ABSTRACT
The L 1 region of bovine adenovirus (BAdV)-3 encodes a multifunctional protein named protein VII. Anti-protein VII sera detected a protein of 26 kDa in transfected or BAdV-3-infected cells, which localizes to nucleus and nucleolus of infected/transfected cells. Analysis of mutant protein VII identified four redundant overlapping nuclear/nucleolar localization signals as deletion of all four potential nuclear/nucleolar localization signals localizes protein VII predominantly to the cytoplasm. The nuclear import of protein VII appears to use importin α (α-1), importin-ß (ß-1) and transportin-3 nuclear transport receptors. In addition, different nuclear transport receptors also require part of protein VII outside nuclear localization sequences for efficient interaction. Proteomic analysis of protein complexes purified from recombinant BAdV-3 expressing protein VII containing Strep Tag II identified potential viral and cellular proteins interacting with protein VII. Here, we confirm that protein VII interacts with IVa2 and protein VIII in BAdV-3-infected cells. Moreover, amino acids 91-101 and 126-137, parts of non-conserved region of protein VII, are required for interaction with IVa2 and protein VIII, respectively.
Subject(s)
Host-Pathogen Interactions , Mastadenovirus , Viral Proteins , Animals , Cattle , Humans , Active Transport, Cell Nucleus , Cell Line , Cell Nucleus/metabolism , Cell Nucleus/virology , Mastadenovirus/metabolism , Mastadenovirus/genetics , Mastadenovirus/physiology , Nuclear Localization Signals , Protein Binding , Proteomics/methods , Viral Proteins/metabolism , Viral Proteins/geneticsABSTRACT
INTRODUCTION: Human mastadenovirus (HAdV) types 8, 37, 64 have been considered the major contributors in Epidemic keratoconjunctivitis (EKC) epidemics, but recent surveillance data have shown the involvement of emerging recombinants, including HAdV-53, HAdV-54, and HAdV-56. In our initial work, positive samples for adenovirus revealed that our strains were closer to HAdV-54 than HAdV-8. Hence, the current study aimed to use whole genome technology to identify the HAdV strain correctly. METHODOLOGY: Oxford Nanopore technique was used, wherein a Targeted sequencing approach using long-range PCR amplification was performed. Primers were designed using HAdV-54 (AB448770.2) and HAdV-8 (AB897885.1) as reference sequences. Amplicons were sequenced on the GridION sequencer. Sequences were annotated using Gatu software, and similarities with standard reference sequence was calculated using Bioedit software. The phylogenetic tree was built after alignment in MEGA v7.0 using Neighbour joining method for each of the genes: Penton, Hexon, and Fiber. The effect of novel amino acid changes was evaluated using the PROVEAN tool. The Recombination Detection Program (RDP) package Beta 4.1 was used to identify recombinant sequences. RESULTS: Of the five samples sequenced, OL450401, OL540403, and OL540406 showed nucleotide similarity to HAdV-54 in the penton region. Additionally, OL450401 showed a statistically significant recombination event with HAdV-54 as minor and HAdV-8 as major parents. This was further supported by phylogenetic analysis as well. CONCLUSIONS: In the present study, we have found evidence of a shift from HAdV-8 towards HAdV-54, thus stressing the need for surveillance of HAdVs and to stay updated on the rise of new recombinants.
Subject(s)
Adenovirus Infections, Human , Adenoviruses, Human , Keratoconjunctivitis , Mastadenovirus , Humans , Adenovirus Infections, Human/epidemiology , Adenovirus Infections, Human/genetics , Adenoviruses, Human/genetics , Genome, Viral , India/epidemiology , Keratoconjunctivitis/epidemiology , Mastadenovirus/genetics , Phylogeny , Sequence Analysis, DNAABSTRACT
Adenoviruses (AdVs) have been detected in a wide variety of animals. To date, eight types of AdVs in sheep and two types in goats have been identified, which belong to two distinct genera, Mastadenovirus and Atadenovirus. Typically, the term pneumo-enteritis is used to describe adenovirus-induced disease in small ruminants, which has been associated with both enteric and respiratory symptoms of varying severity. The aim of this study was to detect and identify AdVs of small ruminants belonging to the genera Mastadenovirus and Atadenovirus. For this purpose, diagnostic samples (47 lung, 27 intestine, and two pooled tissue samples including intestine and lung) from 49 small ruminants (39 sheep and 10 goats) were used. Following the viral DNA extraction, PCR was carried out by using the primers targeting the hexon gene in order to detect both mast- and atadenoviruses. Sequencing the amplified fragments revealed the presence of three types of ovine adenovirus (OAdV): OAdV-3, OAdV-4, and OAdV-8. Specifically, OAdV-3 was detected in two sheep and a goat while OAdV-4 and OAdV-8 were found in only one sheep each. There is still limited data on the interaction between the viruses in different adenovirus genera and the detected disease, as well as the genetic diversity of adenoviruses, especially in small ruminants. In conclusion, the detection of AdVs in lung and intestinal tissues of small ruminants in this study suggests that these viruses may have contributed to the disease and/or predisposed to other agents.
