Nutritional composition of ultra-processed plant-based foods in the out-of-home environment: a multi-country survey with plant-based burgers.

Ultra-processed plant-based foods, such as plant-based burgers, have gained in popularity. Particularly in the out-of-home (OOH) environment, evidence regarding their nutritional profile and environmental sustainability is still evolving. Plant-based burgers available at selected OOH sites were randomly sampled in Amsterdam, Copenhagen, Lisbon and London. Plant-based burgers (patty, bread and condiment) (n 41) were lab analysed for their energy, macronutrients, amino acids and minerals content per 100 g and serving and were compared with reference values. For the plant-based burgers, the median values per 100 g were 234 kcal, 20·8 g carbohydrates, 3·5 g dietary fibre and 12·0 g fat, including 0·08 g TFS and 2·2 g SFA. Protein content was 8·9 g/100 g, with low protein quality according to amino acid composition. Median Na content was 389 mg/100 g, equivalent to 1 g salt. Compared with references, the median serving provided 31% of energy intake based on a 2000 kcal per day and contributed to carbohydrates (17-28%), dietary fibre (42%), protein (40%), total fat (48%), SFA (26%) and Na (54%). One serving provided 15-23% of the reference values for Ca, K and Mg, while higher contributions were found for Zn, Mn, P and Fe (30-67%). The ultra-processed plant-based burgers provide protein, dietary fibre and essential minerals and contain relatively high levels of energy, Na and total fats. The amino acid composition indicated low protein quality. The multifaceted nutritional profile of plant-based burgers highlights the need for manufacturers to implement improvements to better support healthy dietary habits, including reducing energy, Na and total fats.

Developing a Feline Immunodeficiency Virus Subtype B Vaccine Prototype Using a Recombinant MVA Vector.

The feline immunodeficiency virus (FIV) is a retrovirus with global impact and distribution, affecting both domestic and wild cats. This virus can cause severe and progressive immunosuppression culminating in the death of felids. Since the discovery of FIV, only one vaccine has been commercially available. This vaccine has proven efficiency against FIV subtypes A and D, whereas subtype B (FIV-B), found in multiple continents, is not currently preventable by vaccination. We, therefore, developed and evaluated a vaccine prototype against FIV-B using the recombinant viral vector modified vaccinia virus Ankara (MVA) expressing the variable region V1-V3 of the FIV-B envelope protein. We conducted preclinical tests in immunized mice (C57BL/6) using a prime-boost protocol with a 21 day interval and evaluated cellular and humoral responses as well the vaccine viability after lyophilization and storage. The animals immunized with the recombinant MVA/FIV virus developed specific splenocyte proliferation when stimulated with designed peptides. We also detected cellular and humoral immunity activation with IFN-y and antibody production. The data obtained in this study support further development of this immunogen and testing in cats.

Validation of three qPCR for the detection of Burkholderia mallei in equine tissue samples.

Burkholderia mallei is the causative agent of glanders, a zoonosis listed by the World Organization for Animal Health as of mandatory notification. In this work, a comparison of three qPCR protocols was made, two of them based on articles by other authors and one standardized in house, this last one aiming at a genomic region that does not exist in other species of the Burkholderia genus. All qPCRs showed high efficiency and good repeatability. However, reactions with Cq between 36 and 40 were considered suspicious and unreliable, requiring greater clinical criteria to analyze the results.

Indirect ELISA (iELISA) standardization for the diagnosis of bovine enzootic leukosis / Padronização de um ELISA indireto (iELISA) para diagnóstico da leucose enzoótica bovina

Enzootic bovine leukosis (EBL) is an infectious disease caused by bovine leukemia virus (BLV) that affects cattle worldwide. Agar gel immunodiffusion (AGID) was the reference test for EBL diagnosis for many years, but enzyme-linked immunosorbent assay (ELISA) showed higher sensitivity, was faster to perform, and resulted in an objective reading. However, the importation of ELISA kits is lengthy and expensive, and currently, no AGID kits are available in Brazil. The aim of this work was to standardize an indirect ELISA (iELISA) for EBL diagnosis using BLV antigens produced in Tadarida brasiliensis lung (Tb1Lu) cells, which are Bovine viral diarrhea virus (BVDV) free, unlike fetal lamb kidney (FLK) cells, currently used for this purpose. Following standardization, iELISA results were compared with those obtained by AGID and the commercial Chekit Leucose-Serum ELISA. Compared to AGID, iELISA had 94,44% sensitivity, 75.68% specificity, 79.10% positive predictive value (PPV) and 93.30% negative predictive value (NPV), with 84% concordance and a Kappa index of 0.699. Compared to the Chekit Leucose-Serum ELISA, iELISA showed 92.60% sensitivity, 87.09% specificity, 90.27% PPV and 90,00% NPV, with 90.27% concordance and a Kappa index of 0.801. Taking into account the high agreement with the traditional tests and the absence of non-specific reactions with BVDV, the developed assay could be used as diagnostic method to control EBL in Brazil.(AU)

