Polymorphisms in the Prion Protein Gene of cattle breeds from Brazil / Polimorfismos no gene da proteína priônica em bovinos no Brasil

One of the alterations that occur in the PRNP gene in bovines is the insertion/deletion (indel) of base sequences in specific regions, such as indels of 12-base pairs (bp) in intron 1 and of 23- bp in the promoter region. The deletion allele of 23 bp is associated with susceptibility to bovine spongiform encephalopathy (BSE) as well as the presence of the deletion allele of 12 bp. In the present study, the variability of nucleotides in the promoter region and intron 1 of the PRNP gene was genotyped for the Angus, Canchim, Nellore and Simmental bovine breeds to identify the genotype profiles of resistance and/or susceptibility to BSE in each animal. Genomic DNA was extracted for amplification of the target regions of the PRNP gene using polymerase chain reaction (PCR) and specific primers. The PCR products were submitted to electrophoresis in agarose gel 3% and sequencing for genotyping. With the exception of the Angus breed, most breeds exhibited a higher frequency of deletion alleles for 12 bp and 23 bp in comparison to their respective insertion alleles for both regions. These results represent an important contribution to understanding the formation process of the Brazilian herd in relation to bovine PRNP gene polymorphisms.(AU)

Uma das mudanças que ocorrem no gene PRNP em bovinos é a inserção e/ou deleção (indels) de sequências de bases, em determinadas regiões como, por exemplo, as indels de 12 pares de bases (pb) no íntron 1 e 23pb na região promotora. O alelo de deleção de 23pb está relacionado com a suscetibilidade à Encefalopatia Espongiforme Bovina (EEB), assim como a presença do alelo de deleção de 12pb. Neste estudo foi genotipada a variabilidade de nucleotídeos da região promotora e íntron 1 do gene PRNP em bovinos das raças Angus, Canchim, Nelore e Simental, para identificar os perfis genotípicos de resistência e/ou suscetibilidade à EEB de cada animal. Foi realizada a extração de DNA genômico para amplificação das regiões alvo do gene PRNP, por meio da reação em cadeia de polimerase (PCR) utilizando-se primers específicos. Os produtos da PCR foram submetidos à eletroforese em gel de agarose a 3%, e sequenciamento para a realização da genotipagem. Com exceção da raça Angus, a maioria das raças apresentaram maiores frequências do alelo de deleção tanto para 12pb como 23pb, em comparação com seus respectivos alelos de inserção, para as duas regiões. Esses resultados abrem caminhos para o conhecimento de como o rebanho brasileiro está sendo formado com relação aos polimorfismos do gene PRNP bovino.(AU)

An Australian Process that Assesses Country BSE Food Safety Risk.