Subject(s)
Adenoviridae Infections , Goat Diseases , Goats , Mastadenovirus , Phylogeny , Sheep Diseases , Animals , Goats/virology , Sheep/virology , Sheep Diseases/virology , Goat Diseases/virology , Adenoviridae Infections/veterinary , Adenoviridae Infections/virology , Mastadenovirus/genetics , Mastadenovirus/isolation & purification , Mastadenovirus/classification , Turkey , DNA, Viral/genetics , Sequence Analysis, DNA , Atadenovirus/genetics , Atadenovirus/isolation & purification , Atadenovirus/classification , Lung/virology , Adenoviridae/genetics , Adenoviridae/isolation & purification , Adenoviridae/classification , Adenoviridae/pathogenicityABSTRACT
Bat-borne viruses may affect public health and the global economy. These mammals have a wide geographical distribution and unique biological, physiological, and immunogenic characteristics, allowing the dissemination of many known and unknown viruses. Enteric viruses, such as adeno (AdV) and rotaviruses, are recognized as the main causative agents of disease and outbreaks. In the present study, the presence of viruses from Adenoviridae and Reoviridae families was evaluated in molossid, phyllostomid, and vespertilionid bats captured in Rio Grande do Sul, Southern Brazil, between September 2021 and July 2022. Sixty bat rectal swabs were analyzed by PCR. Eight (13.3%) samples were positive for adenovirus and classified as human mastadenovirus C (HAdV-C) (three samples) and HAdV-E (five samples) by sequencing followed by phylogenetic analysis. All samples were negative in rotavirus specific RT-PCR. This is the first study to describe the presence of HAdV in samples of Glossophaga soricina, Eptesicus brasiliensis, and Histiotus velatus. Furthermore, the presence of HAdV-E in bats was reported, which is unusual and may suggest that other HAdV genotypes, in addition to HAdV-C, may also be harbored by wild animals. The data generated in the present study reinforces the importance of eco-surveillance of viral agents related to diseases in humans and wild animals. In addition, it is essential to identify possible new hosts or reservoirs that increase the risk of spillover and dissemination of infectious pathogens, helping to prevent and control zoonotic diseases.
Subject(s)
Chiroptera , Mastadenovirus , Phylogeny , Rotavirus , Animals , Brazil/epidemiology , Chiroptera/virology , Rotavirus/genetics , Rotavirus/classification , Rotavirus/isolation & purification , Mastadenovirus/classification , Mastadenovirus/genetics , Mastadenovirus/isolation & purification , Adenoviridae Infections/veterinary , Adenoviridae Infections/virologyABSTRACT
A 7-mo-old farmed white-tailed deer fawn (Odocoileus virginianus) died after several weeks of progressive deterioration associated with endoparasitism and respiratory signs. A field autopsy was performed, and lung tissue was submitted for histologic examination. The findings were consistent with necrosuppurative bronchointerstitial pneumonia with intranuclear viral inclusions. Immunofluorescence using fluorescently labeled polyclonal antibodies to bovine adenovirus 3 and 5 was positive. To rule out cross-reactivity with other adenoviruses, formalin-fixed, paraffin-embedded tissue sections were submitted for genome sequence analysis, which revealed a 99.6% match to Deer mastadenovirus B (formerly Odocoileus adenovirus 2, OdAdV2). To our knowledge, natural clinical disease associated with OdAdV2 has not been reported previously.
