From laboratory to point of entry: development and implementation of a loop‐mediated isothermal amplification (LAMP)‐based genetic identification system to prevent introduction of quarantine insect species

BACKGROUND

Rapid genetic on‐site identification methods at points of entry, such as seaports and airports, have the potential to become important tools to prevent the introduction and spread of economically harmful pest species that are unintentionally transported by the global trade of plant commodities. This paper reports the development and evaluation of a loop‐mediated isothermal amplification (LAMP)‐based identification system to prevent introduction of the three most frequently encountered regulated quarantine insect species groups at Swiss borders, Bemisia tabaci, Thrips palmi and several regulated fruit flies of the genera Bactrocera and Zeugodacus.

RESULTS

The LAMP primers were designed to target a fragment of the mitochondrial cytochrome c oxidase subunit I gene and were generated based on publicly available DNA sequences. Laboratory evaluations analysing 282 insect specimens suspected to be quarantine organisms revealed an overall test efficiency of 99%. Additional on‐site evaluation at a point of entry using 37 specimens performed by plant health inspectors with minimal laboratory training resulted in an overall test efficiency of 95%. During both evaluation rounds, there were no false‐positives and the observed false‐negatives were attributable to human‐induced manipulation errors. To overcome the possibility of accidental introduction of pests as a result of rare false‐negative results, samples yielding negative results in the LAMP method were also subjected to DNA barcoding.

CONCLUSION

Our LAMP assays reliably differentiated between the tested regulated and non‐regulated insect species within <1 h. Hence, LAMP assays represent suitable tools for rapid on‐site identification of harmful pests, which might facilitate an accelerated import control process for plant commodities. © 2018 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Clinical management of canine babesiosis

Objective – To review and summarize current information regarding epidemiology, pathogenesis, and pathophysiology leading to the various clinical syndromes associated with canine babesiosis. Diagnosis, treatment, preventative strategies, and zoonotic implications are discussed.
Etiology – Babesiosis is caused by hemoprotozoa of the genus Babesia . Numerous species of Babesia exist worldwide. An increased incidence of babesiosis is described, especially in North America. The babesial organism spends the majority of its life cycle within the erythrocyte of the definitive host, resulting in hemolysis, with or without systemic complications.
Diagnosis – Definitive diagnosis depends on direct visualization of the organism on blood smear or polymerase chain reaction. A positive serologic antibody test indicates exposure with or without active infection.
Therapy – Antiprotozoal drugs, antimicrobials, and supportive care are the mainstays of babesiosis therapy.
Prognosis – Prognosis depends on the severity of disease, which in turn depends on both organism and host factors. Clinical syndromes associated with a poorer prognosis include red biliary syndrome, acute renal failure, acute respiratory distress syndrome, neurologic dysfunction, acute pancreatitis, cardiac dysfunction, and hypoglycemia.     

Epidemiology,molecular virology and diagnostics of Schmallenberg virus,an emerging orthobunyavirus in Europe

After the unexpected emergence of Bluetongue virus serotype 8 (BTV-8) in northern Europe in 2006, another arbovirus, Schmallenberg virus (SBV), emerged in Europe in 2011 causing a new economically important disease in ruminants. The virus, belonging to the Orthobunyavirus genus in the Bunyaviridae family, was first detected in Germany, in The Netherlands and in Belgium in 2011 and soon after in the United Kingdom, France, Italy, Luxembourg, Spain, Denmark and Switzerland. This review describes the current knowledge on the emergence, epidemiology, clinical signs, molecular virology and diagnosis of SBV infection.     

