Research advances in the interspecies transmission of hepatitis E virus / 临床肝胆病杂志

‍Hepatitis E virus （HEV） is a pathogen for viral hepatitis and can be transmitted across species via animals. The incidence rate of hepatitis E caused by HEV infection ranks first among acute viral hepatitis worldwide， and now it has become a major public health issue that threatens human health. In recent years， more and more species have been identified as the natural hosts of HEV， and there are more routes for the interspecies transmission of HEV， which greatly inhibits the prevention and control of this virus. Therefore， this article reviews and discusses the advances in the interspecies transmission of HEV， so as to provide a reference for the prevention and control of HEV.

Infección natural por SARS-CoV-2 en gatos y perros domésticos de personas con diagnóstico de COVID-19 en el Valle de Aburrá, Antioquia / Natural SARS-CoV- 2 infection in domestic cats and dogs of humans diagnosed with COVID-19 in Valle de Aburrá, Antioquia

Introducción. El síndrome respiratorio agudo grave causado por el nuevo coronavirus SARS-CoV-2 es causa de la emergencia sanitaria por la pandemia de COVID-19. Si bien el humano es el principal huésped vulnerable, en estudios experimentales y reportes de infección natural, se han encontrado casos de zoonosis inversa de SARS-CoV-2 en animales. Objetivo. Evaluar la infección natural por SARS-CoV-2 en gatos y perros de propietarios con diagnóstico de COVID-19 en el Valle de Aburrá, Antioquia, Colombia. Materiales y métodos. La circulación del SARS-CoV-2 se evaluó por RT-qPCR y RT-PCR en muestras de frotis nasofaríngeos y orofaríngeos de gatos y perros cuyos propietarios se encontraban dentro del periodo de los 14 días de aislamiento. Los casos positivos se verificaron amplificando fragmentos de los genes RdRp, N y E; se secuenció el gen RdRp y se analizó filogenéticamente. Resultados. De 80 animales evaluados, seis gatos y tres perros fueron casos confirmados de infección natural por SARS-CoV-2. Los animales no presentaron signos clínicos y sus propietarios, que padecían la infección, reportaron únicamente signos leves de la enfermedad sin complicaciones clínicas. En el análisis de una de las secuencias, se encontró un polimorfismo de un solo nucleótido (SNP) con un cambio en la posición 647, con sustitución del aminoácido serina (S) por una isoleucina (I). Los casos se presentaron en los municipios de Caldas, Medellín y Envigado. Conclusiones. Se infiere que la infección natural en los gatos y perros se asocia al contacto directo con un paciente con COVID-19. No obstante, no es posible determinar la virulencia del virus en este huésped, ni su capacidad de transmisión zoonótica o entre especie.

Introduction: The severe acute respiratory syndrome caused by the new coronavirus SARS-CoV-2 is the cause of the health emergency due to the COVID-19 pandemic. Although humans are the main susceptible host, experimental studies and reported cases of natural infection have evidenced scenarios of SARS-CoV-2 reverse zoonosis in animals. Objective: To evaluate the natural infection of SARS-CoV-2 in cats and dogs with owners diagnosed with COVID-19 in the Valle de Aburrá subregion in Antioquia, Colombia. Materials and methods. The circulation of SARS-CoV-2 was evaluated by RT-qPCR and RT-PCR in samples of nasopharyngeal and oropharyngeal smears from cats and dogs whose owners presented latent COVID-19 infection. Positive cases were verified through amplification of N, E and RdRp gene fragments; with the latter being sequenced and the phylogenetically analyzed. Results. From 80 tested animals, 6 cats and 3 dogs resulted positive for natural SARS-CoV-2 infection. These animals did not show any clinical signs; and their infected owners only reported mild signs of COVID-19, without clinical complications. Regarding analysis of one of the sequences, a single nucleotide polymorphism (SNP) was found, with a substitution in position 647, resulting in the change of the amino acid serine (S) for isoleucine (I). The cases occurred in the municipalities of Caldas, Medellín and Envigado. Conclusions. It is inferred that natural infection in cats and dogs is associated with direct contact with a positive COVID-19 patient.

