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2.
One Health ; 17: 100617, 2023 Dec.
Article in English | MEDLINE | ID: mdl-38024258

ABSTRACT

The health of humans, domestic and wild animals, plants, and the environment are inter-dependent. Global anthropogenic change is a key driver of disease emergence and spread and leads to biodiversity loss and ecosystem function degradation, which are themselves drivers of disease emergence. Pathogen spill-over events and subsequent disease outbreaks, including pandemics, in humans, animals and plants may arise when factors driving disease emergence and spread converge. One Health is an integrated approach that aims to sustainably balance and optimize human, animal and ecosystem health. Conventional disease surveillance has been siloed by sectors, with separate systems addressing the health of humans, domestic animals, cultivated plants, wildlife and the environment. One Health surveillance should include integrated surveillance for known and unknown pathogens, but combined with this more traditional disease-based surveillance, it also must include surveillance of drivers of disease emergence to improve prevention and mitigation of spill-over events. Here, we outline such an approach, including the characteristics and components required to overcome barriers and to optimize an integrated One Health surveillance system.

5.
Transbound Emerg Dis ; 69(2): 903-907, 2022 Mar.
Article in English | MEDLINE | ID: mdl-33590723

ABSTRACT

Porcine reproductive and respiratory syndrome (PRRS) is a viral disease characterized by reproductive impairment or failure in breeding animals, and a respiratory disease in pigs of any age. Brazil is the fourth largest pork producer and exporter globally, and PRRS virus (PRRSV) infection has never been reported in the country. This study aimed to investigate the status of porcine biological samples from commercial swine herds, quarantined imported boars, wild boars and feral pigs to update PRRS information in Brazil. A total of 14,382 samples were collected from 2008 to 2020, including sera (n = 12,841), plasma (n = 1,000) and oral fluids (n = 541), comprehending 137 herds and free-living pigs in eight Brazilian states. One out of 1,000 (0.1%) plasma and 15 out of 12,841 (0.11%) serum samples tested positive for PRRSV antibodies through ELISA. Upon ELISA retesting, only the plasma sample, from one 8-day-old piglet remained positive. All sixteen previously PRRSV antibody-positive samples were tested through RT-PCR and found to be negative. The presence of false-positive or singleton reactors are quite expected. Thus, the use of different/alternative diagnostic tests is indicated for an efficient PRRSV detection. Taken together, our findings demonstrated no conclusive evidence of PRRSV infection in the tested pigs, highlighting the importance to reinforce the surveillance program to prevent the introduction and eventual dissemination of PRRSV in Brazil.


Subject(s)
Porcine Reproductive and Respiratory Syndrome , Porcine respiratory and reproductive syndrome virus , Swine Diseases , Animals , Antibodies, Viral , Brazil/epidemiology , Enzyme-Linked Immunosorbent Assay/veterinary , Porcine Reproductive and Respiratory Syndrome/epidemiology , Retrospective Studies , Swine
6.
Zoonoses Public Health ; 66(1): 125-132, 2019 02.
Article in English | MEDLINE | ID: mdl-30485723

ABSTRACT

Backyard pig populations are not monitored for influenza A virus (IAV) in Brazil and there are limited data about seroprevalence and risk factors in these populations. Our goal was to assess possible factors associated with IAV seroprevalence in backyard pig populations using an indirect ELISA protocol based on a recombinant nucleoprotein. Following the IAV screening using NP-ELISA, subtype-specific serology based on hemagglutination inhibition (HI) assay of the ELISA-positive pigs was conducted. The survey comprised a total of 1,667 sera samples collected in 2012 and 2014 in 479 holdings and the estimated seroprevalence was 5.3% (3.84%-7.33%) and 2.3% (1.34%-3.71%) in the respective years. In both years, H1N1pdm09 was the most prevalent subtype. The multivariable analysis showed main factors such as "age," "sex," "number of suckling pigs" and "neighbours raising pigs" that presented the greatest effect on IAV seroprevalence in these pig populations. These factors may be associated with the low biosecurity measures and management of backyard holdings. In addition, the low IAV seroprevalences found in these backyard pig populations could be related to a low number of animals in each pig holding and low animal movement/replacement that do not favour IAV transmission dynamics. This low frequency of H1N1pdm09 seropositive pigs could also be due to sporadic human-to-pig transmission of what is now a human seasonal influenza A virus; however, these factors should be explored in future studies. Herein, these results highlight the importance of IAV continued surveillance in backyard pig holdings, since it is poorly known which IAVs are circulating in these populations and the risk they could pose to public health and virus transmission to commercial farms.


