Isolation, Serovar Identification, and Antimicrobial Susceptibility of Avibacterium paragallinarum from Chickens in China from 2019 to 2020

Infectious coryza is an acute infectious respiratory disease in chickens that is caused by Avibacterium paragallinarum (A. paragallinarum). Infectious coryza has major economic effects due to decreased egg production in growing birds and slowed growth in broilers. In this study, we isolated and identified 40 strains of A. paragallinarum from chickens that showed typical clinical signs of coryza in part of China from 2019 to 2020. Using a hemagglutination-inhibition test, 11 isolates were identified as serovar A, 10 isolates were identified as serovar B, and 19 isolates were identified as serovar C. Antimicrobial sensitivity tests showed that high minimum inhibitory concentration (MIC) values were encountered for compounds sulfamethoxine sodium and oxytetracycline hydrochloride. Especially, of the 40 A. paragallinarum isolates, 30% had an MIC value of compound sulfamethoxine sodium of 64 μg/mL, 10% of 128 μg/mL, and 15% of 256 μg/mL. For oxytetracycline hydrochloride, 85% of isolates showed MIC values of 64 μg/mL or more. Excitingly, the MIC values of β-lactamase (amoxicillin, ampicillin, and ceftiofur) were low, with 77.5%, 70%, and 92.5% of isolates having an MIC value of ≤1 μg/mL, respectively. Our results may provide a reference for the treatment of infectious coryza.

Pathogenesis of Velogenic Genotype VII.1.1 Newcastle Disease Virus Isolated from Chicken in Egypt via Different Inoculation Routes: Molecular, Histopathological, and Immunohistochemical Study

Newcastle disease virus (NDV) remains a constant threat to the poultry industry. There is scarce information concerning the pathogenicity and genetic characteristics of the circulating velogenic Newcastle disease virus (NDV) in Egypt. In the present work, NDV was screened from tracheal swabs collected from several broiler chicken farms (N = 12) in Dakahlia Governorate, Egypt. Real-time reverse transcriptase polymerase chain reaction (RRT-PCR) was used for screening of velogenic and mesogenic NDV strains through targeting F gene fragment amplification, followed by sequencing of the resulting PCR products. The identified strain, namely, NDV-CH-EGYPT-F42-DAKAHLIA-2019, was isolated and titrated in the allantoic cavity of 10 day old specific pathogen-free (SPF) embryonated chicken eggs (ECEs), and then their virulence was determined by mean death time (MDT) and intracerebral pathogenicity index (ICPI). The pathogenicity of the identified velogenic NDV strain was also assessed in 28 day old chickens using different inoculation routes as follows: intraocular, choanal slit, intranasal routes, and a combination of both intranasal and intraocular routes. In addition, sera were collected 5 and 10 days post inoculation (pi) for the detection of NDV antibodies by hemagglutination inhibition test (HI), and tissue samples from different organs were collected for histopathological and immunohistochemical examination. A series of different clinical signs and postmortem lesions were recorded with the various routes. Interestingly, histopathology and immunohistochemistry for NDV nucleoprotein displayed widespread systemic distribution. The intensity of viral nucleoprotein immunolabeling was detected within different cells including the epithelial and endothelium lining, as well as macrophages. The onset, distribution, and severity of the observed lesions were remarkably different between various inoculation routes. Collectively, a time-course comparative pathogenesis study of NDV infection demonstrated the role of different routes in the pathogenicity of NDV. The intranasal challenge was associated with a prominent increase in NDV lesions, whereas the choanal slit route was the route least accompanied by severe NDV pathological findings. Clearly, the present findings might be helpful for implementation of proper vaccination strategies against NDV.

