Hindawi Veterinary Medicine International Volume 2021, Article ID 6639701, 18 pages https://doi.org/10.1155/2021/6639701

Review Article Rotavirus in Calves and Its Zoonotic Importance

Umer Seid Geletu ,1 Munera Ahmednur Usmael,2 and Fufa Dawo Bari3

1College of Agriculture, Animal Science Department, Oda Bultum University, P.O. Box 226, Chiro, Ethiopia 2West Hararghe Zone, Chiro Wereda, Animal Health Worker, P.O. Box 226, Chiro, Ethiopia 3Department of Microbiology, Immunology and Veterinary Public Health, College of Veterinary Medicine and Agriculture, Addis Ababa University, P.O. Box 34, Bishoftu, Ethiopia

Correspondence should be addressed to Umer Seid Geletu; [email protected]

Received 21 December 2020; Revised 29 March 2021; Accepted 2 April 2021; Published 21 April 2021

Academic Editor: Francesca Mancianti

Copyright © 2021 Umer Seid Geletu et al. )is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Rotavirus is a major pathogen responsible for diarrheal disease in calves, resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by rotavirus in calves in the world are inadequately understood, considering that diarrheal disease caused by rotavirus is a vital health problem in calves that interrupts production benefits with reduced weight gain and increased mortality, and its potential for zoonotic spread. )e pathological changes made by rotavirus are almost exclusively limited to the small intestine that leads to diarrhea. It is environmentally distributed worldwide and was extensively studied. Reassortment is one of the important mechanisms for generating genetic diversity of rotaviruses and eventually for viral evolution. So, the primary strategy is to reduce the burden of rotavirus infections by practicing early colostrum’s feeding in newborn calves, using vaccine, and improving livestock management. Rotaviruses have a wide host range, infecting many animal species as well as humans. As it was found that certain animal rotavirus strains had antigenic similarities to some human strains, this may be an indication for an animal to play a role as a source of rotavirus infection in humans. Groups A to C have been shown to infect both humans and animals. )e most commonly detected strains in both human and animals are G2, G3, G4, and G9, P [6]. )erefore, this review was made to get overview epidemiology status and zoonotic importance of bovine rotavirus.

