Lack of serological and molecular evidences of Zika virus circulation in non-human primates in three states from Brazil

BACKGROUND Zika virus (ZIKV) was discovered in 1947 with the virus isolation from Rhesus monkey (Macaca mulatta) in Uganda forest, Africa. Old World Primates are involved in a sylvatic cycle of maintenance of this arbovirus, however a limited knowledge about the role of New World primates in ZIKV transmission cycles has been established. OBJECTIVE This work aimed to investigate the presence of enzootic circulation of ZIKV in New World Primates from three Brazilian states: São Paulo, Paraíba, and Paraná. METHODS We analyzed 100 non-human primate samples collected in 2018 and 2020 from free-ranging and captive environments from São Paulo (six municipalities belonging to Sorocaba region), Paraíba (João Pessoa municipality), and Paraná (Foz do Iguaçu municipality) using reverse transcriptase quantitative polymerase reaction (RT-qPCR) assays, indirect enzyme-linked immunosorbent assay (ELISA), and plaque reduction neutralization test (PRNT). FINDINGS All samples (n = 141) tested negative for the presence of ZIKV genome from tissue and blood samples. In addition, all sera (n = 58) from Foz do Iguaçu' non-human primates (NHPs) were negative in serological assays. MAIN CONCLUSION No evidence of ZIKV circulation (molecular and serological) was found in neotropical primates. In addition, the absence of antibodies against ZIKV suggests the absence of previous viral exposure of NHPs from Foz do Iguaçu-PR.

Serosurvey of anti-treponema pallidum (syphilis), antihepatitis C virus and anti-HIV antibodies in homeless persons of São Paulo city, southeastern Brazil

ABSTRACT Homeless persons have been considered as one of the most susceptible populations to sexually transmitted infections. In Brazil, these population experienced an increase of 140% from 2012 to 2020. Accordingly, the present study aimed to assess the seroprevalence of anti-Treponema pallidum, anti-HCV, anti-HIV antibodies, and the risk factors associated with homeless persons in a daytime attendance shelter of São Paulo city during the syphilis epidemic in Brazil. Blood samples of 116 volunteers and epidemiological data were conveniently collected in the shelter from June through August 2018. Detection of syphilis, HCV, and HIV antibodies was performed by chemiluminescent microparticle immunoassay (CMIA). CMIA-reagent samples for anti-T. pallidum antibodies were confirmed by Venereal Disease Research Laboratory (VDRL) non-treponemal test. VDRL non-reagent samples were confirmed by treponemal rapid immunochromatographic test. A rapid immunoblot assay confirmed seropositivity to HIV. Overall, anti-T. pallidum antibodies were observed in 29/116 (25.0%), anti-HCV antibodies in 4/116 (3.4%), and anti-HIV antibodies in 2/116 (1.7%) individuals, both co-infected with anti-T. pallidum antibodies. Associated risk factors for syphilis in homeless persons were being born or previously living in another city (p = 0.043) and becoming homeless due to family conflicts (p = 0.035). Besides homeless vulnerability, worldwide shortage of benzathine penicillin supply and increasing of syphilis testing access through rapid testing in primary health care services may have also impacted disease spreading at the time. The prevalence of syphilis found herein is the highest worldwide to date in this population.

Molecular detection of Leishmania infantum in donkeys and mules under semiarid conditions in Brazil / Detecção molecular de Leishmania infantum em asininos e muares sob condições semiáridas

Abstract Visceral leishmaniasis is a parasitic zoonosis that mainly affects poorest and most vulnerable populations, and domestic dogs are considered to be the main source of infection to the vector and therefore humans. However, several studies have investigated the role of other vertebrate hosts in the disease cycle. In this context, the aim of the present study was to conduct a survey of Leishmania infantum infection in donkeys and mules living in a semiarid region of Brazil. Whole blood sampled from 72 equids (65 donkeys and 7 mules) was used to perform molecular diagnosis using the real-time polymerase chain reaction (qPCR) technique. A total of 25% of the samples (18/72) were positive through qPCR, but there were no significant differences between the species (donkeys or mules), sex (male or female) and abandonment situation of the animals (yes or no). Donkeys and mules living under semiarid conditions have high frequency of L. infantum infection. It is therefore worth assigning importance to these species in the epidemiological cycle of visceral leishmaniasis, either as potential reservoirs or just as an abundant food source for vectors.

