Canine Adenovirus Research Articles

A canine adenovirus type 2 vaccine vector confers protection against foot-and-mouth disease in guinea pigs

Vaccination is a key element in the control of foot-and-mouth disease (FMD). The majority of the antigenic sites that induce protective immune responses are localized on the FMD virus (FMDV) capsid that is formed by four virus-encoded structural proteins, VP1 to VP4. In the present study, recombinant canine adenovirus type 2 (CAV2)-based FMD vaccines, Cav-P1/3C R° and Cav-VP1 R°, respectively expressing the structural P1 precursor protein along with the non-structural 3C protein or expressing the structural VP1 protein of the FMDV strain O/FRA/1/2001, were evaluated as novel vaccines against FMD. A strong humoral immune response was elicited in guinea pigs (GP) following immunization with Cav-P1/3C R°, while administration of Cav-VP1 R° did not induce a satisfying antibody response in GP or mice. GP were then used as an experimental model for the determination of the protection afforded by the Cav-P1/3C R° vaccine against challenge with the FMDV strain O1 Manisa/Turkey/1969. The Cav-P1/3C R° vaccine protected GP from generalized FMD to a similar extent as a high potency double-oil emulsion O1 Manisa vaccine. The results of the present study show that CAV2-based vector vaccines can express immunogenic FMDV antigens and offer protection against generalized FMD in GP. This suggest that Cav-P1/3C R° FMDV vaccine may protect natural host species from FMD. In combination with an appropriate diagnostic test, the Cav-P1/3C R° FMDV vaccine may also serve as a marker vaccine to differentiate vaccinated from infected animals.

IMMUNOLOGICAL AND CLINICAL STUDIES ON VACCINATED DOGS WITH THE LIVE ATTENUATED CANINE ADENO-1 VIRUS VACCINE

The immunogenicity and clinical safety of a local live attenuated canine adenovirus-1 vaccine (CAV-1) were evaluated in experimentally vaccinated native breed 3-4 months puppies. All vaccinated puppies did not show any abnormalities post-vaccination. Studying the blood picture revealed no changes in RBCs and haemoglobin values while there was an increase in the levels of total WBCs count, lymphocyte %, total serum protein and serum globulin. Liver and kidney function tests results were found to be within the normal values. These findings ensure vaccine safety. On the other side, the serum neutralization test was carried out on sera of vaccinated puppies and showed that these animals exhibited good levels of specific CAV-1 serum neutralizing antibodies able to protect them against virus infection. So, it could be said that the local CAV-1 vaccine is immunogenic vaccine suitable for dog protection against infectious canine hepatitis.

A multiplex TaqMan real-time PCR for detection and differentiation of four antigenic types of canine parvovirus in China

Canine parvovirus (CPV) is an important pathogen in domestic dogs, and the original antigenic types CPV-2 and its variants, CPV-2a, 2b and 2c, are prevalent worldwide. A multiplex TaqMan real-time PCR method was developed for the detection and differentiation of four antigenic types of CPV. A set of primers and probes, CPV-305F/CPV-305R and CPV-2-305P (for CPV-2)/CPV-2a-305P (for CPV-2a, 2b and 2c), was able to differentiate CPV-2 and its variants (CPV-2a, 2b and 2c). Another set of primers and probes, CPV-426F/CPV-426R and CPV-2-426P (for CPV-2 and 2a)/CPV-2b-426P (for CPV-2b)/CPV-2c-426P (for CPV-2c), was able to differentiate CPV-2a (2), CPV-2b, and CPV-2c. With these primers and probes, the multiplex TaqMan real-time PCR assay detected effectively and differentiated CPV-2, 2a, 2b and 2c by two separate real-time PCRs. No cross reactivity was observed with canine distemper virus, canine adenovirus, and canine coronavirus. The detection limit of the assay is 101 genome copies/μL for CPV-2, CPV-2a, CPV-2b, and 102 copies/μL for CPV-2c. The multiplex real-time PCR has 100% agreement with DNA sequencing. We provide a sensitive assay that simultaneously detects and differentiate four antigenic types of CPV and the method was also used for quantification of CPVs viral genome.

Detection and genetic characterization of Mamastrovirus 5 from Brazilian dogs.

