Achievements and Challenges of Classical Swine Fever Eradication in Brazil.

Classical swine fever virus (CSFV) causes one of the most critical diseases in the porcine industry worldwide. In Brazil, the first description of the infection was reported in 1888, and the national recognition of the first free zone (FZ) occurred in 2001. Brazil has been recently recognized (2015-2016) by the World Organisation for Animal Health (OIE) with an FZ involving 15 states and the Federal District, corresponding to 95% of the industrial production of pigs in the country, and a non-free zone (NFZ), comprised by the North and Northeast regions of the country, with approximately 18% of the national pig herd and 5% of industrial production. This review aims to describe the history, the control and eradication actions, the recent occurrence of outbreaks in the NFZ, and the results obtained by the surveillance systems' action in the FZ for CSF in Brazil since its creation. In the passive surveillance system, the notification of the suspect cases of classical swine fever (CSF) is mandatory while in the active surveillance system adopted in the FZ consists of serological monitoring of certified swine breeding farms (CSBFs), intensive pig farming (IPF), non-technified pig herds (NTPig), surveillance in slaughterhouses and monitoring the populations of wild pigs. In this region, the last outbreaks of the disease occurred in 1998, while in the NFZ, 28 outbreaks were detected from 2005 to 2017, with an apparent lethality rate of 93.96% (840/894). However, in 2018 and 2019, 68 new outbreaks were registered with an apparent lethality rate of 75.05% (1095/1459). Therefore, in 2019, the Brazil CSF-Free Strategic Plan was created to eradicate the infection from the country's NFZ, since outbreaks in this region present a risk of reintroducing the disease FZ. Finally, differences in characteristics between the regions show factors that still need to be considered for the construction of a robust surveillance system in the NFZ and some improvements in the FZ. Thus, the control of CSF throughout the Brazilian territory requires strict sanitary guidelines, promoting animal health and, consequently, the national production chain's competitiveness.

No effects of noncytopathic bovine viral diarrhea virus type 2 on the reproductive tract of experimentally inoculated boars.

Bovine viral diarrhea virus (BVDV) infections in pigs may result in transient leukopenia, chronic gastroenteritis, septicemia, and hemorrhagic lesions. Both classical swine fever virus (CSF) and the atypical porcine pestivirus (APPV) are shed in the semen of infected boars. Because these viruses share conserved regions and present antigenic similarity, they may not be the only species belonging to the genus Pestivirus that can be shed in the semen of infected pigs. The objective of this study was to evaluate the testicular and epididymal changes, seminal parameters, and viral shedding in the reproductive tract of boars experimentally inoculated with noncytopathic BVDV-2. Six males were selected, and samples of blood, semen, and preputial swabs were collected every four days until the 52nd day after inoculation. The samples were tested for the presence of viral RNA by RT-PCR. An aliquot of whole blood was used to perform hematological analyses, which showed a significant reduction in monocyte counts and a significant increase in lymphocyte counts when comparing the pre- and postinoculation periods. The neutralizing antibody titers were determined by the virus neutralization test. None of the animals presented clinical signs or worsening of the seminal parameters that were evaluated. Moreover, BVDV-2 shedding by the reproductive route was not observed.

Coxiella burnetii associated with BVDV (Bovine Viral Diarrhea Virus), BoHV (Bovine Herpesvirus), Leptospira spp., Neospora caninum, Toxoplasma gondii and Trypanosoma vivax in reproductive disorders in cattle.

This is a cross-sectional study to assess the presence of antibodies in ruminants against selected pathogens associated with reproductive disorders in cattle in four Brazilian states, including the zoonotic agent Coxiella burnetii. The used tests were Virus Neutralization Assay for IBR and BVD, Microscopic Agglutination Test for Leptospira spp., Indirect Fluorescent Antibody Test (IFAT) for C. burnetii and Toxoplasma gondii, and Enzyme-Linked Immunosorbent Assay for Neospora caninum and Trypanosoma vivax. Seropositivity for C. burnetii was 13.7% with titers from 128 to 131,072; 57.8% for BoHV-1, with titers between 2 and 1,024; 47.1% for BVDV-1a, with titers from 10 to 5,120; 89.2% for N. caninum; 50% for T. vivax; and 52.0% for Leptospira spp., with titers between 100 to 800 (the following serovars were found: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona and Icterohaemorrhagiae); 19.6% for T. gondii with titer of 40. This is the first study that has identified C. burnetii in cattle associated with BoHV and BVDV, N. caninum, Leptospira spp., T. gondii and T. vivax. Thus, future studies should be conducted to investigate how widespread this pathogen is in Brazilian cattle herds.