Background: Bovine spongiform encephalopathy (BSE) is a transmissible, fatal neurodegenerative disease of cattle. Recognised in 1986, the disease causes a spongiform degeneration of the neural network in the brain and spinal cord of infected cattle leading to incoordination, ataxia and ultimately death of the infected animal [1]. The agent causing BSE in cattle is a structurally modified prion protein. The BSE epidemic that started in the United Kingdom (UK) resulted in the destruction of more than 3.3 million cattle in the UK alone [2]. Variant Creutzfeldt-Jacob Disease (vCJD), a fatal neurodegenerative human disease described for the first time in 1996, is putatively linked to the consumption of specified tissues from the carcase of cattle infected with the BSE agent that causes BSE [3]. By June 2014, 184 people have died of vCJD infection and most of these lived in the UK http://www.cjd.ed.ac.uk/documents/worldfigs.pdf. As a result of the worldwide prohibition on processed animal proteins being fed to cattle, BSE is no longer a major threat to food and feed safety provided that appropriate control measures are effectively implemented. This paper discusses Australia’s approach to conducting country assessments to determine the food safety risk posed by the classical form of BSE but does not discuss the atypical forms of BSE, i.e. the H-type BSE and L-type BSE, identified more recently [4,5]. Australia has not recorded a case of BSE. In recognition of Australia’s effective BSE surveillance and control measures it has been assigned by its trading partners and the World Organisation for Animal Health (the OIE) the most favourable BSE risk status. In response to the identification of the linkage between BSE and vCJD in the BSE inquiry report [6], the Australian Government in 2001 introduced measures that prohibited the importation of beef and beef products from all countries that had reported cases of BSE. The Australia New Zealand Food Standards Code was amended in 2002 to ensure that beef and beef products sold in Australia were only derived from animals free of BSE. Some products were exempted from this requirement including: (a) collagen and gelatine sourced from bovine skins and hides; (b) bovine fat or bovine tallow at no more than 300 g/kg in a food product; and (c) dairy products sourced from bovines. Countries without BSE cases and wishing to export beef or beef products to Australia at the time were assessed by Food Standards Australia New Zealand (FSANZ) for country BSE risk status using a method based on the geographical BSE risk assessment methodology [7] between 2001 and 2003. As a result, retorted beef products were permitted for importation into Australia from 27 countries that included Argentina, Brazil, Chile, Croatia, Latvia, Lithuania, Mexico, New Zealand, Sweden, and Vanuatu. In view of the updated scientific information on BSE, the Australian Government announced a revised BSE food safety policy in 2009 that permitted the importation of beef and beef products from any country, providing that the country had been assessed by FSANZ as having appropriate and effective BSE controls in place. Countries wishing to export fresh beef (chilled or frozen) to Australia need to apply to the Australian Department of Agriculture for assessment of a broader range of animal health and quarantine risks. Since the announcement of the revised BSE food safety policy, FSANZ received submissions from 16 countries requesting country BSE food safety assessment and determination of their country’s BSE food safety risk status. This extended abstract describes an Australian process developed and applied by FSANZ for assessing country BSE food safety risk. Aims: To describe the features of a process developed and applied by FSANZ for assessing country BSE food safety risk. Study Design: The Australian process that assesses country BSE food safety risk is comprised of: 1) a food safety risk assessment across the beef supply chain; 2) a framework to assure the quality of the assessment outcomes; and 3) a set of arrangements to deliver transparent risk communication. Place and Duration of Study: FSANZ, Canberra, Australia, between April 2010 and December 2014. Methods: The Australian process to assess country BSE food safety risk was developed in accordance with the 2009 Australian Government’s BSE food safety policy http://www.foodstandards.gov.au/industry/bse/bseimports/documents/BSE%20Policy%2025%20Se ptember2009.pdf, and the principles described in the BSE chapter of the Terrestrial Animal Health Code published by the OIE. http://www.oie.int/index.php?id=169&L=0&htmfile=chapitre_bse.htm. Results: The BSE food safety assessment: The food safety assessment across the beef supply chain for BSE risk is comprised of: (a) a desk-based assessment that evaluates information provided by the applicant country; and (b) an in-country verification assessment that verifies the effectiveness of the key BSE control measures implemented in the applicant country. The deskbased assessment evaluates the applicant country’s response to the Australian Questionnaire to Assess BSE Risk (the Questionnaire), http://www.foodstandards.gov.au/industry/bse/bsequestionnaire/pages/default.aspx, information provided as appendices to the applicant country’s response to the Questionnaire, and any relevant information that is publicly accessible. The latter may include data and information published by the applicant country, relevant statistics and audit reports published by the OIE, the European Commission, the United States of America and others, and articles in relevant scientific journals. In addition to undertaking a desk-based assessment for each applicant country, FSANZ risk assessors conducted in-country inspections of all applicant countries that have been assessed to date, to verify the effectiveness of BSE-related controls. The in-country verification inspection assesses the competent authority’s oversight of BSE control and prevention measures, verifies the effectiveness of BSE related control measures implemented on beef and/or dairy farms, in feed producing establishments, and at slaughtering and rendering establishments in the applicant country. The adequacy of the BSE-related food safety control measures developed by the applicant country and the effectiveness of their implementation are assessed against the following key areas: 1) The likelihood of the introduction and release of the BSE agent through importation of live cattle, bovine commodities and animal feed products; 2) The likely exposure of domestic cattle herds to the BSE agent via potential recycling of the BSE agent within the animal feed system; 3) The specific food safety controls around beef and beef products produced for human consumption; 4) The adequacy of BSE control and prevention related infrastructure including an animal identification and traceability system, and the competent authority’s oversight of BSE prevention and control measures; and 5) BSE notification, laboratory diagnostic and surveillance activities. A detailed BSE food safety assessment report is prepared to describe the BSE food safety controls established by the applicant country and the effectiveness of their implementation. The report recommends a BSE food safety risk category for the applicant country as part of the overall conclusion. This BSE food safety risk category then determines the trading conditions for beef products that may be exported from the applicant country to Australia. Governance and quality assurance: The FSANZ country BSE food safety assessment process is supervised by the Australian BSE Food Safety Assessment Committee comprised of experts in the fields of food safety and risk assessment, animal health, animal and agricultural production systems, international trade, and animal identification and traceability. The assessment report prepared by FSANZ is peer reviewed by food safety and veterinary experts, and comments are also invited from the competent authority of the applicant country. The assessment outcomes including the recommended BSE risk status are reviewed and endorsed by the Australian BSE Food Safety Assessment Committee and subsequently approved by the Chief Executive Officer of FSANZ prior to notification to the applicant country and the Australian Department of Agriculture. The Australian Department of Agriculture establishes the export certification required from the competent authority of the applicant country based on the BSE risk status assigned. Risk communication and transparency: Once a country’s status is finalised, FSANZ communicates the assessment outcome to the applicant country and relevant stakeholders including the OIE. The full country BSE food safety assessment report is subsequently published at http://www.foodstandards.gov.au/industry/bse/bsestatus/Pages/default.aspx. Consistency with established international risk assessment framework: The Australian process to assess country BSE food safety risk is consistent with the risk assessment framework applied by the OIE [8] in determining a country’s BSE risk status for animal health purposes. The OIE framework is comprised of: (1) release assessment; (2) exposure assessment; (3) BSE notification and investigation assessment; (4) BSE diagnosis assessment; and (5) BSE surveillance assessment. The Australian country BSE food safety assessment, based on the above OIE framework, addresses additional elements around food safety systems and controls in the applicant country aimed at preventing the contamination of beef and beef products for human consumption with the BSE agent and their tracing within the human food supply chain. Consequently, slaughterhouse operations, cattle identification and traceability, meat traceability and recall systems in the applicant country are examined for their effectiveness to ensure the safety and traceability of exported products of bovine ori