Subject(s)
Influenza A virus/isolation & purification , Orthomyxoviridae Infections/veterinary , Swine Diseases/virology , Animals , Brazil/epidemiology , Enzyme-Linked Immunosorbent Assay , Female , Male , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Seroepidemiologic Studies , Swine , Swine Diseases/epidemiology
7.
Braz. j. microbiol ; 49(2): 351-357, Apr.-June 2018. graf
Article in English | LILACS | ID: biblio-889245

ABSTRACT

Abstract Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.


Subject(s)
Animals , Circovirus/chemistry , Capsid Proteins/chemistry , Protein Conformation , Swine , Swine Diseases/virology , Brazil , Models, Molecular , Circovirus/isolation & purification , Circovirus/genetics , Circoviridae Infections/veterinary , Circoviridae Infections/virology , Amino Acid Substitution , Capsid Proteins/genetics
8.
Braz J Microbiol ; 49(2): 351-357, 2018.
Article in English | MEDLINE | ID: mdl-29128395

ABSTRACT

Economic losses with high mortality rate associated with Porcine circovirus type 2 (PCV2) is reported worldwide. PCV2 commercial vaccine was introduced in 2006 in U.S. and in 2008 in Brazil. Although PCV2 vaccines have been widely used, cases of PCV2 systemic disease have been reported in the last years. Eleven nursery or fattening pigs suffering from PCV2 systemic disease were selected from eight PCV2-vaccinated farms with historical records of PCV2 systemic disease in Southern Brazil. PCV2 genomes were amplified and sequenced from lymph node samples of selected pigs. The comparison among the ORF2 amino acid sequences of PCV2 isolates revealed three amino acid substitutions in the positions F57I, N178S and A190T, respectively. Using molecular modeling, a structural model for the capsid protein of PCV2 was built. Afterwards, the mutated residues positions were identified in the model. The structural analysis of the mutated residues showed that the external residue 190 is close to an important predicted region for antibodies recognition. Therefore, changes in the viral protein conformation might lead to an inefficient antibody binding and this could be a relevant mechanism underlying the recent vaccine failures observed in swine farms in Brazil.


Subject(s)
Capsid Proteins/chemistry , Circovirus/chemistry , Amino Acid Substitution , Animals , Brazil , Capsid Proteins/genetics , Circoviridae Infections/veterinary , Circoviridae Infections/virology , Circovirus/genetics , Circovirus/isolation & purification , Models, Molecular , Protein Conformation , Swine , Swine Diseases/virology
9.
Trop Anim Health Prod ; 49(5): 1071-1075, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28326486

ABSTRACT

Porcine circovirus type 2 (PCV2) has been identified in pig population in Brazil since 2000, but scarce studies involving wild boars with PCV2 infection are reported in the country. This study aimed to perform the genetic characterization of PCV2 detected in clinically healthy captive wild boars from farms located in Southern Brazil. Bronchial and mesenteric lymph nodes from 129 clinically healthy captive wild boars were tested by nested PCR for PCV2 detection. Six out of 38 positive samples (29.5%) were submitted to a quantitative real time PCR (qPCR) and genetic sequencing. Viral load up to 1.19 × 109 viral DNA copies/uL was detected in lymph nodes samples by qPCR. According to the ORF2 gene sequence analysis, all PCV2 samples were classified into PCV2b genotype. Comparisons based on a 702 nt region of the ORF2 of all six isolates revealed a high degree of similarity between these isolates. The ORF2 sequences characterized here share 97.1-100% of nucleotide identity and 95.7-100% of amino acid identity with other PCV2b isolated in Brazil from wild boars and feral pigs. This study reports the first detection and genetic characterization of PCV2b in captive wild boars in Brazil and provides important information on PCV2 infection in this domesticated species.