A leucose enzoótica bovina (LEB) é uma doença infecciosa natural dos bovinos com distribuição mundial causada pelo "bovine leukemia virus" (BLV). A imunodifusão em gel de ágar (IDGA) foi considerada por muitos anos o teste de eleição, porém ensaios imunoenzimáticos (ELISA) apresentam sensibilidade mais elevada e leitura mais rápida e objetiva. No entanto, a importação de kits de ELISA é um processo dispendioso e demorado, e atualmente não há kits de IDGA comercialmente disponíveis no Brasil. Desta forma, o objetivo deste trabalho foi padronizar um ELISA indireto (iELISA) para diagnóstico da LEB utilizando antígenos produzidos a partir do cultivo do BLV em linhagem celular Tadarida brasiliensis "lung" (Tb1Lu) livre de "bovine viral diarrhea virus" (BVDV), diferentemente do que acontece com as linhagens "fetal lamb kidney" (FLK) atualmente utilizadas na produção desses antígenos para uso em ensaios sorológicos. Após a padronização do iELISA, os resultados foram comparados com aqueles obtidos por IDGA e pelo ELISA comercial "Chekit Leucose-Serum". Comparado ao IDGA, o iELISA apresentou 94,44% de sensibilidade, 75,68% de especificidade, valor preditivo positivo (VPP) de 79,1% e valor preditivo negativo (VPN) de 93,3%, com concordância entre os testes de 84% e o índice Kappa 0,699. Quando comparado ao ELISA "Chekit Leucose-Serum", o iELISA apresentou sensibilidade de 92,6%, especificidade de 87,09%, VPP de 90,27% e VPN de 90%, com concordância de 90,27% e o índice Kappa 0,801. Portanto, devido à alta concordância com os testes tradicionais e ausência da ocorrência de reações inespecíficas com BVDV, o ensaio desenvolvido pode ser utilizado como ferramenta diagnóstica para o controle da LEB no Brasil.(AU)

Australian long-finned pilot whales (Globicephala melas) emit stereotypical, variable, biphonic, multi-component, and sequenced vocalisations, similar to those recorded in the northern hemisphere.

While in the northern hemisphere, many studies have been conducted on the vocal repertoire of long-finned pilot whales (Globicephala melas), no such study has been conducted in the southern hemisphere. Presented here, is the first study on the vocalisations of long-finned pilot whales along the southern coast of mainland Australia. Multiple measures were taken of 2028 vocalisations recorded over five years in several locations. These vocalisations included tonal sounds with and without overtones, sounds of burst-pulse character, graded sounds, biphonations, and calls of multiple components. Vocalisations were further categorised based on spectrographic features into 18 contour classes. Altogether, vocalisations ranged from approximately 200 Hz to 25 kHz in fundamental frequency and from 0.03 s to 2.07 s in duration. These measures compared well with those from northern hemisphere pilot whales. Some call types were almost identical to northern hemisphere vocalisations, even though the geographic ranges of the two populations are far apart. Other call types were unique to Australia. Striking similarities with calls of short-finned pilot whales (Globicephala macrorhynchus) and sometimes sympatric killer whales (Orcinus orca) were also found. Theories for call convergence and divergence are discussed.

Song variation of the South Eastern Indian Ocean pygmy blue whale population in the Perth Canyon, Western Australia.

