Reference values of the IBGN.
\r\n\t
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Dr. Yu is a holder of 90 journal papers, with an h index of 21, is a member of A& WA (USA) and AAAR (USA), and is the holder of 24 registered patents.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"188972",title:"Prof.",name:"Mingzhou",middleName:null,surname:"Yu",slug:"mingzhou-yu",fullName:"Mingzhou Yu",profilePictureURL:"https://mts.intechopen.com/storage/users/188972/images/system/188972.jpg",biography:"Mingzhou Yu is now a Professor at China Jiliang University and a Guest Professor at Key Laboratory of Aerosol Chemistry and Physics, Chinese Academy of Science. He received his PhD degree from Zhejiang University in 2008 with the major fluid mechanism. During the time period between 2009 and 2012, he moved to Karlsruhe Institute of Technology, Germany, as a Alexander von Humboldt researcher where he worked with Prof. Gerhard Kasper and Dr. Martin Seipenbusch. Since 2013, he joined Prof. Junji Cao's research group as a guest Professor at Key Laboratory of Aerosol Chemistry and Physics, Chinese Academy of Science. During the time period between 2013 and 2016, he worked in The Hongkong Polytechnic University and Universidad Autónoma de Madrid, Spain, as a research associate or postdoc researcher. He is now leading a Aerosol Science and Technology Laboratory supported by Zhejiang Special Provincial Support in CJLU. 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The wetland of Naâma classified as Ramsar, situated in the arid region of Alegria, offers an important fauna and flora diversity due to its geographical location it constitutes the main resting place in north Africa for migratory birds, animal and plant species, including many endemic species.
Bioindicators are species used to appraise the health conditions of the environment or ecosystem and they are capable of determining the environmental integrity using their functions and populations. Wetlands are fragile ecosystems that perform major functions, such as storage and the restitution of water as well as the natural filtering of mineral and organic matter, they also host a rich biodiversity and particularly adapted to this environment [1].
The insects are responsible for many processes in the ecosystem and its loss can have negative effects on entire ecosystem, Insects are used as bioindicators, because of their sensitivity to environmental conditions which, due to its ecological peculiarities, gives information on the characteristics of the environment in which it is present or on the evolution of this environment under the influence of certain practices are used to detect changes in the environment and the presence of pollution.
Insects are the most abundant animals in almost all ecosystems and can be used to evaluate the impact of environmental change.
Entomofauna studies to furnish information about ecosystems conservation status their productivity and levels of water contamination and pollution. Therefore, bioindicator species identification is essential, due to the important role that these organisms have as transformers and regulators of ecosystems [2].
The aim of this study is:
study the entomofauna structure in the wetland of Naâma (SW Algeria).
identify bioindicator species and assess the quality of aquatic environments in the wetland.
The resort is a wetland listed by Ramsar, localized at (longitude 0°west and latitude 33°north). The water of wetland concerned two hundred hectares surrounded by several units or peripheral areas; immediate area of water is characterized by
Satellite images and photos of the wetland of Naâma region.
We studied the insects in 5 sampling sites: 3 sites on the water bodies of the hawdh edaira wetland and 2 sites along the lake. The plots covered several types of environmental gradients (water body and wild natural areas (wooded, grassy, rocky and forest areas) Land cover of the sampling sites was classified into 5 types based on Google Earth images and field observation.
Insect field collections were carried out continuously for 36 months from September 2017 to September 2020 to obtain baseline data on insect composition. Inventories were carried out in the middle of each month. The transect method (300 m × 200 m) was used to study insects in open areas.
Extrapolating sample data from these methods to the square metre could lead to the overestimation of species abundance and diversity at the site due to the generally patchy distribution of invertebrates in streams [3].
Two qualitative sampling tools (D-frame net and square net) and a quantitative sampling tool (Surber sampler) were used to collect the aquatic insects the wetland of Naâma. A D-frame net with a 1.2-m-long handle and a 60 cm long cone-shaped net with 0.3 mm mesh and a diameter of 0.38 m was used in this study. A square frame net with a 0.5 m × 0.5 m opening, a 90 cm cone shaped net with 0.3 mm mesh, and a 1.2 m long handle were used to collect samples. A sampling tool’s efficiency is also determined by the time required to process the samples [4].
Other techniques have been used namely; − Butterfly net - Catching insects by hand - Pheromone insect trap.
The species inventory is essential for the structural analysis of a station.
The harvest of the plant in the field is the prospecting which aims to know the totality of the flora of the region of Naâma. In this case, it is essential to visit the sites during all seasons to collect the maximum number of plants of different species. In the course of surveys, plastic bags are used. The collected samples are then dried and placed in paper folders with a label, mentioning the date, place, and other interesting observations. All the species have been kept in a herbarium.
The determination of the plant species was carried out using the two guides; Nouvelle Flore d’Algérie of QUÉZEL and SANTA S and the Flore du Sahara of Paul Ozenda.
ni: number of individuals of a given species, i ranging from 1 to S (total number of species).
n: total number of individuals.
n: collection set; m: the average number of individuals in each sample; x: number of individuals from each sample. If: S2 = 0: the distribution is uniform or regular; S2 < m: the distribution is contagious .
F (i): Relative frequency of the species contained in the statement as a percentage.
ni: The number of times the insect (i) are present.
N: Total number of individuals.
Descriptive statistics are the first pieces of information used to understand and represent a dataset. Their goal, in essence, is to describe the main features of numerical and categorical information with simple summaries. These summaries can be presented with a single numeric measure, using summary tables, or via graphical representation. Here, I illustrate the most common forms of descriptive statistics for numerical data but keep in mind there are numerous ways to describe and illustrate key features of data.
The t test tells you how significant the differences between groups are; In other words it lets you know if those differences (measured in means) could have happened by chance.
Factorial correspondence analysis is a descriptive method. It aims at the representation with the minimum loss of information in a space with n dimension [2]. The purpose of this analysis is to realize several graphs from data table. The observation of the graph can give an idea of the interpretation of the factors and show which variables are responsible for the proximity between this or that observation.
The key to determining insects order Plecoptera, Ephemera and Odonata, is performed by the software Xper3, This key is based on a list of taxa and associated descriptors, such as morphological characters.
Factorial correspondence analysis (CFA) was studied by minitab version 19 software.
Student’s t test for single sample, Descriptive statistics of a numeric variable and The Shapiro–Wilks test have been studied by R++ statistics software.
Benthic insects are one of the most famous organism groups used for rivers biomonitoring surveys.
The use of IBGN is especially indicated for disturbances that induce a modification of the nature of the substrate and the organic quality of the water: rejection urban predominantly organic, pollution by suspended matter, side effects of certain types of rejection (organic, metallic) and eutrophication. In addition, the IBGN reflecting the structure of a biocenosis made up of organisms long-term integrators are especially sensitive to chronic disturbances or well to disturbances of the intermittent type but sufficiently intense to cause a immediate mortality.