Sequence variations of the bovine prion protein gene (PRNP) in native Korean Hanwoo cattle

Bovine spongiform encephalopathy (BSE) is one of the fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs) caused by infectious prion proteins. Genetic variations correlated with susceptibility or resistance to TSE in humans and sheep have not been reported for bovine strains including those from Holstein, Jersey, and Japanese Black cattle. Here, we investigated bovine prion protein gene (PRNP) variations in Hanwoo cattle [Bos (B.) taurus coreanae], a native breed in Korea. We identified mutations and polymorphisms in the coding region of PRNP, determined their frequency, and evaluated their significance. We identified four synonymous polymorphisms and two non-synonymous mutations in PRNP, but found no novel polymorphisms. The sequence and number of octapeptide repeats were completely conserved, and the haplotype frequency of the coding region was similar to that of other B. taurus strains. When we examined the 23-bp and 12-bp insertion/deletion (indel) polymorphisms in the non-coding region of PRNP, Hanwoo cattle had a lower deletion allele and 23-bp del/12-bp del haplotype frequency than healthy and BSE-affected animals of other strains. Thus, Hanwoo are seemingly less susceptible to BSE than other strains due to the 23-bp and 12-bp indel polymorphisms.

Prion diseases: A challenge to animal health.

Prion diseases are known as transmissible spongiform encephalopathies (TSE), a group of rare, rapidly progressive, and fatal neurologic diseases. The agents responsible for human and animal prion diseases are abnormal proteins (prion or proteinaceous infectious particle) that can trigger chain reactions causing normal proteins in the brain to change to the abnormal protein. These abnormal proteins are resistant to enzymatic breakdown, and they accumulate in the brain, leading to damage. All have long incubation periods followed by chronic neurological disease and fatal outcomes, have similar pathology limited to the CNS include convulsions, dementia, ataxia (balance and coordination dysfunction), and behavioral changes, and are experimentally transmissible to some other species.