Sea noise collected over 2003 to 2017 from the Perth Canyon, Western Australia was analysed for variation in the South Eastern Indian Ocean pygmy blue whale song structure. The primary song-types were: P3, a three unit phrase (I, II and III) repeated with an inter-song interval (ISI) of 170-194 s; P2, a phrase consisting of only units II & III repeated every 84-96 s; and P1 with a phrase consisting of only unit II repeated every 45-49 s. The different ISI values were approximate multiples of each other within a season. When comparing data from each season, across seasons, the ISI value for each song increased significantly through time (all fits had p << 0.001), at 0.30 s/Year (95%CI 0.217-0.383), 0.8 s/Year (95%CI 0.655-1.025) and 1.73 s/Year (95%CI 1.264-2.196) for the P1, P2 and P3 songs respectively. The proportions of each song-type averaged at 21.5, 24.2 and 56% for P1, P2 and P3 occurrence respectively and these ratios could vary by up to ± 8% (95% CI) amongst years. On some occasions animals changed the P3 ISI to be significantly shorter (120-160 s) or longer (220-280 s). Hybrid song patterns occurred where animals combined multiple phrase types into a repeated song. In recent years whales introduced further complexity by splitting song units. This variability of song-type and proportions implies abundance measure for this whale sub population based on song detection needs to factor in trends in song variability to make data comparable between seasons. Further, such variability in song production by a sub population of pygmy blue whales raises questions as to the stability of the song types that are used to delineate populations. The high level of song variability may be driven by an increasing number of background whale callers creating 'noise' and so forcing animals to alter song in order to 'stand out' amongst the crowd.

Serological diagnosis of equine infectious anemia in horses, donkeys and mules using an ELISA with a gp45 synthetic peptide as antigen.

Equine infectious anemia (EIA) is a disease caused by a Lentivirus that is currently controlled exclusively by identification of seropositive animals. In most countries, including Brazil, the official diagnostic test for EIA is the agar gel immunodiffusion test (AGID). Although this assay has a high specificity it can produce false negative reactions or equivocal results due to weak precipitation lines, especially in samples from donkeys, mules or newly infected equids. In this pioneering study, it was used overlapping synthetic peptide pools to map and identify a consensus, widely recognised antibody epitope within env encoding the EIAV envelope proteins. A 20-mer soluble peptide encompassing this epitope (pgp45) was then synthesized and tested in an indirect ELISA test. Using a panel of 859 EIA positive and negative equid serum samples, the pgp45 ELISA had 96.1% concordance, 98.6% sensitivity and 95.6% specificity respectively, when compared to AGID. The sensitivity and specificity of the pgp45 ELISA was also >90% when tested in individual equid species including horses (Equus caballus), donkeys (Equus asinus) and mules (Equus caballus x Equus asinus). Moreover, in a horse experimentally infected with the pathogenic Wyoming EIAV strain viral-specific antibodies were detected at 10 days post-infection (dpi) whereas in AGID no specific antibody was detected until 18 days of experimental infection. This peptide can now be used as an antigen in serological tests, especially for rapid screening of large numbers of equids, where it may contribute significantly in the control of EIA, especially at sites with high populations of donkeys and mules.

Serological Evidence of Orthopoxvirus Circulation Among Equids, Southeast Brazil.

Since 1999 Vaccinia virus (VACV) outbreaks involving bovines and humans have been reported in Brazil; this zoonosis is known as Bovine Vaccinia (BV) and is mainly an occupational disease of milkers. It was only in 2008 (and then again in 2011 and 2014) however, that VACV was found causing natural infections in Brazilian equids. These reports involved only equids, no infected humans or bovines were identified, and the sources of infections remain unknown up to date. The peculiarities of Equine Vaccinia outbreaks (e.g., absence of human infection), the frequently shared environments, and fomites by equids and bovines in Brazilian farms and the remaining gaps in BV epidemiology incited a question over OPV serological status of equids in Brazil. For this report, sera from 621 equids - representing different species, ages, sexes and locations of origin within Minas Gerais State, southeast Brazil - were examined for the presence of anti-Orthopoxvirus (OPV) antibodies. Only 74 of these were sampled during an Equine Vaccinia outbreak, meaning some of these specific animals presented typical lesions of OPV infections. The majority of sera, however, were sampled from animals without typical signs of OPV infection and during the absence of reported Bovine or Equine Vaccinia outbreaks. Results suggest the circulation of VACV among equids of southeast Brazil even prior to the time of the first VACV outbreak in 2008. There is a correlation of OPVs outbreaks among bovines and equids although many gaps remain to our understanding of its nature. The data obtained may even be carefully associated to recent discussion over OPVs history. Moreover, data is available to improve the knowledge and instigate new researches regarding OPVs circulation in Brazil and worldwide.

From conservation genetics to conservation genomics: a genome-wide assessment of blue whales (Balaenoptera musculus) in Australian feeding aggregations.