Subject(s)
Adenoviridae Infections , Cattle Diseases , Deer , Mastadenovirus , Pneumonia , Cattle , Animals , Mastadenovirus/genetics , Adenoviridae Infections/veterinary , Pneumonia/veterinaryABSTRACT
Although the occurrence of three fiber genes in monkey adenoviruses had already been described, the relatedness of the "extra" fibers have not yet been discussed. Here we report the genome analysis of two simian adenovirus (SAdV) serotypes from Old World monkeys and the phylogenetic analysis of the multiple fiber genes found in these and related AdVs. One of the newly sequenced serotypes (SAdV-2), isolated from a rhesus macaque (Macaca mulatta), was classified into species Human mastadenovirus G (HAdV-G), while the other serotype (SAdV-17), originating from a grivet (Chlorocebus aethiops), classified to Simian mastadenovirus F (SAdV-F). We identified unique features in the gene content of these SAdVs compared to those typical for other members of the genus Mastadenovirus. Namely, in the E1B region of SAdV-2, the 19K gene was replaced by an ITR repetition and a copy of the E4 ORF1 gene. Among the 37 genes in both SAdVs, three genes of different lengths, predicted to code for the cellular attachment proteins (the fibers), were found. These proteins exhibit high diversity. Yet, phylogenetic calculations of their conserved parts could reveal the probable evolutionary steps leading to the multiple-fibered contemporary HAdV and SAdV species. Seemingly, there existed (a) common ancestor(s) with two fiber genes for the lineages of the AdVs in species SAdV-B, -E, -F and HAdV-F, alongside a double-fibered ancestor for today's SAdV-C and HAdV-G, which later diverged into descendants forming today's species. Additionally, some HAdV-G members picked up a third fiber gene either to the left-hand or to the in-between position from the existing two. A SAdV-F progenitor also obtained a third copy to the middle, as observed in SAdV-17. The existence of three fiber genes in these contemporary AdVs brings novel possibilities for the design of optimised AdV-based vectors with potential multiple target binding abilities.
Subject(s)
Adenoviruses, Simian , Mastadenovirus , Animals , Humans , Chlorocebus aethiops , Adenoviridae , Macaca mulatta , Phylogeny , Adenoviruses, Simian/genetics , Mastadenovirus/geneticsABSTRACT
Bovine adenoviruses (BAdV) are known to cause respiratory and/or intestinal disease in calves. Infection can manifest as acute outbreaks, but more often only sporadic cases occur. Here we describe the PCR detection and partial sequence characterization of several BAdVs found in sick or dead calves on different farms in Western Hungary. Intermittent diarrhoeal illnesses occurred after weaning among calves on several farms located up to 40 km apart. A high-sensitivity, broad-spectrum nested PCR, developed for the general detection of adenoviruses, gave positive results in four independent cases. Direct sequencing of PCR products showed clear results from only two samples, whereas sequences from the other two amplicons were mixed. Molecular cloning of these heterogeneous PCR products was performed to separate each DNA fragment therein. By sequencing several plasmid clones from both mixed samples, we were able to detect the simultaneous presence of two different BAdV types, namely types 6 and 10 classified into two separate (Atadenovirus and Mastadenovirus) genera. The sequence of one homogenous sample was identified as being derived also from BAdV-10, whereas the other sample contained a novel type, proposed to be BAdV-11. We demonstrated, for the very first time, the occurrence of the two latter virus types in continental Europe. Their appearance in Hungary marks a significant shift in the types of BAdVs actually circulating in the country. Considering the similarity of the pathological findings to those, attributed to BAdV-10 infections reported to date, the causative role of the viruses in these cases seems to be plausible. Phylogeny reconstruction further confirmed that BAdVs represent multiple genetic lineages.
Subject(s)
Adenoviridae , Mastadenovirus , Cattle , Animals , Base Sequence , Adenoviridae/genetics , Mastadenovirus/genetics , Polymerase Chain Reaction/methods , Polymerase Chain Reaction/veterinary , Europe/epidemiologyABSTRACT
The Adenoviridae family is composed by a high diversity of viruses that are extremely resistant in environment and are frequently excreted in animal reservoir feces for long periods. The knowledge of adenovirus (AdV) diversity among wild species may be important for the understanding of the epidemiology of putative emerging diseases. Cavia aperea aperea, commonly known as wild guinea pigs, wild cavies, or preas, are small herbivorous rodents widely distributed throughout South America and classified in Caviidae family, as well as domestic guinea pigs and capybaras. In order to investigate their potential role as reservoir of zoonotic agents, the present study aimed to verify the presence of AdV in fecal samples of 14 preas from Northeast Brazil. When submitted to nested PCR, two out of 14 samples (14.28%) were positive for AdV and classified as human Mastadenovirus C (HAdV-C) using DNA sequencing and phylogenetic analysis. Wild guinea pigs are synanthropic rodents that live in close contact with humans. The investigation of viral agents in rodents is important due to their potential role as reservoirs of human and animal pathogens. Moreover, the present work presents the first known evidence of HAdV in wild guinea pig stool samples, which may represent both the impact of anthropogenic pollution to wild animals and an important knowledge in terms of human health.
