Droplet digital PCR as a useful tool for the quantitative detection of Enterovirus 71

Hand, foot and mouth disease (HFMD) is a contagious viral disease that frequently affects infants and children and present with blisters and flu-like symptoms. This disease is caused by a group of enteroviruses such as enterovirus 71 (EV71) and coxsackievirus A16 (CA16). However, unlike other HFMD causing enteroviruses, EV71 have also been shown to be associated with more severe clinical manifestation such as aseptic meningitis, brainstem and cerebellar encephalitis which may lead to cardiopulmonary failure and death. Clinically, HFMD caused by EV71 is indistinguishable from other HFMD causing enteroviruses such as CA16. Molecular diagnosis methods such as the use of real-time PCR has been used commonly for the identification of EV71. In this study, two platforms namely the real-time PCR and the droplet digital PCR were compared for the detection quantitation of known EV71 viral copy number. The results reveal accurate and consistent results between the two platforms. In summary, the droplet digital PCR was demonstrated to be a promising technology for the identification and quantitation of EV71 viral copy number.

Forme atipiche di malattia mani-piedi-bocca: La nostra esperienza

Introduzione: La malattia mani-piedi-bocca è una patologia infettiva tipica della prima infanzia causata dagli enterovirus, in particolare i sierotipi Coxsackievirus A16 ed Enterovirus 71. A partire dal 2008, ne è stata descritta un’epidemia da Coxsackievirus A6, in genere accompagnata da febbre elevata e che si distingue per lo sviluppo di piccole vescicole che progrediscono a lesioni vescicolo-bollose e a bolle vere e proprie. Inoltre, nei pazienti affetti da dermatite atopica, le lesioni tendono a localizzarsi sulle aree eczematose. Abbiamo quindi svolto uno studio osservazionale prospettico per descrivere le caratteristiche cliniche delle forme atipiche di malattia mani-piedi-bocca. Metodi: Sono stati arruolati i pazienti affetti da una forma atipica di malattia mani-piedi-bocca giunti consecutivamente presso l’Ambulatorio di Dermatologia Pediatrica del Policlinico S.Orsola-Malpighi di Bologna tra gennaio 2012 e febbraio 2014. Abbiamo valutato distribuzione, tipologia ed estensione delle lesioni. In un gruppo di pazienti è stata inoltre eseguita l’identificazione virale sul fluido vescicolare. Risultati: Abbiamo arruolato 47 pazienti (68% maschi) con un’età mediana di 22 mesi. Sono stati individuati 3 aspetti clinici principali: 1) forma acrale (distribuzione delle lesioni prevalentemente acrale), nel 62% dei soggetti; 2) eczema coxsackium (lesioni localizzate su aree eczematose), nel 23% dei soggetti; 3) forma diffusa (estensione delle lesioni al tronco), nel 15% dei soggetti. Inoltre, nell’80% circa dei pazienti si riscontravano macule purpuriche acrali, circa la metà dei pazienti aveva lesioni con aspetto purpurico e il 72% un interessamento dei glutei. In 9 su 11 soggetti genotipizzati è stato isolato il Coxsackievirus A6. Conclusioni: Questo studio ha permesso di descrivere tre fenotipi di malattia mani-piedi-bocca atipica, che deve essere correttamente identificata al fine di non porre diagnosi errate quali varicella, eczema herpeticum, vasculiti, impetigine, e di potere così procedere ad una gestione appropriata della patologia.

Atypical findings in delayed presentation of unilateral acute idiopathic maculopathy

Unilateral acute idiopathic maculopathy is a rare disease affecting young healthy patients resulting in moderate to severe unilateral visual loss preceded by prodromal flu-like illness. We present a case of delayed presentation of unilateral acute idiopathic maculopathy associated with hand, foot and mouth disease.