Genetic datasets of tens of markers have been superseded through next-generation sequencing technology with genome-wide datasets of thousands of markers. Genomic datasets improve our power to detect low population structure and identify adaptive divergence. The increased population-level knowledge can inform the conservation management of endangered species, such as the blue whale (Balaenoptera musculus). In Australia, there are two known feeding aggregations of the pygmy blue whale (B. m. brevicauda) which have shown no evidence of genetic structure based on a small dataset of 10 microsatellites and mtDNA. Here, we develop and implement a high-resolution dataset of 8294 genome-wide filtered single nucleotide polymorphisms, the first of its kind for blue whales. We use these data to assess whether the Australian feeding aggregations constitute one population and to test for the first time whether there is adaptive divergence between the feeding aggregations. We found no evidence of neutral population structure and negligible evidence of adaptive divergence. We propose that individuals likely travel widely between feeding areas and to breeding areas, which would require them to be adapted to a wide range of environmental conditions. This has important implications for their conservation as this blue whale population is likely vulnerable to a range of anthropogenic threats both off Australia and elsewhere.

Equine infectious anemia prevalence in feral donkeys from Northeast Brazil.

Equine infectious anemia virus (EIAV) is an important cause of morbidity and mortality throughout the world. Although the virus infects all members of the Equidae the vast majority of studies have been conducted in horses (Equus caballus) with comparatively little information available for other equid species. Brazil has one of the most abundant donkey (E. asinus) populations of any nation although the economic importance of these animals is declining as transportation becomes increasingly mechanized. As a result, considerable numbers of donkeys especially in the Northeast of the country have been released and allowed pursue an almost feral existence. Consequently, this large and growing population constitutes a significant risk as a reservoir for the maintenance and transmission of important equine infectious diseases such as glanders and equine arteritis virus in addition to EIAV. This study examines the prevalence of EIA in a semi-wild donkey population from Mossoró city, in Northeast Brazil, using AGID followed by cELISA, rgp90 ELISA and immunoblot (IB). Serum samples were collected from 367 donkeys without obvious EIA clinical signs. Subsequent testing revealed seropositive rates of 1.6% (6/367) in officially approved AGID tests, 3.3% (12/367) in cELISA and 14.4% (53/367) in the rgp90 ELISA. However, 88.7% (47/53) of the rgp90 ELISA positive samples were almost certainly false reactions because they failed to react with two or more antigens in IB. Consequently, the rpg90 ELISA has a similar sensitivity to AGID with donkey serum samples. Such high false positive rates have not been observed previously with serum samples from horses. Another highly significant finding is that 56.9% (33/58) of the donkey serum samples tested in IB had reactivity to EIAV p26 only. Although this could result from recent infection with the virus, it has been found that in some equids p26 only reactivity persists for extensive periods of time suggesting exposure to antigens possessing cross-reactive determinants or EIAV strains with envelope glycoproteins that are different from any that have been previously characterized and so undetectable by current IB techniques.

Equine infectious anaemia in equids of Southern Pantanal, Brazil: seroprevalence and evaluation of the adoption of a control programme / Anemia infecciosa equina em equídeos do Pantanal Sul, Brasil: soroprevalência e avaliação da adoção de um programa de controle

The working equid population in Corumbá, Southern Pantanal, is very large and has a crucial role in the main economic activity of the State of Mato Grosso do Sul, the beef cattle industry. The aim of the present study was to estimate the prevalence of equine infectious anaemia (EIA) in working equids of ranches in the municipality of Corumbá, by the official agar gel immunodiffusion (AGID) test, and evaluate the adoption of the Programme for the Prevention and Control of Equine Infectious Anaemia proposed by Embrapa Pantanal and official entities in the 1990s. From September to November 2009, forty ranches distributed through the area of the municipality were visited, and serum samples were obtained from 721 equines and 232 mules. According to previous publications and the present data, it was concluded that the prevalence of EIA in this population has increased from 18.17% to 38.60%, which represents at this time approximately 13,000 infected animals. There was no significant difference between the apparent prevalence of equines and mules. It was also verified that the control programme was not known by the greater part of the interviewed ranch owners, managers and foremen and, in their perception, EIA is not a primary threat to address. Among the studied variables, the serologic testing practice significantly reduced the risk for the presence of EIA seropositivity, as well as the separation of riding equipment and segregation of seropositives.(AU)