1. Introduction Eight percent of diarrheic calves tested were positive for at least one of the target enteric pathogens (like bovine Bovine rotavirus is the most recognized pathogen causing coronavirus, bovine rotavirus, bovine norovirus, and bovine acute diarrhea in calves under one month of age worldwide torovirus), suggesting that the infectious factor is still a [1, 2]. It has also been recognized as the major pathogen of major cause of calf diarrhea [6]. )e majority of diarrheic acute diarrhea in both humans and animals. So, it has the cases were identified among 0 to 4-week-old calves. A potential of zoonotic and economic impact [3]. Infection successful dairy and beef farm operation requires that a large appears and spreads rapidly, causing extensive damage to the percentage of cows wean a live healthy calf every year. intestinal lining which results in rapid fluid loss and de- Rearing healthy dairy calves to weaning time requires hydration [4]. Genetic reassortment is one of the important maximizing the calf’s level of immunity against disease, mechanisms for generating genetic diversity of rotaviruses while minimizing its exposure to infectious agent. However, and eventually for viral evolution. )ere is no treatment for among the factors that have been hindering success of dairy BRV, but early and confirmatory diagnosis helps to make and beef industry, morbidity and mortality of calves are the appropriate prevention and control measures, which could ones that cause major concern. Morbidity and mortality are prevent the great economic losses to farmers and the live- important causes of economic losses on dairy farms stock industry [5]. worldwide. In spite of advancement made in dairy and beef 2 Veterinary Medicine International husbandry practices, clinical medicine, and diagnostic a gene, numbered 1 to 11 by decreasing size. )e segmented techniques, the morbidity and mortality rates of dairy and genome can be separated by polyacrylamide gel electro- beef calves are still unacceptably high even on many ad- phoresis (PAGE) to reveal an RNA migration pattern or vanced dairy farms in developed countries [7]. )us, it is electropherotype. )e RNA pattern is both constant and necessary to identify risk factors that are responsible for characteristic for a particular strain and has been widely used dairy and beef calf morbidity and mortality in order to in epidemiological studies for monitoring the transmission design and implement preventive measures. and spread of rotavirus [19]. Rotavirus is one of highly distributed disease agents worldwide and has been extensively studied [8]. In different studies, BRV infection rates of 20–60% in samples of di- 2.1.2. Proteins. )e nomenclature of the viral proteins arrhea have been reported [9]. Prevalence of rotavirus ranges designates the structural proteins as VP and nonstructural from 11.8% to 26.8% in India among diarrheic calves [10]. proteins as NSP followed by sequential numbering from 1 to Also, in European countries rotavirus infection was widely 6 [20]. Analysis of gene encoding segments shows that there examined. In Sweden between 1993 and 2006, estimated are six structural proteins (VP1 to VP4, VP6, and VP7) and prevalence was 24–47% [11], 42% in diarrheal outbreaks in six nonstructural proteins (NSP1 to NSP6). )e structural the UK [12], and 37 to 47.4% in France [13]. In Asian proteins build up the viral particle (Figure 1) and the NSPs countries like Bangladesh, prevalence of rotavirus infection have function in either the viral replication cycle or inter- in calf feces varied from 0 to 7% [14]. In Ethiopia, the action with host proteins to influence the pathogenesis or prevalence of rotavirus was 16.7% [15]. immune response. Each of the 11 segments of dsRNA en- To know the epidemiology status, zoonotic importance, codes a single viral protein except segment 11 which encodes and other related information about rotavirus in calves is two proteins [21]. Figure 1 summarizes the six structural very important to develop different strategies for control and (VP) and six nonstructural protein (NSP). )e functions of prevention of rotavirus infection of calves and humans. )is each protein are summarized in Figure 1 and Table 1. review provides an overview of epidemiological status and )e proteins encoded by the rotavirus genes are well zoonotic importance of bovine rotavirus. )is is needed for established. Except for segment 11, which encodes for two planning a proper control and preventive measure in the proteins NSP5 and NSP6, the rest of the segments encode a country. single protein. )e six viral proteins (VP1, 2, 3, 4, 6, and 7) form the virus particle (virion). VP1 is the RNA-dependent, 2. Rotavirus: Overview RNA polymerase for rotavirus, located in the core of the virus particle [24]. VP2 is a replication intermediate, forms Rotavirus was initially reported in 1972 in Australia [16]. )e the core layer of the virion, and binds the RNA genome while virus was recognized by direct electron microscopy visu- VP3 is an enzyme guanylyl transferase that catalyses the alization in the duodenal biopsies of a child with acute formation of the 5′ cap in the posttranscriptional modifi- diarrhea and named duovirus. )e virus was named rota- cation of mRNA. VP4 determines the rotavirus P serotype as virus because of its characteristic wheel-shaped (rota is a well as host specificity, virulence, and protective immunity; Latin word which means wheel) morphology when seen it also binds to molecules on the surface of cells called re- under an electron microscope [17]. ceptors and drives the entry of the virus into the cell [25]. VP6 is highly antigenic and can be used to identify rotavirus 2.1. Virology of Rotavirus species and it also determines the A-G groupings, and I, II subgroupings of rotavirus. VP7 is a glycoprotein that de- 2.1.1. Structure and Its Genome. Bovine rotavirus (BRVs) is termines the G serotype and that is involved in immunity to a primary etiological agent of calf diarrhea. Rotaviruses are infection [26]. double-stranded RNA (dsRNA) held in the inner core of the )e six nonstructural proteins (NSP1, 2, 3, 4, 5, and 6) three-layered virus. Rotavirus is a nonenveloped virion are only produced in cells infected by rotavirus [21]. NSP1 possessing 11 dsRNA segments which a size range 16∼21 kilo binds interferon regulatory factor 3 and may inhibit in- base pairs within the family Reoviridae and is very stable terferon response during rotavirus infection. In conjunction over a wide pH range with heat liability. )ere are seven with NSP5, NSP2 is involved in the synthesis and packaging serogroups (A-G) of rotaviruses based on antigenic and of viral RNA and creation of viroplasms and is required for genetic similarities of the intermediate capsid protein of genome replication. NSP3 binds viral mRNA at the 3′ end, VP6. Group A rotaviruses are the major cause of rotavirus promotes viral protein synthesis, and is responsible for the infection in domestic animals and, initially known as neo- shutdown of host cell protein synthesis. NSP4 is a viral natal calf diarrhea virus, were one of the first identified viral enterotoxin and induces diarrhea during infection. NSP6 is causes of diarrhea [4]. Most BRVs (95%) belong to group A, an RNA binding protein encoded by gene 11 from an out-of- although groups B and C rotaviruses have also been iden- phase open reading frame [27]. tified in field cases [18]. In comparison to most cellular mRNAs, rotavirus Genome segments code for structural proteins found in mRNAs are unique in that they contain 5′-terminal caps but the virus particle and the nonstructural proteins found in lack 3′-terminal poly (A) tails. During replication, the viral infected cells but not part of the mature particles. )e ge- mRNAs serve two functions: (i) direct synthesis and (ii) nome consists of 18,555 nucleotides in total. Each segment is acting as templates for the synthesis of minus-strand RNAs Veterinary Medicine International 3

Genome Encoded Virion schematic segments proteins (protein locations)