Rational selection of hemagglutinin variants of H3N2 influenza viruses in preparation of live influenza vaccine strains to optimize growth characteristics

BACKGROUND: Up to date Russian live attenuated influenza vaccines are produced in developing chicken embryos. During passaging in embryos, the virus isolated from the human respiratory tract undergoes adaptation to the receptors in embryos. The population of the virus, at any passage in chicken embryos, is heterogeneous and contains variants of viruses with one or another set of adaptive substitutions. Before preparing the vaccine strain, the population of the epidemic virus is cloned and the genetic sequence of the hemagglutinin and neuraminidase clones is analyzed. The growth characteristics of the vaccine strain and its antigenic properties depend on the correct choice of the variant of the virus. AIM: The aim of the study was to select the variant of the H3N2 subtype virus for the preparation of a vaccine reassortant based on data on the composition of the population and an assessment of its growth properties. MATERIALS AND METHODS: Viruses were cloned in developing chicken embryos, sequencing of the hemagglutinin and neuraminidase genes of the clones was performed. On the basis of the clones selected based on the results of the analysis of the population, strains of a live influenza vaccine were obtained by the reassortment in the chicken embryos. The growth characteristics of the strains, the phenotype in eggs, and the antigenic properties by hemagglutination inhibition test were evaluated. RESULTS: The influenza virus A/Kansas/14/2017 recommended by WHO for the epidemic season 2019-2020 acquired a pair of D190N + N246T substitutions dominating in the population at the 7th passage in eggs. From the population of A/Kansas/14/2017-like strain A/Brisbane/34/2018, from the third passage in the eggs, it was possible to obtain a variant of the virus with substitutions G186V + S219Y in hemagglutinin. The growth characteristics of the strain based on A/Kansas/14/2017 (passage E7) were significantly inferior to the characteristics of the strain based on A/Brisbane/34/2018 (passage E3), in the absence of differences in antigenic properties. CONCLUSIONS: The variant of egg adaptation of hemagglutinin G186V in strains of clade 3c.3a is preferable for the preparation of live influenza vaccine strains; variant N246T is not optimal. When preparing strains, it is necessary to analyze the composition of the virus population by cloning and choose the most optimal option for preparing strains. The persistence of egg-adaptive substitutions in passaged variants of the virus is not always optimal for strains of live influenza vaccine, and therefore it is preferable to use the population as close as possible to the initial variant to start work on the strain.

In Vitro Antiviral Activity of Green Tea Polyphenon-60 against Avian Paramyxoviruses

Avian paramyxoviruses (APMVs) have caused an economically significant drop in global domestic poultry production because of their high morbidity and mortality rates. Polyphenols are the major components of green tea that have great antiviral effects. This study aimed to evaluate the anti-APMV activities of polyphenon-60. Twelve APMV-1 strains representing three different pathotypes, two strains of APMV-2, one strain of APMV-3, and one strain of APMV-7 were propagated in chicken embryos. To determine the cytotoxic effect, chicken embryo fibroblasts were treated with the test compound in various concentrations. To assess the antiviral properties, time-dependent, dose-dependent, and virulence-dependent experiments were conducted in both cell and chicken embryo models. A reduction in virus titers was measured by the hemagglutination test. The inhibitory effect on virus adsorption to the chicken red blood cell (RBC) surface was examined by the hemagglutination inhibition test. The results showed that lentogenic and mesogenic APMV-1 strains, APMV-3 strain, and APMV-7 strain were significantly inhibited ( p < 0.05 ) by polyphenon-60 at 50 μg/ml, while the 50% cytotoxic concentration of the compound was 345 μg/ml. Polyphenon-60 also exhibited the inhibitory activity against hemagglutination by NDV. Taken together, the results suggest that polyphenon-60 has shown promise as an antiviral agent that has wide safety margins against APMVs, and challenge studies to evaluate its efficacy in chickens are necessary.