A população de equídeos de serviço em Corumbá, Pantanal Sul, é muito numerosa e tem um papel crucial na principal atividade econômica do estado de Mato Grosso do Sul, a pecuária de corte extensiva. O objetivo deste trabalho foi estimar a prevalência atual da anemia infecciosa equina (AIE) em equídeos de serviço em fazendas do município de Corumbá, pelo teste oficial de imunodifusão em gel de ágar (IDGA), e avaliar a adoção do Programa de Prevenção e Controle da Anemia Infecciosa Equina proposto pela Embrapa Pantanal e entidades oficiais nos anos 1990. De setembro a novembro de 2009, quarenta fazendas distribuídas na área do município foram visitadas, e amostras de soro obtidas de 721 equinos e 232 muares. De acordo com publicações anteriores e os dados obtidos neste trabalho, concluiu-se que a prevalência da AIE nesta população aumentou de 18.17% para 38,60%, o que representa atualmente cerca de 13.000 animais infectados. Não houve diferença significativa entre as prevalências aparentes de equinos e muares. Verificou-se, também, que o programa de controle era desconhecido pela maior parte dos produtores, gerentes e capatazes entrevistados e, na percepção dos mesmos, a AIE não é uma ameaça importante a ser enfrentada. Dentre as variáveis estudadas, a prática da realização de testes sorológicos reduziu significantemente o risco para a presença de soropositividade para AIE, assim como a separação dos equipamentos de montaria e a segregação dos soropositivos.(AU)

Multicentric lymphoma in buffaloes in the Amazon region, Brazil.

BACKGROUND: The presence of lymphoma in buffaloes was first reported in India in the 1960s. The disease is similar to Enzootic Bovine Leucosis (EBL) caused by Bovine leukemia virus (BLV) in cattle; however, according to our results and those of other studies, the etiology of these lymphomas in buffalo do not appear to be associated with BLV. The objectives of this study are to describe four cases of the disease in buffaloes belonging to the same herd in the Amazon region of Brazil and to perform a clinical-anatomopathological, immunohistochemical, and etiological study of the lymphomas. RESULTS: Over a period of ten years, four buffaloes were observed presenting progressive weight loss, swelling of peripheral lymph nodes, and nodules in the subcutaneous tissue. Upon necropsy, whitish-colored tumor masses were observed in the form of nodules in the subcutaneous tissue, along with miliary nodules on the serosal surfaces of abdominal and thoracic organs and tumors in lymph nodes and other organs. Neoplastic lymphocyte proliferation was observed through histopathology. An immunohistochemical study revealed that the neoplasias were formed by proliferation of predominantly B lymphocytes. The presence of BLV genome was not detected in the lymphomas when using the real-time PCR technique, nor was it detected through immunohistochemical staining using monoclonal antibodies against two viral proteins. Bovine herpesvirus 6 was not detected in the tumors. However, Bovine immunodeficiency virus (BIV) was detected in samples of lymphoma and in the lymph nodes and kidneys of one of the animals. CONCLUSIONS: The occurrence of lymphoma in buffaloes is reported for the first time in Brazil and is characterized by B-cell multicentric lymphoma. The etiology of the disease does not appear to be associated with BLV; however, the detection of BIV in samples of lymphoma from one sick animal deserves further study, considering the oncogenic potential of this virus.

Evolutionary Diversity of Suid Herpesvirus 1 Based on Ul44 Partial Sequences.

OBJECTIVE: The aim of this study was to use partial Ul44 sequences (glycoprotein C) of Suid herpesvirus 1 to examine the evolution and dynamics of the virus in different periods and hosts. METHODS: Phylogenetic trees were constructed using the software MrBayes after analysis in the software jModelTest to evaluate the best phylogenetic models. The software SplitsTree 4.0 was used to create phylogenetic networks, and the BEAST program was used to generate data on phylogeography. Replication kinetics and serum neutralization tests were applied to tree strains from different phylogenetic groups. RESULTS: Ul44 sequences derived from domestic swine and wild swine clustered in different clades and had different selective pressures depending on the host. We found no differences in replication kinetics and serum neutralization tests in the strains tested. Data show that the evolution of herpesviruses is complex, and different genetic groups may be evolving at different rates. Ul44 is an important marker for molecular evolution and epidemiology studies, but it is not useful for biological information.

Absence of Bovine leukemia virus (BLV) infection in buffaloes from Amazon and southeast region in Brazil.