Cultural awareness in veterinary practice: student perceptions

Australian veterinary classrooms are increasingly diverse and their growing internal diversity is a result of migration and large numbers of international students. Graduates interact with other students and increasingly with clients whose attitudes, beliefs, values, and behaviors differ from their own. An understanding and respect for these differences has an impact on client communication and health care outcomes. The present study explored how students understand and are likely to deal with issues of cultural diversity in veterinary professional practice as well as the educational needs that students feel should be met in regard to preparation to engage productively with diversity in professional practice. The present study also explored the extent to which the rich diversity of the undergraduate student population constitutes an educational resource. A class of final-year veterinary students was invited to participate in a workshop exploring intercultural confidence in veterinary consultation. Twelve groups of six to eight students discussed a fictitious scenario involving a challenging clinical encounter with a client from a different culture. Students were reticent to see the scenario in terms of cultural difference, although they generally recognized that awareness of cultural issues in veterinary practice was important. They also tended to not see their own ethnicity as relevant to their practice. While some felt that veterinary practice should be culture blind, most recognized a need to orient to cultural difference and to respond sensitively. Their suggestions for curricular improvements to address these issues are also included.     

Dental microwear of sympatric rodent species sampled across habitats in southern Africa: Implications for environmental influence

Dental microwear textures have proven to be a valuable tool for reconstructing the diets of a wide assortment of fossil vertebrates. Nevertheless, some studies have recently questioned the efficacy of this approach, suggesting that aspects of habitat unrelated to food preference, especially environmental grit load, might have a confounding effect on microwear patterning that obscures the diet signal. Here we evaluate this hypothesis by examining microwear textures of 3 extant sympatric rodent species that vary in diet breadth and are found in a variety of habitat types: Mastomys coucha, Micaelamys namaquensis and Rhabdomys pumilio. We sample each of these species from 3 distinct environmental settings in southern Africa that differ in rainfall and vegetative cover: Nama‐Karoo shrublands (semi‐desert) and Dry Highveld grasslands in the Free State Province of South Africa, and Afromontane (wet) grasslands in the highlands of Lesotho. While differences between habitat types are evident for some of the species, inconsistency in the pattern suggests that the microwear signal is driven by variation in foods eaten rather than grit‐level per se. It is clear that, at least for species and habitats sampled in the current study, environmental grit load does not swamp diet‐related microwear signatures.     

Inherited defects in pedigree dogs. Part 2: Disorders that are not related to breed standards

Recent debate concerning health problems in pedigree animals has highlighted gaps in current knowledge of the prevalence, severity and welfare implications of deleterious inherited traits within the pedigree-dog population. In this second part of a two-part review, inherited disorders in the top 50 UK Kennel Club registered breeds were researched using systematic searches of existing databases. A set of inclusion and exclusion criteria, including an evidence strength scale (SEHB), were applied to search results. A total of 312 non-conformation linked inherited disorders was identified, with German shepherd dogs and Golden retrievers associated with the greatest number of disorders. The most commonly reported mode of inheritance was autosomal recessive (71%; 57 breed-disorder combinations), and the most common primarily affected body system was the nervous sensory system. To provide a true assessment of the scale of inherited disorders in the pedigree dogs studied more effort is required to collect accurate prevalence data.     

Controversies related to red blood cell transfusion in critically ill patients

Objective – To review the evolution of and controversies associated with allogenic blood transfusion in critically ill patients. Data sources – Veterinary and human literature review. Human Data Synthesis – RBC transfusion practices for ICU patients have come under scrutiny in the last 2 decades. Human trials have demonstrated relative tolerance to severe, euvolemic anemia and a significant outcome advantage following implementation of more restricted transfusion therapy. Investigators question the ability of RBCs stored longer than 2 weeks to improve tissue oxygenation, and theorize that both age and proinflammatory or immunomodulating effects of transfused cells may limit efficacy and contribute to increased patient morbidity and mortality. Also controversial is the ability of pre‐ and post‐storage leukoreduction of RBCs to mitigate adverse transfusion‐related events. Veterinary Data Synthesis – While there are several studies evaluating the transfusion trigger, the RBC storage lesion and transfusion‐related immunomodulation in experimental animal models, there is little research pertaining to clinical veterinary patients. Conclusions – RBC transfusion is unequivocally indicated for treatment of anemic hypoxia. However, critical hemoglobin or Hct below which all critically ill patients require transfusion has not been established and there are inherent risks associated with allogenic blood transfusion. Clinical trials designed to evaluate the effects of RBC age and leukoreduction on veterinary patient outcome are warranted. Implementation of evidence‐based transfusion guidelines and consideration of alternatives to allogenic blood transfusion are advisable.