Subject(s)
Circoviridae Infections/veterinary , Circovirus/genetics , Genome, Viral , Swine Diseases/virology , Animals , Brazil , Circoviridae Infections/virology , Circovirus/classification , Phylogeny , Sequence Analysis, DNA , Swine
10.
mSphere ; 1(6)2016.
Article in English | MEDLINE | ID: mdl-27981236

ABSTRACT

The H1 subtype of influenza A viruses (IAVs) has been circulating in swine since the 1918 human influenza pandemic. Over time, and aided by further introductions from nonswine hosts, swine H1 viruses have diversified into three genetic lineages. Due to limited global data, these H1 lineages were named based on colloquial context, leading to a proliferation of inconsistent regional naming conventions. In this study, we propose rigorous phylogenetic criteria to establish a globally consistent nomenclature of swine H1 virus hemagglutinin (HA) evolution. These criteria applied to a data set of 7,070 H1 HA sequences led to 28 distinct clades as the basis for the nomenclature. We developed and implemented a web-accessible annotation tool that can assign these biologically informative categories to new sequence data. The annotation tool assigned the combined data set of 7,070 H1 sequences to the correct clade more than 99% of the time. Our analyses indicated that 87% of the swine H1 viruses from 2010 to the present had HAs that belonged to 7 contemporary cocirculating clades. Our nomenclature and web-accessible classification tool provide an accurate method for researchers, diagnosticians, and health officials to assign clade designations to HA sequences. The tool can be updated readily to track evolving nomenclature as new clades emerge, ensuring continued relevance. A common global nomenclature facilitates comparisons of IAVs infecting humans and pigs, within and between regions, and can provide insight into the diversity of swine H1 influenza virus and its impact on vaccine strain selection, diagnostic reagents, and test performance, thereby simplifying communication of such data. IMPORTANCE A fundamental goal in the biological sciences is the definition of groups of organisms based on evolutionary history and the naming of those groups. For influenza A viruses (IAVs) in swine, understanding the hemagglutinin (HA) genetic lineage of a circulating strain aids in vaccine antigen selection and allows for inferences about vaccine efficacy. Previous reporting of H1 virus HA in swine relied on colloquial names, frequently with incriminating and stigmatizing geographic toponyms, making comparisons between studies challenging. To overcome this, we developed an adaptable nomenclature using measurable criteria for historical and contemporary evolutionary patterns of H1 global swine IAVs. We also developed a web-accessible tool that classifies viruses according to this nomenclature. This classification system will aid agricultural production and pandemic preparedness through the identification of important changes in swine IAVs and provides terminology enabling discussion of swine IAVs in a common context among animal and human health initiatives.

11.
Vet Microbiol ; 180(1-2): 118-22, 2015 Oct 22.
Article in English | MEDLINE | ID: mdl-26345257

ABSTRACT

Influenza A virus (FLUAV) infections are endemic in pork producing countries worldwide but in Brazil it was not considered an important pathogen in pigs. Since the emergence of 2009 pandemic H1N1 (H1N1pdm) FLUAV, many outbreaks of respiratory disease were observed in pig herds. The aim of this study was to evaluate FLUAV infection in swine in 48 pig farms located in seven Brazilian states with previous reports of influenza-like signs by clinical, serological and virological cross-sectional studies. Serological results showed that pigs from all farms had anti-influenza antibodies by NP-ELISA. Antibodies to H3N2, H1N2 and H1N1pdm were detected by HI in pigs from 24 farms. Co-infection with two or more FLUAV subtypes was detected in pigs in seven of those 24 farms. Detection of FLUAV in nasal swabs and oral fluids by RT-qPCR indicated a global concordance >81% for the two biological samples. Moreover, our results show that H1N1pdm, H1N2 and H3N2 viruses are widespread in Brazilian pig herds. The monitoring of FLUAV emergence and evolution in pigs is urgent, as well the study of the pathogenesis of Brazilian isolates, aiming to control influenza in pigs.