Class 1A blue in color which indicates excellent water quality
Class 1B green in color which indicates good quality water (with pollution moderate)
Class 2 yellow in color which indicates average water quality (with a clear Pollution)
Class 3 orange color which indicates poor quality water (with pollution important)
Excluding class 4 red which indicates poor quality (with pollution excessive)
After identifying the macroinvertebrates, a list faunistic is established, listing all the taxa found by faunistic groups, and indicating the total number of taxa. The index is calculated from the table “IBGN values according to type and the taxonomic variety of macrofauna. We first determine the taxonomic variety (Σt), i.e. the total number of taxa identified (the number of individuals per taxon is not taken into account). Then look for the faunistic indicator group (GI) in the list provided and select the taxon that has the highest degree of pollutant sensitivity of the full sample of the station studied.
The index can then be read in the table of values of the IBGN: it is at the intersection of the column for the taxonomic variety and the row for the indicator faunistic group (Table 1).
IBGN | Class 1A > ou = à17 | Class 1B 16–13 | Class 2 12–19 | Class 3 8–5 | Class 4 < ou = à 4 |
---|---|---|---|---|---|
Reference values of the IBGN.
The insect species recorded are divided into 9 orders; Ephemeroptera, Coleoptera, Orthoptera, Hymenoptera, Lepidoptera, Hemiptera, Odonata, Diptera and Plecoptera. From this present work, 27 insect families were found. The most represented order is Coleoptera with six families and 11 species Orthoptera represent 5 families and 20 species. The Hemiptera, Plecoptera and Ephemeroptera represent only one family and one species for each order (Table 2).
Class | Order | Family | Species | Relative frequencies | Dispersion index |
---|---|---|---|---|---|
Insect | Coleoptera | Coccinellidae | |||
Tenebrionidae | |||||
Carabidae | |||||
Cantharidae | |||||
Geotrupidae | |||||
Meloidae | |||||
orthoptera | Acrididae | ||||
Pyrgomorphidae | |||||
Pamphagidae | |||||
Gryllidae | |||||
Tettigonidae | |||||
Hymenoptera | Vespidae | ||||
Pompilidae | |||||
Apidae | |||||
Formicidae | |||||
Lepidoptera | Nymphalidae | ||||
Pieridae | |||||
Sphingidae | |||||
Noctuidae | |||||
Diptera | Muscidae | ||||
Calliphoridae | |||||
Sarcophagidae | |||||
Odonata | Libellulidae | ||||
Coenagrionidae | |||||
Hemiptera | Pyrrhocoridae | ||||
Plecoptera | Leuctridae | ||||
Ephemeroptera | Baetidae |
Summary of insect species identified in the wetland of Naâma.
The Shannon-Weaver diversity index obtained in the study area is 2.24bit, maximum diversity is 1.89. Equitability is of the order of 0.56. These values indicate that the wetland is characterized by a very important entomofauna diversity, these results show a great diversity of insects in the naama wetland (Figure 2).
Distribution of insect orders by families and species in the study area.
The study of the Dispersion index and The relative frequency of each species identified in the hawdh ed. daira wetland of Naâma allowed to know the frequency and the type of distribution of each species;distribution uniform, regular or contagious (Table 2). This information is necessary for the monitoring and control of these species over time (Figure 3).
Dispersion index and The relative frequency of insect species order in the wetland of Naâma.
In this study the calculation of the Shapiro-Wilk normality test is equal:
this value means, the p-value is greater than 0.05, the data of this stady are normally distributed.
the calculation of the Descriptive statistics of a numeric variable of insect species in the Naâma wetland show that the population insects are normally distributed with a Standard deviation equal 3.4345 (Table 3).
Minimum | Quartile 1 | Median | Mean | Quartile 3 | Maximum | Standard deviation |
---|---|---|---|---|---|---|
1 | 1 | 1 | 3.065 | 3 | 12 | 3.4345 |
Descriptive statistics of a numeric variable of insect species in the Naâma wetland.
The calculation of the student test for the identified insect population in the Naama wetland shows that the test statistic would follow a normal distribution. The sample mean is equal to the population mean with a Standard deviation equal 3.4345 and Student’s t test equal 4.26 (Table 4).
t | Df | p-value | Confidence interval, 95% |
---|---|---|---|
4.268 | 51 | 2.55e-05 | [1.8047, 4.3243] |
Variable | 51 | 3.0645 | 3.4345 |
Student’s t test for single sample of insect species in the Naâma wetland.
The initial table (1) corresponding to 20 surveys show the presence of species in the stations according to the type of environment; plants environment, rocky environment and aquatic environment An AFC conducted on this matrix allowed to build a hierarchical classification calculated from the coordinates of species. Dendrogram clearly differentiates three groups of species of unequal size:
Factorial analysis of the correspondence of insect species in the wetland of Naâma.
From the Euclidean distances based on the scores of the three factors A.F.C (Figure 2), it is possible to recognize three groups:
The ascending hierarchical classification (C.H.A) confirm our results of Correspondence factor analysis (Figure 5).
Hierarchical ascending classification of insects species in the wetland of Naâma.
In the plants and rocky environment., very high faunal diversity of orthoptera (locust)species was recorded; this is supported by the fact that high environmental quality usually correlates with the greatest species richness and diversity.
The first entity in the right of the projection is the largest as it includes 75% of species. It represents the species caught in plants environment (
Factorial analysis of the correspondence of Orthoptera species of Naâma (Algeria).
The IBGN is organized in rows 9 faunal indicator groups and in columns 14 classes of taxonomic varieties. For this, we successively determine:
The taxonomic variety of the sample (Σt) which is equal to the total number of taxa collected even if they are represented by only 1 individual.
The faunistic indicator group (GI) by taking into account only the indicator taxa represented in the sample by 3 individuals or 10 individuals depending on the taxa. The determination of GI is carried out by prospecting the columns of the table from top to bottom and by selecting the taxon which represents the highest degree of pollutant sensitivity of the entire sample of the station studied. Then the IBGN value be read by crossing the taxonomic variety column and the indicator faunal group row. Depending on the taxonomic diversity of the OglatedDaira station and the presence or absence of indicator taxa, a variant hydrobiological quality score is assigned from 1 to 20.
We see that the study area has good hydrobiological quality with moderate pollution (IBGN = 14: the taxonomic variety ST = 51 and an indicator group (GI = 9) (Table 1).
The diversity analysis by Shannon index (H’), shows that the most important diversity is marked in the wetland of Ain Ben Khelil with (H’ = 2.43). the maximum diversity is equal 3.13.
Equitability is equal 0.77, which is usually taken as an indicator of a balanced population and which reflects the stability of the environment.
In the wetland of naama,
15 families are present in the wetland of Naama, the most representative families are Asteraceae with a relative abundance of 17.39%, followed by the Amaranthaceae and Poaceae with 13.04%.
The biological type leads to the natural form of the plant that is one of the basic criteria for classifying species in biological types, composed of perennial species, woody or herbaceous and annual species. The specific aspect of the form obtained is dependent on environmental variations.
In our study area, therophytes dominate other biological types with 40.62% followed by chamaephytes with31.25%, hemicryptophytes remain in third with 15.62%.