Specific detection of enterovirus 71 directly from clinical specimens using real-time RT-PCR hybridization probe assay

Enterovirus 71 (EV71) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. Thus, rapid detection of the virus is required to enable measures to be implemented in preventing widespread transmission. Based on primers and probes targeting at the VP1 region, a real-time reverse-transcriptase polymerase chain reaction (RT-PCR) hybridization probe assay was developed for specific detection of EV71 from clinical specimens. Quantitative analysis showed that the assay was able to detect as low as 5 EV71 viral copies and EV71 was detected from 46 of the 55 clinical specimens obtained from pediatric patients suffering from HFMD during the period from 2000 to 2003 in Singapore. This study showed that the single tube real-time RT-PCR assay developed in this study can be applied as a rapid and sensitive method for specific detection of EV71 directly from clinical specimens. (c) 2005 Elsevier Ltd. All rights reserved.

Spatial and Functional Organization of Pig Trade in Different European Production Systems: Implications for Disease Prevention and Control

Understanding the complexity of live pig trade organization is a key factor to predict and control major infectious diseases, such as classical swine fever (CSF) or African swine fever (ASF). Whereas the organization of pig trade has been described in several European countries with indoor commercial production systems, little information is available on this organization in other systems, such as outdoor or small-scale systems. The objective of this study was to describe and compare the spatial and functional organization of live pig trade in different European countries and different production systems. Data on premise characteristics and pig movements between premises were collected during 2011 from Bulgaria, France, Italy, and Spain, which swine industry is representative of most of the production systems in Europe (i.e., commercial vs. small-scale and outdoor vs. indoor). Trade communities were identified in each country using the Walktrap algorithm. Several descriptive and network metrics were generated at country and community levels. Pig trade organization showed heterogeneous spatial and functional organization. Trade communities mostly composed of indoor commercial premises were identified in western France, northern Italy, northern Spain, and north-western Bulgaria. They covered large distances, overlapped in space, demonstrated both scale-free and small-world properties, with a role of trade operators and multipliers as key premises. Trade communities involving outdoor commercial premises were identified in western Spain, south-western and central France. They were more spatially clustered, demonstrated scale-free properties, with multipliers as key premises. Small-scale communities involved the majority of premises in Bulgaria and in central and Southern Italy. They were spatially clustered and had scale-free properties, with key premises usually being commercial production premises. These results indicate that a disease might spread very differently according to the production system and that key premises could be targeted to more cost-effectively control diseases. This study provides useful epidemiological information and parameters that could be used to design risk-based surveillance strategies or to more accurately model the risk of introduction or spread of devastating swine diseases, such as ASF, CSF, or foot-and-mouth disease.

Salmon stocks and fisheries in England and Wales, 2001. Preliminary assessment prepared for ICES, April 2002

This report presents a preliminary assessment of the state of salmon stocks and fisheries in England and Wales in 2001 to assist ICES in providing scientific advice to NASCO and to provide early feedback to fishery managers and anglers. The chief indicators of the state o f salmon stocks are normally the catches taken by rod and net fisheries. However, in 2001 angling was affected by the outbreak of foot and mouth disease (FMD), which restricted angling opportunities and access to rod fisheries in many parts of the country for lengthy periods. It is impossible to quantify the impact that FMD had on rod catches, although these were undoubtedly significantly reduced; net fisheries were unaffected by FMD. The declared salmon catch for 2001 (including those fish released alive by anglers) is provisionally estimated at 209 tonnes, representing some 57,000 fish, and comprising 153 tonnes (-43,000 fish) by nets and fixed engines and 56 tonnes (-14,000 fish) by rods. For direct comparison with previous years, it should be noted that the declared catch prior to the issue of a second reminder was about 49 tonnes (see below). An estimated 26 tonnes (43%) of the rod catch was released alive. These figures do not take account of catches of salmon which go unreported (including those taken illegally), and it is estimated that there may have been a total of 33 tonnes of additional fish caught in 2001; approximately 15% of all fish killed.

Future novel threats and opportunities facing UK biodiversity identified by horizon scanning

1. Horizon scanning is an essential tool for environmental scientists if they are to contribute to the evidence base for Government, its agencies and other decision makers to devise and implement environmental policies. The implication of not foreseeing issues that are foreseeable is illustrated by the contentious responses to genetically modified herbicide-tolerant crops in the UK, and by challenges surrounding biofuels, foot and mouth disease, avian influenza and climate change.