Mamastrovirus 5 (MAstV5), belonging to the Astroviridae (AstV) family, previously known as canine astrovirus or astrovirus-like particles, has been reported in several countries to be associated with viral enteric disease in dogs since the 1980s. Astroviruses have been detected in fecal samples from a wide variety of mammals and birds that are associated with gastroenteritis and extra enteric manifestations. In the present study, RT-PCR was used to investigate the presence of MAstV5 in 269 dog fecal samples. MAstV5 was detected in 26% (71/269) of the samples. Interestingly, all MAstV5-positive samples derived from dogs displaying clinical signs suggestive of gastroenteritis, other enteric viruses were simultaneously detected (canine parvovirus, canine distemper virus, canine coronavirus, canine adenovirus and canine rotavirus). Based on genomic sequence analysis of MAstV5 a novel classification of the species into four genotypes, MAstV5a-MAstV5d, is proposed. Phylogenetic analyses based on the ORF2 amino acid sequences, samples described herein grouped into the putative genotype ‘a’ closed related with Chinese samples. Other studies are required to attempt the clinical and antigenic implications of these astrovirus genotypes in dogs.

Genetic dissection of the neuro-glio-vascular machinery in the adult brain

The adult brain actively controls its metabolic homeostasis via the circulatory system at the blood brain barrier interface. The mechanisms underlying the functional coupling from neuron to vessel remain poorly understood. Here, we established a novel method to genetically isolate the individual components of this coupling machinery using a combination of viral vectors. We first discovered a surprising non-uniformity of the glio-vascular structure in different brain regions. We carried out a viral injection screen and found that intravenous Canine Adenovirus 2 (CAV2) preferentially targeted perivascular astrocytes throughout the adult brain, with sparing of the hippocampal hilus from infection. Using this new intravenous method to target astrocytes, we selectively ablated these cells and observed severe defects in hippocampus-dependent contextual memory and the metabolically regulated process of hippocampal neurogenesis. Combined with AAV9 targeting of neurons and endothelial cells, all components of the neuro-glio-vascular machinery can be simultaneously labeled for genetic manipulation. Together, we demonstrate a novel method, which we term CATNAP (CAV/AAV Targeting of Neurons and Astrocytes Perivascularly), to target and manipulate the neuro-glio-vascular machinery in the adult brain.

A screening for canine distemper virus, canine adenovirus and carnivore protoparvoviruses in Arctic foxes (Vulpes lagopus) and red foxes (Vulpes vulpes) from Arctic and sub-Arctic regions of Norway

ABSTRACTCanine distemper virus (CDV), canine adenovirus (CAdV) and canine parvovirus type 2 (CPV-2) cause disease in dogs (Canis familiaris). These, or closely related viruses, may also infect wild...

Oncolytic Viruses: A Gene Therapy for Treatment of Cancer in Companion Animals

Cancer is the leading cause of disease-related death in companion animals such as dogs and cats. Oncolytic viruses refer to those viruses that will selectively eliminate and lysis cancerous tissues without causing harm to normal tissues. Oncolytic viruses can kill infected cancer cells in many different ways, ranging from direct virusmediated cytotoxicity through a variety of cytotoxic immune effector mechanisms. Treatment of pet cancer with advanced disease stages is very poor prognosis. Therefore, developing unique cancer therapies is essential for work synergistically in combination with the conventional treatment options. Several oncolytic viruses including canine distemper virus, adenovirus strains, and vaccinia virus strains have been used for canine cancer therapy in preclinical studies. This review is focused on the probable use of oncolytic viral agents for canine cancer therapy in the future.

Incidence and Sero-survey of Canine Adenovirus Type 2 in Various Animal Species

Incidence and Sero-survey of Canine Adenovirus Type 2 in Various Animal Species

New vaccination protocols for the dog and cat: the challenge of today for a better and safer prevention

Vaccinations are an integral part of a comprehensive preventive health care program targeting to minimize the incidence of major canine and feline infectious diseases. Currently, vaccination practices are re-evaluated globally towards a twofold objective: to strengthen "herd immunity", which depends on the percentage of vaccinated animals in a population, and to reduce the "vaccine load" per animal in order to minimize the vaccine-associated adverse reactions. To this end, the updated canine and feline vaccination guidelines, encourage the vaccination of as many animals as possible, while at the same time classify the vaccines into core, non-core and not recommended. Core vaccines should be administered, if possible, to every dog and cat.Canine parvovirus-2, canine adenovirus-2, canine distemper virus, feline parvovirus, feline calicivirus/herpesvirus-1 and rabies vaccines fall into this category. Non-core vaccines are selectively given to dogs and cats after assessing the risk/benefit ratio. There are also vaccines for which there is currently no sufficient scientific evidence to justify their use. Importantly, after the one-year booster inoculation that follows the completion of the initial puppy/kitten vaccination series, core vaccines should be given no more frequently than every three years, as the duration of the protective immunity far exceeds this time interval. This review focuses on the updated canine and feline vaccination guidelines pertaining to the individual animal as well as to those living in groups. Important questions related to vaccination programs and to relevant adverse reactions are also answered. An effort has been made to align these guidelines according to what is considered a "norm" among the small amimal practitioners in Greece.

