**6. Eukaryotes**

Eukaryotes are capable of several different types of DNA repair process:

well-studied processes. These include mismatch repair (MMR), nucleotide excision repair (NER) and base excision repair (BER) [14]. These processes appear to have arisen in the archaea [59], but are most well understood in the eukaryotes. This option was not available to organisms with ssRNA genomes because the double-stranded state exists only transiently during replication. In any case the enzymes that carry out such repair processes in organisms with DNA genomes are not known to be encoded in the ssRNA virus genomes. Thus this type of mechanism was not likely present during the early evolutionary stages in ssRNA genome containing organisms.

During meiosis homologous chromosomes originating from different parents align intimately with each other. This is followed by transfer of sequence information between homologs, homologous recombination. The main mechanism is SDSA (copy-choice recombination), a central characteristic of meiosis (see Section 6.1). Less frequent homologous recombination by breakage and exchange of chromosomes also occurs during meiosis.

Copy-choice recombination is also an important general mechanism for dealing with DNA damages that block the movement of the DNA polymerase during DNA replication (see Section 6.2).

#### **6.1 Meiotic and mitotic recombination**

The results of numerous studies in a wide range of eukaryotes indicate that during meiosis a variety of DNA damages are repaired by recombinational repair (reviewed in [62]). In somatic cells, mitotic recombination also facilitates DNA repair. Molecular models of recombination have been revised over the years as relevant evidence accumulated. Our current understanding of recombination reflects the work of several groups of investigators that have provided evidence that SDSA is a major mechanism of recombination [11, 63–65]. Furthermore, SDSA is a type of copy-choice mechanism since it involves switching from one template to another during strand synthesis and the return to the original template after a short distance (compare **Figure 5** to **Figure 2**).

**Figure 5** illustrates the series of steps that occur by the meiotic SDSA process in the repair of a double-strand break (DSB) in one chromosome using information

**33**

**Figure 5.**

from an adjacent undamaged homologous chromosome. As shown in the figure, the steps include strand invasion by a broken strand to form a D-loop, the further extension of the strand by DNA synthesis, and then the reassociation of the transferred strand with its original pairing partner. These strand-switching and DNA synthesis events associated with repair of a damage are similar to the copy-choice

*Synthesis-dependent strand annealing (SDSA) in the repair of a double-strand break.*

*Origin of DNA Repair in the RNA World DOI: http://dx.doi.org/10.5772/intechopen.93822* *DNA - Damages and Repair Mechanisms*

ssRNA genomes.

somes also occurs during meiosis.

**6.1 Meiotic and mitotic recombination**

replication (see Section 6.2).

(compare **Figure 5** to **Figure 2**).

stages in ssRNA genome containing organisms.

well-studied processes. These include mismatch repair (MMR), nucleotide excision repair (NER) and base excision repair (BER) [14]. These processes appear to have arisen in the archaea [59], but are most well understood in the eukaryotes. This option was not available to organisms with ssRNA genomes because the double-stranded state exists only transiently during replication. In any case the enzymes that carry out such repair processes in organisms with DNA genomes are not known to be encoded in the ssRNA virus genomes. Thus this type of mechanism was not likely present during the early evolutionary

c.Double-strand damages in double-stranded DNA, such as double-strand breaks, can be repaired without the presence of an homologous template by such processes as non-homologous end joining (NHEJ) and microhomology mediated end joining (MMEJ). These processes depend on the duplex nature of DNA but not on strict homology. NHEJ can be accurate if the ends of the DNA in double-strand breaks do not need processing. However, if the ends need processing before rejoining then mutations are very likely to be introduced [60]. MMEJ is inaccurate and is always associated with a DNA deletion [61]. Thus these processes are inaccurate and generate mutations and are not applicable to

d.Homologous recombinational repair is possible when two templates are present and adjacent. Such repair may occur for various types of DNA damage. For double-strand breaks in mitosis, homologous recombinational repair, either by the less common breakage and exchange mechanism or by the more frequently used SDSA (copy-choice) mechanism [11], are the only accurate forms of repair available. Template switching can occur during mitosis when two sister chromatids are present and adjacent after DNA synthesis and before cell division.

During meiosis homologous chromosomes originating from different parents align intimately with each other. This is followed by transfer of sequence information between homologs, homologous recombination. The main mechanism is SDSA (copy-choice recombination), a central characteristic of meiosis (see Section 6.1). Less frequent homologous recombination by breakage and exchange of chromo-

Copy-choice recombination is also an important general mechanism for dealing with DNA damages that block the movement of the DNA polymerase during DNA

The results of numerous studies in a wide range of eukaryotes indicate that during meiosis a variety of DNA damages are repaired by recombinational repair (reviewed in [62]). In somatic cells, mitotic recombination also facilitates DNA repair. Molecular models of recombination have been revised over the years as relevant evidence accumulated. Our current understanding of recombination reflects the work of several groups of investigators that have provided evidence that SDSA is a major mechanism of recombination [11, 63–65]. Furthermore, SDSA is a type of copy-choice mechanism since it involves switching from one template to another during strand synthesis and the return to the original template after a short distance