Resumo A leishmaniose visceral é uma zoonose parasitária que afeta principalmente populações mais pobres e vulneráveis, e os cães domésticos são considerados as principais fontes de infecção para o vetor e, portanto, para os humanos. Porém diversos estudos têm pesquisado o papel de outros hospedeiros vertebrados no ciclo da doença. Neste contexto, objetivou-se realizar um levantamento da infecção por Leishmania infantum em asininos e muares, vivendo em região semiárida do Brasil. Foi utilizado sangue total de 72 equídeos (65 asininos e 7 muares) para a realização de diagnóstico molecular por meio da técnica de Reação em Cadeia de Polimerase em Tempo Real (qPCR). Um total de 25% das amostras (18/72) resultaram positivas na qPCR, porém não houve diferença significativa entre as espécies (asininos e muares), sexo (macho e fêmea) e situação de abandono dos animais (sim ou não). Asininos e muares, vivendo em condições semiáridas, apresentam alta frequência de infecção por L. infantum, sendo válido atribuir importância a essas espécies no ciclo epidemiológico da leishmaniose visceral, seja como um reservatório em potencial, seja apenas como uma fonte alimentar abundante para os vetores.

Leptospira interrogans infection in tegu lizard (Tupinambis merianae), Brazil / Infecção por Leptospira interrogans em lagarto teiú (Tupinambis merianae), Brasil

ABSTRACT: We used 12 tegu lizards (Tupinambis merianae) from northeastern Brazil, and we reported nine (75%) Leptospira sp. PCR-positive animals and six (50%) seropositive. Leptospira sp. DNA sequencing revealed 99% similarity with L. interrogans. Our findings indicated that this species may play a role in the transmission of human leptospirosis.

RESUMO: Foram utilizados 12 lagartos Teiús (Tupinambis merianae) do Nordeste do Brasil. Encontramos nove animais positivos (75%) para Leptospira sp. na PCR e seis (50%) soropositivos. O sequenciamento de DNA de Leptospira sp. revelou 99% de semelhança com L. interrogans. Os resultados indicam que esta espécie pode desempenhar um papel importante na transmissão da leptospirose humana.

Serosurvey of anti- Leptospira sp. and anti- Toxoplasma gondii antibodies in capybaras and collared and white-lipped peccaries

Abstract INTRODUCTION: Brazilian native species are reemerging as increasingly free-ranging populations. METHODS: Sera from 31 capybaras (Hydrochoerus hydrochaeris) and 28 peccaries (Pecari tajacu and Tayassu pecari) were tested for anti-Leptospira and anti-Toxoplasma gondii antibodies using microscopic seroagglutination test. RESULTS: Nineteen percent of free-ranging and 10.0% of captive capybaras, along with 31.8% of collared peccaries, were seropositive for T. gondii. None was seropositive for Leptospira sp. CONCLUSIONS: The present findings indicated low risk of disease, particularly among capybaras and white-lipped peccaries; however, active surveillance programs are important for monitoring wildlife health and public health once they are in public parks around cities.

Mycobacterium genavense infection in two species of captive snakes

Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.