Hepatitis E virus infection in buffaloes in South China / [Hepatite E infecção viral em búfalos no Sul da China]

Hepatitis E virus (HEV) infection is an important global public health issue. HEV infections are recognized as a zoonotic disease. Swine are believed to be the main reservoir of HEV. Recently, yaks, cows, and yellow cattle have been reported as new reservoirs of HEV. However, whether other species of cattle and buffaloes are sensitive to HEV infection is unknown. To investigate the prevalence of HEV infection in buffaloes, enzyme-linked immunosorbent assay (ELISA) and reverse transcription-nested polymerase chain reaction (RT-nPCR) were performed. Only one buffalo was positive to anti-HEV IgM antibody (1/106, 0.94%), and none were positive for anti-HEV IgG antibody. To our surprise, five serum (5/106, 4.72%) and three milk samples (3/40, 7.50%) from buffaloes were positive to HEV RNA. All strains of HEV isolated from buffaloes belong to genotype 4. Results indicate that buffaloes may be a new reservoir of HEV.(AU)

Infecção com o vírus Hepatite E (HEV) é uma importante questão de saúde pública global. Infecções HEV são reconhecidas como doença zoológica. Acredita-se que suínos são o principal reservatório de HEV. Recentemente iaques, vacas, e gado amarelo foram reportados como novos reservatórios do HEV. Porém, não se sabe se outras espécies de gado e búfalo são sensíveis a infecção HEV. Para investigar a prevalência de infecção HEV em búfalos, foram realizados prova de imunoabsorção enzimática e polimerização em cadeia inversa ancorada em transcrição. Apenas um búfalo foi positivo para o anticorpo anti-HEV IgM (1/106, 0,94%), e nenhum foi positivo para o anticorpo anti-HEV IgG. Para nossa surpresa cinco (5/106, 4,72%) e três amostras de leite (3/40, 7,50%) de búfalos foram positivos para HEV RNA. Todas as estirpes de HEV isoladas de búfalos pertencem ao genótipo 4. Resultados indicam que búfalos podem ser um reservatório de HEV.(AU)

Identification of a novel coronavirus causing severe pneumonia in human: a descriptive study / 中华医学杂志(英文版)

Background: Human infections with zoonotic coronaviruses (CoVs), including severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV, have raised great public health concern globally. Here, we report a novel bat-origin CoV causing severe and fatal pneumonia in humans. Methods: We collected clinical data and bronchoalveolar lavage (BAL) specimens from five patients with severe pneumonia from Jin Yin-tan Hospital, Wuhan, Hubei province, China. Nucleic acids of the BAL were extracted and subjected to next-generation sequencing. Virus isolation was carried out, and maximum-likelihood phylogenetic trees were constructed. Results: Five patients hospitalized from December 18 to December 29, 2019 presented with fever, cough, and dyspnea accompanied by complications of acute respiratory distress syndrome. Chest radiography revealed diffuse opacities and consolidation. One of these patients died. Sequence results revealed the presence of a previously unknown β-CoV strain in all five patients, with 99.8–99.9% nucleotide identities among the isolates. These isolates showed 79.0% nucleotide identity with the sequence of SARS-CoV (GenBank NC_004718) and 51.8% identity with the sequence of MERS-CoV (GenBank NC_019843). The virus is phylogenetically closest to a bat SARS-like CoV (SL-ZC45, GenBank MG772933) with 87.6–87.7% nucleotide identity, but is in a separate clade. Moreover, these viruses have a single intact open reading frame gene 8, as a further indicator of bat-origin CoVs. However, the amino acid sequence of the tentative receptor-binding domain resembles that of SARS-CoV, indicating that these viruses might use the same receptor. Conclusion: A novel bat-borne CoV was identified that is associated with severe and fatal respiratory disease in humans.

Characterization, prevalence and antibiogram study of Staphylococcus aureus in poultry

Objective To reveal the presence of methicillin resistant Staphylococcus aureus (S. aureus) (MRSA) in poultry samples and to determine the antibiogram pattern against five antibiotics. Methods Samples from different poultry farm of Chittagong city, Bangladesh were examined for S. aureus by different biochemical tests and confirmed as MRSA by identifying the presence of mecA gene using PCR. Antibiotic resistance pattern in S. aureus was determined by antibiotic disk diffusion method. Results In this study, a total of 60 samples (30 from nasal swabs and 30 from cloacal swabs) were used, of which 54 were confirmed as S. aureus by different biochemical tests. Among these, 12 were confirmed as MRSA by detecting mecA gene using PCR. During antibiogram study, both nasal and cloacal samples showed the highest resistance against penicillin-G and the lowest resistance was observed against neomycin. Conclusions Based on the present study, it can be said that different antibiotics are used extensively in poultry that leads to MRSA and is alarming for human health.