Subject(s)
Disease Outbreaks/veterinary , Influenza A Virus, H1N1 Subtype/isolation & purification , Orthomyxoviridae Infections/veterinary , Swine Diseases/virology , Animals , Brazil/epidemiology , Cross-Sectional Studies , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Real-Time Polymerase Chain Reaction , Swine , Swine Diseases/epidemiology
12.
Virus Res ; 210: 327-36, 2015 Dec 02.
Article in English | MEDLINE | ID: mdl-26362524

ABSTRACT

A genome-wide association study for immune response to influenza vaccination in a crossbred swine population was conducted. Swine influenza is caused by influenza A virus (FLUAV) which is considered one of the most prevalent respiratory pathogens in swine worldwide. The main strategy used to control influenza in swine herds is through vaccination. However, the currently circulating FLUAV subtypes in swine are genetically and antigenically diverse and their interaction with the host genetics poses a challenge for the production of efficacious and cross-protective vaccines. In this study, 103 pigs vaccinated with an inactivated H1N1 pandemic virus were genotyped with the Illumina PorcineSNP60V2 BeadChip for the identification of genetic markers associated with immune response efficacy to influenza A virus vaccination. Immune response was measured based on the presence or absence of HA (hemagglutinin) and NP (nucleoprotein) antibodies induced by vaccination and detected in swine sera by the hemagglutination inhibition (HI) and ELISA assays, respectively. The ELISA test was also used as a measurement of antibody levels produced following the FLUAV vaccination. Associations were tested with x(2) test for a case and control data and using maximum likelihood method for the quantitative data, where a moderate association was considered if p<5×10(-5). When testing the association using the HI results, three markers with unknown location and three located on chromosomes SSCX, SSC14 and SSC18 were identified as associated with the immune response. Using the response to vaccination measured by ELISA as a qualitative and quantitative phenotype, four genomic regions were associated with immune response: one on SSC12 and three on chromosomes SSC1, SSC7, and SSC15, respectively. Those regions harbor important functional candidate genes possibly involved with the degree of immune response to vaccination. These results show an important role of host genetics in the immune response to influenza vaccination. Genetic selection for pigs with better response to FLUAV vaccination might be an alternative to reduce the impact of influenza virus infection in the swine industry. However, these results should to be validated in additional populations before its use.


Subject(s)
Genome-Wide Association Study , Host-Pathogen Interactions , Influenza A Virus, H1N1 Subtype/immunology , Influenza Vaccines/immunology , Animals , Antibodies, Viral/blood , Enzyme-Linked Immunosorbent Assay , Female , Hemagglutination Inhibition Tests , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Influenza Vaccines/administration & dosage , Male , Nucleocapsid Proteins , RNA-Binding Proteins/immunology , Swine , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/immunology , Viral Core Proteins/immunology
13.
Emerg Infect Dis ; 21(8): 1339-47, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26196759

ABSTRACT

The evolutionary origins of the influenza A(H1N1)pdm09 virus that caused the first outbreak of the 2009 pandemic in Mexico remain unclear, highlighting the lack of swine surveillance in Latin American countries. Although Brazil has one of the largest swine populations in the world, influenza was not thought to be endemic in Brazil's swine until the major outbreaks of influenza A(H1N1)pdm09 in 2009. Through phylogenetic analysis of whole-genome sequences of influenza viruses of the H1N1, H1N2, and H3N2 subtypes collected in swine in Brazil during 2009-2012, we identified multiple previously uncharacterized influenza viruses of human seasonal H1N2 and H3N2 virus origin that have circulated undetected in swine for more than a decade. Viral diversity has further increased in Brazil through reassortment between co-circulating viruses, including A(H1N1)pdm09. The circulation of multiple divergent hemagglutinin lineages challenges the design of effective cross-protective vaccines and highlights the need for additional surveillance.