Molecular detection of Mycoplasma suis in captive white-lipped peccaries (Tayassu pecari) and wild boars (Sus scrofa) in Brazil.

Mycoplasma suis, the etiological agent of swine hemoplasmosis, is an epicellular bacterium that adheres to the surface of pig erythrocytes leading to deformations of the target cells. Little is known about the occurrence of M. suis in wild swine populations around the world, its economic impact on swine herds, and the risk of human infection. The aim of this study was to investigate, by quantitative real-time PCR (qPCR) based on the 16S rRNA gene, the occurrence of M. suis in a captive population of white-lipped peccaries (100 Tayassu pecari) and in free-living wild boars (14 Sus scrofa) in Brazil. None of the white-lipped peccaries were positive for M. suis, whereas seven (50%) wild boars were positive in qPCR assays. The quantification of M. suis-16S rRNA copies/µL ranged from 1.42 × 10° to 3.906 × 101 in positive animals, indicating a low bacteremia and a chronic carrier status in free-living wild boars. In conclusion, M. suis might be a non-frequent pathogen in wild suids maintained in captivity. Despite the low bacteremia, the prevalence of M. suis in wild boar population in Brazil seems to be high.

High frequency and molecular characterization of porcine hemotrophic mycoplasmas in Brazil.

Mycoplasma suis and Mycoplasma parvum are the two hemotrophic mycoplasmas species described in pigs. M. suis is involved in infectious anemia, while M parvum infection is commonly subclinical. The objectives of this study were twofold: (i) to investigate the prevalence of porcine hemotrophic mycoplasmas in sows from the southern region of Brazil by quantitative real-time PCR (qPCR) and (ii) to genetically characterize a subset of the samples based on the 16S rRNA gene. A total of 429 blood samples were evaluated from 53 different farm sites. Porcine hemoplasmas was detected at all the 53 tested sites and in 79.72% of the samples (342/429). Two sequences were obtained for Mycoplasma spp. The phylogenetic analysis based on the 16S rRNA gene (900 bp) showed that the Mycoplasma sequences were closely related to the M. suis cluster and that one sequence was positioned in the M. parvum cluster. In conclusion, porcine hemoplasmas have a high rate of prevalence in sows from commercial farms in the southern region of Brazil. This study demonstrated the first molecular detection and characterization of partial 16S rRNA gene of M. parvum in Brazil.

Coxiella burnetii associated with BVDV (Bovine Viral Diarrhea Virus), BoHV (Bovine Herpesvirus), Leptospira spp, Neospora caninum, Toxoplasma gondii and Trypanosoma vivax in reproductive disorders in cattle / Coxiella burnetii associada ao BVDV (Vírus da Diarreia Viral Bovina), BoHV (Vírus do Herpes Bovino), Leptospira spp, Neospora caninum, Toxoplasma gondii e Trypanosoma vivax em distúrbios reprodutivos em bovinos

Abstract This is a cross-sectional study to assess the presence of antibodies in ruminants against selected pathogens associated with reproductive disorders in cattle in four Brazilian states, including the zoonotic agent Coxiella burnetii. The used tests were Virus Neutralization Assay for IBR and BVD, Microscopic Agglutination Test for Leptospira spp., Indirect Fluorescent Antibody Test (IFAT) for C. burnetii and Toxoplasma gondii, and Enzyme-Linked Immunosorbent Assay for Neospora caninum and Trypanosoma vivax. Seropositivity for C. burnetii was 13.7% with titers from 128 to 131,072; 57.8% for BoHV-1, with titers between 2 and 1,024; 47.1% for BVDV-1a, with titers from 10 to 5,120; 89.2% for N. caninum; 50% for T. vivax; and 52.0% for Leptospira spp., with titers between 100 to 800 (the following serovars were found: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona and Icterohaemorrhagiae); 19.6% for T. gondii with titer of 40. This is the first study that has identified C. burnetii in cattle associated with BoHV and BVDV, N. caninum, Leptospira spp., T. gondii and T. vivax. Thus, future studies should be conducted to investigate how widespread this pathogen is in Brazilian cattle herds.