Enzootic bovine leucosis is an infectious disease caused by Bovine leukemia virus (BLV) and is well described in bovines. The majority of infected animals are asymptomatic, one to five percent develop lymphoma and from 30 to 50% present a persistent lymphocytosis. The virus occurs naturally in cattle and experimentally in buffaloes, capybaras and rabbits. The occurrence of lymphoma in buffaloes has been attributed to BLV infection by some authors in India and Venezuela, but not confirmed by other studies and little information on natural BLV infection in buffaloes is available. The aim of this study was to evaluate the occurrence of BLV in a sub-sample of buffalo from Amazon and southeast regions in Brazil. Three hundred and fifteen serum samples were negative using commercial AGID and ELISA (ELISA-gp51) which detect anti-BLV glycoprotein gp51 antibodies. The same samples were also evaluated for antibodies to whole virus through a commercial ELISA (ELISA-BLV) in which 77 (24.44%) were found seropositive and two (0.63%) inconclusive. On the other hand, all animals were negative by PCR to BLV targeted to the env and tax genes. These results suggest that ELISA-BLV produces false positive results in buffalo serum (p<0.001). In addition, one buffalo lymphoma sample was negative in both PCR assays used in this study. BLV was not detected in buffaloes from the Amazon basin and the southeast region of Brazil. Serological tests, like ELISA-BLV, usually used for cattle may produce false-positive results for BLV in buffaloes and direct detection tests such as PCR should be chosen in these surveys. The occurrence of lymphoma in buffalo was not associated with BLV infection in the one case analyzed in this work and the etiology and pathogenesis of this disease should be clarified.

Estudo comparativo e validação de três técnicas de PCR em tempo real (qPCR) para diagnóstico de Peste Suína Africana / Comparative study and validation of three quantitative PCR (qPCR) techniques for the diagnosis of African Swine Fever

Este estudo verificou o desempenho de três técnicas de PCR quantitativa (Real-Time) para o diagnóstico de Peste Suína Africana, uma doença exótica no Brasil, a partir de amostras de tecidos. As três técnicas escolhidas baseiam-se na amplificação de sequências do gene da proteína viral VP72 e são preconizadas, cada uma, por laboratórios oficiais da OIE (PSA-OIE), dos Estados Unidos (PSA-USDA) e da União Europeia (PSA-EU), respectivamente. Oligonucleotídeos iniciadores e sondas de hidrólise marcadas com fluoróforos foram sintetizados conforme a literatura de referência consultada. Sequências-alvo do DNA viral foram inseridos em plasmídeo sintético, os quais serviram de controle positivo para a padronização das técnicas e otimização de reagentes, determinação dos limites de detecção e testes de verificação de desempenho. Para aferição de repetibilidade e reprodutibilidade das técnicas, as técnicas padronizadas foram repetidas em dias diferentes, por um segundo analista, com alteração no mix comercial de reagentes utilizado e em um equipamento diferente, e também por outro laboratório. Realizaram-se, ainda, provas de sensibilidade analítica com amostras de DNA viral de referência e especificidade analítica e diagnóstica, com amostras negativas. As técnicas de PSA-EU e PSA-USDA apresentaram-se mais vantajosas quanto ao consumo de iniciadores. Não houve diferenças significativas nos resultados quantitativos variando-se os dias dos ensaios, os analistas, os equipamentos e o mix de reagentes. As três técnicas apresentaram alta especificidade analítica e diagnóstica e sensibilidade diagnóstica. As três técnicas de qPCR mostraram-se eficazes para serem adotadas por um mesmo laboratório para emissão de diagnósticos oficiais de Peste Suína Africana.(AU)

This study evaluated the performance of three real time PCR techniques (qPCR) for the diagnosis of African Swine Fever in tissue samples. The three chosen techniques are based on amplification of viral protein VP72 gene sequences and are recommended by OIE (PSA-OIE), the United States official laboratories (PSA-USDA) and the European Union (PSA-EU). Target sequences of the viral DNA were inserted into synthetic plasmid, which served as a positive control for the standardization of techniques and optimization of reagents, determination of limits of detection and performance verification testing. To gauge repeatability and reproducibility of techniques, standard procedures were repeated on different days by two analysts and by changing mix reagents and equipment, and also by another laboratory. Analytical sensitivity tests were done with reference samples provided by an OIE reference laboratory and analytical and diagnostic specificity were tested with negative samples. The PSA-EU and PSA-USDA techniques were more advantageous to use because of lower concentration of oligos used. There were no significant differences in quantitative results varying the days of tests, analysts, equipment and the mix of reagents. The three techniques had high analytical and diagnostic specificity and sensitivity. The three qPCR techniques were considered equivalent and effective and can be adopted by any laboratory for issuing official diagnosis of African Swine Fever.(AU)

Equine infectious anemia on Marajo Island at the mouth of the Amazon river / Anemia infecciosa equina na Ilha de Marajó na foz do Rio Amazonas