Recombinant canine adenovirus type 2 expressing rabbit hemorrhagic disease virus VP60 protein provided protection against RHD in rabbits

Rabbit hemorrhagic disease virus (RHDV) is responsible for rabbit hemorrhagic disease (RHD), which is an acute, lethal and highly contagious disease in both wild and domestic rabbits. Although current vaccines are highly effective for controlling RHD, they are derived from infected rabbit livers and their use is thus associated with safety and animal-welfare concerns. In this study, we generated a recombinant lentogenic canine adenovirus type 2 (CAV2) vector expressing the RHDV vp60 gene, named rCAV2-VP60. rCAV2-VP60 expressed VP60 protein in Madin–Darby canine kidney cells as demonstrated by western blot and immunofluorescence assay. Polymerase chain reaction confirmed that the vp60 gene was successfully inserted into rCAV2-VP60 and was still detectable after 20 passages, indicating its stable genetic character. We evaluated the feasibility of rCAV2-VP60 as a live-virus-vectored RHD vaccine in rabbits. rCAV2-VP60 significantly induced specific antibodies to RHDV and provided effective protection against RHDV lethal challenge. These results suggest that rCAV2 expressing RHDV VP60 could be a safe and efficient candidate vaccine against RHDV in rabbits.

Long-lived immunity to canine core vaccine antigens in UK dogs as assessed by an in-practice test kit.

To determine the utility of an in-practice test kit to detect protective serum antibody against canine distemper virus, canine adenovirus and canine parvovirus type 2 in a sample of the UK dog population. Serum samples from 486 dogs, last vaccinated between less than 1 month and 124 months previously, were tested with the VacciCheck™ test kit for protective antibodies against distemper, adenovirus and parvovirus type 2. A high proportion of the dogs tested (93·6%) had protective antibody against all three of the core vaccine antigens: 95·7% of the dogs were seropositive against canine distemper virus, 97·3% against canine adenovirus and 98·5% against canine parvovirus type 2. The small number of dogs that were seronegative for one or more of the antigens (n = 31) may have had waning of previous serum antibody or may have been rare genetic non-responders to that specific antigen. UK veterinarians can be reassured that triennial revaccination of adult dogs with core vaccines provides long-lived protective immunity. In-practice serological test kits are a valuable tool for informing decision-making about canine core revaccination.

European surveillance of emerging pathogens associated with canine infectious respiratory disease

Canine infectious respiratory disease (CIRD) is a major cause of morbidity in dogs worldwide, and is associated with a number of new and emerging pathogens. In a large multi-centre European study the prevalences of four key emerging CIRD pathogens; canine respiratory coronavirus (CRCoV), canine pneumovirus (CnPnV), influenza A, and Mycoplasma cynos (M. cynos); were estimated, and risk factors for exposure, infection and clinical disease were investigated.CIRD affected 66% (381/572) of the dogs studied, including both pet and kennelled dogs. Disease occurrence and severity were significantly reduced in dogs vaccinated against classic CIRD agents, canine distemper virus (CDV), canine adenovirus 2 (CAV-2) and canine parainfluenza virus (CPIV), but substantial proportions (65.7%; 201/306) of vaccinated dogs remained affected.CRCoV and CnPnV were highly prevalent across the different dog populations, with overall seropositivity and detection rates of 47% and 7.7% for CRCoV, and 41.7% and 23.4% for CnPnV, respectively, and their presence was associated with increased occurrence and severity of clinical disease. Antibodies to CRCoV had a protective effect against CRCoV infection and more severe clinical signs of CIRD but antibodies to CnPnV did not.Involvement of M. cynos and influenza A in CIRD was less apparent. Despite 45% of dogs being seropositive for M. cynos, only 0.9% were PCR positive for M. cynos. Only 2.7% of dogs were seropositive for Influenza A, and none were positive by PCR.

Evaluation of protective efficacy of three novel H3N2 canine influenza vaccines.