This study was carried out in the arid region of Naâma, this region characterized by steppe formations dominated by
The insect species recorded are divided into 9 orders; Ephemeroptera, Coleoptera, Orthoptera, Hymenoptera, Lepidoptera, Hemiptera, Odonata, Diptera and Plecoptera. From this present work, 27 insect families were found. The most represented order is Coleoptera with six families and 11 species Orthoptera represent 5 families and 20 species. The Hemiptera, Plecoptera and Ephemeroptera represent only one family and one species for each order. Invertebrates are more severely and quickly affected than other taxa by changes in the landscape. The insects are responsible for many processes in the ecosystem and its loss can have negative effects on entire communities. Thus, a strong understanding of insect responses to human activity is necessary both to support policy decisions for conservation and to evaluate functional consequences of human disturbance on ecosystems [6].
many diversity indices have been developed to describe responses of a community to environment variation, combining the three components of community structure, namely richness (number of species present), Shannon-Wiener Index [9].
The Shannon-Weaver diversity index obtained in the study area is 2.24bit, maximum diversity is 1.89. Equitability is of the order of 0.56. These values indicate that the wetland is characterized by a very important fauna diversity.
Orthoptera (locust) are able to threaten arid ecosystems, human health and resistant to pesticides, a study was carried by (Brahimi et al., 2020) on the chemical mechanism of locust resistance in the arid region of Naama. Hardersen [10] reported the potential of aquatic insects as indicators of water quality. Several other species of the families Gyrinidae, Dytiscidae, Hydrophilidae (Coleoptera), Notonectidae, Veliidae (Heteroptera) and Plecoptera and Ephemeroptera Orders have high adaptive capacity, colonizing most of the environments and occurring throughout the year, reflecting ecological and geographical changes, and hence their conservation status.
Davis [11] confirm beetles species (Coleoptera: Scarabaeidae) have a high potential as environmental indicators in forest area .
In this study an important diversity of beetles (coleoptera) was recorded in this area, beetles play an essential role as decomposers of organic matter in the balance of ecosystems. Beetles from Order Coleoptera and Family Carabidae are important predators. They participate of biological control, biological monitoring of pollution from oil, sulfur, herbicides, CO2, insecticides and radioactive phosphorus. The moths and butterflies (Lepidoptera), besides having basic requirements, have ecological faithfulness in temperate and tropical regions and are very sensitive to changes in the environment [12]. the trapping methods used have a great importance to better collect the species in quantity and quality. This explains the differentiation from other orders in number of species.
According to Rizo-Patrón et al. [13], group of macroinvertebrates (
In this study we noticed a scarcity of pollinating species, this scarcity is due to the uncontrolled and anarchic use of phytosanitary products and insecticides in the action of the Locust control. Pollinators, especially honeybees (
According to Eggleton et al. [16], termites are important decomposers in land ecosystems. Its activity increases soil infiltration capacity, leading to water retention and soil productivity. Read and Anderson [17] showed The value of ants as bio-warning indicators in the Australian arid rangelands.
According to Nummelin et al. [18], A study of the heavy metal concentrations of different predatory insects (Gerridae), dragon fly larvae (Odonata), anteater larvae (Myrmeleontidae) and ants (Formicidae) showed higher metal concentrations and that these groups of insects can be used as indicators of heavy metals.
Goncharov et al. [19] indicates that using the density index of Ephemeroptera Plecoptera-Trichoptera (EPT); showed a capacity for regeneration of these indicator groups under unfavorable conditions. Cortelezzia [20] indicates that In some taxa of chironomids, their potential as a bioindicator increased as the taxonomic level decreased (eg, Chironominae). However, in other taxa, this potential as a bio-indicator of water quality remains at the subfamily level. The Ephemeroptera and Plecoptera larvae are recognized as good bioindicators of eutrophication in running water due to their sensitivity to oxygen depletion. The pollution indicators can be attributed by the disappearance of certain more or less sensitive species or, on the contrary, by the appearance of other so-called resistant species. The specific river environment sampled may influence the proliferation of certain taxa and the specific behaviours of those taxa in certain habitats [21].
The insect species recorded are divided into 9 orders; Ephemeroptera, Coleoptera, Orthoptera, Hymenoptera, Lepidoptera, Hemiptera, Odonata, Diptera and Plecoptera. From this present work, 27 insect families were found. The most represented order is Coleoptera with six families and 11 species Orthoptera represent 5 families and 20 species. The Hemiptera, Plecoptera and Ephemeroptera represent only one family and one species for each order. by this study we noticed a scarcity of pollinating species, this scarcity is due to the uncontrolled and anarchic use of phytosanitary products and insecticides in the action of the Locust control.
The Shannon-Weaver diversity index obtained in the study area is 2.24bit, maximum diversity is 1.89. Equitability is of the order of 0.56. These values indicate that the wetland is characterized by a very important fauna diversity. Statistical study of the entomofauna identified in the Naama wetland show that the population insects are normally distributed in this environment.
The study of the hydrobiological quality in the wetland of Naama, assessed by the IBGN method, showed good hydrobiological quality with moderate pollution (IBGN = 14). This pollution is precisely marked by the requirement of Ephemeroptera and Plecoptera. Ephemeroptera are polluo-resistant species in polluted aquatic environments, unlike stoneflies which are polluo-sensitive species. These two orders are used as good biological indicators of polluted aquatic environments.
This study concluded that the Class Insecta has many potential representatives that can be used as environmental bioindicators, among which are some species from the Coleoptera, Diptera, Lepidoptera, Hymenoptera, Hemiptera, Isoptera Orders and others.
These data constitute a first database on the structure of the entomofauna in the wetland classified by Ramsar Hawdh ed. daira of arid region ofAlgeria, the results obtained also make it possible to control and monitor pollution in this wetland, in order to protect these fragile and arid ecosystems. Threatened by desertification and human actions .
This work was supported by the General Directorate of Scientific Research and Technological Development DGRSDT of Algeria republic.
The authors declare that they have no conflict of interest.
Canine parvovirus (CPV-2) is a member of the
CPV-2 causes 100 percent morbidity and mortality rate of 10 percent and 91 percent in adult and young dogs respectively [9]. However, a mortality of 91 percent was reported in experimentally infected dogs that were not treated [10]. CPV-2 affects predominately the younger dogs between 6 weeks and 6 months [8] with an increased susceptibility to puppies less than 6 months. In dogs over the age of 6 months, sexually intact males are more likely (twice) to develop canine parvovirus enteritis (CPVE) in comparison to intact females [11]. The CPV-2 antibody titer transmitted to the newborn via absorbed colostral antibody is 50–60% of the mother’s titer. The half-life of paroviral maternal antibodies is around 10 days [12]. Therefore, puppies are highly susceptible to the CPV-2 infection as the maternal antibody titres start declining. CPVE affects dogs of all ages, although it is more severe in puppies. Puppies can succumb to shock and die within two days after being sick. The most striking symptom of CPV-2 myocarditis is the abrupt mortality in young puppies, generally around the age of 4 weeks [13].