Subject(s)
Disease Transmission, Infectious , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Influenza A virus/genetics , Influenza, Human/epidemiology , Phylogeny , Swine Diseases/epidemiology , Animals , Brazil/epidemiology , Humans , Swine
14.
J Virol Methods ; 219: 14-17, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25813599

ABSTRACT

Porcine parvovirus 4 (PPV4) is a DNA virus, and a member of the Parvoviridae family within the Bocavirus genera. It was detected recently in swine, but its epidemiology and pathology remain unclear. A TaqMan-based real-time PCR (qPCR) targeting a conserved region of the ORF3 gene of PPV4 was developed. The qPCR detection limit was 9.5 × 10(1) DNA copies/µL. There was no cross-reaction with porcine parvovirus, torque teno virus 1, torque teno virus 2, porcine circovirus type 1, porcine circovirus type 2, or with pseudorabies virus. Two hundred and seventy-two samples, including serum, semen, lungs, feces, ovarian follicular fluids, ovaries and uterus, were evaluated by qPCR and PPV4 was detected in 36 samples (13.2%). When compared with a conventional PCR (cPCR), the qPCR assay was 10 times more sensitive and the detection of PPV4 DNA in field samples was increased 2.5 times. Partial sequencing of PPV4 ORF3 gene, obtained from two pooled samples of uterus and ovaries, revealed a high nucleotide identity (98-99%) with a reference PPV4 sequence. The qPCR can be used as a fast and accurate assay for the detection and quantification of PPV4 in field samples and for epidemiological studies in swine herds.


Subject(s)
Parvoviridae Infections/veterinary , Parvovirus, Porcine/genetics , Real-Time Polymerase Chain Reaction , Swine Diseases/diagnosis , Swine Diseases/virology , Animals , DNA, Viral , Parvovirus, Porcine/classification , Polymerase Chain Reaction , Reproducibility of Results , Sensitivity and Specificity , Sequence Analysis, DNA , Swine , Viral Proteins/genetics
15.
Ann Agric Environ Med ; 22(1): 11-6, 2015.
Article in English | MEDLINE | ID: mdl-25780820

ABSTRACT

INTRODUCTION AND OBJECTIVE: Recently, investigations in a swine herd identified evidence of the existence of a novel member of the Hepadnavirus family endemic in swine. The aim of this study was to investigate the serological and molecular markers of Hepadnavirus circulation in Brazilian domestic swine and wild boar herds, and to evaluate the identity with HBV and other Hepadnaviruses reported previously. MATERIALS AND METHODS: For the study, 376 swine were screened for hepatitis B virus serological markers. Analyses were performed in serum samples using commercial enzyme-linked immunosorbent assay (ELISA) kits (DiaSorin®) for anti-HBc, HBsAg and anti-HBs. Reactive and undetermined swine serum samples were selected to perform DNA viral extraction (QIAamp DNA Mini Kit, Qiagen®), partial genome amplification and genome sequencing. RESULTS: From 376 swine samples analysed, 28 (7.45%) were reactive to anti-HBc, 3 (0.80%) to HBsAg and 6 (1.6%) to anti-HBs. Besides, more 17 (4.52%) swine samples analyzed were classified in the grey zone of the EIA test to anti-HBc and 2 (0.53%) to HBsAg. From 49 samples molecularly analyzed after serological trial, 4 samples showed a positive result for the qualitative PCR for Hepadnavirus. Phylogenetic reconstruction using partial genome sequencing (360 bp) of 3 samples showed similarity with HBV with 90.8-96.3% of identity. CONCLUSIONS: Serological and molecular data showed evidence of the circulation of a virus similar to hepatitis B virus in swine.


Subject(s)
Hepatitis B virus/classification , Hepatitis B virus/isolation & purification , Hepatitis B/veterinary , Swine Diseases/epidemiology , Animals , Brazil/epidemiology , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Hepatitis B/epidemiology , Hepatitis B/virology , Hepatitis B Antibodies/analysis , Hepatitis B Surface Antigens/analysis , Hepatitis B virus/genetics , Male , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction/veterinary , Sequence Analysis, DNA/veterinary , Swine , Swine Diseases/virology , Viral Proteins/genetics
16.
Influenza Other Respir Viruses ; 9(3): 161-7, 2015 May.
Article in English | MEDLINE | ID: mdl-25648743