Canine influenza virus (CIV) has the potential risk to spread in different areas and dog types. Thus, there is a growing need to develop an effective vaccine to control CIV disease. Here, we developed three vaccine candidates: 1) a recombinant pVAX1 vector expressing H3N2 CIV hemagglutinin (pVAX1-HA); 2) a live attenuated canine adenovirus type 2 expressing H3N2 CIV hemagglutinin (rCAV2-HA); and 3) an inactivated H3N2 CIV (A/canine/Guangdong/01/2006 (H3N2)). Mice received an initial intramuscular immunization that followed two booster injections at 2 and 4 weeks post-vaccination (wpv). The splenic lymphocytes were collected to assess the immune responses at 6 wpv. The protective efficacy was evaluated by challenging H3N2 CIV after vaccination (at 6 wpv). Our results demonstrated that all three vaccine candidates elicited cytokine and antibody responses in mice. The rCAV2-HA vaccine and the inactivated vaccine generated efficient protective efficacy in mice, whereas limited protection was provided by the pVAX1-HA DNA vaccine. Therefore, both the rCAV2-HA live recombinant virus and the inactivated CIV could be used as potential novel vaccines against H3N2CIV. This study provides guidance for choosing the most appropriate vaccine for the prevention and control of CIV disease.

Novel adenoviruses detected in British mustelids, including a unique Aviadenovirus in the tissues of pine martens (Martes martes).

Several adenoviruses are known to cause severe disease in veterinary species. Recent evidence suggests that canine adenovirus type 1 (CAV-1) persists in the tissues of healthy red foxes (Vulpes vulpes), which may be a source of infection for susceptible species. It was hypothesized that mustelids native to the UK, including pine martens (Martes martes) and Eurasian otters (Lutra lutra), may also be persistently infected with adenoviruses. Based on high-throughput sequencing and additional Sanger sequencing, a novel Aviadenovirus, tentatively named marten adenovirus type 1 (MAdV-1), was detected in pine marten tissues. The detection of an Aviadenovirus in mammalian tissue has not been reported previously. Two mastadenoviruses, tentatively designated marten adenovirus type 2 (MAdV-2) and lutrine adenovirus type 1 (LAdV-1), were also detected in tissues of pine martens and Eurasian otters, respectively. Apparently healthy free-ranging animals may be infected with uncharacterized adenoviruses with possible implications for translocation of wildlife.

Evaluation of a multicomponent vaccine in dogs

The concept of combined viral and bacterial vaccines has been tried in cattle, canine and humans. Use of multicomponent vaccines has numerous advantages over individual vaccine. In this study, apparently healthy 8-9 weeks old 41 puppies were divided into 2 groups. 31 puppies (test group) were vaccinated subcutaneously with a commercially available multicomponent vaccine containing live attenuated strains of Canine Distemper virus, Canine Adenovirus type 2, Canine Parvo virus and Canine Parainfluenza virus and inactivated antigens of Leptospira canicola and Leptospira icterohemorrhagiae. 10 puppies were kept unvaccinated (control). Test group puppies were given a booster after 4 weeks of primary vaccination. Serum samples from both the groups were collected on 4 occasions (0th, 28th, 42nd and 90th day) and were tested for antibody titres against Canine Distemper virus and Canine Adenovirus type 2 by Serum Neutralization test, against Canine Parvovirus and Canine Parainfluenzavirus by Haemagglutination Inhibition test and against Leptospira canicola and L. icterohemorrhagiae by Microscopic Agglutination Test. A significant rise (P 0.5) in antibody titres was observed against all the antigens after vaccination. The control group did not show significant variation in the antibody titres. All puppies vaccinated subcutaneously with this vaccine did not evoke any local/ adverse reactions.

SURVEILLANCE FOR ANTIBODIES AGAINST SIX CANINE VIRUSES IN WILD RACCOONS (PROCYON LOTOR) IN JAPAN.

Raccoons (Procyon lotor) are found worldwide. They are frequently seen in crowded inner cities as well as in forests or wooded areas, often living in proximity to humans and their pets. We examined sera from 100 wild raccoons in Japan for antibodies to six canine viruses with veterinary significance to assess their potential as reservoirs. We also aimed to understand the distribution of potentially infected wildlife. We found that 7% of samples were seropositive for canine distemper virus (CDV), 10% for canine parvovirus type 2, 2% for canine adenovirus type 1, 6% for canine adenovirus type 2, and 7% for canine coronavirus. No samples were found to be seropositive for canine parainfluenza virus. Seropositivity rates for canine distemper virus and canine parvovirus type 2 were significantly different between areas, and younger raccoons (<1 yr old) were more frequently seropositive than older raccoons. Because raccoons belong to the suborder Caniformia, similar to dogs (Canis lupus familiaris), our results suggest that they can act as reservoirs for some of these important canine viruses and might be involved in viral transmission. Further study should include isolation and analysis of canine viruses in wild raccoons from a wider area.