In recent years, CPVE outbreaks caused by multiple CPV-2 variants have been recorded in diverse geographical locations throughout the world. Previously, CPV-2, which could not infect cats, has been replaced by CPV-2 variants that can now infect cats, suggesting that CPV-2 may be capable of spreading between species [14]. Since CPV-2 infects a wide range of wild animals in the order Carnivora, subclinical infection appears to be prevalent. As a result, significant CPV-2 reservoirs in wildlife appear to exist, and transmission of virus between domestic dogs and wildlife appears to be common and bidirectional [15]. Despite the availability of a wide range of immunoprophylactic and antiviral agents to control CPV-2 infections in dogs, many outbreaks have been reported throughout the world, and the disease has remained a major veterinary and economic concern due to the presence of unvaccinated dogs, intervention of active immunization by maternally derived antibodies, and the emergence of a different antigenic variants of CPV-2.
Canine parvovirus infection is caused by
Schematic representation of
The virus is nonenveloped having icosahedral symmetry and is 25 nm in diameter. The CPV virus is made up of the sixty protein subunits containing VP1 (5–6 units) and VP2 (54–55 units). The protein structure is made up of antiparallel β-barrel (8-stranded) capsid. The viral replication occurs inside the nucleus of multiplying cells and therefore the intranuclear inclusion bodies are formed during the infection. The viral capsid structure is made up of spike at the three-fold axes of the icosahedral unit, a 15-Å depression around the five-fold axes and two-fold axes is formed. Antigenic determinant regions have been plotted to the three-fold protrusion and the two-fold depression are related to the host cell features [17]. The surface of the capsid is composed of four loops inserted between the strands, resulting in spike-like protrusions around threefold axes of approximately 22 Å. The antigen neutralization site, also known as epitope A, is composed of loops 1 and 2 of one VP2 and loop 4 of a threefold related molecule [21]. The molecular weight (MW) is around 5.5 to 6.2 × 106 Da. There is an equal ratio of protein to nucleic acid.
NS1 is the largest non structural protein in CPV-2, and it is primarily involved in viral replication and pathogenicity [22]. NS1 is a key mediator of cytotoxicity of CPV and can selectively cause tumor cell lysis by inducing an antitumor immune response in different tumor models [23]. A recent study demonstrated the amino acid residues of T598 and T601 in the C-terminal phosphorylation sites of NS1 protein, involved in replication and pathogenicity of CPV-2 [24].
In the 1970s, CPV-2 emerged as a novel pathogen in dogs. Since then, CPVE has been reported across all the continents [25, 26]. Other related viruses such as Feline panleukopenia virus (FPV), Mink enteritis virus (MEV), Raccoon parvovirus (RPV) are closely related to the CPV-2 [27]. Mutations in the canine transferrin receptor (TfR) type-1 lead to adaptation of CPV-2 in different species 2 [28, 29]. There is more than 98% genome homology reported in the CPV and FPV nonetheless infect different species and have typical antigenic capsid and haemagglutination (HA) properties [28, 30]. The mutations in different amino acid positions have led to the effective adaptation in the new hosts [30]. There are over five to six mutations in the VP2 residue of the CPV-2 and FPV and also 375 and 323 amino acid position regulates the pH functionality of HA [31, 32]. CPV-2a (Asn CPV-2a) replaced CPV 2 in 1980s in the USA and various European countries. CPV 2a can infect the cats which was not a feature of CPV 2. CPV 2a has been displaced by the CPV-2b (426Asp) which was first reported in USA in 1982 and CPV-2c (426Glu) variant in Italy [31, 33]. Although two variants, CPV-2a and 2b had been identified much earlier, however, the third variant CPV-2c had been recognized in early 2000 [33]. Thereafter it has been reported frequently from many different countries. In addition, new CPV-2a and new CPV-2b have also been documented due to non-synonymous substitution at 297 residues (Ser to Ala) of VP2 protein [34]. In India, CPV-2a has recently become the most prevailing antigenic type among all variants. Recent emergence of new antigenic variants that differ significantly from the current vaccine strains is a matter of concern for efficacy of vaccine [35].
The CPV is of two types: CPV-1, commonly known as minute virus of canine and accountable for gastrointestinal and respiratory infection of dogs whereas, CPV-2, most pathogenic type and is responsible for severe gastroenteritis/hemorrhagic gastroenteritis, in young puppies as well as adult dogs.
It is quite difficult to distinguish the clinical diseases caused by CPV-2 variants owing to its overlapping nature of signs and symptoms. These variants are believed to produce similar pathogenicity however; some studies showed that severity of clinical manifestations is influenced by variants of CPV-2 based on clinical, hematological, serological and histopathological examinations [36, 37].
Although puppies under 6 months of age are highly susceptible, adult dogs with insufficient immunity are also considered as high risk to the CPVE. CPV-2 can persist in the environment for more than a year, enabling susceptible dogs to pick up infection from CVP-2 contaminated feces, vomitus, or fomites. Although the feco-oral route is considered as primary path of disease transmission, infection through the oro-nasal route is also common in naive or under-immunized dogs due to ingestion of viruses shed in the vomitus or feces of CPV-2-infected animals [38]. However, direct contact or environmental contamination may also play a role [39]. Breed predisposition and seasonal prevalence of the disease are subject to considerable variations in wide geographical areas [40, 41].
Doberman, Rottweiler, and German shepherd (GS) dogs have been reported to be more susceptible to CPVE than other breeds [42]. Due to inherited immunodeficiency, the exotic breeds, German Sphered and Doberman, are more susceptible than the other breeds [43]. German shepherd has the highest CPV infection rate (70%) followed by the Doberman (55%) [44]. A cytokine bioassay revealed that the magnitude of TNF-α production by peripheral blood monocytes was greatest in dogs with a breed-related risk for CPVE. When compared to mixed breeds, highly susceptible breeds such as Rottweiler and Doberman Pinscher produce more TNF-α in response to LPS stimulation [45]. Increased TNF activity is predictive of mortality in naturally occurring CPVE infection in veterinary medicine [46]. Therefore, it has been hypothesized that dogs with a breed-related risk of developing CPVE, a disease associated with sepsis, would have a greater pro-inflammatory cytokine response to endotoxin [45].
The incubation period of CPV-2 infection ranges from 4 to 14 days. The infected dogs start to shed virus few days prior to the visible clinical signs and shedding of virus gradually declines 3–4 weeks postexposure [47]. Following entry into the body, the CPV-2 rapidly multiply in oropharyngeal lymph node, thymus and mesenteric lymph node, resulting in viremia within one week of exposure. After that, the virus attacks rapidly multiplying cells of crypts of intestine, epithelium of the tongue, oral cavity, bone marrow, and cardiac myocytes, besides lung, spleen, liver, and kidneys [48]. The key pathogenic event in CPV-2 infection is the virus-induced destruction of enterocyte, leading to mucosal barrier disruption, and villous atrophy. This causes profuse vomiting and hemorrhagic diarrhea, nutrient malabsorption, dehydration/hypovolemia, metabolic acidosis and/or alkalosis. The disruption of mucosal barrier allows bacterial translocation from intestinal compartment to systemic circulation, resulting in septicemia, endotoxemia, systemic inflammatory response syndrome as well as hypercoagulability [49]. The CPV-2 infection in the thymus and bone marrow precursor cells results in loss of thymic cortex and profound leucopenia, respectively [48]. Death may occur due to multi-organ failure when the affected dogs remain unattended [40, 49]. Previously, myocarditis was thought to be the acute cause of death in young puppies however, this form nowadays occurs rarely because of widespread CPV vaccination of dogs. The concurrent infections with parasitic, virus, or bacterial intestinal pathogens or stressors may aggravate the disease [50, 51, 52].