1 2 3 VP1 VP1 4 VP2 VP2 VP3 VP3 VP4 VP4

5

6 NSP1

7 VP6 VP6 8 VP7 VP7 9

NSP2 NSP3 10 NSP4 NSP5 11 NSP6 (not visible, off bottom of gel) Figure 1: Diagrammatic representation of the rotavirus particle and its genome coding. (I) )e migration pattern of 11 dsRNA genome segments of rotavirus on a polyacrylamide gel. (II) Virus proteins encoded by specific genome segments in Section 1. )e proteins were blotted onto a cellulose membrane and detected with rotavirus-specific antibodies. (III) Schematic diagram of rotavirus particle showing the cross section arrangement of viral proteins through the three capsid layers namely: outer (VP4, red; VP7, yellow), inner (VP6, blue) and the inner core (VP2, green). Source: [23].

Table 1: Rotavirus proteins, genome segments, and structural localization. dsRNA Location in virus Numbers of Protein Function segment no capsid molecules/virion dsRNA synthesis (RNA-dependent RNA VP1 1 Core 12 polymerase) VP2 2 Core Inner shell protein 120 VP3 3 Core Capping enzyme 12 VP4 (cleaved to VP5 4 Outer capsid Viral attachment, P-type neutralization antigen 120 and VP8) VP6 6 Inner capsid Middle shell protein 780 VP7 9 Outer capsid G type neutralization antigen 780 NSP1 5 INF antagonist — NSP2 8 Viroplasm formation — Enhance viral mRNA synthesis, associated with NSP3 7 — systemic spread Outer capsid assembly, regulate calcium NSP4 10 — homeostasis, enterotoxin NSP5 11 Viroplasm formation — NSP6 11 Viroplasm formation — Source: [22]. to produce dsRNAs [28]. )e synthesis of dsRNAs is an of the 11 species of viral mRNAs into dsRNAs must be a event that follows or occurs simultaneously with the highly coordinated process [29]. packaging of mRNA templates, as naked dsRNA cannot be Both outer capsid proteins VP7 and VP4 (the spike detected in infected cells. Likewise, the absence of free protein) are targets for neutralizing antibodies. VP4, VP6, dsRNA in the infected cell indicates that dsRNA remains and VP7 play a major role in maintaining viral structure, particle associated once synthesized. Given that the 11 ge- virus attachment, and antigenicity. Although early studies nomic dsRNAs are present in equimolar concentration in implicated VP7 in the cell entry process, subsequent studies both infected cells and virions, the packaging and replication increasingly have indicated that VP4 is the major player in 4 Veterinary Medicine International this process. VP4 is susceptible to proteolysis and viral CxMx-Ax-Nx-Tx-Ex-Hx, where x is an integer, defines the infectivity is increased several folds when VP4 is proteo- genotype of the VP7-VP4-VP6-VP1-VP2-VP3-NSP1- lytically cleaved and facilitates virus entry into cells [22]. NSP2-NSP3-NSP4-NSP5 genes of a given rotavirus strain. During proteolysis, VP4 is cleaved into VP8 ∗ (amino acids 1 Following the advent of hybridization techniques, re- to 247) and VP5 ∗ (amino acids 248 to 776), and the cleavage searchers could investigate the occurrence of reassortment products remain associated with the virion [30]. events between human strains that belong to different genogroups or between human and animal strains which frequently lead to generation of novel rotavirus strains. 2.1.3. Classification and Serogroups. Based on the group Human rotaviruses were classified into two major (repre- specific epitopes localized in an immunodominant site of sented by the Wa and DS-1 reference strains) genogroups VP6 between amino acid residue 48 and 75, rotaviruses have and one minor (represented by the AU-1 reference strain) been divided into five serological species (A-E) and two genogroup [37]. additional tentative species (F and G) according to the )e Wa-like strains are characterized by non-G/P ge- International Committee on Taxonomy of Viruses (ICTV) notypes (I1-R1-C1-M1-A1-N1-T1- E1-H1) and tend to have [31]. )ese rotavirus species are commonly referred to as G/P genotypes G1P [8], G3P [8], G4P [8], or G9P [8] [38]. In rotavirus groups. Rotaviruses belonging to groups A, B, C, contrast, the DS-1-like strains are characterized by non-G/P and H (RVA, RVB, RVC, and RVH, respectively) have been genotypes (I2-R2-C2-M2- A2-N2-T2-E2-H2) and tend to associated with acute gastroenteritis in humans and animals, have G/P genotype G2P [4]. )e third minor AU-1-like whereas groups D, E, F, and G (RVD, RVE, RVF, and RVG, strains are characterized by non-G/P genotypes (I3-R3-C3- respectively) rotaviruses are known to infect only animals, M3-A3-N3-T3E3-H3) and tend to have G/P genotype G3P mostly birds [32]. A novel tentative group I was recently [9]. Whole genome-based analysis is a reliable method for described in Hungarian sheltered dogs [33]. Table 2 is obtaining conclusive data on the origin of an RVA strain and summarized the rotavirus group with respective host species. for tracing its evolutionary pattern [36]. RVA of VP7 and Group A rotaviruses (RVA) can be further classified into VP4 genotypes with their respective host species are sum- P or G types based on genetic and antigenic similarities of marized in Table 3. VP4 and VP7. VP4 (P protein for “protease-sensitive” due to Rotavirus surveillance also generates valuable data on its trypsin mediated cleavage required for virus adsorption the circulating rotavirus strains (Table 4). )ese data are vital into cells) determines the P serotypes. VP7 (G protein for to improving vaccine development tracking emergent types, “glycoprotein” forming the matrix of the capsid) defines G and helping to assess vaccine effectiveness, and changes in serotypes [26]. For G types, serotypes (determined by strain diversity after vaccines are introduced. Globally, G1, neutralization assay) and genotypes (determined by RT- G2, G3, G4, and G9 are the most prevalent VP7 serotypes; P PCR) are largely identical, thereby allowing the use of the [4], P [6], and P [8] are the most common VP4 genotypes, same numbering system. For P types, more genotypes than and G1P [8], G2P [4], G3P [8], G4P [8], and G9P [8] serotypes have been identified, owing to lack of monospe- comprise 70–90% of circulating rotavirus strains [39, 41]. In cific P antisera. As a result, P types are identified as serotypes Taiwan, G1 (40%), G3 (27%), G9 (18%), and G2 (8%) are the by Arabic numbers and as genotypes by Arabic numbers in most common VP7 serotypes [42]. G6 and G10 type are square brackets. )us, the serotype of prototype human reported to be the most prevalent in cattle [43]. )e geo- rotavirus strain Wa is described as G1P [8]. To date, at least graphic distribution of rotavirus serotypes is summarized in 27 G types and 37 P types have been found in humans and Table 4. animals [31, 35]. Unlike P types, correlation between G serotypes and genotypes is complete. )erefore, where available, P serotypes and genotypes are designated jointly 2.1.4. Reassortment and Antigenic Variation. Reassortment is with genotypes in square brackets, for instance, RVA/Hu- one of the important mechanisms for generating genetic man-tc/USA/DS-1/1976/G2P1B [4] [31]. diversity of rotaviruses and eventually for viral evolution. Although the dual typing system has been widely used in Although host species barriers and host range restriction most epidemiological and molecular characterization exist in rotavirus, reassortment can result in interspecies studies, its use is primarily limited to classifying rotavirus transmission, which also contributes to the diversity and strains. )e dual typing system cannot determine factors that evolution of rotavirus. A crucial factor in the generation of are involved in viral tropism and virulence of rotavirus reassortant viruses is the frequency of coinfection. In de- strains. Furthermore, some evolutionary pathways like veloping countries, the rate of RV coinfection can be as high reassortment and recombination followed by all the 11 as 20%, while in developed countries, the rate is typically less genome segments of rotaviruses cannot be studied because than 5% [44]. It may be because of the high rate of coin- the dual classification is restricted only to outer capsid fection that the genetic diversity of viruses in developing encoding genome segments [36]. countries can be so much higher than in developed countries In addition to the G and P genotyping of rotavirus, a [44]. Due to the high frequency of coinfection, large ge- whole genome-based genotyping system was recently pro- netically distinct RV clades may not be detectable in some posed based on the assignment of genotypes to all the 11 developing countries [44]. gene segments (i.e., G/P and non-G/P genes) [36]. In the Sequence analysis has shown that the antigenic epitopes new genotyping system, the acronym Gx-P [x]-Ix-Rx- of VP7 and VP4 proteins assigned to the same G and P type, Veterinary Medicine International 5