**Figure 5** illustrates the series of steps that occur by the meiotic SDSA process in the repair of a double-strand break (DSB) in one chromosome using information

**32**

from an adjacent undamaged homologous chromosome. As shown in the figure, the steps include strand invasion by a broken strand to form a D-loop, the further extension of the strand by DNA synthesis, and then the reassociation of the transferred strand with its original pairing partner. These strand-switching and DNA synthesis events associated with repair of a damage are similar to the copy-choice

recombination described above for ssRNA viruses. Thus a central feature of eukaryotic recombination in meiosis and mitosis, strand-switching copy-choice recombinational repair, may have evolved from the simpler repair-related copy-choice events postulated above for ssRNA protocells based on the known processes in ssRNA viruses. Experimental evidence demonstrating that SDSA is a major recombination pathway in meiosis was presented by McMahill et al. [64].

The process of SDSA can accurately repair genome damage by copying the information lost in a damaged template strand from another intact homologous template strand without the need for physical breakage and exchange of DNA. Evidence bearing on the role of SDSA during meiotic recombination was reviewed by Bernstein et al. [66]. An alternative mechanism for recombinational repair termed the Double-Strand Break Repair (DSBR) model also explains some types of recombination events, but in contrast to SDSA recombination, the DSBR model does require physical breakage and exchange of DNA strands [67]. However,

#### **Figure 6.**

*Bypassing a DNA damage during replication. This mechanism involves reversal of the replication fork, where the newly replicated strands dissociate from their previous templates and anneal to form a cruciform intermediate, known as the "chicken foot" structure. Further replication of the previously blocked strand can then continue, leading to the bypass of the damaged site.*

**35**

*Origin of DNA Repair in the RNA World DOI: http://dx.doi.org/10.5772/intechopen.93822*

elements of copy-choice recombination.

simpler copy-choice processes in ssRNA protocells.

**6.2 DNA replication**

**7. Conclusions**

ribozyme polymerases.

reassortment.

would be similar.

have occurred.

both the SDSA and DSBR models include a step in a which a DNA strand switches at a site of damage from one complementary partner strand to another and then continues synthesis with the new partner as template. Thus both models have

With respect to mitotic recombination in somatic cells, Andersen and Sekelsky [11] reviewed evidence that DSBR is a minor pathway for recombinational repair, and that the SDSA model appears to describe mitotic repair more accurately.

During DNA replication, a DNA damage in a template strand may be present and act as roadblock to the movement of the DNA polymerase as it extends synthesis of a new complementary strand. A blocked replication fork may be accurately bypassed by the mechanism illustrated in **Figure 6** [12, 13]. When movement of the replicative polymerase is blocked by a damage, the polymerase can switch template strands (mediated by a helicase) [12, 13] to form a structure referred to as a "chickenfoot" intermediate. As synthesis of the new strand proceeds along the alternate template it synthesizes the DNA region that is complementary to the damaged site in its original partner strand. The newly forming strand may then unwind and then re-associate with its original partner to continue synthesis along its original track. Polymerase-mediated strand-switching to deal with a damaged template during DNA synthesis appears to be an important general mechanism in eukaryotic cells [64]. This mechanism can be regarded as a type of copy-choice recombinational repair, and it too may have evolved from

Given the copy-choice genomic repair mechanism present in today's ssRNA viruses, it appears that copy choice as a repair process may have emerged as early as 3.5 to 2.5 billion years ago when RNA was apparently the only genetic material. It is possible that the capability for strand-switching was a property of the earliest

In early protocells, the ssRNA genomes may have been segmented, as some ssRNA viruses are in the present day. Two protocells with damaged segmented genomes could have been able to generate undamaged progeny after fusion and then reassortment of segments. Present day ssRNA segmented genome viruses can repair damage in their genomes through both copy choice and segment

The early stages of the evolution of genome repair proposed here are based on known capabilities of extant RNA viruses. Currently it is not known if these RNA viruses are the actual evolutionary descendants of early RNA life forms, or if they arose later. It has only been assumed here that the problem of dealing with damage to an RNA genome arises in the two cases, and that the solutions to this problem

The earliest ssRNAs that formed folded structures that acted as ribozymes can be designated plus (+) strands. Such a ribozyme strand could have had polymerase activity and acted as an RdRp. The progeny ssRNAs that it synthesizes would be complementary to the corresponding parental (+) strands, and can be designated minus (−) strands. During the synthesis of (−) strands template-switching may

both the SDSA and DSBR models include a step in a which a DNA strand switches at a site of damage from one complementary partner strand to another and then continues synthesis with the new partner as template. Thus both models have elements of copy-choice recombination.

With respect to mitotic recombination in somatic cells, Andersen and Sekelsky [11] reviewed evidence that DSBR is a minor pathway for recombinational repair, and that the SDSA model appears to describe mitotic repair more accurately.