The degree of clinical manifestations may vary with age, breed, and immune status, duration of illness and virulence of virus. The clinical signs of dogs with CPV infection are nonspecific in nature and resembles to gastritis and enteritis. The most notable clinical signs of CPVE are lethargy, depression, weakness, lack of appetence, bouts of vomiting, and diarrhea. The diarrhea is characterized by foul-smell and mucoid to purely hemorrhagic because slugging of intestinal mucosa and bleeding. The excessive loss of fluid during vomiting and diarrhea causes marked dehydration that results in development of hypovolemic shock. Occasionally, intussusception occurs due to intestinal dysmotility. Neurologic signs in puppies with CPVE may result from hypoxia secondary to myocarditis, hypoglycemia, or intracranial thrombosis or hemorrhages [52]. The bacterial translocation from intestine to systemic circulation can cause fever, systemic inflammatory response syndrome and septic shock with hypotension and organ failure [40, 48]. Apart from diarrhea, respiratory distress, pulmonary congestion and edema, alveolar and bronchiolar hemorrhage and convulsions are also occasionally manifested due to hypovolemia, endotoxic and septicemic shock [8, 53]. The malabsorbtion of nutrients and inadequate storage of glycogen in muscle and liver result in hypoglycemic encephalopathy which leads to seizures. On hospital admission, the prognosis is poor in CPVE dogs with intussusception, systemic inflammatory response syndrome and severe leucopenia.
Virus isolation is considered as a gold standard for any viral disease diagnosis. In case of CPV-2 different cell lines like CRFK (Crandell Rees feline kidney), MDCK (Madin-Darby canine kidney) and A-72 are used for the isolation and propagation of the virus. The adapted virus causes distinct cytopathic effect in infected cell lines as cell rounding, aggregation, and necrosis of the affected cells. This requires the presence of special laboratory and is laborious [54].
It is an expensive technique for the detection of the virions by negative staining in the stool samples or culture isolated virus. Immunoelectron microscopy can also be done by using CPV-specific antibodies. The need of expensive electron microscope makes it out of reach for regular usage [55].
The property of the CPV to cause agglutination of the pig, cat or rhesus monkey red blood cells at 4°C is used for detection of the CPV. The reciprocal of the maximum dilution of virus exhibiting ample agglutination of erythrocytes (mat formation) is designated as HA titer. The HA titer of more than 1:32 is usually considered as specific for CPV-2 [56].
The use of electric current allows the rapid movement of antigen and antibody towards each other resulting into the formation of precipitation line quicker than simple diffusion reaction. This technique is not commonly used but have been utilized for the prevalence of CPV infection in clinically suspected dogs [57].
In this test, an antibody tagged with fluorescent dye is employed for detection of specific CPV antigen. Mostly it is used as direct FAT for the diagnosis of CPVE but is not used routinely for diagnostic purpose [58].
This is a commonly used test utilizing antigen–antibody interactions employing specific antigen or antibody and is mostly useful under the field conditions. Here, the property of agglutination of polystyrene beads coated with either specific antigen or antibody on their surface is used with anti-CPV monoclonal and polyclonal antibody to detect CPV-2 in the stool samples. Earlier it has been used for both qualitative and quantitative evaluation of CPV in suspected dog feces. Also, a recombinant VP2 protein-based LAT for determination of immune status in dogs against CPV-2. Besides LAT, a slide agglutination inhibition test has been used to detect the presence of CPV-specific antibodies by utilizing the agglutination property of CPV-2 [59].
This method is developed for the detection of CPV-2. SIT is an antibody typing system based on the ability of viral antibodies to bind with the virus and prevents the virus from binding to RBC. SAT is used for antigen detection by serially diluting the clinical sample and then incubating it with a fixed amount of RBC containing virus surface receptors. The virus particles in the sample bind to the RBC and form a lattice that can be seen visually [60].
It is an enzyme-based immunoassay involving antigen–antibody interactions to screen a large number of samples at a time. Recombinant VP2 protein-based indirect ELISAs has been developed to detect and quantify antibodies against CPV-2. Novel polyclonal antibody-based antigen capture ELISA using rabbit anti-CPV hyperimmune sera as capture antibody and guinea pig anti-CPV hyperimmune sera as detector antibody has been also developed. IgY-based ELISA comprising of the chicken egg yolk-derived has been developed for the detection of both antigen and antibodies. Different commercial ELISA kits are currently available for CPV-2 antigen and antibody detection [61].
IC assays or Lateral flow assays are strip-based devices utilized for the detection of a target analyte in test samples. Colloidal gold nanoparticles are commonly used in synthesis of the probe (conjugate) in majority of these strip-based points of care assays. Different components used are the sample pad, conjugate pad, nitrocellulose membrane, absorbent pad and a plastic cassette. These tests are now used routinely for the parvovirus diagnosis in affected dogs. A number of lateral flow assay-based commercial kits are available for rapid detection of both CPV-2 antigen in feces and antibodies in serum, which are also available in the market. These are helpful in the field and gives rapid results within 10–15 mins. Recombinant VP2 protein based immunochromatography tests has also been developed based on the rapid detection of CPV-2 [62].
It is an immunological test which uses charging of test antigen on to a nitrocellulose or PVDF membrane followed by detection using specific antibody against the antigen and an enzyme labeled secondary antibody which forms a color on addition of an insoluble substrate. It is helpful as on the spot assay for CPV diagnoses. It has been developed for detection of CPV-2 using hyperimmune sera raised against the whole virus/recombinant VP2 protein. Commercial dot ELISA kits are also available for evaluating IgM response against CPV-2 after vaccination or infection [63].
PCR is a molecular diagnostic assay which is used for the detection of viral nucleic acid and is relatively more sensitive than other conventional tests. Diverse antigenic types of the CPV can be distinguished by employing strain-specific primer or nested PCR or restriction enzyme analysis of the PCR. Also strain differentiation may be carried out with the help of oligonucleotide sequencing of the amplified gene [64].
This has also been reported for the detection of CPV nucleic acid. Here hybridization with CPV-specific biotin or radiolabelled probe is carried out onto the CPV nucleic acid charged nitrocellulose paper or nylon membrane from suspected samples and then formation of color and band in the radiograph indicates the presence of the virus [65].
It uses an isotopic-labeled probe for both the detection and tracking of CPV nucleic acid in affected morbid tissue specimens thus,using more incubation time for development of the positive reaction [66].
This technique can be employed to quantitate CPV-2 in samples using either TaqMan probe technology or SYBR Green method. It is used for strain differentiation of concurrent infection using Multiplex Real-time PCR; and also, to differentiate vaccine strain from wild CPV strains. Different multiplex assays real-time PCR has been validated for the presence of CPV, FPV and PPV [67].