Subject(s)
Animals, Wild , Mastadenovirus , Humans , Guinea Pigs , Animals , Phylogeny , Feces , Sequence Analysis, DNA , Mastadenovirus/geneticsABSTRACT
Over one hundred Mastadenovirus types infect seven orders of mammals. Virus-host coevolution may involve cospeciation, duplication, host switch and partial extinction events. We reconstruct Mastadenovirus diversification, finding that while cospeciation is dominant, the other three events are also common in Mastadenovirus evolution. Linear motifs are fast-evolving protein functional elements and key mediators of virus-host interactions, thus likely to partake in adaptive viral evolution. We study the evolution of eleven linear motifs in the Mastadenovirus E1A protein, a hub of virus-host protein-protein interactions, in the context of host diversification. The reconstruction of linear motif gain and loss events shows fast linear motif turnover, corresponding a virus-host protein-protein interaction turnover orders of magnitude faster than in model host proteomes. Evolution of E1A linear motifs is coupled, indicating functional coordination at the protein scale, yet presents motif-specific patterns suggestive of convergent evolution. We report a pervasive association between Mastadenovirus host diversification events and the evolution of E1A linear motifs. Eight of 17 host switches associate with the gain of one linear motif and the loss of four different linear motifs, while five of nine partial extinctions associate with the loss of one linear motif. The specific changes in E1A linear motifs during a host switch or a partial extinction suggest that changes in the host molecular environment lead to modulation of the interactions with the retinoblastoma protein and host transcriptional regulators. Altogether, changes in the linear motif repertoire of a viral hub protein are associated with adaptive evolution events during Mastadenovirus evolution.
Subject(s)
Adenovirus E1A Proteins , Evolution, Molecular , Host-Pathogen Interactions , Mastadenovirus , Adenovirus E1A Proteins/chemistry , Adenovirus E1A Proteins/genetics , Amino Acid Motifs , Animals , Mammals/virology , Mastadenovirus/chemistry , Mastadenovirus/genetics , Protein Interaction MappingABSTRACT
Human mastadenovirus (HAdV), a linear double-stranded DNA (dsDNA) virus, is the causal agent of several diseases, including pharyngoconjunctival fever, epidemic keratoconjunctivitis, and hemorrhagic cystitis, in immunocompromised individuals. There are more than 100 reported types of adenoviruses, but the pathogenicity of many HAdVs remains unknown. Brincidofovir (BCV) is a hexadecyloxypropyl lipid conjugate of cidofovir (CDV) that is active against dsDNA viruses. Clinical effectiveness of BCV against certain HAdV species has been reported; however, its activity against novel HAdV types remains unknown. We investigated the half-maximal inhibitory concentration (IC50) values of BCV for novel HAdV types and found that the epidemic keratoconjunctivitis-associated HAdV-D54 prevalent in the Asian region was the most susceptible. The mean overall IC50 value of BCV was lower than that of CDV, indicating that BCV is effective against HAdVs, including the novel types. IMPORTANCE We investigated the IC50 values of BCV for novel HAdV types and found that the epidemic keratoconjunctivitis-associated HAdV-D54 prevalent in the Asian region was the most susceptible. In addition, the mean overall IC50 value of BCV was lower than that of CDV, indicating that BCV is effective against HAdVs.
Subject(s)
Adenoviridae Infections/virology , Adenovirus Infections, Human/virology , Cytosine/analogs & derivatives , Keratoconjunctivitis/virology , Mastadenovirus/drug effects , Organophosphonates/pharmacology , Adenoviridae Infections/immunology , Adenovirus Infections, Human/immunology , Cystitis , Cytosine/pharmacology , Humans , Immunocompromised Host , Keratoconjunctivitis/immunology , Mastadenovirus/classification , Mastadenovirus/genetics , Mastadenovirus/physiologyABSTRACT
Here, we report a novel bat adenovirus strain isolated from apparently healthy bats of the species Rhinolophus cornutus in Japan. The genome of the isolate was 36,506 bp in length and encoded at least 33 proteins. Phylogenetic analysis of the DNA polymerase amino acid sequence, which provides one demarcation criterion for adenoviral species, indicated that the isolate belongs to the species Bat mastadenovirus C in the genus Mastadenovirus. Most of the encoded proteins shared high sequence similarity with those of known bat adenovirus C strains detected in different species of Rhinolophus, whereas the fiber protein and some E3- and E4-related proteins shared moderate similarity, and only the large E3 protein, which contains several host immune-suppression-related motifs, showed considerably lower similarity.