### **6.2 DNA replication**

*DNA - Damages and Repair Mechanisms*

recombination described above for ssRNA viruses. Thus a central feature of eukaryotic recombination in meiosis and mitosis, strand-switching copy-choice recombinational repair, may have evolved from the simpler repair-related copy-choice events postulated above for ssRNA protocells based on the known processes in ssRNA viruses. Experimental evidence demonstrating that SDSA is a major recom-

The process of SDSA can accurately repair genome damage by copying the information lost in a damaged template strand from another intact homologous template strand without the need for physical breakage and exchange of DNA. Evidence bearing on the role of SDSA during meiotic recombination was reviewed by Bernstein et al. [66]. An alternative mechanism for recombinational repair termed the Double-Strand Break Repair (DSBR) model also explains some types of recombination events, but in contrast to SDSA recombination, the DSBR model does require physical breakage and exchange of DNA strands [67]. However,

*Bypassing a DNA damage during replication. This mechanism involves reversal of the replication fork, where the newly replicated strands dissociate from their previous templates and anneal to form a cruciform intermediate, known as the "chicken foot" structure. Further replication of the previously blocked strand can*

bination pathway in meiosis was presented by McMahill et al. [64].

**34**

**Figure 6.**

*then continue, leading to the bypass of the damaged site.*

During DNA replication, a DNA damage in a template strand may be present and act as roadblock to the movement of the DNA polymerase as it extends synthesis of a new complementary strand. A blocked replication fork may be accurately bypassed by the mechanism illustrated in **Figure 6** [12, 13]. When movement of the replicative polymerase is blocked by a damage, the polymerase can switch template strands (mediated by a helicase) [12, 13] to form a structure referred to as a "chickenfoot" intermediate. As synthesis of the new strand proceeds along the alternate template it synthesizes the DNA region that is complementary to the damaged site in its original partner strand. The newly forming strand may then unwind and then re-associate with its original partner to continue synthesis along its original track. Polymerase-mediated strand-switching to deal with a damaged template during DNA synthesis appears to be an important general mechanism in eukaryotic cells [64]. This mechanism can be regarded as a type of copy-choice recombinational repair, and it too may have evolved from simpler copy-choice processes in ssRNA protocells.


**3. Discussion**

In Brazil, first cases of DHF occurred after the introduction of DENV-2 in the State of Rio de Janeiro. The cases of DHF by DENV-2 occurred after an epidemic by DENV-1 in Rio de Janeiro four years ago (Dias et al. 1991; Zagne et al., 1994). The same pattern was seen in Cuba during the 1981 epidemic with sequential infection by two serotypes (DENV-1 and DENV-2), and an interval of six months to five years or so (Kouri et al., 1986).The dynamics of epidemic of dengue in the Amazon is similar to that of other regions of Brazil and America. In Manaus – Amazonas, the first cases of dengue were registered in March 1998 by serological studies during the first epidemic of dengue (Figueiredo et al., 2004). In 2001, it was possible to identify the DENV-1 as the causative agent of that epidemic (Figueiredo et al., 2002). That same year, cases of dengue hemorrhagic fever were registered in the State of Amazonas, with the viral isolation of DENV-2 (Figueiredo et al., 2002). In 2002, DENV3 was first isolated from a patient coming from Bahia. Since then, DENV-3 was diagnosed by viral isolation from several patients with DF (Figueiredo et al., 2003).The virus DENV-4 was detected during the first epidemic reported in Brazil in Boa Vista-Roraima in 1981-1982. At

Molecular Characterization of Dengue Virus

associated with DHF in the Asian continent.

**5. Recognition**

**6. References**

(2): 122.

hemorrhagic dengue fever in other regions.

DF. Most of them are associated with mild DF.

Circulating in Manaus, the Capital City of the State of Amazonas, Brazil 87

emergence of new serotypes followed by cases of dengue hemorrhagic. The emergence of DENV-4 in 2008 did not just caused a major epidemic with severe and fatal cases. Other serotypes were also observed. It is possible due to competition between with other serotypes, DENV-4 took some time to become established and the population might have been protected by antibodies obtained in heterologous infections by heterologous serotypes. The viral strain should be studied in detail since it is an Asian genotype that has been

Laboratory of Virology at the Foundation for Tropical Medicine Dr. Hector Vieira Gold (FMT/HVD); Dr. RajendranathRamasawmy for review of the manuscript; ConselhoNacional

Araújo et al. Concurrent infection with dengue virus type-2 and DENV-3 in apatient from Ceará, Brazil.*MemInstOswaldo Cruz,* Rio de Janeiro, 2006. 101(8): 925-928. Burke DS, Nisalak A, Johnson De, Scott RM, 1988. A prospective study of dengue infections

Castro, M.G.; Nogueira, R.M.; Schatzmayr, H.G.; Miagostovich, M.P.; Lourenco-De-Oliveira,

Cunha, R.V. & Nogueira, R.M. Dengue E Dengue Hemorrágico. In: *Dinâmica das*

Dash, P.K. et al. Reemergence of dengue vírus type-3 (subtype-III) in Índia.Implications for