Anti-leptospirosis agglutinins in Brazilian capybaras (hydrochoerus hydrochaeris)

The interest in commercial use of wild animals is increasing, especially regarding raising of capybaras. Although this wild species is potentially lucrative for the production of meat, oil and leather, it is suggested as a probable reservoir of leptospires. Methods Due to the economic importance of this species and the lack of studies concerning leptospirosis, the presence of anti-leptospirosis agglutinins was assayed in 55 serum samples of capybaras (Hydrochoerus hydrochaeris) from commercial and experimental breeding flocks located in São Paulo state, Paraná state, and Rio Grande do Sul state, Brazil. Samples were obtained through cephalic or femoral venipunction (5 to 10 mL). Microscopic agglutination test was used according to the Brazilian Health Ministry considering as cut-off titer of 100. Results Out of the 55 samples analyzed, 23 (41.82 %) tested positive. The most prevalent serovar was Icterohaemorrhagiae (56.52 %) in 13 samples, followed by Copenhageni in nine samples (39.13 %), Pomona in four samples (17.39 %), Djasiman and Castellonis in three samples each (13.04 %), Grippotyphosa, Hardjo, Canicola, and Cynopteri in two samples each (8.7 %), and Andamana and Bratislava in one sample each (4.34 %). Conclusions These results suggest the evidence of exposure toLeptospira spp. and the need of new studies to evaluate a higher number of capybaras from different regions to better understand the importance of leptospirosis infection in these animals and verify the zoonotic role of this species as a possible source of infection to humans and other animals.

Mycobacterium genavense infection in two species of captive snakes

Background: Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.(AU)

Anti-leptospirosis agglutinins in Brazilian capybaras (hydrochoerus hydrochaeris)

Abstract Background The interest in commercial use of wild animals is increasing, especially regarding raising of capybaras. Although this wild species is potentially lucrative for the production of meat, oil and leather, it is suggested as a probable reservoir of leptospires. Methods Due to the economic importance of this species and the lack of studies concerning leptospirosis, the presence of anti-leptospirosis agglutinins was assayed in 55 serum samples of capybaras (Hydrochoerus hydrochaeris) from commercial and experimental breeding flocks located in São Paulo state, Paraná state, and Rio Grande do Sul state, Brazil. Samples were obtained through cephalic or femoral venipunction (5 to 10 mL). Microscopic agglutination test was used according to the Brazilian Health Ministry considering as cut-off titer of 100. Results Out of the 55 samples analyzed, 23 (41.82 %) tested positive. The most prevalent serovar was Icterohaemorrhagiae (56.52 %) in 13 samples, followed by Copenhageni in nine samples (39.13 %), Pomona in four samples (17.39 %), Djasiman and Castellonis in three samples each (13.04 %), Grippotyphosa, Hardjo, Canicola, and Cynopteri in two samples each (8.7 %), and Andamana and Bratislava in one sample each (4.34 %). Conclusions These results suggest the evidence of exposure toLeptospira spp. and the need of new studies to evaluate a higher number of capybaras from different regions to better understand the importance of leptospirosis infection in these animals and verify the zoonotic role of this species as a possible source of infection to humans and other animals.

Seroprevalence and seroincidence of Leptospira infection in dogs during a one-year period in an endemic urban area in Southern Brazil

INTRODUCTION: Leptospirosis is a zoonosis that affects both humans and animals. Dogs may serve as sentinels and indicators of environmental contamination as well as potential carriers for Leptospira. This study aimed to evaluate the seroprevalence and seroincidence of leptospirosis infection in dogs in an urban low-income community in southern Brazil where human leptospirosis is endemic. METHODS: A prospective cohort study was designed that consisted of sampling at recruitment and four consecutive trimestral follow-up sampling trials. All households in the area were visited, and those that owned dogs were invited to participate in the study. The seroprevalence (MAT titers ≥100) of Leptospira infection in dogs was calculated for each visit, the seroincidence (seroconversion or four-fold increase in serogroup-specific MAT titer) density rate was calculated for each follow-up, and a global seroincidence density rate was calculated for the overall period. RESULTS: A total of 378 dogs and 902.7 dog-trimesters were recruited and followed, respectively. The seroprevalence of infection ranged from 9.3% (95% CI; 6.7 - 12.6) to 19% (14.1 - 25.2), the seroincidence density rate of infection ranged from 6% (3.3 - 10.6) to 15.3% (10.8 - 21.2), and the global seroincidence density rate of infection was 11% (9.1 - 13.2) per dog-trimester. Canicola and Icterohaemorraghiae were the most frequent incident serogroups observed in all follow-ups. CONCLUSIONS: Follow-ups with mean trimester intervals were incapable of detecting any increase in seroprevalence due to seroincident cases of canine leptospirosis, suggesting that antibody titers may fall within three months. Further studies on incident infections, disease burden or risk factors for incident Leptospira cases should take into account the detectable lifespan of the antibody. .