Characterization, prevalence and antibiogram study of Staphylococcus aureus in poultry

Objective: To reveal the presence of methicillin resistant Staphylococcus aureus (S. aureus) (MRSA) in poultry samples and to determine the antibiogram pattern against five antibiotics. Methods: Samples from different poultry farm of Chittagong city, Bangladesh were examined for S. aureus by different biochemical tests and confirmed as MRSA by identifying the presence of mecA gene using PCR. Antibiotic resistance pattern in S. aureus was determined by antibiotic disk diffusion method. Results: In this study, a total of 60 samples (30 from nasal swabs and 30 from cloacal swabs) were used, of which 54 were confirmed as S. aureus by different biochemical tests. Among these, 12 were confirmed as MRSA by detecting mecA gene using PCR. During antibiogram study, both nasal and cloacal samples showed the highest resistance against penicillin-G and the lowest resistance was observed against neomycin. Conclusions: Based on the present study, it can be said that different antibiotics are used extensively in poultry that leads to MRSA and is alarming for human health.

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

Prevalence of hepatitis E virus antibodies in individuals exposed to swine in Mato Grosso, Brazil

This study was conducted to estimate the prevalence of hepatitis E antibodies (anti-HEV) among individuals exposed to swine in the rural areas of the state of Mato Grosso (MT) in Brazil. The study included 310 participants who had an average age of 39 years. Fifty-one per cent of the participants were female and 26 (8.4%) were anti-HEV-positive. Concomitantly, we studied 101 blood donors from the urban area of the state capital who had never lived in a rural area or handled swine. Four per cent (4%) of these individuals were anti-HEV-positive (p = 0.206). When we compared the anti-HEV-positive participants who had been exposed to swine with the anti-HEV-negative participants, we noticed associations between the presence of anti-HEV and increased age, a history of blood transfusions and contact with other farm animals. However, after a multivariate analysis was performed, this association was not confirmed. Finally, the ratio of anti-HEV-positive individuals who had been exposed to swine in rural MT was similar to that found in previous studies in Brazil. This prevalence did not characterise this type of exposure as a risk factor for HEV infection in this region.

Identification of human Norovirus (HNoV) in domestic pig stool samples / Identificación de Norovirus Humano (HNoV) en muestras de estiércol de cerdos domésticos / Identificação de Norovírus Humano (HNoV) em amostras de suínos domésticos

Identificación de Norovirus Humano (HNoV) en muestras de estiércol de cerdos domésticos. Objetivo: determinar la presencia de NoVs como posible agente zoonótico causal de diarrea aguda entre cerdos y humanos. Materiales y métodos: se recolectaron un total de 77 muestras diarreicas provenientes de niños menores de cinco años y de cerdos menores de dos meses de la población La Chamba en el Tolima, Colombia. Estas muestras fueron transportadas al Laboratorio de Virología de la Pontificia Universidad Javeriana en Bogotá, donde inicialmente se les realizó extracción con Trizol-reagent, siguiendo las instrucciones del fabricante. Una vez obtenido el RNA, el siguiente paso fue hacer la RT-PCR para obtener el producto de amplificacion esperado para NoVs de 213 pb. Finalmente, las muestras positivas obtenidas en la RT-PCR fueron secuenciadas y analizadas mediante métodos bioinformáticos. Resultados: se obtuvieron seis muestras positivas de diarrea de niños y una muestra positiva de diarrea de cerdos, las cuales se evidenciaron en una banda de 231 pb. Cinco de las seis muestras positivas en niños y la muestra positiva en cerdos fueron secuenciadas y analizadas. Conclusiones: dada la estrecha relación genética que se evidencia entre las secuencias del cerdo y el humano, este podría ser un indicio de que exista la posibilidad de un animal en común como reservorio para cepas de humano u otras cepas de animales...