Biochemical Characterization and Bio-evaluation of Collagen and Collagen Peptides Extracted and Purified from Fish Skin: In vitro and In vivo Studies on Antiarthritic and Wound Healing Properties

In fish processing plants, there is huge amount of skin that is left as the waste. When this skin is taken and processed into fish collagen, it will save large amount of money that is used for extraction of collagen from other animal s.Fish collagen can be used as an alternative to replace mammalian collagen, especially collagen extracted from bovine, when we consider the outbreak of bovine spongiform encephalopathy (BSE), transmissible spongiform encephalopathy (TSE) and the foot - and-mouth disease (FMD) issues. BSE and TSE are progressive neurological disorders affecting cattles caused by proteinacious infectious particles called prions.The study aims in producing collagen that has been extracted from fish skin to replace other animal collagen so as to overcome the problem of other animal collagen issues. Also the study utilized the abandoned fish waste produced by fish processing industry since bone, skin, fin and scales of fish can be a useful source of collagen.

An exploration of the drivers to bio-security collective action among a sample of UK cattle and sheep farmers

At present, collective action regarding bio-security among UK cattle and sheep farmers is rare. Despite the occurrence of catastrophic livestock diseases such as bovine spongiform encephalopathy (BSE) and foot and mouth disease (FMD), within recent decades, there are few national or local farmer-led animal health schemes. To explore the reasons for this apparent lack of interest, we utilised a socio-psychological approach to disaggregate the cognitive, emotive and contextual factors driving bio-security behaviour among cattle and sheep farmers in the United Kingdom (UK). In total, we interviewed 121 farmers in South-West England and Wales. The main analytical tools included a content, cluster and logistic regression analysis. The results of the content analysis illustrated apparent 'dissonance' between bio-security attitudes and behaviour.(1) Despite the heavy toll animal disease has taken on the agricultural economy, most study participants were dismissive of the many measures associated with bio-security. Justification for this lack of interest was largely framed in relation to the collective attribution or blame for the disease threats themselves. Indeed, epidemic diseases were largely related to external actors and agents. Reasons for outbreaks included inadequate border control, in tandem with ineffective policies and regulations. Conversely, endemic livestock disease was viewed as a problem for 'bad' farmers and not an issue for those individuals who managed their stock well. As such, there was little utility in forming groups to address what was largely perceived as an individual problem. Further, we found that attitudes toward bio-security did not appear to be influenced by any particular source of information per se. While strong negative attitudes were found toward specific sources of bio-security information, e.g. government leaflets, these appear to simply reflect widely held beliefs. In relation to actual bio-security behaviours, the logistic regression analysis revealed no significant difference between in-scheme and out of scheme farmers. We concluded that in order to support collective action with regard to bio-security, messages need to be reframed and delivered from a neutral source. Efforts to support group formation must also recognise and address the issues relating to perceptions of social connectedness among the communities involved. (c) 2008 Elsevier B.V. All rights reserved.

Latin America and the Caribbean in the World Economy 2005-2006

Includes bibliography

Bovine abortion associated with Neospora caninum: Diagnosis and epidemiological aspects of a dairy cattle herd in the northeast region of São Paulo state, Brazil