It is used for the detection and typing of the known point mutations/single nucleotide polymorphism based on variable size of PCR-amplified products specific to a particular allele. In this PCR basically 2 pairs of primers are used (2 inner and 2 outer specific primers matching to individual allele type) in a single PCR tube and there are no post-PCR protocols used as restriction enzyme digestion (PCR-RFLP) and sequencing therefore they provide an economical confirmation. ARMS-PCR is a well-known technique frequently employed for phenotypic association and single nucleotide polymorphism (SNP) studies. This has been used for CPV detection and its antigenic typing [54].
It contains a stable electrically neutral peptide backbone and the PNA-DNA hybridization assay are relatively more sensitive and specific than TaqMan-based real-time PCR for CPV differentiation [68].
The assay is a sensitive and rapid technique used for amplification of DNA and thereby pathogen detection in an hour by using the DNA polymerase by autocycling strand displacement action by boiling at persistent temperature (60–65°C) in water bath. Usually, 2 sets of primers bind to 4 to 6 different regions of target viral DNA. LAMP has field application as there is no need for any thermocycler to carry out the target gene amplification. The amplification of VP2 gene of CPV-2 by LAMP assay has been developed. LAMP assay along with lateral flow dipstick (LFD) and LAMP-ELISA are also used for CPV DNA detection [69].
It is a convection-based method using a hydrolysis probe for detection of CPV-2 and its antigenic variants. The reaction mixture is sequentially allowed to pass in an automatic manner through variable temperature zones in a capillary tube which undergoes thermocyclic phase to amplify the DNA and the probe hydrolysis produces optical output providing the result within an hour [70].
This technique makes use of both conventional PCR and isothermal amplification as in LAMP and is completed within one and a half hour. Here mostly an exogenous sequence from an unrelated species or of botanical origin is incorporated at the 5′ end into the primer sequences used in PSR if a human or veterinary pathogen is targeted. PSR has been successfully used to detect all CPV antigenic variants with ten-fold higher sensitivity than traditional PCR [71].
It is a probe-based assay that uses melting curve analysis to detect and differentiate between CPV-2 variants. This assay consists of 2 TaqMan probes namely FAM labeled and HEX labeled. The FAM-labeled probe sequence is perfectly complementary to CPV-2a, with a 1 bp mismatch to CPV-2b and a 2 bp mismatch to CPV-2c. The HEX-labeled probe has complete complementarity with the original CPV-2 and a 1-bp mismatch with the other variants. This method is also capable of detecting samples containing more than one variant without sequencing [72].
Aptamers emerged as a good alternative to antibodies as affinity reagents. Recently, ssDNA aptamers that specifically bind with the recombinant VP2 (rVP2) protein of CPV-2 with affinity in the nanomolar range have been reported. The ssDNA aptamers specific to CPV-2 (rVP-2) were selected by the Systematic evolution of ligands through exponential enrichment (SELEX) method and their target binding was assessed by dot blot and enzyme-linked oligonucleotide assay (ELONA). Aptamers with high binding affinity and specificity against rVP-2 could be employed in diagnostics for rapid detection of CPV-2 [73].
It is primarily used for most viral genome identification and confirmation. Thus, considered as a gold standard for the antigenic typing of CPV variants. The amplified PCR product is either directly sequenced or cloned which is sequenced in a sequencer utilizing apt primers. The sequence data is analyzed using the appropriate bioinformatics database. Either nucleotide or amino acid sequence data or even both could be employed to recognize the evolutionary analysis of CPV-2 isolates from different geographical sites [74].
It is an analytical device which detects the DNA/RNA/protein/enzymes and alters it to the detectable electrical signals. A biosensor for CPV detection has been established by means of quartz crystal microbalance biosensor and ProLinker B [75]. Summary of different types of diagnostic assays are listed in the Table 1.
Diagnosis | Specimen | Diagnostic assay used | Feature | Remarks |
---|---|---|---|---|
CPV antigen | Feces or rectal swab | ELISA | High specificity Low sensitivity | Feces or rectal swab |
Haemagglutination assay | Low-cost and rapid. | Sensitivity and specificity vary | ||
Tissues or morbid samples | Necropsy specimens | Histopathology | Different histopathological techniques and IHC may be used. | Differential diagnosis with other enteric infections |
Viral DNA | Feces or rectal swab or any tissue | Polymerase chain reaction (PCR);qPCR | Efficient in diagnosing even minute amount of viral genome, can be quantified; Antigenic typing | Sensitivity and specificity vary. Vaccine virus shedding occurs upto weeks after immunization leading to false positives results. Inhibitory components may lead to false negative results. |
Virus | Feces or rectal swab or any tissue | Virus isolation | Confirmatory diagnosis | Requires special facility |
Virus particles | Feces or rectal swab or any tissue | Electron microscopy | Confirmatory diagnosis | Requires special facility, expensive |
Summary of the different types of diagnostic assays for CPVE diagnosis.
Commercially available kits are mostly based on antigen–antibody reactions, such as ELISA, dot ELISA, and immunochromatographic strip-based assays (Table 2).
Sl. No. | Test | Company | Principle | Reference |
---|---|---|---|---|
1 | SNAP parvo antigen test | IDEXX, United States | ELISA | [76] |
2. | Rapid Immunochromatographic (IC) strip test | ADDBIO, Korea | Immunochromatography test | [43, 77, 78] |
3. | Witness Parvo Test Kit | Zoetis, United states | Rapid Immuno Migration (RIM™) technology. | [79] |
4. | Fassisi® Parvo | Fassisi, Gottingen, Germany | Lateral flow immunoassays | [80] |
5. | FASTest parvo card | Vet lab, UK | Lateral flow immunoassays | [55] |
6. | 4 CPV Antigen Rapid Test Kit | Ubio Biotechnology systems Pvt. Ltd., India | Lateral flow immunoassays | [79] |
7. | Anigen Rapid CPV Ag Test Kit® | Bionote, Dongtan, South Korea | Lateral flow immunoassays | [80] |
8. | ImmunoRun CPV antigen detection kit | Biogal- Galed labs, Israel | Immunochromatographic assay | [79] |
9. | Primagnost® Parvo H + K | Dechra, Aulendorf, Germany | Lateral flow immunoassays | [80] |
10. | Canine Parvovirus & Distemper IgMAntibody Test Kit | Biogal Galed Laboratories Acs Ltd., Israel | Immunocomb | [79] |
11. | Vetexpert Rapid Test CPV Ag® | Vetexpert, Vienna, Austria | Lateral flow immunoassays | [80] |
List of commericially available kits for CPV-2 detection.
In absence of effective and appropriate antiviral drugs, the most universal therapeutic regimen for CPVE is supportive and symptomatic care until vomiting and diarrhea have resolved. Because of long-term illness of CPVE infected dogs, the challenges faced by the pet owners are cost of treatment and hospitalization. In private practice settings, the treatment cost may be huge, indicating that financial constraints may be a factor in disease-related euthanasia [81]. Therefore, fatality of CPVE is documented more in socioeconomically underprivileged areas, where level of education and financial opportunity for care and vaccination are not adequate [82]. Although the survival rate of CPVE in hospitalized and outpatient dogs is debatable, a recent prospective, randomized trial found no significant differences in survival (90% vs. 80%, P = 0.66) or duration of hospitalization (4.6d vs. 3.8d, P = 0.20) between inpatient and outpatient dogs [83]. However, given the possible risks of long-term hypoglycemia and leukopenia, aspiration pneumonia, edema, and intussusception in CPVE dogs, hospitalization appears to be the better option over outpatient treatment [84].