Subject(s)
Chiroptera , Mastadenovirus , Animals , Genome, Viral , Japan , Mastadenovirus/genetics , PhylogenyABSTRACT
Using a broad-range nested PCR assay targeting the DNA-dependent DNA polymerase (pol) gene, we detected adenoviruses in 17 (20.48%) out of 83 fecal samples from small Indian mongooses (Urva auropunctata) on the Caribbean island of St. Kitts. All 17 PCR amplicons were sequenced for the partial pol gene (~300 bp, hereafter referred to as Mon sequences). Fourteen of the 17 Mon sequences shared maximum homology (98.3-99.6% and 97-98.9% nucleotide (nt) and deduced amino acid (aa) sequence identities, respectively) with that of bovine adenovirus-6 (species Bovine atadenovirus E). Mongoose-associated adenovirus Mon-39 was most closely related (absolute nt and deduced aa identities) to an atadenovirus from a tropical screech owl. Mon-66 shared maximum nt and deduced aa identities of 69% and 71.4% with those of atadenoviruses from a spur-thighed tortoise and a brown anole lizard, respectively. Phylogenetically, Mon-39 and Mon-66 clustered within clades that were predominated by atadenoviruses from reptiles, indicating a reptilian origin of these viruses. Only a single mongoose-associated adenovirus, Mon-34, was related to the genus Mastadenovirus. However, phylogenetically, Mon-34 formed an isolated branch, distinct from other mastadenoviruses. Since the fecal samples were collected from apparently healthy mongooses, we could not determine whether the mongoose-associated adenoviruses infected the host. On the other hand, the phylogenetic clustering patterns of the mongoose-associated atadenoviruses pointed more towards a dietary origin of these viruses. Although the present study was based on partial pol sequences (~90 aa), sequence identities and phylogenetic analysis suggested that Mon-34, Mon-39, and Mon-66 might represent novel adenoviruses. To our knowledge, this is the first report on the detection and molecular characterization of adenoviruses from the mongoose.
Subject(s)
Adenoviridae/classification , Adenoviridae/genetics , Adenoviridae/isolation & purification , Herpestidae/virology , Adenoviridae Infections/veterinary , Adenoviridae Infections/virology , Amino Acid Sequence , Animals , Atadenovirus/classification , Atadenovirus/genetics , Atadenovirus/isolation & purification , DNA-Directed DNA Polymerase , Feces/virology , Lizards/virology , Mastadenovirus/classification , Mastadenovirus/genetics , Mastadenovirus/isolation & purification , Phylogeny , Polymerase Chain Reaction , Turtles/virology , West IndiesABSTRACT
Although a canine adenovirus (CAdV)-based oncolytic virus (OV) candidate targeting canine tumors has been reported, its oncolytic effect could be attenuated by CAdV vaccine-induced neutralizing antibodies in dog patients. To circumvent this issue, we focused on the bat adenovirus (BtAdV) strain, which was previously isolated from healthy microbats. We previously showed that this virus replicated efficiently in canine cell lines and did not serologically cross-react with CAdVs, suggesting that it may offer the possibility of an OV candidate for canine tumors. Here, we tested the growth properties and cytotoxicity of the BtAdV Mm32 strain in a panel of canine tumor cells and found that its characteristics were equivalent to those of CAdVs. To produce an Mm32 construct with enhanced tumor specificity, we established a novel reverse genetics system for BtAdV based on bacterial artificial chromosomes, and generated a recombinant virus, Mm32-E1Ap + cTERTp, by inserting a tumor-specific canine telomerase reverse transcriptase promoter into its E1A regulatory region. The growth and cytotoxicity of this recombinant were superior to those of wild-type Mm32 in canine tumor cells, unlike in normal canine cells. These data suggest that Mm32-E1Ap + cTERTp could be a promising OV for alternative canine cancer therapies.
Subject(s)
Chiroptera/virology , Dog Diseases/therapy , Mastadenovirus , Neoplasms , Oncolytic Virotherapy , Oncolytic Viruses , Animals , Cell Line, Tumor , Chromosomes, Artificial, Bacterial/genetics , Dogs , Madin Darby Canine Kidney Cells , Mastadenovirus/genetics , Mastadenovirus/metabolism , Neoplasms/therapy , Neoplasms/veterinary , Oncolytic Viruses/genetics , Oncolytic Viruses/metabolismABSTRACT
With the arrival of coronavirus disease 2019 (COVID-19) in Brazil in February 2020, several preventive measures were taken by the population aiming to avoid severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection including the use of masks, social distancing, and frequent hand washing then, these measures may have contributed to preventing infection also by other respiratory viruses. Our goal was to determine the frequencies of Influenza A and B viruses (FLUAV/FLUBV), human mastadenovirus C (HAdV-C), Enterovirus 68 (EV-68), and rhinovirus (RV) besides SARS-CoV-2 among hospitalized patients suspect of COVID-19 with cases of acute respiratory disease syndrome (ARDS) in the period of March to December 2020 and to detect possible coinfections among them. Nucleic acid detection was performed using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) in respiratory samples using naso-oropharyngeal swabs and bronchoalveolar lavage. A total of 418 samples of the 987 analyzed (42.3%) were positive for SARS-CoV-2, 16 (1.62%) samples were positive for FLUAV, no sample was positive for FLUBV or EV-68, 67 (6.78%) samples were positive for HAdV-C, 55 samples were positive for RV 1/2 (26.3%) and 37 for RV 2/2 (13.6%). Coinfections were also detected, including a triple coinfection with SARS-CoV-2, FLUAV, and HAdV-C. In the present work, a very low frequency of FLUV was reported among hospitalized patients with ARDS compared to the past years, probably due to preventive measures taken to avoid COVID-19 and the high influenza vaccination coverage in the region in which this study was performed.