ABSTRACT

BACKGROUND: Swine influenza virus (SIV) is the cause of an acute respiratory disease that affects swine worldwide. In Brazil, SIV has been identified in pigs since 1978. After the emergence of pandemic H1N1 in 2009 (H1N1pdm09), few studies reported the presence of influenza virus in Brazilian herds. OBJECTIVES: The objective of this study was to evaluate the serological profile for influenza virus in farrow-to-finish pig farms in Minas Gerais state, Brazil. METHODS: Thirty farms with no SIV vaccination history were selected from the four larger pig production areas in Minas Gerais state (Zona da Mata, Triângulo Mineiro/Alto Paranaíba, South/Southwest and the Belo Horizonte metropolitan area). At each farm, blood samples were randomly collected from 20 animals in each production cycle category: breeding animals (sows and gilts), farrowing crate (2-3 weeks), nursery (4-7 weeks), grower pigs (8-14 weeks), and finishing pigs (15-16 weeks), with 100 samples per farm and a total of 3000 animals in this study. The samples were tested for hemagglutination inhibition activity against H1N1 pandemic strain (A/swine/Brazil/11/2009) and H3N2 SIV (A/swine/Iowa/8548-2/98) reference strain. RESULTS: The percentages of seropositive animals for H1N1pdm09 and H3N2 were 26.23% and 1.57%, respectively, and the percentages of seropositive herds for both viruses were 96.6% and 13.2%, respectively. CONCLUSIONS: The serological profiles differed for both viruses and among the studied areas, suggesting a high variety of virus circulation around the state, as well as the presence of seronegative animals susceptible to influenza infection and, consequently, new respiratory disease outbreaks.


Subject(s)
Antibodies, Viral/blood , Influenza A Virus, H1N1 Subtype/immunology , Influenza A Virus, H3N2 Subtype/immunology , Orthomyxoviridae Infections/veterinary , Swine Diseases/epidemiology , Agriculture , Animals , Brazil , Hemagglutination Inhibition Tests , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Seroepidemiologic Studies , Swine , Swine Diseases/prevention & control , Swine Diseases/virology
17.
J Gen Virol ; 95(Pt 12): 2618-2626, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25127710

ABSTRACT

Interactions between the viral surface glycoprotein haemagglutinin (HA) and the corresponding receptors on host cells is one important aspect of influenza virus infection. Mutations in HA have been described to affect pathogenicity, antigenicity and the transmission of influenza viruses. Here, we detected polymorphisms present in HA genes of two pandemic 2009 H1N1 (H1N1pdm09) isolates, A/California/04/2009 (Ca/09) and A/Mexico/4108/2009 (Mx/09), that resulted in amino acid changes at positions 186 (S to P) and 194 (L to I) of the mature HA1 protein. Although not reported in the published H1N1pdm09 consensus sequence, the P186 genotype was more readily detected in primary infected and contact-naïve pigs when inoculated with a heterogeneous mixed stock of Ca/09. Using reverse genetics, we engineered Ca/09 and Mx/09 genomes by introducing Ca/09 HA with two naturally occurring variants expressing S186/I194 (HA-S/I) and P186/L194 (HA-P/L), respectively. The Ca/09 HA with the combination of P186/L194 with either the Ca/09 or Mx/09 backbone resulted in higher and prolonged viral shedding in naïve pigs. This efficiency appeared to be more likely through an advantage in cell surface attachment rather than replication efficiency. Although these mutations occurred within the receptor-binding pocket and the Sb antigenic site, they did not affect serological cross-reactivity. Relative increases of P186 in publicly available sequences from swine H1N1pdm09 viruses supported the experimental data, indicating this amino acid substitution conferred an advantage in swine.