The principal components of supportive and symptomatic therapy include 1) fluid therapy and oncotic support, 2) antibiotics, 3) antiemetics, and 4) nutritional support. A wide range of other treatment measures including, though not limited to, antiviral treatments and pain management have been assessed in the past or are currently under investigation regarding their potential utility in CPVE.
The development of severe hypovolemia is the first impact of pathophysiology in dogs with CPVE, hence re-establishment of the circulating volume is the utmost need [85]. The hypokalemia, hypochloremic metabolic alkalosis, hypoglycemia, hypoproteinemia and loss of oncotic pressure in circulation are the major fluid and electrolyte abnormalities during episode of diarrhea and vomition in acute CPVE [86]. The most aggressive therapies consisting of administration of intravenous (IV) fluids to restore intravascular fluid volume status, replenish interstitial fluid losses, maintenance of hydration and oncotic support. A balanced isotonic crystalloid solution (eg, Lactated Ringers) should be used for initial restoration of intravascular volume and rehydration, with a rate titrated to improve perfusion parameters such as capillary refill time, mucosal color, pulse character, and mean arterial pressure or lactate concentrations. Apart from fluid administration, potassium need to be supplemented in hypokalemic patients whereas, 25% dextrose at the dose rate of 1-2 mL/Kg body weight followed by addition of 2.5–5% dextrose in the crystalloid fluids will be required for hypoglycemic patients with blood glucose level < 60 mg/dL. Initially, the fluid is administered at the dose rate of 80–90 mL/kg with a boluses of 15–20 mL/kg over 15–20 minutes to counter the hypovolemic shock and, to improve the fluid perfusion. After that, the maintenance dose for daily fluid depends on the body weight (kg) and percent of dehydration. The volume (L) required to correct the daily fluid loss is calculated as body weight (Kg) × % dehydration. Generally, 40–60 mL fluid for each kg body weight is considered as ideal maintenance dose. Since fluid absorption through subcutaneous route is impaired in hypovolemic patients, intravenous access is considered as choice of fluid treatment. However, intraosseous or jugular catheter are considered as appropriate option in severe hypovolemic or interstitially dehydrated patients [87].
In CPVE, protein loosing enteropathy attributes to pronounced hypoalbuminemia (<2 g/dL) and/or hypoproteinemia (<4 g/dL) resulting in peripheral edema, pleural or abdominal effusions [88]. In that case, provision of oncotic support in the form of either natural or synthetic colloids are very important to minimize the morbidity and mortality of patients [89]. For correction of hypoalbuminemia, fresh plasma (20 mL/kg) or fresh-frozen plasma (6.6–11 mL/kg IV or 3 doses administered intraperitoneally 12 hours apart) and canine-specific albumin concentrate are used [90]. The concentrated human albumin products can also be used but the risk of immune reaction is the major limitation. If further oncotic support is required, hydroxyethyl starch (20–30 mL/kg/d) can be given, depending on clinician choice [6]. Sometimes, administrations of whole blood (20 mL/kg, within 4 hours) or packed RBCs are needed in severe anemic dogs with CPVE.
Apart from fluid and electrolyte imbalance, emesis is another clinical manifestation in CPVE. So, antiemetic treatment is warranted in CPVE otherwise persistent vomition may enhance the duration of hospital stay and further aggravates the condition of patient. The clinical efficacy of number of antiemetics in CPVE had been investigated with varying degree of results. The earlier studies showed that metoclopramide, a dopaminergic antagonist, was found to be effective in reducing episode of vomition by exerting a prokinetic effect in the upper intestinal tract and blocking the chemoreceptor trigger zone when administered as a bolus or as a constant-rate infusion in dogs. The ondasetron or dolasetron, the serotonin receptor antagonists, are also found effective in reducing the number of vomiting events [85]. Recently, a substantial antiemetic effect of maropitant, an antagonist of neurokinin1 receptors, by stimulation of either central or peripheral emetic pathways has been reported in dogs however, the efficacy of maropitant in CPVE has yet to be thoroughly investigated [91]. The administration of maropitant once daily, singly or in combination with metoclopramide, is very effective in reducing vomition in CPVE [5].
Translocation of bacteria from intestinal compartment to systemic circulation is very common in CPVE because of villous collapse and disruption of the mucosal barrier. The translocation with concurrent marked neutropenia leads to a high risk of septicemia and endotoxemia. Additionally, hypotension from fluid loss and sepsis make dogs with CPVE at high risk of developing acute kidney injury. Therefore, parenteral administration of broad-spectrum bactericidal antibiotics is necessary in dogs with CPVE. Ampicillin and cefoxitin as single-agent treatments or in combination with enrofloxacin are the choice antimicrobials against Gram-positive and negative bacteria [85]. Aminoglycosides may also be considered in well-hydrated animals otherwise it may be avoided due to its inherent risk of nephrotoxicity. Puppies with CPVE often have comorbidities, including gastrointestinal parasitism. Hence, antiparasite therapy should be initiated once the puppy can tolerate oral therapies [6].
Restoration of early mucosal integrity and prevention of bacterial translocation from gut compartment to systemic circulation are very important for faster recovery of dogs with CPVE. Enteral feeding is reported to improve the mucosal integrity and faster repair, resulting in lower possibilities for bacterial translocation [8]. In earlier study, it was demonstrated that early enteral nutrition via nasoesophageal catheter starting 12 hours post-admission led to clinical improvement, significant weight gain, and improved gut barrier function was more early as compared to withholding of the traditional food until cessation of vomiting for 12 hours [92].
Severe vomition, enteritis, and or concurrent intussusception in CPVE are the possible reasons for abdominal pain. Hence, analgesic treatment to reduce visceral pain is one the important aspect in therapeutic management in CPVE. Partial mu-agonists such as buprenorphine (0.01–0.02 mg/kg IV every 8 hours) or an agonist–antagonist such as butorphanol (0.1–0.2 mg/kg/h) are the preferred analgesics over the pure mu agonists as opioid analgesics can promote ileus and vomiting. The α-2 agonists that promote extreme vasoconstriction and limit gastrointestinal perfusion, and non-steroidal anti-inflammatory drugs that impair gastrointestinal and renal perfusion, both are not indicated [93].