Objective. To determine the presence of NoVs as a possible causal zoonotic agent of acute diarrhea in pigs and humans. Materials and methods. We collected a total of 77 samples from diarrheal children under 5 years and pigs under 2 months from La Chamba town in Tolima, Colombia. These samples were transported to the Laboratory of Virology of the Pontificia Universidad Javeriana in Bogotá, and extraction with Trizol-reagent was done following the manufacturer’s instructions. After obtaining the RNA, the next step was to perform RT-PCR for obtaining the expected amplification product of 213- bp NoVs. Finally, the positive samples obtained in the RT-PCR were sequenced and analyzed by bioinformatics methods. Results. Six positive diarrheic samples from children and a positive diarrheic sample from pigs were detected by a band of 231 bp. Five of the six positive samples in children and the positive pig sample were sequenced and analyzed. Conclusion. Given the close genetic relationship between pig and human sequences, this could be an indication of the potential existence of a common animal acting as a reservoir for human or other animal strains...

Identificação de Norovírus Humano (HNoV) em amostras de suínos domésticos. Objetivo. Determinar a presença de NoVs como possível agente zoonótico causal de diarréia aguda entre porcos e seres humanos. Materiais e métodos. Foram coletadas um total de 77 amostras de crianças diarréicas menores de cinco anos e porcos com menos de dois meses da população “La Chamba” Tolima-Colômbia. Estas amostras foram transportadas ao laboratório de virologia da Pontifícia Universidade Javeriana - Bogotá, onde foram inicialmente submetidas à extração com Trizol reagment e seguindo as instruções do fabricante, após a obtenção do RNA o próximo passo foi realizar a RT-PCR para obter o produto de amplificação esperado para NoVs de 213 bp. Finalmente as amostras positivas obtidas no RT-PCR foram seqüenciadas e analisadas por métodos de bioinformática. Resultados. Foram obtidas seis amostras positivas de diarréia nas crianças e uma amostra positiva de diarréia em suínos, as que foram representadas em uma banda de 231 pb. Cinco das seis amostras positivas em crianças e a amostra positiva em suínos foram seqüenciadas e analisadas. Conclusões. Dada a estreita relação genética que se manifesta entre as seqüências de suínos e humanos, isso poderia ser uma indicação de que existe a possibilidade de um animal comum como reservatório para o humano ou outras cepas de animais...

Evidence of zoonotic Chlamydophila psittaci transmission in a population at risk in Zulia state, Venezuela / Evidencia de transmisión zoonótica de Chlamydophila psittacci en una población de riesgo del Estado Zulia, Venezuela

Objective This study was aimed at investigating the frequency of infection by Cp. psittaci and determining its genotype in individuals at potential risk of exposure to the bacteria. Methodology The study involved 170 individuals: a risk group (n= 96) and a low-risk control group (n=74). Cp. psittaci was detected and genotyped by single-tube nested PCR and ompA gene sequencing. Results Eight (8.3 %) positive cases were detected in the risk group and 1 (1.4 %) in the control group (p<0.04). Cp. psittaci was found in 16.7 % of pigeons' fecal samples. Cp. psittaci infection with was more frequent in symptomatic (17.7 %) than asymptomatic (6.3 %) individuals in the risk group. Analysing the genomes isolated from human and bird specimens revealed the presence of genotype B. Conclusion The presence of Cp. psittaci genotype B in the population being evaluated could have been attributed to zoonotic transmission from pigeons to humans, an underestimated potential public health problem in Venezuela requiring the health authorities' involvement.

Objetivo El objetivo de este estudio fue investigar la frecuencia de infecciones por Cp. psittaci y determinar su genotipo en individuos con potencial riesgo de exposición a la bacteria. Metodología Se incluyeron 170 individuos, un grupo de riesgo (n=96) y un grupo control (n=74). La detección y genotipificación de Cp. psittaci se llevó a cabo por PCR anidada y secuenciación del gen ompA. Resultados Se detectaron ocho (8,3 %) casos positivos en el grupo de riesgo y 1 (1,35 %) en el grupo control (p<0,04). Cp. psittaci fue detectada en 16,7 % muestras fecales de palomas. En el grupo de riesgo, la frecuencia de infección por Cp. psittaci fue 17,7 % en individuos sintomáticos y 6,3% en asintomáticos. El análisis de los genomas aislados de muestras humanas y aves, revelaron la presencia del genotipo B. Conclusión La presencia de Cp. psittaci genotipo B en la población evaluada podría ser atribuida a transmisión zoonótica de palomas a humanos, un potencial problema de salud pública en nuestra región que requiere la intervención de autoridades sanitarias.

Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil

An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD) technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20 percent similarity to each primer and 60 percent when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8 percent) and the oral cavities (66 percent) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.