Resumo Este é um estudo transversal para avaliar a presença de anticorpos em ruminantes contra patógenos selecionados e associados a distúrbios reprodutivos em bovinos de quatro estados brasileiros, incluindo o agente zoonótico Coxiella burnetii. Os testes utilizados foram Teste de Vírus-Neutralização para BoHV e BVDV, teste de Aglutinação Microscópica para Leptospira spp., Reação de Imunofluorescência Indireta for C. burnetii e Toxoplasma gondii, e Ensaio de Imunoabsorção Enzimática para Neospora caninum e Trypanosoma vivax. A soropositividade para C. burnetii foi de 13,7% com títulos de 128 a 131.072; 57,8% para BoHV-1, com títulos entre 2 a 1.024; 47,1% para BVDV-1a, com títulos de 10 a 5.120; 89,2% para N. caninum; 50% para T. vivax; e 52,0% para Leptospira spp., com títulos entre 100 a 800 (sorovares encontrados: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona e Icterohaemorrhagiae) 19,6% para T. gondii com título de 40. Este é o primeiro estudo que evidencia a participação de C. burnetii em bovinos associada ao Vírus da Rinotraqueíte bovina infecciosa e da diarreia viral bovina, N. caninum, Leptospira spp., T. gondii e T. vivax em bovinos. Desta forma, futuros estudos devem ser conduzidos a fim de investigar o quão disseminado se encontra este patógeno em rebanhos bovinos brasileiros.

Atypical Porcine Pestivirus (APPV) as a New Species of Pestivirus in Pig Production.

The genus Pestivirus, which belongs to the family Flaviviridae, includes ssRNA+ viruses responsible for infectious diseases in swine, cattle, sheep, goats, and other domestic and wild animals. Recently, several putative pestiviruses species have been discovered and characterized in mammalian species (giraffe pestivirus, antelope pestivirus, HoBi virus, Bungowannah virus, and Linda virus); one of these is a genetically distinct pestivirus, named atypical porcine pestivirus (APPV), discovered using the next-generation sequencing technology. APPV has been detected in piglets with congenital tremor (CT) from four different continents, including North America, South America, Europe, and Asia. There is strong evidence that experimental inoculation and in field outbreaks involving APPV induce CT in piglets. Additionally, splay leg (SL) syndrome has been observed concurrently with CT, and it was induced by APPV in experimental studies and some field cases. Animals with a persistent and/or chronic infection condition can shed the virus over time. Viral-RNA is frequently detected in different tissues from CT-piglets; however, high loads of APPV are detected most consistently in central nervous tissue. Moreover, the APPV genome has been recently detected in semen and preputial swabs from boar studs, as well as in serum and tissue samples from wild boars and domestic adult pigs, all known to be clinically healthy. Phylogenetic analysis revealed that the APPV sequence (complete or partial polyprotein) exhibits high genetic diversity between viral strains detected in different countries and formed independent clusters according to geographic location. Additional studies are needed to evaluate the molecular detection and sero-prevalence of APPV around the world. Lastly, more research is needed to understand clinical presentations associated with APPV infection, as well as the economic losses related to the virus in pig production worldwide.

Ultrasonography for lymph nodes metastasis identification in bitches with mammary neoplasms.