Equine infectious anemia (EIA) is a transmissible and incurable disease caused by a lentivirus, the equine infectious anemia virus (EIAV). There are no reports in the literature of this infection in Equidae on Marajo Island. The objective of this study was to diagnose the disease in the municipalities of Cachoeira do Arari, Salvaterra, Santa Cruz do Arari and Soure, on Marajó Island, state of Pará, Brazil. For serological survey samples were collected from 294 horses, over 5-month-old, males and females of puruca and marajoara breeds and from some half-breeds, which were tested by immunodiffusion in Agar gel (AGID). A prevalence of 46.26% (136/294) positive cases was found. EIA is considered endemic in the municipalities studied, due to the ecology of the region with a high numbered population of bloodsucking insect vectors and the absence of official measures for the control of the disease...

A anemia infecciosa equina (EIA) é uma importante enfermidade, transmissível e incurável causada por um lentivírus, equine infectious anemia vírus (EIAV), e não há relatos na literatura desta infecção em equinos da Ilha de Marajó. O objetivo deste estudo foi diagnosticar a anemia infecciosa equina nos municípios de Cachoeira do Arari, Salvaterra, Santa Cruz do Arari e Soure, Ilha de Marajó, no bioma amazônico do estado do Pará, Brasil. Para a pesquisa sorológica foram coletadas 294 amostras de animais da espécie equina, acima de cinco meses de idade, de ambos os sexos, das raças puruca, marajoara e de mestiços, testadas pela imunodifusão em gel de Agar (IDGA). Foi verificada uma prevalência de 46.26% (136/294) de casos positivos para EIA. A doença é considerada endêmica nos municípios estudados, tanto pelos aspectos ecológicos da região que propiciam a manutenção da população de insetos hematófagos vetores, quanto pela ausência de medidas oficiais de controle da doença...

Low genetic diversity in pygmy blue whales is due to climate-induced diversification rather than anthropogenic impacts.

Unusually low genetic diversity can be a warning of an urgent need to mitigate causative anthropogenic activities. However, current low levels of genetic diversity in a population could also be due to natural historical events, including recent evolutionary divergence, or long-term persistence at a small population size. Here, we determine whether the relatively low genetic diversity of pygmy blue whales (Balaenoptera musculus brevicauda) in Australia is due to natural causes or overexploitation. We apply recently developed analytical approaches in the largest genetic dataset ever compiled to study blue whales (297 samples collected after whaling and representing lineages from Australia, Antarctica and Chile). We find that low levels of genetic diversity in Australia are due to a natural founder event from Antarctic blue whales (Balaenoptera musculus intermedia) that occurred around the Last Glacial Maximum, followed by evolutionary divergence. Historical climate change has therefore driven the evolution of blue whales into genetically, phenotypically and behaviourally distinct lineages that will likely be influenced by future climate change.

Migratory movements of pygmy blue whales (Balaenoptera musculus brevicauda) between Australia and Indonesia as revealed by satellite telemetry.

In Australian waters during the austral summer, pygmy blue whales (Balaenoptera musculus brevicauda) occur predictably in two distinct feeding areas off western and southern Australia. As with other blue whale subspecies, outside the austral summer their distribution and movements are poorly understood. In order to describe the migratory movements of these whales, we present the satellite telemetry derived movements of eleven individuals tagged off western Australia over two years. Whales were tracked from between 8 and 308 days covering an average distance of 3,009±892 km (mean ± se; range: 832 km-14,101 km) at a rate of 21.94±0.74 km per day (0.09 km-455.80 km/day). Whales were tagged during March and April and ultimately migrated northwards post tag deployment with the exception of a single animal which remained in the vicinity of the Perth Canyon/Naturaliste Plateau for its eight day tracking period. The tagged whales travelled relatively near to the Australian coastline (100.0±1.7 km) until reaching a prominent peninsula in the north-west of the state of Western Australia (North West Cape) after which they travelled offshore (238.0±13.9 km). Whales reached the northern terminus of their migration and potential breeding grounds in Indonesian waters by June. One satellite tag relayed intermittent information to describe aspects of the southern migration from Indonesia with the animal departing around September to arrive in the subtropical frontal zone, south of western Australia in December. Throughout their migratory range, these whales are exposed to impacts associated with industry, fishing and vessel traffic. These movements therefore provide a valuable tool to industry when assessing potential interactions with pygmy blue whales and should be considered by conservation managers and regulators when mitigating impacts of development. This is particularly relevant for this species as it continues to recover from past exploitation.

