%]); all 21 Greyhounds with OSA were identified as having retired from racing. Sex was not identified as a risk factor for OSA in these breeds, but in all 3 breeds, risk of OSA increased with age. Greyhounds were significantly older at the time of OSA diagnosis (mean, 9.9 years) than were Rottweilers (8.3 years) and Great Danes (7.8 years). Rottweilers and Great Danes were more likely to have OSA involving the forelimbs than the hind limbs. The most frequent lesion sites for all 3 breeds were the proximal end of the humerus and distal end of the radius. The proximal end of the femur was also a common site for the Greyhounds. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the present study suggested that Greyhounds, Rottweilers, and Great Danes had an increased risk of developing OSA, compared with mixed-breed dogs.     

Strengthening understanding and perceptions of mineral fertilizer use among smallholder farmers: evidence from collective trials in western Kenya

It is widely recognized that mineral fertilizers must play an important part in improving agricultural productivity in western Kenyan farming systems. This paper suggests that for this goal to be realized, farmers’ knowledge must be strengthened to improve their understanding of fertilizers and their use. We analyzed smallholder knowledge of fertilizers and nutrient management, and draw practical lessons from empirical collective fertilizer-response experiments. Data were gathered from the collective fertilizer-response trials, through focus group discussions, by participant observation, and via in-depth interviews representing 40 households. The collective trials showed that the application of nitrogen (N) or phosphorous (P) alone was insufficient to enhance yields in the study area. The response to P on the trial plots was mainly influenced by incidences of the parasitic Striga weed, by spatial variability or gradients in soil fertility of the experimental plots, and by interactions with N levels. These results inspired farmer to design and conduct experiments to compare crop performance with and without fertilizer, and between types of fertilizers, or responses on different soils. Participating farmers were able to differentiate types of fertilizer, and understood rates of application and the roles of respective fertilizers in nutrient supply. However, notions were broadly generated by unsteady yield responses when fertilizers were used across different fertility gradients, association with high cost (especially if recommended rates were to be applied), association of fertilizer use with hybrids and certain crops, historical factors, among other main aspects. We identified that strengthening fertilizer knowledge must be tailored within existing, albeit imperfect, systems of crop and animal husbandry. Farmers’ perceptions cannot be changed by promoting more fertilizer use alone, but may require a more basic approach that, for example, encourages farmer experimentation and practices to enhance soil properties such as carbon build-up in impoverished local soils.     

不同碳氮有机物料对有机菜田土壤细菌多样性的影响

通过模拟试验,利用聚合酶链式反应-变性梯度凝胶电泳法(PCR-DGGE),研究了施入不同碳氮有机物料(秸秆、苜蓿、有机肥、尿素)56d后,有机生产系统菜田土壤细菌群落结构的特征。结果表明,常规和有机生产系统土壤的细菌群落结构有明显差异。由DGGE图谱ShannonWiener多样性指数分析得知,有机背景处理的细菌多样性整体高于常规背景处理,且有机生产系统土壤加秸秆处理(OS)多样性最高,加入尿素后细菌多样性降低,相反,加入苜蓿后细菌多样性升高。非加权组平均法(UPGMA)聚类分析将常规和有机背景土壤分为两大族群。DGGE条带测序和系统进化树表明,30个条带归属为Proteobacteria、Acidobacteria、Actinobacteria、Firmicutes、Verrucomicrobia。常规土壤加苜蓿(CA)处理出现的特征性条带B13与有机背景处理的共有条带B28分别与Bacillus属和Pseudomonas假单胞菌属同源性最高。     

Use of a vessel sealant device for splenectomy in dogs

Objective: To (1) describe a technique for splenic vessel hemostasis and (2) report complications and outcome after use of bipolar sealant device during splenectomy in dogs. Study Design: Case series. Animals: Dogs (n=27) with naturally occurring splenic disease. Methods: Between January 2006 and March 2008, splenectomy was performed using a vessel sealant device in 27 dogs with naturally occurring splenic disease. Number of sutures needed for splenectomy and complications were recorded. Splenic artery diameter was measured using a caliper. Intraoperative hemostasis, device ease of use, postoperative hemorrhage, and short‐term survival were evaluated. Results: Splenectomy was performed successfully in 27 dogs with the vessel sealant device; none of the dogs required vessel ligation with suture. The splenic artery was dissected and adequately sealed in each dog. One dog was readmitted 4 days after surgery with hemoabdomen. Abdominal exploration revealed splenic pedicle hemorrhage and pancreatitis, the vessel sealant device was used to coagulate splenic pedicle bleeding. The dog was alive at suture removal. Conclusion: In dogs, a vessel sealant device may be used to achieve efficient and safe hemostasis of the splenic vascular pedicle without sutures.     