Table 2: Rotavirus group detected so far in different mammalian shown to be released very early during an infection, first as a and/or avian host species. cleavage product including the toxic region released from Rotavirus group/ infected cells, starting at 4 hours after infection and later Host species species during infection as fully glycosylated NSP4. Based on cell A wide variety of mammalian and avian culture studies, the general steps of rotavirus replication are A species as follows [48] (Figure 2). B Humans, cattle, goats, pigs, rats, and sheep Virus attaches to the cell surface by VP4 or the cleavage Humans, cattle, dogs, goats, juvenile ferrets, product VP8. )e conformational change is protease-de- C and pigs pendent, where VP4 is cleaved into VP8 and VP5. Rotavirus D Chicken and turkey has tropism for mature enterocytes but the exact receptor for E Pigs viral binding in vivo has not yet been identified, although F Chicken sialic acid, integrins, histo-blood group antigens [49, 50], G Chicken and toll-like receptors (TLR) have been suggested. Cell entry, H Humans and pigs by receptor-mediated endocytosis occurs via VP5, thus Source: [34]. indicating that cleavage of VP4 into VP5 and VP8 is re- quired. Calcium dependent endocytosis has also been shown. Nonclathrin, noncaveolin-dependent endocytosis Table 3: Common RVA G and P genotypes found in humans and delivers the virion to the early endosome. It has also been animals. suggested that rotavirus can enter the cell by direct entry or Host species Typical RVA VP7 and VP4 genotypes fusion. Uncoating of the TLP, reduced calcium concentra- tions in the endosome are thought to trigger the uncoating of Cattle G6, G8, G10, P [1], P [5], P [11] Pigs G3-G5, G9, G11, P [6], P [7] VP7 and loss of the outer capsid (VP7, VP5, and VP8). Horses G3, G14, P [12] Double-layered particles (DLP) (core proteins and inner Cats and dogs G3, P [3], P [9] capsid VP6) are released into the cytosol [51]. Humans G1-G4, G9, G12, P [4], P [6], P [8] Transcription and translation take place in the cytoplasm Source: [34]. of the cell. )e internal polymerase complex (PC) (VP1 and VP3) starts to transcribe capped (+) RNAs from each of the eleven dsRNA segments. (+) RNA serves either as mRNA for respectively, will frequently show amino acid variation [45]. direct translation, synthesis of viral proteins by cellular ri- )is has been seen for VP7 and VP4 proteins of viruses bosomes, or as a template for (−) RNA synthesis of viral recovered from different countries in the same year or that genome replication, taking place in viroplasm. Assembly is belong to different cocirculating clades at one site. Such the NSP2 and NSP5 interact to form viroplasms, where amino acid variation may ultimately have an impact on replication and sub-viral particle assembly takes place. DLPs vaccine efficacy, particularly if protection is based chiefly on are formed within the viroplasms. )e assembly process of G and P type specific homotypic responses. )e effective titer the outer capsid is not fully understood but it is thought that of a G type specific neutralizing antiserum is affected by the the transmembrane protein NSP4 recruits DLPs and the amino acid composition of VP7 antigenic epitopes, even if outer capsid protein VP4 to the cytosolic side of the ER the VP7 proteins are of the same G type [46]. membrane. )e NSP4/VP4/DLP-complex then buds into ER. )e removal of the ER membrane and NSP4 takes place in the ER through interaction with ER-resident VP7 and the 2.1.5. Replication. Viruses interact with the host at all stages final TLP is formed. Virus release from the infected cell is of replication: cell entry, viral transcription, translation, through cell lysis or Golgi-independent nonclassical vesic- genome synthesis and packaging, and cell exit. )ese in- ular transport. In the GIT, the virion will be exposed to teractions not only are important for producing new virus trypsin-like proteases, which will cleave the protease-sen- progeny, but also enable the host to recognize the presence sitive VP4 into VP5 and VP8, thus resulting in a fully in- of an infectious agent. As host species have evolved fectious virion [48]. mechanisms to defend against pathogens, viruses have in turn evolved strategies to avoid the host immune response [47]. 2.2. Epidemiology of Rotavirus and Geographical Distribution Rotavirus replication takes place in the cytoplasm of infected cells, in viroplasms, electron dense structures near 2.2.1. Epidemiology of Rotavirus in Animals. Rotavirus can the nucleus and ER [48]. Newly made viruses budded out cause a diarrhea and lead is a serious welfare problem in from viroplasms into ER, through binding to the tail of the calves and a cause of economic loss due to mortality, ER transmembrane viral glycoprotein NSP4. Although the treatment costs, and poor growth. Rotavirus is highly in- virus replication process includes synthesis and transport of fectious because (1) virus particles are present in very large glycoproteins, the Golgi apparatus is not involved in rota- numbers (1010–1012 particles/ml) in infected feces and (2) the virus replication. Instead, rotavirus replication, morpho- virus is resistant to inactivation and can remain infectious genesis, and pathogenesis are regulated by intracellular for 9 months at room temperature or for 1 hr at 60°C. calcium concentrations. )e rotavirus toxin NSP4 has been Furthermore, rotaviruses are not easily inactivated by the 6 Veterinary Medicine International