Genetic characterization of influenza virus circulating in Brazilian pigs during 2009 and 2010 reveals a high prevalence of the pandemic H1N1 subtype.

BACKGROUND: Influenza A viruses circulating in pigs in Brazil are still not characterized, and only limited data are available about swine influenza epidemiology in the country. Therefore, we characterized the hemagglutinin (HA) and neuraminidase (NA) genes of influenza viruses isolated from Brazilian pigs. We also evaluated one case of probable swine-to-human transmission. METHODS: Twenty influenza viruses isolated from pigs during 2009-2010 in five Brazilian states (Minas Gerais, Sao Paulo, Parana, Rio Grande do Sul, and Mato Grosso) were used. One human isolate, from a technician who became ill after visiting a swineherd going through a respiratory disease outbreak, was also used in the study. Phylogenetic analysis for the HA and NA genes and hemagglutinin amino acid sequence alignment were performed. RESULTS: All isolates clustered with pandemic H1N1 2009 (pH1N1) viruses and appeared to have a common ancestor. Genetic diversity was higher in the HA than in the NA gene, and the amino acid substitution S203T in one of HA's antigenic sites was found in most of the samples. The human isolate was more related to swine isolates from the same herd visited by the technician than to other human isolates, suggesting swine-to-human transmission. CONCLUSION: Our results show that pH1N1 was disseminated and the predominant subtype in Brazilian pigs in 2009-2010.

Diagnosis and clinic-pathological findings of influenza virus infection in Brazilian pigs / Diagnóstico, achados clínicos e patológicos da infecção pelo vírus influenza em suínos no Brasil

Influenza A virus (IAV) is a respiratory pathogen of pigs and is associated with the porcine respiratory disease complex (PRDC), along with other respiratory infectious agents. The aim of this study was to diagnose and to perform a clinic-pathological characterization of influenza virus infection in Brazilian pigs. Lung samples from 86 pigs in 37 farrow-to-finish and two farrow-to-feeder operations located in the States of Minas Gerais, São Paulo, Paraná, Rio Grande do Sul, Santa Catarina, and Mato Grosso were studied. Virus detection was performed by virus isolation and quantitative real time reverse-transcription PCR (qRT-PCR). Pathologic examination and immunohistochemistry (IHC) were performed in 60 lung formalin-fixed paraffin-embedded tissue fragments. Affected animals showed coughing, sneezing, nasal discharge, hyperthermia, inactivity, apathy, anorexia, weight loss and growth delay, which lasted for five to 10 days. Influenza virus was isolated from 31 (36.0%) lung samples and 36 (41.9%) were positive for qRT-PCR. Thirty-eight (63.3%) lung samples were positive by IHC and the most frequent microscopic lesion observed was inflammatory infiltrate in the alveoli, bronchiole, or bronchi wall or lumen (76.7%). These results indicate that influenza virus is circulating and causing disease in pigs in several Brazilian states.

O vírus influenza A (IAV) é um patógeno respiratório comum de suínos e faz parte do complexo de doenças respiratórias do suíno (PRDC) junto com outros agentes infecciosos. O objetivo deste estudo foi diagnosticar e realizar a caracterização clínica e patológica de casos/surtos de influenza em suínos brasileiros. Foram utilizadas amostras de tecido pulmonar de 86 suínos de 37 granjas de ciclo completo e duas unidades produtoras de leitões localizadas em Minas Gerais, São Paulo, Paraná, Rio Grande do Sul, Santa Catarina e Mato Grosso. A detecção viral em fragmentos pulmonares frescos foi realizada através do isolamento viral e da transcrição reversa-PCR em tempo real quantitativa (qRT-PCR). Exame patológico e imuno-histoquímica (IHQ) foram realizados em 60 amostras de pulmão fixadas em formalina 10% e embebidas em parafina. As amostras eram de animais apresentando tosse, espirros, secreção nasal, hipertermia, prostração, apatia, anorexia, perda de peso e ganho de peso reduzido, com duração entre cinco e 10 dias. O vírus influenza foi isolado de 31 (36,0%) amostras e 36 (41,9%) foram positivas na qRT-PCR. Na IHQ, 38 (63,3%) amostras foram positivas e a lesão mais frequentemente observada foi a presença de infiltrado inflamatório na parede e lúmen de vias aéreas (76,7%). Estes resultados indicam que o vírus influenza está circulando e causando lesões e doença respiratória em suínos de diversos Estados do Brasil.