Subject(s)
Gene Expression Regulation, Viral/physiology , Hemagglutinin Glycoproteins, Influenza Virus/metabolism , Influenza A Virus, H1N1 Subtype/metabolism , Orthomyxoviridae Infections/veterinary , Swine Diseases/virology , Virus Shedding/genetics , Animals , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Influenza A Virus, H1N1 Subtype/genetics , Nose/virology , Orthomyxoviridae Infections/transmission , Orthomyxoviridae Infections/virology , Polymorphism, Genetic , Swine , Swine Diseases/transmission
18.
Vet Microbiol ; 168(1): 34-40, 2014 Jan 10.
Article in English | MEDLINE | ID: mdl-24238665

ABSTRACT

Influenza is a viral disease that affects human and several animal species. In Brazil, H1N1, H3N2 and 2009 pandemic H1N1 A(H1N1)pdm09 influenza A viruses (IAV) circulate in domestic swine herds. Wild boars are also susceptible to IAV infection but in Brazil until this moment there are no reports of IAV infection in wild boars or in captive wild boars populations. Herein the occurrence of IAV in captive wild boars with the presence of lung consolidation lesions during slaughter was investigated. Lung samples were screened by RT-PCR for IAV detection. IAV positive samples were further analyzed by quantitative real-time PCR (qRRT-PCR), virus isolation, genomic sequencing, histopathology and immunohistochemistry (IHC). Eleven out of 60 lungs (18.3%) were positive for IAV by RT-PCR and seven out of the eleven were also positive for A(H1N1)pdm09 by qRRT-PCR. Chronic diffuse bronchopneumonia was observed in all samples and IHC analysis was negative for influenza A antigen. Full genes segments of H1N2 IAV were sequenced using Illumina's genome analyzer platform (MiSeq). The genomic analysis revealed that the HA and NA genes clustered with IAVs of the human lineage and the six internal genes were derived from the H1N1pdm09 IAV. This is the first report of a reassortant human-like H1N2 influenza virus infection in captive wild boars in Brazil and indicates the need to monitor IAV evolution in Suidae populations.


Subject(s)
Genome, Viral/genetics , Influenza A Virus, H1N2 Subtype/classification , Influenza A Virus, H1N2 Subtype/genetics , Orthomyxoviridae Infections/veterinary , Orthomyxoviridae Infections/virology , Phylogeny , Sus scrofa/virology , Swine Diseases/virology , Animals , Brazil , Genes, Viral/genetics , Humans , Influenza A Virus, H1N2 Subtype/isolation & purification , Lung/virology , Molecular Sequence Data , Swine
19.
Pesqui. vet. bras ; 33(1): 61-73, Jan. 2013. ilus, tab
Article in Portuguese | LILACS | ID: lil-668094

ABSTRACT

Este trabalho descreve a colheita adequada de amostras, as técnicas/procedimentos disponíveis para o diagnóstico de influenza A em suínos, assim como os resultados e suas respectivas interpretações, para auxiliar médicos veterinários de campo na identificação dessa doença. Em suínos vivos, as amostras adequadas são: secreção nasal, fluido oral e sangue (soro). Para suínos mortos, colher preferencialmente amostras de pulmão com consolidação cranioventral. Secreção nasal e fragmentos de pulmão refrigerado são utilizados para detectar partícula viral viável (isolamento viral - IV) ou ácido nucleico viral (RT-PCR convencional e RT-PCR em tempo real). As amostras não devem ser congeladas, pois o vírus é inativado a -20°C. A caracterização molecular dos isolados é feita pela análise filogenética obtida pelo sequenciamento de DNA. O soro é utilizado para a detecção de anticorpos (Acs) por meio do teste da inibição da hemaglutinação e ELISA. O fluido oral pode ser utilizado para detecção de anticorpo (ELISA) ou de vírus. Fragmentos de pulmão fixados em formol a 10% são examinados microscopicamente para identificar pneumonia broncointersticial e para detecção de antígeno viral pela imuno-histoquímica (IHQ). Para o sucesso do diagnóstico, as amostras devem ser colhidas de suínos que estão preferencialmente na fase aguda da doença, para aumentar as chances de detecção viral. As melhores opções para o diagnóstico de influenza A em suínos vivos são RT-PCR e isolamento viral de amostras de swab nasal ou fluido oral. Pulmão para análise por RT-PCR, isolamento viral ou IHQ é a amostra de escolha em suínos mortos. Testes sorológicos têm valor diagnóstico limitado e são utilizados apenas para determinar o estado imune do rebanho, não indicando doença clínica, pois os Acs são detectados 7-10 dias pós-infecção (fase subaguda). O diagnóstico de influenza é importante para avaliar o envolvimento desse agente no complexo de doença respiratória suína. Além disso, o isolamento do vírus influenza é essencial para o monitoramento dos principais subtipos circulantes em uma determinada região ou país, assim como para a detecção de novos rearranjos virais, já que influenza é considerada uma zoonose.