Assessing Knowledge Production for Agrosilvopastoral Systems in South America

In recent decades agroforestry has undergone significant development in Latin America. South America generates the most scientific research on the topic in the region. We conducted a comprehensive review and analysis of knowledge production for South American agroforestry that includes livestock grazing, known as agrosilvopastoralism (AS), examining how different sociopolitical factors such as sources of funding, institutional priorities, and international cooperation can bias the direction and objectives of scientific research. We assessed the major attributes of scientific publications on the topic (25 articles per country; overall n = 210) and the potential factors underlying the processes of research and development in the region. The tree component was the most studied, while the livestock component received less attention. Studies were mainly focused on the production of goods and services (monetary or nonmonetary approaches), except in Brazil, where conservation was the major study objective. Stakeholders were involved in more than half of the studies (60%), and they were mostly ranchers and local producers. More than half (70%) of the studies offered recommendations based on their results, and such recommendations were mostly concerned with the management of agrosilvopastoral system components. In general, studies were led just as often by local as foreign first authors and coauthored by more than three people as part of interinstitutional working groups. Brazil, Argentina, Colombia, and Chile had more frequent cooperation among institutions and countries but mainly used their own funding. In contrast, Bolivia, Ecuador, and Peru had almost 100% of their studies supported by foreign countries (North America and Europe). Countries with greater internal funding of research generated more long-term studies. Besides this, two clear trends were detected: 1) conservation and social aspects were mainly supported by sources from external countries led by foreign principal investigators, and 2) production issues were supported from sources within countries and supported high levels of cooperation among institutions.     

Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Background

There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.

Results

Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.

Conclusions

Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

Electronic supplementary material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.     

Efficacy of a single milbemycin oxime administration in combination with praziquantel against experimentally induced heartworm (Dirofilaria immitis) infection in cats

The efficacy of a combination of milbemycin oxime and praziquantel in preventing the establishment of experimentally induced heartworm (Dirofilaria immitis) infection was investigated in a study involving 24 young domestic short-hair cats. The animals were inoculated with 50 infective larvae on day 0. Subsequently they were divided into two groups of 12 animals each. The animals in group 1 were treated once with medicated tablets containing 4 mg milbemycin (minimum dose 2 mg/kg body weight) and 10 mg praziquantel (MILBEMAX) on day 30 after infection. Cats in group 2 received placebo tablets on the same day. On day 183 post-infection a blood sample was taken from each animal before euthanasia and necropsy. The blood samples were tested for the presence of microfilariae and the necropsied animals were examined for the presence of adult worms. Microfilariae were not found in any of the investigated cats. No heartworms were found in the animals in group 1 (treated with medicated tablets). Out of the 12 placebo-treated cats 1 was heartworm-free, whereas all the others were found to be infected with 1-3 adult heartworms.     

Susceptibility of the Mexican bean beetle <Emphasis Type="Italic">Epilachna varivestis</Emphasis> Mulsant (Coleoptera: Coccinellidae) to endemic isolates of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae)

Entomopathogenic fungi represent excellent candidates for biological insecticides. Among the many entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) is one of the most promising species. Here, we report the results of infection studies performed using four B. bassiana native isolates (Bb37, Bb38, Bb40, and Bb45) compared with the commercial product Mycotrol^® (strain GHA) on adult Mexican bean beetle Epilachna varivestis Mulsant (Coleoptera: Coccinellidae), one of the most important pests of bean production in Mexico. First, single concentration (1 × 10⁸ conidia mL^?1) assays were carried out using all isolates applied using an immersion method. Examinations at 10 days after inoculation indicated that isolates Bb37, Bb38, and Bb40 significantly reduced adult survival, causing 60–75% mortality compared to the commercial strain GHA (33%) and control (29%). Second, using two selected isolates (Bb37 and Bb40), the mean lethal concentration values (LC₅₀) were estimated for third instar larvae and adult E. varivestis. The LC₅₀ values for Bb37 and Bb40 isolates decreased 5.26-fold and 3.19-fold, respectively, in third instars compared with adults. However, such difference for Bb40 isolate was not significant. Finally, an experiment, to compare the median survival time (MST) values between third instar and adult E. varivestis, was conducted using Bb37 and Bb40 at 1 × 10⁹ conidia mL^?1. MST values were significantly reduced against larvae (87 and 100 h) compared with adults (130 and 134 h) by Bb37 and Bb40, respectively. We conclude that native isolates of B. bassiana represent an important alternative for the control of E. varivestis. However, to demonstrate its effectiveness under field conditions, detailed studies are needed.     

Germline mutations in genes within the MAPK pathway cause cardio-facio-cutaneous syndrome

Cardio-facio-cutaneous (CFC) syndrome is a sporadic developmental disorder involving characteristic craniofacial features, cardiac defects, ectodermal abnormalities, and developmental delay. We demonstrate that heterogeneous de novo missense mutations in three genes within the mitogen-activated protein kinase (MAPK) pathway cause CFC syndrome. The majority of cases (18 out of 23) are caused by mutations in BRAF, a gene frequently mutated in cancer. Of the 11 mutations identified, two result in amino acid substitutions that occur in tumors, but most are unique and suggest previously unknown mechanisms of B-Raf activation. Furthermore, three of five individuals without BRAF mutations had missense mutations in either MEK1 or MEK2, downstream effectors of B-Raf. Our findings highlight the involvement of the MAPK pathway in human development and will provide a molecular diagnosis of CFC syndrome.