Seroprevalence of Newcastle Disease Virus in Slaughtered Pigeons from Selected Markets in Zaria, Kaduna State Nigeria

Newcastle disease (ND) outbreak even in the face of vaccination is a common problem in Nigeria. Three Newcastle disease (ND) panzootic were recorded in the past and one of such was caused by feral pigeon fecal contamination of poultry feed materials. The study was designed to investigate the presence of ND antibodies in pigeons slaughtered in three markets in Zaria Nigeria. A total of 1500 Pigeons sera samples were tested across the selected markets, namely: Sabon-gari, Samaru and Tudun-wada markets. Five hundred blood samples were collected from each market. An overall seroprevalence of 22.33% (335/1500) was recorded in this study. Hemagglutination inhibition test revealed that pigeons from Sabon-gari market had the highest seroprevalence of 29% (145/500). Female pigeons had higher seroprevalence of 27.66% (130/470) compared to male pigeons 19.90% (205/1030). Younger pigeons of < 10 weeks old are more prone to ND infection with a seroprevalence of 34.21% (52/152) compared to pigeons > 10 weeks old 20.99% (238/1348). Also, only in Samaru market was there a significant association between ages (P = 0.00) and sexes (P = 0.00) of pigeons and seroprevalence of ND. Therefore, free routine ND vaccination campaign should be launched in the study areas with more emphases targeting the market species in order to mitigate the epidemic cycle of the virus. Moreover, market poultry farmers should be enlightened on the economic significance of the disease and the need to maintain strict biosecurity measures on their market farms.

A Para-Bombay Blood Group Case Associated with a Novel FUT1 Mutation c.361G>A

<b><i>Background:</i></b> Here we report a case of para-Bombay phenotype due to a novel mutation <i>FUT1</i> c.361G&#x3e;A p.(Ala121Thr) and a nonfunctional allele <i>FUT1</i>*<i>01N.13</i>(c.881_882delTT) which showed a discrepancy in the routine ABO blood group typing. <b><i>Materials and Methods:</i></b> The ABO phenotype and the Lewis blood group were typed with serological methods. The ABH antigens in saliva were determined by a hemagglutination inhibition test. The CDS region of <i>ABO, FUT1</i>and <i>FUT2</i> were amplified with polymerase chain reaction and then directly sequenced. The novel mutation was confirmed by cloning and sequencing. Three-dimensional (3-D) structural analysis of the mutant and wild-type Fut1 were performed by the Chimera software. <b><i>Results:</i></b> A, B and H antigens were not detected on the surface of red blood cells (RBCs) by the serological technique, and the B and H blood group substances were detected in the saliva, while the Lewis phenotype was Le(a–b+). Sequencing and cloning analysis showed the presence of a novel <i>FUT1</i> mutation c.361G&#x3e;A and a nonfunctional allele <i>FUT1</i>*<i>01N.13</i>(c.881_882delTT). The ABO genotype was <i>ABO</i>*<i>B.01/ABO</i>*<i>O.01.01</i>. The in silico analysis showed that the mutation p.(Ala121Thr) of <i>FUT1</i>did not change the 3-D structure of the whole enzyme but caused a certain amplitude of turnover in the loop region where Ala121 was located. <b><i>Conclusions:</i></b> A novel <i>FUT1</i> allele (<i>FUT1</i>*c.361G&#x3e;A) was identified in a Chinese individual with para-Bombay B phenotype. The <i>FUT1</i>c.361G&#x3e;A mutation may significantly downregulate the expression of H antigens on RBCs by damaging the enzyme conformation.

Role of B2-adrenoreceptors Polymorphism in the Development of Cardiovascular Complications in Severe Pneumonia with AH1N1/09 B Influenza in 2019 in Zabaykalsky Territory