De Melo FL, Romano CM, De Andrade Zanotto PM. Introduction of Dengue virus 4 (DENV-4) genotype I in Brazil from Asia?PLoSNeglTrop Dis. 2009; 3(4):390. Dias, M.; Zagne, S.M.O.; Pacheco, M.; Stavola, M.S.; Costa, A.J.L. Dengue

Figueiredo, R.M.P.; Bastos, M.S.; Lima, M.L.; Almeida, T.C.; Alecrim, W.D. Dinâmica da

R. Dengue vírus detection by using reverse transcriptionpolymerase chain reaction in saliva and progeny of experimentally infected *Aedesalbopictus* from Brazil.

hemorrágicoemNiterói.*Revista da SociedadeBrasileira de Medicina Tropical*, 1991. 24

sorologia e isolamento viral na epidemia de dengue em Manaus (1998-2001). In:

visitors from around the world, other viruses need to be investigated.

de DesenvolvimentoCientífico e Tecnológico (CNPq) for financial support.

in Bangkok.Am J TropMed Hyg, 1988,38:172– 180.

*doençasInfecciosas e Parasitarias,* 2005. p.1767-1781.

increased incidence of DHF & DSS.*Virology Journal*, 2006.

*MemInstOswaldo Cruz*, 2004.99: 809-814.

that time the DENV-1 was also found (Osanai, 1984). Since then, no isolate of DENV-4 was recorded anywhere else in the country until later in the year 2008 DENV-4 was isolated in Manaus from patients with DF (Figueiredo et al., 2008).

The DENV-4 present in the State of Amazonas and analyzed by nucleotide sequence was shown to belong to genotype I, only present in the Asian continent and never described in the Americas (De Melo et al., 2009).

Dengue fever in uncomplicated cases of co-infection has also been observed by other authors, contradicting the hypothesis that simultaneous infection with dengue virus permits the emergence of a more severe disease (Santos et al., 2003; Araújo et al.,2006). In areas where more than one serotype are transmitted at the same time, clinical cases caused by more than one serotype of dengue fever can be common (Lorono et al., 1999). The high rate of cases occurring during epidemics can result in many infections with multiple serotypes in humans (both clinical and subclinical), and also provide opportunity for mosquitoes to become infected with two or more serotypes (Gubler et al., 1985; Burke et al., 1988). This suggests that co-infection by multiple dengue serotypes may influence the clinical expression of disease and it was initially considered as an explanation for the emergence of DHF (Hammon, 1973).The Amazon is situated in the Northern Region of Brazil, bordered to the north with the State of Roraima, Venezuela and Colombia to the east with the State of Pará, the southeast by the State of Mato Grosso, to the south with the State of Rondônia and southwest with the State of Acre and Peru (Viverde Tourism). The proximity to other countries where endemic DENV-4 is observed as in Venezuela and Colombia may have influenced the detection of this serotype first in the Amazon. Besides the geographical location, being today a city with eco-tourism development, and the Industrial Pole of Manaus, with over 450 factories of large, medium and small size (PORTAL AMAZON), which attracts investors from around the world. Manaus receives many people from these and other states and countries.

It is important to remember that the DENV-3 was first detected in Manaus from a patient coming from Salvador-Bahia (Figueiredo et al., 2003).The analysis of the region the C/prM of the DENV-3 in this study belonged to genotype III strains. All DENV-3 isolated in the Amazon were very close to the Indian strain GWL-60 (N°acessoAY770512) and the Brazilian strain BR-74 886 (N access AY679147). Subtype III is related to outbreaks of DHF in India (Dash et al., 2006). In Brazil, the prevalence of DENV-3 two years after its introduction in 2000 was associated with major epidemics in terms of more severe clinical manifestations, and the number of deaths (Nogueira et al., 2005). Twenty-two isolates were classified as DENV-3 subtype III (Miagostovich et al., 2002). The similarity of these strains to other represented by the same genotype III ranged from 96% to 98% and 98-99% for sequences nucleotides and amino acids, respectively. These data demonstrate that this virus is circulating around the world, again indicating high potential for distribution, adaptation in in various geographic areas of the world. This subtype has been implicated in outbreaks of DHF in Asia, Africa and the Americas, and has high potential to cause a pandemic of dengue (Messer et al., 2003).

#### **4. Conclusions**

The emergence of dengue virus in Manaus follows the same pattern as to what occurs in other Brazilian cities. Every two to three years there are new outbreaks of dengue and the emergence of new serotypes followed by cases of dengue hemorrhagic. The emergence of DENV-4 in 2008 did not just caused a major epidemic with severe and fatal cases. Other serotypes were also observed. It is possible due to competition between with other serotypes, DENV-4 took some time to become established and the population might have been protected by antibodies obtained in heterologous infections by heterologous serotypes. The viral strain should be studied in detail since it is an Asian genotype that has been associated with DHF in the Asian continent.