The aim of this study was to evaluate and compare the diagnostic accuracy of B-mode, Doppler ultrasonography and Acoustic Radiation Force Impulse (ARFI) elastography in the identification of axillary and inguinal lymph nodes metastasis in bitches with mammary neoplasms. The axillary (n = 96) and inguinal (n = 100) lymph nodes of 100 bitches were evaluated using B-Mode, Colour Doppler and ARFI-elastography. After this evaluation, mastectomy and lymph nodes excision were performed and these structures were histologically classified as free, reactive or metastatic. Ultrasonographic parameters were compared by Chi-Square or ANOVA tests and if they are significant, discriminative power analysis according to histopathological classification was performed (ROC analysis). The ARFI-elastography shear wave velocity (SWV) enabled metastasis identification in inguinal (sensitivity 95% specificity 87%) and axillary lymph nodes (sensitivity 100% specificity 94%). While B-Mode ultrasound Short/Long axis ratio evaluation of inguinal and axillary lymph nodes only resulted in a sensitivity around of 71% and specificity of 55%. In conclusion, B-Mode ultrasonography may contribute to diagnosis of metastasis in axillary and inguinal lymph nodes of bitches affected by mammary neoplasm with limited accuracy, while SWV evaluation proved to be an excellent diagnosis tool, which allows differentiation between free, reactive and tumour metastatic lymph nodes.

Ocorrência de dermatófitos em tegumento de bovinos e ovinos hígidos / Occurrence of dermatophytes in healthy cattle and sheep tegument / Ocurrencia de dermatofitos en tegumento de bovinos y ovinos hígidos

O objetivo desta pesquisa foi verificar a prevalência de dermatófitos em tegumento de bovinos e ovinos hígidos e a sua capacidade de transmissão e desenvolvimento da doença. Para isso, foram coletadas amostras de pelo e descamações de 90 bovinos e 90 ovinos hígidos. Essas amostras foram semeadas em meio DTM e ágar Sabouraud Dextrose enriquecido com extrato de levedura, tiamina, antibióticos (estreptomicina e cloranfenicol) e suplementado de cicloheximida, sendo essas incubadas a 35 oC por 10 dias. As culturas positivas foram avaliadas macro e microscopicamente e, os fungos foram identificados por métodos bioquímicos. Verificou-se que as espécies isoladas com maior frequência, nos bovinos, foram T. mentagrophytes e M. gypseum. Nos ovinos, foi constatada maior ocorrência do agente T. verrucosum, seguido por T. mentagrophytes e M. gypseum. Houve também isolamento de M. canis, porém, em um número reduzido de amostras. Assim, concluiu-se que o tegumento de bovinos e de ovinos hígidos apresentou incidência elevada de dermatófitos de diferentes espécies, em amostras coletadas durante o período chuvoso do ano. Por isso, em animais jovens ou naqueles submetidos a elevados níveis de estresse e, consequente queda da resposta imunológica, o risco de desenvolvimento da dermatofitose e da transmissão dos dermatófitos neste período são eminentes.(AU)

The purpose of this study was to determine the prevalence of dermatophytes in healthy cattle and sheep teguments, and their ability to transmit and develop diseases. Samples of fur and flaking from 90 healthy cattle and 90 healthy sheep were collected. These samples were plated on DTM medium and Sabouraud Dextrose agar supplemented with yeast extract, thiamine, antibiotics (streptomycin and chloramphenicol) enhanced with cycloheximide. They were then incubated at 35 °C for 10 days. Positive cultures were macroscopically and microscopically evaluated, and fungi were identified by biochemical methods. It was found that the most frequent species isolated in cattle were T. mentagrophytes and M. gypseum. In sheep, a higher occurrence of the agent T. verrucosum could be observed, followed by T. mentagrophytes and M. gypseum. There was also the isolation of M. canis, but in a small number of samples. Thus, it could be concluded that the tegument of healthy cattle and sheep showed high incidence of dermatophytes from different species in samples collected during the rainy season of the year. Therefore, in young animals, in those subjected to high levels of stress and consequent drop in immune response, there is eminent risk of developing dermatophytosis and transmission of dermatophytes in this period.(AU)