The purpose of the study. Th e aim of this study was to study the polymorphism of the ADRB2 gene, as well as the frequency and nature of cardiovascular complications in its presence in patients with severe pneumonia with infl uenza A/H1N1/09. Material and methods. Th e study included 100 patients with pneumonia with infl uenza A/H1N1 / 09 in 2019 in the Trans-Baikal Territory. All patients were treated in the intensive care unit. In patients, the causative agent-infl uenza virus A/H1N1 / 09 was verifi ed by PCR-detection of the pathogen in nasopharyngeal smears or hemagglutination inhibition test (HIT) with an increase in the antibody titer in paired sera. DNA was isolated from peripheral blood leukocytes; fragments were amplifi ed by polymerase reaction and genotyped by short fragment length polymorphism for Gln27Glu and Arg16Gly ADRB2. Results. The patients were divided into 2 groups: Group I (n=41) – patients with cardiovascular complications; group II (n=59) – patients without cardiovascular complications. Th e groups were comparable in gender and age. Among the cardiovascular complications prevailed: arrhythmias (n=13); decompensation of chronic heart failure (n=13); acute myocardial infarction (n=7); pulmonary embolism (n=5); myocarditis/pericarditis (n=3). When studying the ADRB2 polymorphism, it was found that the most unfavorable variant is the homozygous mutation Arg16Gly (Gly/Gly). This mutation was detected in 6 (14.6 %) patients in group I, but was not detected in group II (P=0.003). In the presence of this mutation in patients with severe secondary viral-bacterial pneumonia with infl uenza A/H1N1 / 09, the risk of developing cardiovascular complications increases (HR=2.6; CI 2.06; 3.49). Conclusion. Patients with severe pneumonia during the A/H1N1/09 infl uenza epidemic in 2019 had a high incidence of cardiovascular complications (41 %). The presence of the homozygous Arg16Gly mutation (Gly/Gly) increases the risk of cardiovascular complications by 2.06 times, and the presence of the heterozygous Arg16Gly mutation (Arg/Gly) reduces this risk by 0.5 times

Study of Arboviruses in Philander opossum, Didelphis marsupialis and Nectomys rattus captured from forest fragments in the municipality of Belém, Pará, Brazil

ABSTRACT: Arboviruses are viruses that maintain their life cycle in the wild and are transmitted to vertebrate hosts by hematophagous diptera. They are zoonotic and can establish an enzootic cycle in the urban areas; in humans, the infection can manifest from being encephalitogenic to hemorrhagic. This study aimed to report the occurrence of arboviruses in mammals of the order Didelphimorphia and Rodentia, captured from the Amazon. Serum samples were subjected to hemagglutination inhibition test using a viral panel of 19 species of arboviruses that are known to occur in the Amazon. Altogether, 14 wild mammals, 12 of Philander opossum, 1 of Didelphis marsupialis, and 1 of Nectomys rattus were captured. Eight of these were reported to be seropositive for arboviruses (57.14%) with monotypic seroprevalence for the Eastern Equine Encephalitis Virus (n=1), the Ilheus Virus (n=2), and the Catu virus (n=4); 4 heterotypic responses were observed for Flavivirus and Orthobunyavirus. In conclusion, several arbovirus species are in active circulation and maintenance, exhibiting enzootic characteristics in the wild mammals of the Amazon region; these animals prove to be potential hosts in the transmission of diseases to humans.

Hemagglutination inhibition antibody titers as a correlate of protection against influenza disease in the 2018/2019 epidemic season in Poland

The aim of this study was to determine the level of antibodies against hemagglutinin of influenza viruses in the sera of people in the seven age groups in the epidemic season 2018/2019 in Poland. The level of anti-hemagglutinin antibodies was determined by hemagglutination inhibition test (HAI). 1050 clinical samples from all over the country were tested. The level of antibodies against influenza viruses was highest in the 10–14 age group for A/Singapore/INFIMH-16-0019/2016 (H3N2) and B/Phuket/3073/2013 Yamagata lineage antigens. These results confirm the circulation of four antigenically different influenza virus strains, two subtypes of influenza A virus – A/Michigan/45/2015 (H1N1)pdm09 and A/Singapore/INFIMH-16-0019/2016 (H3N2) and two lineages of influenza B virus – B/Colorado/06/2017 – Victoria lineage and B/Phuket/3073/2013 Yamagata lineage.