**Abstract**

District heating plays an important role in future sustainable energy system by integrating any available heat source, including waste heat and renewable heat sources such as geothermal or solar heat. The low-temperature district heating system is the latest generation of district heating. It was introduced less than ten years ago in adaption to the need for lower heat demand of energy-efficient buildings. The low-temperature district heating system provides an infrastructure for a higher share of renewable energy sources while reduces heat loss in pipes. Several small-scale projects were commissioned since the introduction of the technology, and many existing district heating systems are in the process of adaptation. The recent progress of low-temperature district heating systems has been discussed here. First, the fundamental knowledge that is required to understand the main advantages of a low-temperature district heating system was explained briefly. Then the most recent and important projects were discussed with emphasis on solar and geothermal district heating systems. The results of case studies show that the low-temperature solution has the lowest capital costs and has a unique position to be the primary source for building heating demand.

**Keywords:** district heating, low-temperature, smart heat networks, microgrid, distributed generation, community energy, solar district heating, geothermal, energy storage


2.2.1. Clan-updating Operator

According to the natural habits of elephants, a matriarch leads the elephants in each clan. Therefore, the new position of each elephant *ci* is influenced by matriarch *ci*. The elephant *j* in clan *ci* can be calculated by Equation (1).

$$\mathbf{x}\_{nm\mu;i,j} = \mathbf{x}\_{ci,j} + a \times (\mathbf{x}\_{best,ci} - \mathbf{x}\_{ci,j}) \times r \tag{1}$$

where *xnew,ci,j* and *xci,j* present the new position and old position for elephant *j* in clan *ci*, respectively. *xbest,ci* is matriarch *ci* which represents the best elephant in the clan. *a* ∈ [0,1] indicates a scale factor, *r* ∈ [0,1]. The best elephant can be calculated by Equation (2) for each clan.

$$\mathbf{x}\_{n\mathbf{w},ci,j} = \boldsymbol{\beta} \times \mathbf{x}\_{c\text{center},ci} \tag{2}$$

where β ∈ [0,1] represents a factor which determines the influence of the *xcenter,ci* on *xnew,ci,j*. *xnew,ci,j* is the new individual. *xcenter,ci* is the center individual of clan *ci*. It can be calculated by Equation (3) for the *d*-th dimension.

$$\mathbf{x}\_{\text{center},ci,d} = \frac{1}{n\_{ci}} \times \sum\_{j=1}^{n\_{ci}} \mathbf{x}\_{ci,j,d} \tag{3}$$

where 1 ≤ *d* ≤ *D* and *nci* indicate the number of elephants in clan *ci*. *xci,j,d* represents the *d*-th dimension of elephant individual *xci,j*. *xcenter,ci* is the center of clan *ci* and it can be updated by Equation (3).


*Edited by Bernhard Resch*

Respiratory syncytial virus (RSV) causes seasonal epidemics during the winter and wet seasons, particularly in young children. Although there is one drug available for prophylaxis, palivizumab, it is indicated for high-risk children only. Therefore a vaccine is urgently needed.

This book provides an overview of RSV and discusses its incidence and presentation in different populations, including preterm infants and children with congenital heart disease. It also discusses future strategies for RSV prophylaxis and developments in and barriers to creating a viable vaccine.

Published in London, UK © 2019 IntechOpen © Dr\_Microbe / iStock

The Burden of Respiratory Syncytial Virus Infection in the Young

The Burden of Respiratory

Syncytial Virus Infection

in the Young

*Edited by Bernhard Resch*


**6.1 Clinical diagnosis**

The identification of dogs infested with *E. granulosus* is extremely important for epidemiological studies and monitoring of this zoonosis in control programs [25]. Compared to other gastrointestinal infections of parasitic origin in dogs, echinococcosis is difficult to detect, even if the parasite load is high, premortem diagnosis appears to be very difficult. Recently, there has been considerable progress in research and development of immunological diagnosis for canine echinococcosis. The coproantigen test, in particular, can be considered to be of good sensitivity to reflect a common infestation and can therefore replace purging with arecoline (**Table 2**).

## **6.2 Parasitological diagnosis**

The diagnosis of cystic echinococcosis in ante-mortem dogs can be made using several techniques:

#### *6.2.1 Coproscopy*

Coproscopy consists of looking for the eggs or proglottis of the adult worm in the feces of the final host. Eggs can be detected in fecal samples by the flotation technique or on the perianal skin by attaching a transparent adhesive paper to the skin and examining it under a magnifying glass under a microscope. However, this microscopic detection of *Echinococcus granulosus* eggs is not recommended because of the morphological nature of the egg which is similar in all taenia species. In addition, the removal of eggs is often irregular. However, proglottis from *Echinococcus granulosus* released spontaneously by dogs and detected on the surface of fecal samples can provide a good diagnosis of parasitosis in dogs [9].