The objective of this study was to report the presence of Neospora caninum-associated abortion in bovines at a farm in the northeast region of São Paulo State. In January 2010, it was sent to the Department of Pathology, UNESP-Jaboticabal, a bovine fetus with an estimated age of seven months, which was natural of a dairy farm with 300 animals and an average daily production of 3,000 liters of milk, nearly 20 liters per cow. The animals were vaccinated against rabies, foot and mouth disease, carbuncle, brucellosis, leptospirosis, bovine herpes virus type I and bovine viral diarrhea virus. The herd consisted of purebred Holstein animals, Jersey, and mostly by crossbred animals 7-8 (gir x holstein). During necropsy, samples of the serosanguineous liquid present at the thoracic cavity and the heart of the fetus were collected for the detection of anti-Neospora caninum antibodies through Indirect Immunofluorescence Assay (IFA). Fragments of brain, cerebellum, tongue, liver, heart and kidneys were collected for the execution of histopathology (HP), immunohistochemistry (IHC) and Polymerase Chain Reaction. In order that IFA could be performed, the owner was requested blood samples without anticoagulants of the mother and other cows in the farm, with or without a history of abortion. At necropsy, it was verified a severe autolysis of the fetus. The serology of the fetus was 1:25, while the serology of the mother was 1:3,200. At HP, it was observed discrete multifocal non-suppurative encephalomyelitis characterized by gliosis and mononuclear inflammatory infiltration associated with cellular debris. DNA amplification of N. caninum was positive in fragments of brain, tongue, cerebellum, heart and kidneys. At IHC, it has been observed immunoreactivity to a cyst located in the tongue. The owner reported that his herd showed endemic episodes of abortion, while 27.69% (18/65) of the 65 animals sampled were seropositive. Although it has not been a significant difference (p>0.05), a higher seropositivity was observed in animals with a history of abortion (10/26) 38.46%, in comparison with animals without previous abortion (8/39) 20.51%. These findings show that the abortion under study was provoked by the protozoan N. caninum, while this is the first report concerning cattle in the northeast region of São Paulo State.

Detection of serum antibodies to parainfluenza type 3 virus, respiratory syncytial virus, bovine viral diarrhea virus, and herpes virus type 1 in sheep in the region of Botucatu, São Paulo - Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Expert Opinion on the Perceived Effectiveness and Importance of On-Farm Biosecurity Measures for Cattle and Swine Farms in Switzerland.

Biosecurity is crucial for safeguarding livestock from infectious diseases. Despite the plethora of biosecurity recommendations, published scientific evidence on the effectiveness of individual biosecurity measures is limited. The objective of this study was to assess the perception of Swiss experts about the effectiveness and importance of individual on-farm biosecurity measures for cattle and swine farms (31 and 30 measures, respectively). Using a modified Delphi method, 16 Swiss livestock disease specialists (8 for each species) were interviewed. The experts were asked to rank biosecurity measures that were written on cards, by allocating a score from 0 (lowest) to 5 (highest). Experts ranked biosecurity measures based on their importance related to Swiss legislation, feasibility, as well as the effort required for implementation and the benefit of each biosecurity measure. The experts also ranked biosecurity measures based on their effectiveness in preventing an infectious agent from entering and spreading on a farm, solely based on transmission characteristics of specific pathogens. The pathogens considered by cattle experts were those causing Bluetongue (BT), Bovine Viral Diarrhea (BVD), Foot and Mouth Disease (FMD) and Infectious Bovine Rhinotracheitis (IBR). Swine experts expressed their opinion on the pathogens causing African Swine Fever (ASF), Enzootic Pneumonia (EP), Porcine Reproductive and Respiratory Syndrome (PRRS), as well as FMD. For cattle farms, biosecurity measures that improve disease awareness of farmers were ranked as both most important and most effective. For swine farms, the most important and effective measures identified were those related to animal movements. Among all single measures evaluated, education of farmers was perceived by the experts to be the most important and effective for protecting both Swiss cattle and swine farms from disease. The findings of this study provide an important basis for recommendation to farmers and policy makers.

Structure and mode of multiplication of animal virus types. IV. Experimental part. Translated from the German.

TT: 67-62423.

Avian flu: the creation of expectations in the interplay between science and the media

This paper examines the emerging cultural patterns and interpretative repertoires in reports of an impending pandemic of avian flu in the UK mass media and scientific journals at the beginning of 2005, paying particular attention to metaphors, pragmatic markers ('risk signals'), symbolic dates and scare statistics used by scientists and the media to create expectations and elicit actions. This study complements other work on the metaphorical framing of infectious disease, such as foot and mouth disease and SARS, tries to link it to developments in the sociology of expectations and applies insights from pragmatics both to the sociology of metaphor and the sociology of expectations.