SEROLOGICAL AND MOLECULAR SURVEY OF Leptospira spp. AMONG CART HORSES FROM AN ENDEMIC AREA OF HUMAN LEPTOSPIROSIS IN CURITIBA, SOUTHERN BRAZIL / Inquérito sorológico e molecular de Leptospira spp. em cavalos carroceiros de área endêmica para leptospirose humana em Curitiba, Sul do Brasil

Introduction: Cart horses are a re-emerging population employed to carry recyclable material in cities. Methods: Sixty-two horses were sampled in an endemic area of human leptospirosis. The microscopic agglutination test (MAT) and real-time polymerase chain reaction (qPCR) were performed. Results: A seropositivity of 75.8% with serovar Icterohaemorrhagiae in 80.8% of the horses was observed. Blood and urine were qPCR negative. MAT showed positive correlations with rainfall (p = 0.02) and flooding (p = 0.03). Conclusions: Although horses may be constantly exposed to Leptospira spp. in the environment mostly because of rainfall and flooding, no leptospiremia or leptospiruria were observed in this study.

Introdução: Cavalos carroceiros são uma população reemergente empregada para transportar materiais recicláveis em cidades. Métodos: Em área endêmica para leptospirose humana foram amostrados 62 cavalos. Soroaglutinação microscópica e reação em cadeia da polimerase em tempo real foram empregadas. Resultados: Observou-se soropositividade em 75,8% com sorovar Icterohaemorrhagiae em 80,8% cavalos. Amostras de sangue e urina foram negativas no qPCR. Observou-se correlação positiva entre SAM e pluviosidade (p = 0,02) e alagamentos (p = 0,03). Conclusão: Embora cavalos possam estar constantemente expostos a Leptospira spp. no ambiente, principalmente por chuvas e inundações, leptospiremia e leptospiruria não foram encontradas neste estudo.

Detection of anti-Toxoplasma gondii antibodies in carthorses in the metropolitan region of Curitiba, Paraná, Brazil / Detecção de anticorpos anti-Toxoplasma gondii em cavalos carroceiros da região metropolitana de Curitiba, Paraná, Brasil

Toxoplasma gondii, the agent for toxoplasmosis, has worldwide distribution. Horses normally play a secondary role in its life cycle, but movement around urban areas, feeding on grass and the increasing use of carthorses for gathering recyclable material in some urban areas of Brazil may increase their exposure to T. gondii infection. The aim of the present study was to investigate the frequency of anti-T. gondii antibodies in carthorses in the metropolitan region of Curitiba, PR. IgG antibodies against T. gondii were detected using the indirect fluorescence antibody test (IFAT) (titers ≥ 64). Seventeen (17.0%) of the 100 horses sampled were seropositive. There were no statistical differences in relation to sex (p = 0.28) or age (p = 0.15). Our findings suggest that carthorses are exposed to T. gondii infections and that no associations with age or sex exist.

Toxoplasma gondii, agente da toxoplasmose, tem distribuição mundial. Geralmente cavalos desempenham papel secundário no ciclo de vida do T. gondii. Entretanto, a circulação em meio urbano, a alimentação com gramíneas e o aumento no uso de cavalos carroceiros para recolhimento de material reciclável em algumas áreas urbanas do Brasil podem aumentar a exposição desses cavalos ao T. gondii. O objetivo deste estudo foi investigar a frequência de anticorpos anti-T. gondii em cavalos carroceiros da região metropolitana de Curitiba, PR. Os anticorpos da classe IgG contra T. gondii foram detectados pela reação de imunofluorescência indireta (RIFI) (títulos ≥ 64). Dezessete (17%) dos 100 cavalos avaliados foram positivos e não houve diferença entre sexos (p = 0,28) ou idade (p = 0,15). Esses achados sugerem que cavalos carroceiros estão expostos a infecções por T. gondii, não existindo associação com a idade ou o sexo dos animais.