El objetivo de esta investigación ha sido verificar la prevalencia de dermatofitos en tegumento de bovinos y ovinos hígidos y su capacidad de transmisión y desarrollo de la enfermedad. Para ello, se recolectó muestras de pelo y descamaciones de 90 bovinos y 90 ovinos hígidos. Esas muestras fueron sembradas en medio DTM y agar Sabouraud Dextrosis enriquecida con extracto de levadura, tiamina, antibióticos (estreptomicina y cloranfenicol) y suplementado de cicloheximida, siendo esas incubadas a 35ºC durante 10 días. Los cultivos positivos fueron evaluados macro y microscópicamente y los hongos fueron identificados por métodos bioquímicos. Se verificó que las especies aisladas con mayor frecuencia, en los bovinos, fueron T. mentagrophytes y M. gypseum. En los ovinos, se constató mayor ocurrencia del agente T. verrucosum, seguido por T. mentagrophytes y M. gypseum. También hubo aislamiento de M. canis, sin embargo, en un número reducido de muestras. Así, se concluyó que el tegumento de bovinos y de ovinos hígidos presentó una elevada incidencia de dermatofitos de diferentes especies, en muestras recogidas durante el período lluvioso del año. Por lo tanto, en animales jóvenes o en aquellos sometidos a altos niveles de estrés y, consecuente caída de la respuesta inmunológica, el riesgo de desarrollo de la dermatofitosis y de la transmisión de los dermatofitos en este período son eminentes.(AU)

Description of risk factors associated with the detection of BVDV antibodies in Brazilian pig herds.

Bovine viral diarrhea virus (BVDV) infects ruminants as primary hosts. However, other animals like pigs are susceptible. This study was conducted to investigate seroprevalence and risk factors associated with the detection of BVDV antibodies in pig herds. A total of 1.705 serum samples of 33 finisher herds, from seven Brazilian states, were collected in slaughterhouses. The samples were tested by virus neutralization (VN) test. In total, 5.35% (91/1.705) were positive and 64% (21/33) of the herds had positive animals. A significant association with "trucks are not cleaned and disinfected" and "visitors do not respect 72-h interval between visits to farms" (P < 0.05) was found in association with detection of BVDV-2 antibodies. This study suggests that important biosecurity gaps are present in Brazilian pig farms, as the presence of BVDV antibodies in pigs suggests (direct or indirect) contact with population(s) of ruminant species. Closing biosecurity gaps prevents spread of BVDV and other pathogens such as foot-and-mouth disease virus (FMDV) between pig and ruminant farms. This data should be taken in account by CSF surveillance programs, once cross-reaction in serologic tests between classical swine fever virus (CSFV) and BVDV antibodies has been shown to occur.

Porcine Astrovirus Type 3 in Central Nervous System of Swine with Polioencephalomyelitis.

Using next-generation sequencing, we identified and genetically characterized a porcine astrovirus type 3 strain found in tissues from the central nervous system of 1 piglet and 3 sows with neurologic signs and nonsuppurative polioencephalomyelitis. Further studies are needed to understand the potential for cross-species transmission and clinical impact.

A cross-sectional study of swine influenza in intensive and extensive farms in the northeastern region of the state of São Paulo, Brazil.

Swine influenza (SI) is a seasonal infectious disease highly important to the world pig industry. Loss of daily weight gain, increased costs for the prevention and treatment of secondary infections are the main economic losses associated with the presence of this disease. However, some epidemiological features of SI remain quite unclear. This study focused on assessing the prevalence of swine influenza virus (SIV) infection in intensive and extensive pig herds and associating risk factors. A set of 601 blood samples of five intensive farrow-to-finish farms and 361 blood samples from 56 extensive farms were analyzed using an indirect ELISA kit CIVTEST SUIS INFLUENZA®, Hipra (Amer, Spain), in order to detect anti-SIV antibodies. In total, 24.13 % of samples from intensive herds were positive, while no positive samples were detected in extensive rearing herds. Sow and weaning piglets had the highest prevalence values. In the intensive rearing system, occurrence of reproductive disorders and exposure to recently introduced animals were positively associated with the disease occurrence in swine herds. The findings highlight the importance of sows in the epidemiology of the disease and bring information about risk factors involved in the occurrence of swine influenza in intensive herds.

Bovine viral diarrhea virus infections in pigs: why is this situation important for Brazilian herds? / A infecção pelo vírus da diarreia viral bovina em suínos: por que essa situação é importante para os rebanhos brasileiros?