#### *6.2.2 Arecoline purging*

Arecoline purging is the standard diagnostic method used for years in the detection of *Echinococcus granulosus* worms in dogs is purging the intestinal contents

#### **Table 2.**

*Comparison of the sensitivity and specificity of different diagnostic methods for canine echinococcosis [12].*

**85**

*Incrimination of Dog Vector of Cystic Echinococcosis and Impact of the Appropriate Dogs'…*

of the host with hydrobromide arecoline. Arecoline is a parasympathomimetic molecule which has an effect on the smooth muscles of the small intestine and at the same time paralyzes adult worms. The purgation removes the paralyzed worms with

The advantage of this technique is the high specificity, which can reach 100%. On the other hand, the sensitivity does not even reach 50% if it is only used once. This test is contraindicated in pregnant dogs, older dogs and puppies. Arecoline should be administered orally at a dose of 4 mg/kg BW. This dose should be carefully calculated since severe undesirable secondary effects may occur [7].

After cutting the intestine into several sections, these must be placed in metal trays, opened with scissors and finally immersed in physiological saline solution. The worms adhering to the mucous membrane are then counted using a magnifying glass or binocular microscope. The disadvantage of this method is that small worms

This technique consists of searching for one of two types of antigens, either antigens extracted raw somatically from the worm or excretory-secretory antigens from the protoscolex in the feces of the host using double sandwich ELISA kits [26]. Positive ELISA results can be collected even in the prepatent period, starting on day 5 post-infestation. The values begin to decrease to negative values 2–4 days after the elimination of *Echinococcus granulosus* worms by treatment with praziquantel. The results of studies using this technique have shown that ELISA values correlate positively with the amount of worms present in the intestine, and that antigen levels are correlated with the amount of worms present in the intestine. This technique has been shown to be important with a sensitivity of 99% and a

Fecal samples can be taken directly from the ground or rectum and can be kept

Specific serum antibodies (IgG, IgA and IgE) can be detected in the serum of dogs infected with *Echinococcus granulosus* using antigenic preparations from the protoscolexes in ELISA kits. These antibodies can be detected 2–3 weeks post-infestation. One study suggests that eggs released in the small intestine of the final host, after proglottis apolysis, can penetrate the intestinal barrier and cause immunologi-

The ELISA kits available have low sensitivity and highly variable specificity. However, a new kit using a newly derived recombinant antigen from the protoscolex showed 100% specificity, but the sensitivity is not comparable to that of older kits. The use of ELISA kits for the detection of serum antibodies is still questionable because of their low sensitivity, the persistence of antibodies in serum after worm

If a seropositive test has been detected but the result is negative for the coproan-

removal and the lack of correlation with infestation pressure [12, 26, 27].

tigens, this is an indication of possible recent exposure [28].

cold (−20°C) for up to 6 months. The test can be used for the identification of infected cases in control program, including pregnant dogs, older dogs and puppies.

*DOI: http://dx.doi.org/10.5772/intechopen.95776*

the feces, which must be inspected afterwards [9].

*6.2.3 Necropsy diagnosis: Sedimentation and counting*

**6.3 Immunological diagnosis by detection of coproantigens**

Three ELISA kits are commercially available today [27].

can escape detection [9].

specificity of 97% [26].

*6.3.1 Serum antibody detection*

cal stimulation in the host [27].

*Incrimination of Dog Vector of Cystic Echinococcosis and Impact of the Appropriate Dogs'… DOI: http://dx.doi.org/10.5772/intechopen.95776*

of the host with hydrobromide arecoline. Arecoline is a parasympathomimetic molecule which has an effect on the smooth muscles of the small intestine and at the same time paralyzes adult worms. The purgation removes the paralyzed worms with the feces, which must be inspected afterwards [9].

The advantage of this technique is the high specificity, which can reach 100%. On the other hand, the sensitivity does not even reach 50% if it is only used once. This test is contraindicated in pregnant dogs, older dogs and puppies. Arecoline should be administered orally at a dose of 4 mg/kg BW. This dose should be carefully calculated since severe undesirable secondary effects may occur [7].

#### *6.2.3 Necropsy diagnosis: Sedimentation and counting*

*Canine Genetics, Health and Medicine*

**6.1 Clinical diagnosis**

**6.2 Parasitological diagnosis**

several techniques:

*6.2.2 Arecoline purging*

*6.2.1 Coproscopy*

(**Table 2**).

dogs (unpublished study) (**Figures 1**–**3**).

**6. Diagnostics of echinococcosis in dogs**

three regions of Morocco revealed that stray dogs were the most infested category by *E. granulosus*, representing a prevalence of 40% in the Middle Atlas Mountains, 33% in Sidi Kacem and 25% in Oulmès, followed by semi-stray dogs and owned

The identification of dogs infested with *E. granulosus* is extremely important for epidemiological studies and monitoring of this zoonosis in control programs [25]. Compared to other gastrointestinal infections of parasitic origin in dogs, echinococcosis is difficult to detect, even if the parasite load is high, premortem diagnosis appears to be very difficult. Recently, there has been considerable progress in research and development of immunological diagnosis for canine echinococcosis. The coproantigen test, in particular, can be considered to be of good sensitivity to reflect a common infestation and can therefore replace purging with arecoline