Seroprevalence of Toxoplasma gondii infection in cats from Curitiba, Paraná, Brazil / Soroprevalência de infecção anti-Toxoplasma gondii em gatos domésticos de Curitiba, Paraná, Brasil

Toxoplasmosis is a worldwide zoonosis caused by the protozoa Toxoplasma gondii which infects all warm-blood vertebrates. This study aimed to assess the seroprevalence of anti-Toxoplasma gondii antibodies in a population of domestic cats seen at a major cat-only veterinary clinic in Curitiba, Paraná State, Southern Brazil. Serum samples were processed by indirect immunofluorescence assay (IIFA) for the detection of IgG. Antibody titers were found in 16.3 percent (46/282) of sera analyzed, with titers to T. gondii of 16 in eight cats, 64 in 23 cats, 256 in 14 cats and 1024 in one cat. Statistical differences were not found regarding the association with age, gender and different areas of the city (p > 0.05). No significant differences were found in any variable when comparing seropositivity with potential risk factors. The seroprevalence was relatively lower when compared to other Brazilian regions, probably due to the fact that the cats studied were owned, domiciled with restricted dietary habits based on processed foods, restricted access to the street and no prey access. In conclusion, low feline toxoplasmosis seroprevalence may be associated to owned cats due to adequate dietary care and restricted outdoor access, as well as low local environmental exposure.

A toxoplasmose é uma zoonose de distribuição mundial, causada pelo protozoário Toxoplasma gondii que infecta os vertebrados de sangue quente. Este estudo objetiva estabelecer a prevalência de anticorpos anti-Toxoplasma gondii e o estudo de associação em uma população de gatos domésticos atendidos em uma clínica veterinária exclusiva de gatos localizada em Curitiba, estado do Paraná, Brasil. As amostras de soro foram obtidas e processadas pela reação de imunofluorescência indireta IgG. Títulos de anticorpos foram encontrados em 46/282 (16,3 por cento) das amostras analisadas, com título de 16 em oito gatos, 64 em 23, 256 em 14 e 1024 em um gato. Diferenças estatísticas não foram encontradas no estudo de associação entre idade, gênero e região da cidade (p > 0,05). Nenhuma diferença significativa foi encontrada para qualquer variável, quando comparando a soropositividade com a ocorrência do possível fator de risco. A soroprevalência foi relativamente menor quando comparada a outras regiões brasileiras, provavelmente por serem gatos de proprietários, que tinham hábito domiciliado e alimentação restrita baseada em alimentos industrializados, com acesso restrito a rua e sem acesso a caça. Em conclusão, a baixa soroprevalência de toxoplasmose felina pode estar relacionada a gatos com proprietários devido aos cuidados alimentares e acesso restrito à rua, e também pode estar associada à baixa exposição ambiental local.

Surveillance of canine visceral leishmaniasis in a disease-free area / Vigilância da leishmaniose visceral canina em área indene

Leishmaniasis is an important re-emergent parasitosis worldwide, particularly in tropical countries. There are no reports of autochthonous disease in the State of Paraná, southern Brazil. No surveillance has been carried out in the most populated areas such as the city of Curitiba and its surroundings. The purpose of the present study was to determine the seroprevalence of visceral leishmaniasis in dogs at the Center for Zoonosis Control of São José dos Pinhais, Paraná, before euthanasia. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence antibody test (IFAT) were used to detect antibody levels against Leishmania sp. in dog sera. Imprints of the popliteal lymph nodes that were also randomly collected from 50 dogs with suspected clinical signs of visceral leishmaniasis, and evaluated under light microscopy for the detection of amastigote forms, were negative. A total of 364 dog samples were tested. The results showed only one positive sample (0.0027 percent) by ELISA test but negative by IFAT, however, the dog had no clinical signs. Random surveillance of dog populations from several districts of a metropolitan area may be a means of preventing Leishmania spreading. Based on our results, the city of Curitiba and its metropolitan area were considered at low risk for visceral leishmaniasis.