Colite necrohemorrágica causada por Entamoeba histolytica em um cão

Background: Intestinal amebiasis with morphological lesions and clinical manifestations is uncommon in dogs. The disease is caused by the protozoan Entamoeba histolytica, which is commonly observed in its natural hosts, humans and some non-human primates. It is occasionally found in the company of animals, usually associated with contact with infected humans. Thus, the objective here is to describe a case of necro-hemorrhagic colitis caused by E. histolytica in a dog infected with the canine distemper virus, in order to characterize the epidemiological and clinicopathological aspects of the disease.Case: An adult, mixed-breed bitch displaying anorexia and ataxia was referred to the veterinary hospital for treatment. Clinical evaluation showed a cachectic animal with 12% dehydration, ocular discharge, and bilateral purulent nasal dis-charge. A clinical diagnosis of distemper was made, and treatment was instituted. The dog’s signs progressed to walking in circles, aimlessly, with lethargy and blindness. After three days of the onset of neurological signs, the dog developed diarrhea with hematochezia. With no improvement noted, we elected to euthanize the dog. At necropsy, edema was present in subcutaneous tissues, and the lungs had yellow areas in the cranio-ventral portions, which the court was flowing purulent discharge. In the large intestine, segmental distention of the distal portion of the descending colon was observed. The segment was approximately 15 cm in length and consisted of irregular reddish areas. There was also slight thickening of the wall with edematous mucosa containing blood clots, fibrin, and multiple areas of ulceration. Microscopically showed necro-hemorrhagic colitis associated with rounded structures, approximately 15 μm in size, containing abundant eosinophilic cytoplasm that was slightly granular or vacuolated. They also contained nuclei and nucleoli that were central or slightly eccentric. These organisms were consistent with amoeba trophozoites. There was also demyelinating encephalitis associated with malacia, corpuscular intranuclear eosinophilic inclusions and / or intracytoplasmic inclusions in ependymal cells, astrocytes, and gemistocytes, characteristic of infection with canine distemper virus. Using immunohistochemistry with polyclonal anti-E. histolytica antibodies in the dilution of 1:1000, trophozoites were immunomarked, confirming the suspected amebiasis.Discussion: The diagnosis of intestinal amebiasis was based on clinical signs and by morphological characteristics on gross and microscopic examination, and was confirmed as E. histolytica by immunohistochemistry. Limited information on theepidemiology and pathological findings of infection with Entamoeba sp. has been reported in the literature, as it is relatively uncommon in pets. Affected animals are usually asymptomatic, but immunosuppression caused by canine distemper virus may have triggered the clinical manifestations of the disease in this dog. Enteritis due Entamoeba sp. should be considered in dogs with chronic weight loss and bloody diarrhea. It should also be included in the differential diagnoses for weight loss and hemorrhagic gastroenteritis, such as canine parvovirus, canine adenovirus 1, Pythium insidiosum, and Giardia sp.

Acute fatal haemorrhagic pneumonia caused by Streptococcus equi zooepidemicus in greyhounds in Ireland with subsequent typing of the isolates

Two cases of acute fatal haemorrhagic pneumonia caused by Streptococcus equi zooepidemicus have been confirmed in Ireland. Both cases involved young greyhounds (17 and 10 months, respectively) from racing kennels...

Detection of canine adenovirus 1 in red foxes ( Vulpes vulpes) and raccoons ( Procyon lotor) in Germany with a TaqMan real-time PCR assay.

We developed a real-time (rt)PCR assay based on TaqMan probe technology for the specific detection of canine adenovirus 1 (CAdV-1). The assay is able to detect three 50% tissue culture infectious dose/mL in CAdV-1-containing cell culture supernatant. Viral genomes were not amplified of canine adenovirus 2 or of several bovine, porcine, and avian adenoviruses. In silico analysis provided no indication of amplification of other heterologous genomes. The sensitivity of the real-time assay exceeded that of a conventional gel-based CAdV-1 PCR by a factor of 100. Following the integration of the novel PCR into the Hessian wildlife-monitoring program, CAdV-1 DNA was detected in none of the tested raccoons ( n = 48) but in 11 of 97 foxes.

Novel sequence variants of viral hexon and fibre genes in two dogs with canine adenovirus type 1-associated disease

There is little information on sequence variation of canine adenovirus type 1 (CAdV-1), the aetiological agent of infectious canine hepatitis (ICH). This study reports hexon and fibre gene sequence variants of CAdV-1 in a dog with systemic ICH and a dog with the ocular form of the disease (‘blue eye’) in Northern Italy in 2013. One of the sequence variants matched a CAdV-1 fox sequence previously detected in Italy.