Enfermedad de Creutzfeldt-Jakob: reporte de dos casos / Creutzfeldt- Jakob disease: report of two cases

INTRODUCCIÓN: La enfermedad de Creutzfeldt-Jakob (ECJ) es una enfermedad priónica neurodegenerativa que afecta el Sistema Nervioso Central (SNC), invariablemente mortal. Clasificada en esporádica, familiar e iatrogénica, se manifiesta por cuadro demencial subagudo, síntomas motores, visuales, y mioclonías. Presenta electroencefalograma (EEG) con actividad espicular pseudoperiódica; resonancia agnética (RM) con hiperintesidad de núcleos estriados y áreas de corteza cerebral; y líquido céfalo raquídeo (LCR) con aumento de proteína 14-3-3. Finalmente, un estudio histopatológico del cerebro establece la encefalopatía espongiforme. PRESENTACIÓN DEL CASO: Se exponen dos casos de ECJ. Ambas mujeres, 44 y 67 años, con alteración de la marcha, deterioro cognitivo y mioclonías, cuadro rápidamente progresivo hasta la dependencia absoluta. EEG en ambos casos compatible con ECJ, además de imágenes que revelan hiperintensidad en núcleos estriados y/o zonas de la corteza. Ambas cursaron con infecciones durante su hospitalización y se les realizó gastrostomía por presentar trastorno deglutorio severo. DISCUSIÓN: El diagnóstico de ECJ se sospecha con la clínica, y se fundamenta con hallazgos característicos en RM, EEG y análisis de LCR. Ante la sospecha de la forma familiar se sugiere estudio genético, sin embargo, en el Hospital Clínico Herminda Martin (HCHM) no se realiza. Al ser una enfermedad invariablemente mortal y sin tratamiento, dificulta la decisión entre realización de estudios e intervenciones, contra el manejo expectante.

INTRODUCTION: Creutzfeldt-Jakob disease (CJD) is a neurodegenerativedisease caused by prions that affect the central nervous system. It’s invariably fatal. It can be classified as sporadic, familial and iatrogenic. It’s clinical features are subacute dementia, associated with motor and visual symptoms, as well as myoclonus. Electroencephalogram (EEG) shows pseudoperiodic spicular activity. Magnetic Resonance Imaging (MRI) presents with hyperintense signals in both striated corpus and some cerebral cortex areas. Cerebrospinal fluid (CSF) shows an increased level of 14-3-3 protein. Finally, histopathological study of brain states spongiform encephalopathy. CASE REPORT: We report two cases of CJD. Two women who started at ages 44 and 67 years old with abnormal gait, cognitive impairment and myoclonus, with rapid progression of symptoms to absolute dependence. EEG was compatible with the disease and imaging tests showed hyperintense signals in striatum and / or some areas of the cortex. Both patients coursed with infections during their hospitalization period and both underwent gastrostomy due to the presence of a severe swallowing disorder. DISCUSSION: The diagnosis of CJD is suspected based on the clinical findings and it is supported by characteristic features on MRI, EEG and CSF analysis. If the familial form of CJD is suspected, a genetic study is suggested however, in Herminda Martin Clinical Hospital (HCHM) this is not available. CJD is an invariably fatal disease and it doesn’t have any treatment. This makes difficult to choose between executing tests and interventions, and having an expectant management.

Encefalopatías espongiformes transmisibles:biología del prion y estado actual de la vigilanciaepidemiológica en Colombia / Encefalopatias espongiformes transmissíveis: biologia do prião e estado atual davigilância epidemiológica na Colômbia / Transmissible spongiform encephalopathies: biology of the Prion and current state of the epidemiologic surveillance in Colombia

Las Encefalopatías Espongiformes Transmisibles, también llamadas enfermedades priónicas, sonun grupo de enfermedades neurodegenerativas, que afectan una gran variedad de mamíferos. El agenteresponsable de estas patologías se ha identificado como una isoforma anormal de una proteína celular, lacual luego de sufrir un cambio conformacional (prion), adquiere la capacidad de comportarse como unagente infeccioso. Se ha demostrado la capacidad de los priones para cruzar la barrera de especies entreel ganado y los seres humanos; lo cual se ha reflejado en un problema de salud pública que ha afectadogravemente a los países donde se han presentado brotes de estas enfermedades. Las EncefalopatíasEspongiformes Transmisibles se han reportado en una gran cantidad de países y Colombia no ha sido ajenaa la presencia de casos esporádicos humanos, no relacionados con el consumo de material contaminadoprocedente de animales enfermos. Con el presente artículo se pretende dar una visión de la historia y elestado actual de Colombia frente a estas enfermedades, las cuales representan una grave amenaza para lasalud pública y la agrocadena ganadera del país.