A leishmaniose é uma importante parasitose re-emergente observada no mundo, particularmente em países tropicais. Não há ainda relatos de casos autóctones no estado do Paraná. Não há até o momento referência de vigilância no reservatório canino, tais como Curitiba e região metropolitana do estado. O objetivo do estudo foi determinar a soroprevalência da leishmaniose visceral em cães entregues ao Centro de Controle de Zoonoses de São José dos Pinhais, Paraná para eutanásia. A detecção sorológica da presença de anticorpos contra Leishmania sp. foi realizada por (ELISA) indireto e pela Reação de Imunofluorescência Indireta (RIFI). Além disso, impressão de linfonodo poplíteo coletadas ao acaso de 50 cães com sinais clínicos suspeitos para leishmaniose visceral e analisados sob microscopia óptica para detecção de formas amastigotas, foram negativas. Amostras de soro de 364 animais foram testadas, e os resultados mostraram somente uma amostra positiva (0,0027 por cento), reagente ao ELISA e negativa à RIFI, entretanto, o cão não apresentava sinais clínicos. A vigilância ao acaso em uma população de vários locais de uma área metropolitana pode ser uma forma de prevenção da disseminação da doença. Com base nos resultados observados, Curitiba e região metropolitana foram consideradas de baixo risco para a leishmaniose visceral.

Surveillance of canine visceral leishmaniasisin a disease-free area / Vigilância da leishmaniose visceral canina em área indene

Leishmaniasis is an important re-emergent parasitosis worldwide, particularly in tropical countries. There are no reports of autochthonous disease in the State of Paraná, southern Brazil. No surveillance has been carried out in the most populated areas such as the city of Curitiba and its surroundings. The purpose of the present study was to determine the seroprevalence of visceral leishmaniasis in dogs at the Center for Zoonosis Control of São José dos Pinhais, Paraná, before euthanasia. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence antibodytest (IFAT) were used to detect antibody levels against Leishmania sp. in dog sera. Imprints of the popliteal lymph nodes that were also randomly collected from 50 dogs with suspected clinical signs of visceral leishmaniasis, and evaluated under light microscopy for the detection of amastigote forms, were negative. A total of 364 dog samples were tested. The results showed only one positive sample (0.0027%) by ELISA test but negative by IFAT, however, the dog had no clinical signs. Random surveillance of dog populations from several districts of a metropolitan area may be a means of preventing Leishmania spreading. Based on our results, the city of Curitiba and its metropolitan area were considered at low risk for visceral leishmaniasis.

A leishmaniose é uma importante parasitose re-emergente observada no mundo, particularmente em países tropicais. Não há ainda relatos de casos autóctones no estado do Paraná. Não há até o momento referência de vigilância no reservatório canino, tais como Curitiba e região metropolitana do estado. O objetivo do estudo foi determinar a soroprevalência da leishmaniose visceral em cães entregues ao Centro de Controle de Zoonoses de São José dos Pinhais, Paraná para eutanásia. A detecção sorológica da presença de anticorpos contra Leishmania sp. foi realizada por(ELISA) indireto e pela Reação de Imunofluorescência Indireta (RIFI). Além disso, impressão de linfonodo poplíteo coletadas ao acaso de 50 cães com sinais clínicos suspeitos para leishmaniose visceral e analisados sob microscopia óptica para detecção de formas amastigotas, foram negativas. Amostras de soro de 364 animais foram testadas, e os resultados mostraram somente uma amostra positiva (0,0027%), reagente ao ELISA e negativa à RIFI, entretanto, o cão não apresentava sinais clínicos. A vigilância ao acaso em uma população de vários locais de uma área metropolitana pode ser uma forma de prevenção da disseminação da doença. Com base nos resultados observados, Curitiba e região metropolitana foram consideradas de baixo risco para a leishmaniose visceral.