Like other viral infections, prophylaxis is the cornerstone for prevention of CPV in dogs. Although, an adequate number of killed and live CPV vaccines are marketed by pharmaceuticals but vaccines sometimes fail to protect completely due to poorly responding breeds (Rottweilers and Doberman pinschers), variation in genetic makeup of field and vaccine viruses, interference by presence of maternal antibodies and adjunct factors [94]. Therefore, development of some suitable antiviral drugs is utmost important for effective management of the CPVE in its acute illness stage. Till now, only few antiviral drugs have been evaluated for its clinical efficacy against CPVE. In an earlier placebo-control study, the therapeutic efficacy of Oseltamivir, a neuraminidase inhibitor, in CPVE had been evaluated and noted that Oseltamivir did not produce any additional benefit in terms of reduction of mortality or duration of hospitalization except some improvements in body weight and hemogram in dogs with CPV-illness [95]. In another study on naturally infected dogs, a promising anti-CPV activity of recombinant feline interferon-ω (rFeIFN-ω) has been recorded as compared to placebo-group. The intravenous administration of rFeIFN-ω at the dose rate of 2.5 mU/kg daily for consecutive three days remarkably reduced the clinical symptoms and mortality [96, 97]. Although the drug is currently available for use in Europe and Australia, the high price and frequent non-availability are major limitations. Recently, another antivital drug, Acyclovir, guanine analogue commonly used to treat herpes simplex virus infection, have been shown to improve the disease conditions [98]. Further, an
Passive immunization with specific antibodies against enteric viral infections in animals confers significant protection, reduces diarrhea and virus shedding and increase survival rates [101]. Thus, the of immunotherapeutics in viral infections is promising treatment approach because of lower adverse effects as well as no chance of any resistance as in antiviral drugs. The passive immunization by means of oral or intravenous administration of IgY specific for CPV-2 shows the protective effect in dogs challenged with the virus [102]. The reduction of clinical scores, duration of symptoms and mortality and improvement of body weight gain has been reported by anti-CPV-2 IgY therapy in experimentally produced CPVE [103]. Recent study reported that chicken IgY- single chain fragment variables (scFv) generated against the virus capsid protein could be a promising therapeutic target against CPV [104, 105]. Aside from IgY, the neutralization of CPV by anti-feline panleukopenia virus antibodies is also reported from an
The key physiopathological alterations of CPVE are destruction of intestinal crypts, neutropenia, secondary bacterial translocation, immunosuppression due to thymus atrophy, sepsis and systemic inflammatory response syndrome in puppies [6, 109]. Therefore, immunomodulators could be an option to enhance therapeutic efficacy of supportive treatment. A recent study demonstrated that subcutaneous administration of human dialyzable leukocyte extract-h (hDLE) along with supportive therapy in puppies with CPVE significantly increased the leukogram and reduced the clinical score, duration of hospitalization, mortality as compared to supportive therapy alone [110].
Leukopenia is one of the most important prognostic indicators of mortality in dogs with CPVE. Hence, stimulation of bone marrow and improvement of leukogram in peripheral circulation are considered as strategic approaches to reduce the CPVE associated mortality. Enhancement of endogenous canine G-CSF (cG-CSF) concentrations by exogenous administration of human G-CSF (hGCSF) and cG-CSF is reported to stimulate bone marrow, resulting in improvement of neutrophil counts in puppies with CPV infection [111]. However, the use of hG-CSF and cG-CSF may not necessarily improve survival [112, 113].
The interferon (IFN)-ω, a type I IFN (similar to IFN-α), is known for its antiviral, anti-proliferation, and antitumor activities. A notable therapeutic effect of rIFN-ω on CPV-infected dogs is reported [114]. Additionally, the promising therapeutic potential of other type I (IFN-α, IFN-β, IFN-ε, and IFN-κ) and III (IFN-λ) IFNs in CPVE has also been reported [115].
Recently, anti-CPV activity of the serum derived transfer factors (TFs), low molecular weight (<5000 daltons) biological response modifiers has been documented. It imparts therapeutic benefit in CPVE by altering the cytokine response of the host [116].
Probiotics, primarily comprised of live microorganisms in fermented foods, protect gut from acute diarrhea through adherence and colonization on gut mucosa [117]. Therapeutic efficacy of probiotics has been verified in dogs with CPV associated illnesses [118]. In an earlier study, oral administration of probiotic preparations as an adjunct therapy to young dogs with CPVE has shown faster resolution of clinical signs, improved leukogram and decreased mortality as compared to supportive treatment alone [119]; whereas, no benefit with respect to length of hospital stay or case fatality was recorded in other study [120].
The disturbance in oxidant/antioxidant equilibrium is evident in CPV-gastroenteritis and oxidative stress is believed to link with pathogenesis of CPVE [121]. Hence, addition of antioxidants in supportive therapy has emerged as a promising therapeutic option to improve the response of treatment in viral diseases. Treatment with
An interest in natural products including herbs, plants and their extracts/metabolites as antiviral drug candidates has increased in the last few decades especially due to rising emergence of antimicrobial resistance globally and potential side-effects of many antimicrobials [123]. Very recently, anti-parvoviral activity of propolis, a traditional Chinese medicine, prepared from honeybee hives has been documented [124]. The
Alteration in the gut microbiome is reported in enteric viral diseases including CPVE and other gastrointestinal diseases in dogs [125]. The disruption of gut microbiota leads to impediment in the enterocyte nutrition, immune regulation, protective barrier function, and gastrointestinal motility [126]. Therefore, restoration or re-establishment of the microbiota could have a good interest therapeutically. Recently, a randomized clinical trial showed that administration of fecal microbiota (10 g feces diluted in 10 mL of sterile 0.9% saline) obtained from healthy donor rectally at 6–12 hours post-admission caused faster resolution of diarrhea, shortened the duration of hospitalization and reduced the mortality in young dogs with CPVE when compared with standard therapy alone [126].
A modified live virus (MLV) and an inactivated vaccine are the two types of CPV-2 vaccines currently available [94]. Administration of the vaccine should start at 6 to 8 weeks of age and then every 2–4 weeks until 16 weeks of age or older. For dogs that are 16 weeks or older, 2 doses of vaccination are recommended with an interval of 2–4 weeks [127]. A recombinant vaccine based on virus-like particles (VLPs) is being developed, which has the advantage of becoming highly immunogenic and safe [128]. Peptide vaccines containing major antigen neutralizing region N terminal of VP2 are also under developmental stage [129]. A single-dose vaccination of Vaccinia virus encoding CPV2-VP2 elicited substantial antibody responses and provided comparable protection for dogs with attenuated CPV2 vaccine. This vaccine could be used as a promising vaccine candidate to prevent CPV-2 infection in dogs [130].
CPV-2 is one of the most significant viral enteropathogens of canines causing high morbidity and mortality and manifested by vomition and severe acute haemorhagic gastroenteritis. Prompt symptomatic therapy will increase survivability of infected puppies but vaccination is best way to prevent the disease in dogs. Despite the pups are protected through vaccination from the pregnant bitch, it is more vulunerable to CPV-2 infection as maternal antibody titers started declining. Despite the availability of high sensitive and specific diagnostic approaches and the effective prophylactics such as modified live virus and inactivated vaccines, a large number of outbreaks are still reported in wide geographical areas across the globe in both vaccinated and unvaccinated dogs. The future studies should be taken up towards vaccination failures, occurrence of CPV-2 in different canine species and the emergence of antigenic variants of the CPV-2 involved in the outbreaks.
All the authors acknowledge and thank to their Institute.
The authors declare no conflict of interest.
This compilation is a book chapter written by its authors and required no substantial funding to be stated.
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