Subject(s)
Adenoviridae Infections/epidemiology , COVID-19/epidemiology , Common Cold/epidemiology , Enterovirus Infections/epidemiology , Influenza, Human/epidemiology , Physical Distancing , Adenoviridae Infections/prevention & control , Adolescent , Adult , Aged , Aged, 80 and over , Brazil/epidemiology , COVID-19/prevention & control , Child , Child, Preschool , Coinfection/epidemiology , Coinfection/virology , Common Cold/prevention & control , Enterovirus D, Human/genetics , Enterovirus D, Human/isolation & purification , Enterovirus Infections/prevention & control , Female , Humans , Infant , Influenza A virus/genetics , Influenza A virus/isolation & purification , Influenza B virus/genetics , Influenza B virus/isolation & purification , Influenza, Human/prevention & control , Male , Masks , Mastadenovirus/genetics , Mastadenovirus/isolation & purification , Middle Aged , Nucleic Acid Amplification Techniques/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Rhinovirus/genetics , Rhinovirus/isolation & purification , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Young AdultABSTRACT
BRD is associated with infectious agents, but management and transport-stress are trigger factors. Metaphylactic administration of antimicrobial reduces colonization of respiratory tract by pathogens, but the development of antibiotic-resistance raises public health concerns leading to propose new control strategies. The study analyzed nasopharyngeal swabs of 231 imported cattle, 10% of 49 trucks, transported from France to southern Italy and, through Real-time PCR identified the prevalence of the involved pathogens speculating on strategies to reduce the impact of BRD. The samples were tested by Real-time PCR, for the detection of bovine coronavirus (BCoV), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus (BPiV), bovine adenovirus (BAdV), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Yates-corrected chi squared, or Fisher's exact test were used to compare both animal-health status and positivity/negativity to pathogens, and the relationship between presence/absence of clinical signs and Real-time PCR-positivity. H. somni and BCoV were the most frequently identified pathogens. In BRD-diagnosed cattle, BAdV was detected in 13.8% (19/138), BRSV in 14.5% (20/138) and BPiV in 4.3% (6/138). Healthy cattle were mostly positive for H. somni (89.2%, 83/93). A statistically significant association was observed between clinical signs and positivity to M. haemolytica (p value = 0.016). Although mass-medication and vaccination are used for BRD control, it still remains a primary health problem. Our results highlight that the nasopharyngeal microbiota could be affected by transport and that strategies to enhance calf immunity for reducing BRD-risk development would be more effective if applied at farm of origin prior to loading.
Subject(s)
Cattle Diseases/epidemiology , Coronavirus, Bovine/isolation & purification , Microbiota , Pasteurellaceae/isolation & purification , Respiratory Tract Diseases/veterinary , Animals , Cattle , Cattle Diseases/microbiology , Cattle Diseases/prevention & control , Coronavirus, Bovine/genetics , Epidemiologic Studies , France/epidemiology , Immunity , Italy/epidemiology , Male , Mastadenovirus/genetics , Mastadenovirus/isolation & purification , Nasopharynx/microbiology , Pasteurellaceae/genetics , Respiratory Syncytial Virus, Bovine/genetics , Respiratory Syncytial Virus, Bovine/isolation & purification , Respiratory System/microbiology , Respiratory Tract Diseases/epidemiology , Respiratory Tract Diseases/microbiology , Respiratory Tract Diseases/prevention & control , Respirovirus/genetics , Respirovirus/isolation & purification , TransportationABSTRACT
The complete genomic sequence along with phylogenetic analyses of an adenovirus (AdV), isolated from a dead captive pygmy marmoset (Callithrix pygmaea) from a Hungarian zoo is reported. Earlier, based on the phylogenetic analysis of the sequence of a PCR-amplified fragment from the DNA polymerase gene, the pygmy marmoset AdV (PMAdV) has been reported to cluster closest to certain chiropteran AdVs. In the following years similar AdVs were discovered in additional mammalian hosts, including a skunk (Mephitis mephitis), African pygmy hedgehogs (Atelerix albiventris), North American porcupines (Erethizon dorsatum) and grey fox (Urocyon cinereoargenteus). After the full genome analysis of the skunk adenovirus (SkAdV-1), a novel species Skunk mastadenovirus A (SkAdV-A) has been established. The AdVs, originating from the African pygmy hedgehogs, have been found to belong to virus species SkAdV-A. Partial gene sequences from the porcupine AdVs have also implied their very close genetic relatedness to SkAdV-A. The complete genomic sequence of PMAdV, examined in this study, was found to share 99.83% nucleotide identity with SkAdV-1, thus unequivocally represents a genomic variant of SkAdV-1. The observation that viruses classifiable as SkAdV-A are able to infect and cause diseases in several, distantly related mammals seems to deserve further studies to elucidate the infection biology of this intriguing AdV.