Table 4: Geographic distribution of rotavirus serotypes. Rotavirus serotypes Region G1P [8] (%) G2P [4] (%) G3P [8] (%) G4P [8] (%) G9 (%) Other (%) North America 73 11 6 1 3 5 South America 34 23 2 9 16 11 Europe 72 9 2 11 4 1.4 Australia 82 14 1 2 0.5 0.1 Asia 34 13 1 20 12 14 Africa 23 2 21 4 7 27 Taiwan 40 80 27 0 18 8 Sources: [39, 40].

Attachment Protease-primed conformational change

Integrin VP7 VP5 I domain VP5 VP7 VP8 Sialo DGE 2 1 v 3 × 2 hsc70 Endocytosis receptor Lipid raft in the cell membrane

Golgi-independent Transcription nonclassical 2+ vesicular [Ca ]i transport

VP7 Loss of Translation envelope and NSP4 VP4 particle Budding and maturation transient enveloped particle

ER Double-layered NSP2, 5, 6 particle VP1, 2, 3, 6 Replication intermediate and double-stranded RNA synthesis LC3 Cell lysis Viroplasm Nucleus Lipid droplet

Figure 2: )e rotavirus replication cycle. Source: [32]. commonly used disinfectants. Rotavirus surviving in a that 28 percent of all under-five diarrheal disease hospi- contaminated environment from one calving season to the talizations in Ethiopia are caused by rotavirus [64]. Also next may therefore be the source of infection in an outbreak. some study showed, among children <5 years of age, ro- However, adults are the major source of infection for calves. tavirus prevalence ranging within 18%–28% of diarrhea Whatever the source of the virus, infection spreads pre- hospitalizations [65]. In a cross-sectional study carried out dominantly by fecal-oral contact [52]. Calves most often in Jima Hospital, Ethiopia, to reveal the prevalence of ro- become infected with rotavirus during the first week of life. tavirus infection among 154 infants and young children, Table 5 summarizes some study of rotavirus in animals in rotavirus was detected in 26.6 % of fecal specimens and different parts of the world. 90.2% (37/41) occurred in children under 2 years. )e highest rate of rotavirus antigen detection was observed among the 7–12 months age group (34%) [66]. 2.2.2. 2e Status of Rotavirus in Human and Animals in In a study to see the epidemiology of rotavirus and Ethiopia. Ethiopia is one of the five countries with the norovirus in Awassa, southern Ethiopia from 200 under-five greatest human rotavirus burden worldwide and accounts children with diarrhea 2008-2009, the prevalence of rota- for 6% of all rotavirus deaths globally [63]. It is estimated virus was 22% and the genotyping showed G3P [6] (48%, Veterinary Medicine International 7

Table 5: Prevalence of rotavirus infection in animals. Country Prevalence rotavirus (%) Reference Western Algeria 14.63 Ammar et al. [53] Northern India 26.8 Jindal et al. [54] Ethiopia 16.7 Abraham et al. [15] India 15.68 Rai et al. [55] Iraq 15.5 Al-Robaiee & Al-Farwachi [56] Brazil 20.2 Alfieri et al. [1] Tunisia 22.8 Zrelli et al. [57] Brazil 25.1 Langoni et al. [58] Algeria 21.84 Kam et al. [59] England 42 Reynolds et al. [12] Scotland 50 Snodgrass et al. [60] Spain 42.7 De la Fuente et al. [61] Australia 79.9 Izzo et al. [62]