The diagnosis of cystic echinococcosis in ante-mortem dogs can be made using

Coproscopy consists of looking for the eggs or proglottis of the adult worm in the feces of the final host. Eggs can be detected in fecal samples by the flotation technique or on the perianal skin by attaching a transparent adhesive paper to the skin and examining it under a magnifying glass under a microscope. However, this microscopic detection of *Echinococcus granulosus* eggs is not recommended because of the morphological nature of the egg which is similar in all taenia species. In addition, the removal of eggs is often irregular. However, proglottis from *Echinococcus granulosus* released spontaneously by dogs and detected on the surface of fecal

Arecoline purging is the standard diagnostic method used for years in the detection of *Echinococcus granulosus* worms in dogs is purging the intestinal contents

**Methods Sensitivity % Specificity % Autopsy** > 90 100 **Arecoline purging** 50–70 100 **Serology** 35–70 > 90 **The coproantigen test** 75–80 > 95

*Comparison of the sensitivity and specificity of different diagnostic methods for canine echinococcosis [12].*

samples can provide a good diagnosis of parasitosis in dogs [9].

**84**

**Table 2.**

After cutting the intestine into several sections, these must be placed in metal trays, opened with scissors and finally immersed in physiological saline solution. The worms adhering to the mucous membrane are then counted using a magnifying glass or binocular microscope. The disadvantage of this method is that small worms can escape detection [9].

#### **6.3 Immunological diagnosis by detection of coproantigens**

This technique consists of searching for one of two types of antigens, either antigens extracted raw somatically from the worm or excretory-secretory antigens from the protoscolex in the feces of the host using double sandwich ELISA kits [26].

Positive ELISA results can be collected even in the prepatent period, starting on day 5 post-infestation. The values begin to decrease to negative values 2–4 days after the elimination of *Echinococcus granulosus* worms by treatment with praziquantel. The results of studies using this technique have shown that ELISA values correlate positively with the amount of worms present in the intestine, and that antigen levels are correlated with the amount of worms present in the intestine. This technique has been shown to be important with a sensitivity of 99% and a specificity of 97% [26].

Fecal samples can be taken directly from the ground or rectum and can be kept cold (−20°C) for up to 6 months. The test can be used for the identification of infected cases in control program, including pregnant dogs, older dogs and puppies. Three ELISA kits are commercially available today [27].

#### *6.3.1 Serum antibody detection*

Specific serum antibodies (IgG, IgA and IgE) can be detected in the serum of dogs infected with *Echinococcus granulosus* using antigenic preparations from the protoscolexes in ELISA kits. These antibodies can be detected 2–3 weeks post-infestation. One study suggests that eggs released in the small intestine of the final host, after proglottis apolysis, can penetrate the intestinal barrier and cause immunological stimulation in the host [27].

The ELISA kits available have low sensitivity and highly variable specificity. However, a new kit using a newly derived recombinant antigen from the protoscolex showed 100% specificity, but the sensitivity is not comparable to that of older kits. The use of ELISA kits for the detection of serum antibodies is still questionable because of their low sensitivity, the persistence of antibodies in serum after worm removal and the lack of correlation with infestation pressure [12, 26, 27].

If a seropositive test has been detected but the result is negative for the coproantigens, this is an indication of possible recent exposure [28].

#### **6.4 Molecular biology diagnosis**

Parasite DNA can be obtained from eggs, proglottis or worm cells and can be detected in feces after PCR amplification. However, no copro-PCR is currently available for the detection of all strains of *Echinococcus granulosus*; PCR primers for G1, G5 and combined G6/7 strains have been developed. This technique, due to its high cost, is only used for confirmation on positive samples in areas where the prevalence of cystic echinococcosis is low [29].


**4. Method of data collection**

The data used in this study is a secondary data that was collected from www.ca shcraft.com under stock trend and analysis. Daily stock price was collected on Zenith bank stock price from October 21st 2004 to May 8th 2017.

The returns was calculated using the formula below

$$R\_t = \ln P\_t - \ln P\_{t-1} \tag{29}$$

**Figure 2.**

**Figure 1.**

*The time plot of the log of zenith Bank returns.*

*Financial Time Series Analysis via Backtesting Approach*

*DOI: http://dx.doi.org/10.5772/intechopen.94112*

**Unit root testing**

**ARCH test**

**Table 1.**

**111**

*The time plot of the removal of possible outliers in the log of zenith Bank returns.*

**Statistic Value** Mean 0.000114 Median 0.000000 Maximum 0.338000 Minimum 0.405850 Std. Dev. 0.027600 Skewness 1.267452 Kurtosis 33.76662 Jarque-Bera 121905.9 (p = 0.00000)

Number of Observation 3070

*Descriptive statistics and unit root testing of zenith Bank stock returns.*

ADF 47.11172 (p = 0.0000) DF-GLS 1.842682 PP 46.52078 (p = 0.0000)

Chi-squared = 123.05, df = 12, p-value <2.2e-16

Where Rt is stock returns; Pt is the present stock price; Pt-1 is the previous stock price and ln is the natural logarithm transformation. Then total observation becomes 3070.