Transmissible Spongiform Encephalopaties, also called prion diseases, are a group of neurodegenerativediseases affecting a variety of mammals. The responsible agent consists of an abnormal isoform of a cellularprotein that suffers a conformational change (prion), acquiring the ability of being transmissible. It hasbeen demonstrated prions capacity to cross the species barrier between the cattle and humans; affectingpublic health in countries with reported cases of bovine spongiform encephalopathy. The transmissiblespongiform encephalitis have been reported in a number of countries and Colombia is not the exceptionwith some human sporadic cases, not related with the intake of contaminated material from sick animals.With this manuscript we pretend to give a view of the history and the current state of prion diseases inColombia, which represent a serious threat for the public health and the cattle industry of the country.

As doenças encefalopatias espongiformes transmissíveis, são do grupo de doenças neurodegenerativas,que afetam uma grande variedade de mamíferos. O agente responsável de estas patologias estáidentificado como uma isoforma anormal de uma proteína celular, a qual logo de apresentar umamudança de conformação (prião), adquire a capacidade de comportar-se como um agente infeccioso.Tem-se demonstrado a capacidade que tem o prião para ultrapassar a barreira de espécies entre o gadoe o ser humano; o qual está refletido gravemente aos países onde tem-se apresentado estas doenças.As encefalopatias espongiformes transmissíveis estão reportadas em uma grande quantidade de países.Na Colômbia tem-se apresentado casos esporádicos em humanos, não relacionados com o consumo dematerial contaminado procedente de animais doentes. O presente artigo, busca dar uma visão da historiae o estado atual da Colômbia frente as doenças, as quais representam uma grave ameaça para a saúdepública e da pecuária do pais.

BSE situation and establishment of Food Safety Commission in Japan

Eight major policies were implemented by Japanese Government since Oct. 2001, to deal with bovine spongiform encephalopathy (BSE). These are; 1) Surveillance in farm by veterinarian, 2) Prion test at healthy 1.3mi cows/yr, by veterinarian, 3) Elimination of specified risk material (SRM), 4) Ban of MBM for production, sale use, 5) Prion test for fallen stocks, 6) Transparent information and traceability, 7) New Measures such as Food Safety Basic Law, and 8) Establish of Food Safety Commission in the Cabinet Office. At this moment, the extent of SRM risk has only been indicated by several reports employing tests with a limited sensitivity. There is still a possibility that the items in the SRM list will increase in the future, and this indiscriminately applies to Japanese cattle as well. Although current practices of SRM elimination partially guarantee total food safety, additional latent problems and imminent issues remain as potential headaches to be addressed. If the index of SRM elimination cannot guarantee reliable food safety, we have but to resort to total elimination of tissues from high risk-bearing and BSE-infected animals. However, current BSE tests have their limitations and can not yet completely detect highrisk and/or infected animals. Under such circumstances, tissues/wastes and remains of diseased, affected fallen stocks and cohort animals have to be eliminated to prevent BSE invading the human food chain systems. The failure to detect any cohort should never be allowed to occur, and with regular and persistent updating of available stringent records, we are at least adopting the correct and useful approach as a reawakening strategy to securing food safety. In this perspective, traceability based on a National Identification System is required.

BSE crisis in Japan: A chronological overview

In recent years, food-related issues have become increasingly more publicised in developed countries. This holds true for Japan where food-related issues have been drawing attention as a socially significant topic, particularly since the appearance of BSE (Bovine Spongiform Encephalopathy). In 2003 a new governmental office, the Food Safety Commission was established in the Cabinet Office of the Japanese Government based on a new law, "the Food Safety Basic Law". This change of administration was raised by the outbreak of BSE, which is considered to be a drastic reformation of food safety policy in Japan.In addition, BSE impact was significant not only on administration but also on the agriculture and food industries. It revealed to the public lots of holes in the food related system which have been concealed for years.In this paper, I would like to show the inadequacy of management before the outbreak of BSE and the subsequent governmental actions and reactions for food safety. Furthermore, problems that still remain after the reformation, such as ban on US beef and policy of blanket testing, will be discussed.