globally uncommon strain), G1P [8] (27%), and G2P [4] decreased absorption of salt and water related to selective (7%) being the strains most commonly identified. Data from infection of the absorptive intestinal villous cells, resulting in hospital-based surveillance of rotavirus gastroenteritis net fluid secretion. )e main place for rotavirus infection is among children less than five years from 2007 to 2011 in brush border of villous epithelial cells in the small intestine. Addis Ababa, Ethiopia, showed that rotavirus was prevalent )e infected cells are rapidly replaced with undifferentiated in 20% of children enrolled from 1,749 diarrheal samples crypt cells, and this results in reducing activity of lactase in collected in the five-year period. Another study showed the villous [77]. prevalence of rotavirus 25% in children less than five years in )e primary mode of transmission of rotavirus is fecal- northwest Ethiopia by Gelaw et al. [67]. Only two reports oral, although some studies have reported low titers of virus were found in Ethiopia by Abraham et al. [15] and Geletu in respiratory tract secretions and other body fluids, indi- et al. [68] that indicated presence of 16.7% and 7.2% in calves cating the possibilities for airborne and waterborne trans- in central Ethiopia, respectively. missions of rotavirus [78]. After ingestion, the rotavirus particles exclusively infect the mature differentiated enterocytes in the middle and upper part of the villi of the 2.3. General Pathophysiology. )e severity and localization small intestine leading to structural changes in the intestinal of rotavirus infection vary among animal species and be- epithelium [69]. )e virus replicates in the cytoplasm of tween studies, but pathological changes are almost exclu- epithelial cells of the mature absorptive and enzyme pro- sively limited to the small intestine. Rotavirus infects the ducing enterocytes of small intestinal villi. Destruction of mature nondividing enterocytes in the middle and top parts mature entrecotes in the villi leads to rupture and sloughing of the villi in the small intestine [69]. At the cellular level, the of the enterocytes with release of virus to infect adjacent infection is characterized by vacuolization, blunting, and cells. Unlike the parvovirus, rotavirus can infect neither the shortening of the villi. Rotavirus also produces the en- immature villous crypt cells nor the colonic enterocytes. terotoxin NSP4, which is thought to play an important role Rotavirus attaches to its cellular receptors (sialoglycoprotein in the pathophysiology and clinical symptom of rotavirus and integrins) via the VP4 protein. )e virus is thought to disease [70–72]. )e incubation time is 24 to 48 hours and invade target cells in two possible ways, by direct entry or illness usually lasts from 3 to 5 days, longer in immune- fusion with enterocytes and through Ca2+-dependent en- compromised individuals [73]. )ere are few pathology docytosis [79]. studies of the duodenal mucosa of infants infected with Rotavirus may cause diarrhoea by three different rotavirus. Biopsies have displayed shortening and atrophy of mechanisms. First, within 12–24 hours after infection, villi, distended endoplasmic reticulum, mononuclear cell enterocytes are intact but the levels of the brush-border infiltration, mitochondrial swelling, and loss of microvilli disaccharidases (sucrase, maltase, and lactase) are greatly [74]. Systemic spread of rotavirus has been reported but is reduced. As a result, disaccharides in the diet cannot be very rare and its clinical importance remains unclear. In a hydrolysed to monosaccharides and thus cannot be absor- few cases, rotavirus RNA has been detected in cerebrospinal bed, leading to osmotic diarrhoea [21]. Second, NSP4 has an fluid (CSF) [75], possibly associated with meningitis, en- effect in opening calcium channels in the enterocytes. )is cephalopathy, and encephalitis [76]. causes an efflux of sodium and water, producing secretory diarrhea [80]. Finally, the higher intraenterocyte calcium 2.4. Pathogenesis of Rotavirus Infection. Bovine rotaviruses concentration causes enterocytes to die by oncosis. )e rate group A are enteropathogenic agents more commonly as- of death of the mature villous tip enterocytes exceeds the rate sociated with neonatal diarrhea in calves up to 30 days old [1]. of growth of immature enterocytes that are regenerated from )e mechanism of rotavirus-induce diarrhea is not com- the stem cells in the crypt, causing villous blunting and thus pletely known. )e major mechanism appears to be a malabsorption [22]. Infection resolves both as the virus runs 8 Veterinary Medicine International out of susceptible mature enterocytes and an immune re- 2.7. Clinical Features of Rotavirus Infection sponse is generated [69]. )e sign of emesis, which is a hallmark of the rotavirus 2.7.1. Symptoms in Animals. Rotavirus diarrhea in calves disease, is caused by serotonin (5-hydroxytryptamine, 5- presents an acute disease having very short incubation HT). 5-HT is secreted by enterochromaffin cells (EC) that period of 12–24 hours or at times ranging within 18–96 can be directly infected with and replicate rotaviruses in hours. Fortunately, most rotavirus infections are mild and humans. )e 5-HT activates vagal afferent nerves connected self-limiting, although there is usually high morbidity. to the nucleus of the solitary tract and area postrema in the Variations in clinical disease observed in calves depend on a brainstem structures associated with nausea and vomiting number of factors, including difference in virulence among [65]. rotavirus strains, age of the host, host immune status, dose of the inoculum, occurrence of mixed infections, environ- mental stress (weather conditions, housing, overcrowding), 2.5. Immune Response to Rotavirus. )e mechanisms re- and nutrition, in addition to systemic consequences of sponsible for immunity to rotavirus infections are not electrolyte imbalances, fluid loss and metabolic acidemia, completely understood. Animal models have been useful in anorexia, profuse watery diarrhea, and various degrees of elucidating the role of antibodies and in exploring the rel- systemic dehydration. In severe cases, death occurs as a ative importance of systemic and local immunity [81]. In result of electrolyte imbalances, dehydration, and cardiac humans, rotavirus infection has been shown to induce a arrest [88]. good humoral immune response and protection increases with each new infection and reduces the severity of the diarrhea [82]. 