Subject(s)
Callithrix/virology , Genome, Viral , Mastadenovirus/genetics , Mephitidae/virology , Animals , Mastadenovirus/classification , Whole Genome Sequencing/veterinaryABSTRACT
Non-human primates (NHPs) are known hosts for adenoviruses (AdVs), so there is the possibility of the zoonotic or cross-species transmission of AdVs. As with humans, AdV infections in animals can cause diseases that range from asymptomatic to fatal. The aim of this study was to investigate the occurrence and diversity of AdVs in: (i) fecal samples of apes and monkeys from different African countries (Republic of Congo, Senegal, Djibouti and Algeria), (ii) stool of humans living near gorillas in the Republic of Congo, in order to explore the potential zoonotic risks. Samples were screened by real-time and standard PCRs, followed by the sequencing of the partial DNA polymerase gene in order to identify the AdV species. The prevalence was 3.3 folds higher in NHPs than in humans. More than 1/3 (35.8%) of the NHPs and 1/10 (10.5%) of the humans excreted AdVs in their feces. The positive rate was high in great apes (46%), with a maximum of 54.2% in chimpanzees (Pan troglodytes) and 35.9% in gorillas (Gorilla gorilla), followed by monkeys (25.6%), with 27.5% in Barbary macaques (Macaca sylvanus) and 23.1% in baboons (seven Papio papio and six Papio hamadryas). No green monkeys (Chlorocebus sabaeus) were found to be positive for AdVs. The AdVs detected in NHPs were members of Human mastadenovirus E (HAdV-E), HAdV-C or HAdV-B, and those in the humans belonged to HAdV-C or HAdV-D. HAdV-C members were detected in both gorillas and humans, with evidence of zoonotic transmission since phylogenetic analysis revealed that gorilla AdVs belonging to HAdV-C were genetically identical to strains detected in humans who had been living around gorillas, and, inversely, a HAdV-C member HAdV type was detected in gorillas. This confirms the gorilla-to-human transmission of adenovirus. which has been reported previously. In addition, HAdV-E members, the most often detected here, are widely distributed among NHP species regardless of their origin, i.e., HAdV-E members seem to lack host specificity. Virus isolation was successful from a human sample and the strain of the Mbo024 genome, of 35 kb, that was identified as belonging to HAdV-D, exhibited close identity to HAdV-D members for all genes. This study provides information on the AdVs that infect African NHPs and the human populations living nearby, with an evident zoonotic transmission. It is likely that AdVs crossed the species barrier between different NHP species (especially HAdV-E members), between NHPs and humans (especially HAdV-C), but also between humans, NHPs and other animal species.
Subject(s)
Adenoviridae Infections/epidemiology , Adenoviridae Infections/veterinary , Mastadenovirus/classification , Mastadenovirus/isolation & purification , Adenoviridae Infections/transmission , Algeria/epidemiology , Animals , Chlorocebus aethiops/virology , Congo/epidemiology , DNA, Viral/genetics , DNA-Directed DNA Polymerase/genetics , Djibouti/epidemiology , Feces/virology , Gorilla gorilla/virology , Humans , Macaca/virology , Mastadenovirus/genetics , Pan troglodytes/virology , Papio hamadryas/virology , Papio papio/virology , Senegal/epidemiology , Viral Zoonoses/epidemiology , Viral Zoonoses/transmissionABSTRACT
CD8 T cells contribute to effective clearance of mouse adenovirus type 1 (MAV-1) and to virus-induced pulmonary inflammation. We characterized effects of a CD8 T cell effector, TNF, on MAV-1 pathogenesis. TNF inhibited MAV-1 replication in vitro. TNF deficiency or immunoneutralization had no effect on lung viral loads or viral gene expression in mice infected intranasally with MAV-1. Absence of TNF delayed virus-induced weight loss and reduced histological evidence of pulmonary inflammation, although concentrations of proinflammatory cytokines and chemokines in bronchoalveolar lavage fluid (BALF) were not significantly affected. BALF concentrations of IL-10 were greater in TNF-deficient mice compared to controls. Our data indicate that TNF is not essential for control of viral replication in vivo, but virus-induced TNF contributes to some aspects of immunopathology and disease. Redundant CD8 T cell effectors and other aspects of immune function are sufficient for antiviral and pro-inflammatory responses to acute MAV-1 respiratory infection.