BSE safety standards: An evaluation of public health policies of Japan, Europe, and USA

Since the advent of bovine spongiform encephalopathy (BSE) in the United Kingdom in 1986, new BSE cases have recently become rare. However, in Japan and the United States, positive cases have started to be seen recently. The rise in BSE cases paved the way for the human form of this disease, the variant Creutzfeldt-Jakob disease (vCJD). The observed trends in the UK may be attributed to effective implementation of public health policies coupled with increased vigilance through advancement in science and technology, or they may well be a reflection of the natural disease progression. We aim to discuss the BSE chronology of events, and compare examination methods, costs and cost-efficiency, management, and public policies of Japan, Europe, and the USA.

BSE Safety Standards: An Evaluation of Public Health Policies of Japan, Europe, and USA

Since the advent of bovine spongiform encephalopathy (BSE) in the United Kingdom in 1986, new BSE cases have recently become rare. However, in Japan and the United States, positive cases have started to be seen recently. The rise in BSE cases paved the way for the human form of this disease, the variant Creutzfeldt-Jakob disease (vCJD). The observed trends in the UK may be attributed to effective implementation of public health policies coupled with increased vigilance through advancement in science and technology, or they may well be a reflection of the natural disease progression. We aim to discuss the BSE chronology of events, and compare examination methods, costs and cost-efficiency, management, and public policies of Japan, Europe, and the USA.

BSE Crisis in Japan: A Chronological Overview

In recent years, food-related issues have become increasingly more publicised in developed countries. This holds true for Japan where food-related issues have been drawing attention as a socially significant topic, particularly since the appearance of BSE (Bovine Spongiform Encephalopathy). In 2003 a new governmental office, the Food Safety Commission was established in the Cabinet Office of the Japanese Government based on a new law, “the Food Safety Basic Law”. This change of administration was raised by the outbreak of BSE, which is considered to be a drastic reformation of food safety policy in Japan. In addition, BSE impact was significant not only on administration but also on the agriculture and food industries. It revealed to the public lots of holes in the food related system which have been concealed for years. In this paper, I would like to show the inadequacy of management before the outbreak of BSE and the subsequent governmental actions and reactions for food safety. Furthermore, problems that still remain after the reformation, such as ban on US beef and policy of blanket testing, will be discussed.

Immunohistochemical detection of Prion protein (PrP-Sc) and epidemiological study of BSE in Korea

Though the aetiology of transmissible spongiform encephalopathies (TSEs) remains uncertain, proteinase resistant prion protein (PrP-Sc), a converted form of the normal cellular prion protein (PrP-C), accumulates in the lysosome of cells of the nervous systems of animals with TSEs. In this study, clinical and epidemiological examinations of bovine spongiform encephalopathy (BSE) were conducted in Korea. During the investigated period, none of the cattle exhibited typical clinical signs of BSE, such as behavioral disturbances, high sensitivity, and abnormal locomotion. Immunohistochemical analysis and western immunoblotting were established to detect PrP-Sc in the brain tissue using monoclonal antibody (MAb) F89/160.1.5, produced by immunizing mice with a synthetic peptide which corresponds to bovine PrP residues 146-159, NH2-SRPLIHFGSDYEDRC-COOH. Although some BSE-like spongiform changes were observed in bovine brains randomly collected from Korean slaughterhouses from 1996 to 1999, no PrP-Sc was detected in those brains with the established immunohistochemistry and western immunoblotting assay. Also, no positive reaction was observed in bovine brains infected with rabies. These immunohistochemical and western immunoblotting methods using MAbs, specifically reactive with conserved epitopes on ruminant PrP, can be used for postmortem diagnosis of BSE. Further, the method can be applied to antemortem and the preclinical diagnosis of ovine scrapie by detecting PrP-Sc in lymphoid tissues, such as the tonsils, third eyelid or peripheral lymph nodes.