This article is intended to describe the adequate sample collection, the laboratory procedures/techniques, the expected results and their interpretation for diagnosis of influenza infection in swine, serving as a support for field veterinarians. In live pigs, the samples to be taken are nasal secretions, oral fluids and blood. For dead pigs, preference should be given to samples of cranioventral lung consolidation. Nasal discharge and chilled lung fragments are used for detection of virus (virus isolation - VI) or viral nucleic acids (conventional RT-PCR and real-time RT-PCR). Samples should not be frozen, because the virus is inactivated at -20°C. Molecular characterization of isolates is performed by phylogenetic analysis of gene sequences obtained by DNA sequencing. Serum is used for the detection of antibodies using hemagglutination inhibition (HI) test and ELISA. Oral fluid may be used for either antibody (ELISA) or viral detection. Fragments of lung fixed in 10% formaldehyde are used for histopathological analysis to identify bronchointerstitial pneumonia, and for immunohistochemistry (IHC) for antigens. For a successful diagnosis, sampling should be preferably performed in the acute phase of the disease to improve chances of virus detection. The best options to perform the diagnosis of influenza A in a swine herd are RT-PCR and VI from nasal swabs or oral fluid in live pigs and/or lung tissue for RT-PCR, VI or IHC in dead pigs. Serological tests are of very limited diagnostic value and are useful only to determine the immune status of the herd, not indicating clinical disease, because antibodies are detected after 7-10 days post infection (subacute phase). The diagnosis of influenza is important to evaluate the involvement of this agent in the complex of respiratory diseases in pigs. Furthermore, the isolation of influenza virus is essential for monitoring the main subtypes circulating in a given region or country, as well as for the detection of potential new viral reassortants, because influenza is considered a zoonosis.


Subject(s)
Animals , Alphainfluenzavirus/isolation & purification , Specimen Handling , Swine/virology , Diagnostic Techniques and Procedures/veterinary , Polymerase Chain Reaction , Saliva
20.
Curr Top Microbiol Immunol ; 370: 113-32, 2013.
Article in English | MEDLINE | ID: mdl-22266673

ABSTRACT

The 2009 pandemic H1N1 infection in humans has been one of the greatest concerns for public health in recent years. However, influenza in pigs is a zoonotic viral disease well-known to virologists for almost one century with the classical H1N1 subtype the only responsible agent for swine influenza in the United States for many decades. Swine influenza was first recognized clinically in pigs in the Midwestern U.S. in 1918 and since that time it has remained important to the swine industry throughout the world. Since 1988, however, the epidemiology of swine influenza changed dramatically. A number of emerging subtypes and genotypes have become established in the U.S. swine population. The ability of multiple influenza virus lineages to infect pigs is associated with the emergence of reassortant viruses with new genomic arrangements, and the introduction of the 2009 pandemic H1N1 from humans to swine represents a well-known example. The recent epidemiological data regarding the current state of influenza A virus subtypes circulating in the Canadian and American swine population is discussed in this review.


Subject(s)
Influenza A virus/isolation & purification , Orthomyxoviridae Infections/veterinary , Reassortant Viruses/isolation & purification , Swine Diseases/epidemiology , Swine Diseases/virology , Animals , Humans , Influenza A Virus, H1N1 Subtype/genetics , Influenza A Virus, H1N1 Subtype/isolation & purification , Influenza A virus/genetics , Influenza, Human/epidemiology , Influenza, Human/virology , North America/epidemiology , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Reassortant Viruses/genetics , Swine
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