Subject(s)
Adenoviridae Infections/virology , Mastadenovirus/physiology , Pneumonia/immunology , Tumor Necrosis Factor-alpha/immunology , Virus Replication , Adenoviridae Infections/genetics , Adenoviridae Infections/immunology , Animals , CD8-Positive T-Lymphocytes/immunology , Humans , Interleukin-10/genetics , Interleukin-10/immunology , Lung/immunology , Lung/virology , Male , Mastadenovirus/genetics , Mice , Mice, Inbred C57BL , Pneumonia/genetics , Pneumonia/virology , Tumor Necrosis Factor-alpha/geneticsABSTRACT
Bovine adenovirus type 3 (BAdV-3) is an important pathogen causing bovine respiratory disease complex (BRDC). From Jun 2016 to Jun 2018, 108 nose swab samples were collected from cattle with BRDC from 11 farms in five cities, and 78.7% (85/108) samples were detected as BAdV-3 positive by Real-time PCR. Interestingly, the sequences of 7/10 fiber (852 bp) and hexon (785 bp) fragments cloned from 10 positive samples from eight farms were clustered into a single branch of the evolutionary tree. A BAdV-3 strain (BO/YB24/17/CH) was successfully isolated. The isolate caused pathological changes of lung, trachea and spleen in BALB/c mice. Notably, 79 amino acid deletions in the shaft domain and 74 unique amino acid mutations were found in the fiber gene of the isolate compared with the available complete sequences for fiber genes in the GenBank database. These characteristics indicated that the isolate may represent a novel fiber genotype of BAdV-3. A pair of specific primers covering the deletion region in the fiber gene was designed to screen the prevalence of BAdV-3 encoding the novel fiber gene. The results showed that 7 of the 10 strains possessed the novel fiber gene, and these novel fiber strains were detected from six farms in which calves were just imported from five provinces, indicating that this BAdV-3 with the natural deletion fiber gene has a wide geographical distribution in China. In conclusion, our results reveal that BAdV-3 is widespread in China and a pathogenic BAdV-3 strain with a novel fiber gene has been detected at high frequency, which is beneficial to understand the prevalence and genetic evolution of BAdV-3.
Subject(s)
Adenoviridae Infections/virology , Cattle Diseases/virology , Mastadenovirus/genetics , Viral Proteins/genetics , Adenoviridae Infections/epidemiology , Adenoviridae Infections/veterinary , Animals , Cattle , Cattle Diseases/epidemiology , China/epidemiology , Gene Deletion , Mastadenovirus/isolation & purification , Mastadenovirus/pathogenicity , Mice, Inbred BALB C , Molecular Epidemiology , Phylogeny , Prevalence , Real-Time Polymerase Chain ReactionABSTRACT
Bovine adenovirus-3 (BAdV-3) is a non enveloped, icosahedral DNA virus containing a genome of 34446 bps. The intermediate region of BAdV-3 encodes pIX and IVa2 proteins. Here, we report the characterization of BAdV-3 IVa2. Anti-IVa2 serum detected a 50 kDa protein at 24-48 h post infection in BAdV-3 infected cells. The IVa2 localizes to nucleus and nucleolus of BAdV-3 infected cells. Analysis of mutant IVa2 demonstrated that amino acids 1-25 and 373-448 are required for nuclear and nucleolar localization of IVa2, respectively. The nuclear import of IVa2 utilize importin α -1 of importin nuclear import pathway. Although deletion/substitution of amino acids 4-18 is sufficient to abrogate the nuclear localization of IVa2, amino acids 1-25 are required for nuclear localization of a cytoplasmic protein. Furthermore, we demonstrate that amino acids 1-25 and 120-140 of IVa2 interact with importin α-1 and pV proteins, respectively in BAdV-3 infected cells.