Swine can be infected by the bovine viral diarrhea virus (BVDV) under natural conditions. For this reason, further information and divulgation are needed regarding the pathogenicity of this virus in swine. This infection is practically unknown in the realm of pig farming, and, as it shares some similarities with the classical swine fever virus (CSFV), its diagnosis becomes a challenge for official sanitary programs. Studies have shown the absence of clinical signs in piglets and reproductive problems in sows due to BVDV infections. There is little research on the prevalence, risk factors, preventive measures and control of BVDV in pigs around the world. And in Brazil, the data is practically non-existent. At the time of diagnosis, comparing the most efficient laboratory tests such as virus neutralization, ELISA, RT-PCR, and immunofluorescence so as to minimize the risk of cross serological reactions when dealing with a persistent or transient infection, can be an important tool. Moreover, the practical implications for CSFV eradication programs are a main reason for the development of further research against this infection. Therefore, this paper aims to review various aspects of BVDV infection in pigs, and how this information can be important for Brazilian herds.(AU)

O suíno pode ser infectado pelo vírus da diarreia viral bovina (BVDV) em condições naturais, por isso são necessárias maiores informações e mais divulgação sobre a ação deste vírus nos suínos. Esta infecção é praticamente desconhecida na suinocultura e, devido a algumas semelhanças com vírus da peste suína clássica (VPSC), torna-se um desafio para os programas sanitários oficiais. Estudos revelam a ausência de sinais clínicos em leitões ao mesmo tempo em que evidenciam problemas reprodutivos em porcas devido à infecção do BVDV. Poucas são as pesquisas sobre a prevalência, fatores de riscos, medidas de prevenção e controle do BVDV em suínos no mundo e, no Brasil, os dados são praticamente inexistentes. No diagnóstico, comparar os exames laboratoriais mais eficientes como a virusneutralização, ELISA, RT-PCR e imunofluorescência, diante de uma infecção persistente ou transitória, e assim minimizar o risco de reações sorológicas cruzadas pode ser uma ferramenta fundamental. Ademais, as implicações práticas em programas de erradicação da PSC são um grande motivo para o desenvolvimento de mais pesquisas frente a esta infecção. Portanto, este trabalho pretende revisar diversos aspectos da infecção do BVDV em suínos evidenciando o quanto essa situação pode ser importante para os rebanhos brasileiros.(AU)

A cross-sectional study to estimate the frequency of anti-bovine viral diarrhea virus-1 antibodies in domestic pigs of Mossoró region in the state of Rio Grande do Norte, Brazil / Estudo transversal para estimar a frequência de anticorpos anti-vírus da diarreia viral bovina-1 em suínos domésticos da região de Mossoró, no estado do Rio Grande do Norte, Brasil

ABSTRACT: This study investigated the occurrence of antibodies for BVDV-1 in swine herds located in the region of Mossoró city of the state of Rio Grande do Norte, Brazil. A sample size of 412 animals was estimated assuming unknown prevalence (set at 50%). Virus neutralization assay was used to the detect the presence of antibodies for BVDV-1 and the results found were analysed using multivariable logistic regression model. The obtained prevalence was 4% at animal level and 45% at the animal and herd level. The titers were highly variable between animals and within farms. The multivariable logistic regression analysis showed an association between being housed outside and exposure to BVDV-1 (OR=0.24, 95% CI:0.06, 0.96, P=0.04). Highly correlated data and low prevalence of antibodies at the animal level resulted in insufficient power to detect significant differences with other selected risk factors. In conclusion, the prevalence is within the range reported for other countries.

RESUMO: Este estudo investigou a ocorrência de anticorpos contra o BVDV-1 em rebanhos suínos localizados na região da cidade de Mossoró do Estado do Rio Grande do Norte, Brasil. Um tamanho de amostra de 412 animais foi estimado assumindo uma prevalência desconhecida (fixada em 50%). O teste de virusneutralização foi utilizado para detectar a presença de anticorpos ao BVDV-1 e os resultados obtidos foram analisados usando o modelo de regressão logística multivariada. A prevalência obtida foi de 4% em nível animal e de 45% dos rebanhos. Os títulos foram muito variáveis entre os animais e dentro de fazendas. A análise de regressão logística multivariada apontou associação entre animais criados soltos e a exposição ao BVDV-1 (OR=0,24; 95% IC: 0,06; 0,96; P=0.04). A alta correlação entre os dados junto com a baixa prevalência de anticorpos a nível animal pode ter sido insuficiente para que as diferenças reais fossem detectadas. Em conclusão, a prevalência está dentro do intervalo referido em outros países.