Acknowledgements

This work was supported by the National Science Centre in Poland by grant no. 2015/19/B/ST8/02636 (A. Chrobak, G. Ziółkowski). D. Chrobak acknowledges support from the National Science Centre of Poland, grant no. 2016/21/B/ST8/02737.


*2.4. Cueing Action Sequences*

In previous work, we explored the relationship between LEDs and the state of objects in a simple problem-solving exercise [33]. The objective is to ensure that four boxes had satisfied their goals (Figure 9). The goal of each box was defined by a set of rules known to the box, and defined by the position of the box and its proximity to other boxes. Each box has three LEDs representing its state: one to indicate if the goal has been satisfied, one to indicate 'communication status' (in terms of connection with the table), and one to indicate 'proximity'. We were interested in whether people would try to learn the 'rules' that the boxes were using or whether they would find it easier to learn the pattern, or arrangement of the boxes, and whether the rules or patterns could generalize to new configurations. The argument for this comparison was that the patterns could be considered in terms of 'affordance' [34–37]. Not only were the patterns easier to understand (which suggests that the visual cues provides useful semantic information) but also participants found it easier to generalize patterns than the rules (contrary to our expectations). *Sensors* **2017**, *17*, 2308 11 of 16

**Figure 9.** Visual feedback on networked objects [38]. **Figure 9.** Visual feedback on networked objects [38].

### 2.4.1. Procedure 2.4.1. Procedure

drawer and stirring before pouring from the jug.

to obtain timings. The following timings are obtained:

green, spoon lit, spoon stirring cup.

drawer is moved.

spoon picked up.

cup.

debrief.

The trials used the same participants as trial 1. Cues for stirring and pouring were tested with a combination of lights and sounds (from vibration motor). A sequence of actions similar to making a cup of tea was performed purely from the objects' cues. For a 'team making' task, the experimenter raised the handle of the jug and, once the participant had lifted the jug, the experimenter turned the LED on one of the cups to green. When the participant had poured the jug to the cup, the experimenter turned the LED in the drawer handle green and turned on the vibration motor in the handle of the spoon (inside the drawer). The participant opened the drawer and lifted the spoon, and the experimenter turned the LED on the cup (that had been previously used to pour into) green. The trials used the same participants as trial 1. Cues for stirring and pouring were tested with a combination of lights and sounds (from vibration motor). A sequence of actions similar to making a cup of tea was performed purely from the objects' cues. For a 'team making' task, the experimenter raised the handle of the jug and, once the participant had lifted the jug, the experimenter turned the LED on one of the cups to green. When the participant had poured the jug to the cup, the experimenter turned the LED in the drawer handle green and turned on the vibration motor in the handle of the spoon (inside the drawer). The participant opened the drawer and lifted the spoon, and the experimenter

activations but had them appear in an order that did not feel correct, e.g., taking the spoon from the

Each sequence (logical or illogical) was followed by a memory test which entailed placing cards in the same sequence that was performed. The number of mistakes was recorded and participants were not told that they will perform a memory test at the beginning the sequence. Seven cards were used: jug with handle raised, jug tilted (pouring), cup lit green, drawer opened, drawer handle lit

Video footage from experiment three (with participant's permission) was analysed using ELAN

• Time taken to open drawer: Timer begins when light on the handle is on and timer ends when

• Time taken to pick up spoon: Timer begins when drawer fully opened and timer ends when

• Time taken to stir: Timer begins when spoon is picked up till head of the spoon is inside the

Midway through the experiment, only one cup lit up. There were no other audible or visual cues. In the experiment with lights on the jug, the majority of participants performed no action when the cup lit up. On the other hand, in the no lights experiment, the majority of participants picked up the jug and poured into the cup even when the jug's handle was not raised, so participants picked up the jug with two hands and poured. As with the previous trials, participants were interviewed at turned the LED on the cup (that had been previously used to pour into) green. The participant put the spoon into the cup (perhaps making a stirring motion) and the task was completed. This constituted the 'logical' sequence of tea making, and was repeated four times. Additionally, an 'illogical' sequence was also repeated four times: this employed the same object activations but had them appear in an order that did not feel correct, e.g., taking the spoon from the drawer and stirring before pouring from the jug.

Each sequence (logical or illogical) was followed by a memory test which entailed placing cards in the same sequence that was performed. The number of mistakes was recorded and participants were not told that they will perform a memory test at the beginning the sequence. Seven cards were used: jug with handle raised, jug tilted (pouring), cup lit green, drawer opened, drawer handle lit green, spoon lit, spoon stirring cup.

Video footage from experiment three (with participant's permission) was analysed using ELAN to obtain timings. The following timings are obtained:

Midway through the experiment, only one cup lit up. There were no other audible or visual cues. In the experiment with lights on the jug, the majority of participants performed no action when the cup lit up. On the other hand, in the no lights experiment, the majority of participants picked up the jug and poured into the cup even when the jug's handle was not raised, so participants picked up the jug with two hands and poured. As with the previous trials, participants were interviewed at debrief.