2.8. Transmission. Rotaviruses are highly contagious, Primary rotavirus infections induce production of ro- ubiquitous in the environment, and relatively resistant to tavirus-specific memory B and T cells [82]. Since the im- disinfectants. )e adult animals are the main source of munity against severe diarrhea in humans resulting from infection in newborn animals, and serological surveys series of childhood rotavirus infections often wanes with age, revealed that 50–100% of adult animals might show immune elderly persons become more susceptible to rotavirus re- response against RVA. Young calves, especially aged 1–3 infection [83]. )e significance of the systemic presence of weeks, are most vulnerable to the rotavirus infection and IgA, IgG, and IgM antibodies towards protection against infection rate declines as age of calf increases [89]. )e rotavirus infection in both humans and animals remains to infectious dose is low (as few as 10 particles) [90]; and the be understood [81, 84]. However, it is known that maternal virus is shed in large quantities (as many as 1011 particles per IgG antibodies may play a role in protecting infants under gram of stool) both before the onset of symptoms and for the age of three months from developing severe diarrhoea several weeks afterward. )e virus transmits through a fecal- caused by rotavirus infections as evidenced by the neu- oral route and calves are most often infected by contact with tralizing activity of antibodies detected from transitional other calves, primarily or secondarily through objects, feed, milk and colostrum specimens [85]. Protection of neonates and water. It has been proposed that calves can also be against rotavirus infection appears to be conferred by both infected by virus shed by the dam at birth. )e infected transplacental acquired maternal antibodies and by anti- calves shed virus through the feces from the second day of bodies and other factors in breast milk. Interestingly, ro- infection and the shedding may last for 7-8 days. )e virus tavirus infection in neonates often results in asymptomatic primarily affects neonatal individuals, and calves more than infection unless novel serotypes emerge, and rotavirus can 3 months of age are usually not affected. Rotavirus that circulate silently in neonatal units [86]. infects calves causes often severe and sometimes life- threatening diarrhea [77]. 2.6. Factors Affecting Disease Severity. )e factors that in- Transmission to susceptible individuals occurs mainly by fluence the severity of the disease as well as pathogenesis are the fecal-oral route through direct contact with the rotavirus, reduced intake of colostrum, age and health status of the including children and adults with asymptomatic illness and calves, immune status of the dam, degree of exposure and contact with contaminated fomites, food, water, and envi- virulence of virus, and the presence of secondary pathogens ronmental surfaces [91, 92]. It has been reported that im- [87]. If rotavirus infection occurs in combination with E. coli provements in hand hygiene in hospitals can decrease the or coronavirus, the mortality rate could be high. Several other incidence in healthcare-associated rotavirus infections. It factors like dehydration, unhygienic environment, tempera- has also been suggested that aerosol transmission might be ture variations or chilling during winter, and high population important. Evidence of the airborne spread of rotavirus density in farms may also enhance disease severity. However, gastroenteritis is primarily circumstantial, including the the major stress factors that potentiate the infection have been short incubation period (1–3 days) and the fact that the virus found to be cold climate and marked fluctuations in the often presents in explosive outbreaks [78]. Rotavirus has also ambient temperature between day and night. An age-related been detected in the respiratory secretions from a small resistance has also been observed. As there is competition number of patients, and cases of pneumonia have been between the rate of replication of rotavirus and replacement of described. Rotavirus epidemics exhibit a seasonal pattern enterocytes in older animals, highly virulent strains can only [93]. In temperate climates, rotavirus infections peak in the cause diarrhea in adult calves [77]. winter months. Seasonality is less marked closer to the Veterinary Medicine International 9 equator, but the disease is more common during drier and generally used in a thin-section EM on fixed tissues. In cooler months. Recent data suggest that the seasonality of comparison, IEM has a higher sensitivity and specificity than rotavirus could have been changed by the introduction of direct EM as a specimen is incubated with antibody specific rotavirus vaccines [94, 95]. for the target virus in order to agglutinate the virus before staining. )e visualization of viruses, particularly non- cultivatable ones, is a major advantage of EM with rapid 2.9. Diagnosis of Rotavirus. Laboratory diagnosis of rota- turnaround. Most of bovine enteric viruses, such as BRV, virus is very important for management and control of BToV, and BCV, are difficult to isolate or propagate in cell outbreak of disease related with rotavirus infection in calves. culture, but these viruses can be differentiated by their Viral gastroenteritis is caused by different types of viral morphology under an electron microscope. )e cost of antigens like coronavirus, noroviruses, astroviruses, and electron microscopes and requirement of skilled laboratory adenoviruses. It is very difficult to diagnose specific causal personnel are still a challenge for the EM test being used as a agents by clinical examination, so laboratory diagnosis is routine diagnostic test [6]. vital for confirmatory diagnosis. )is can be carried out by using various tests [5]. Rapid and accurate detection of the etiological agent is important to further contain the spread of 2.9.3. Isolation of Virus in Cell Culture. Virus isolation test is infection in animals. Rotavirus is shed in high concentration a confirmatory diagnostic test that is still considered as “gold in the stool (∼1012 viruses/gram) of children with gastro- standard” for detecting the presence of viral pathogens in enteritis. )erefore, measurement of rotavirus antigen in the specimens [6]. Cell culture techniques are commonly used stool has been used to identify rotavirus infected patients. for virus isolation for diagnostic purpose, as well as virus Generally, the diagnosis of rotavirus is based on isolation propagation for vaccine production or further virus char- and identification of the virus in intestinal contents or feces acterization such as antigenic variation or gene sequencing [88]. Isolation of rotavirus has been performed in rotavirus- [100]. )e isolation of rotavirus in cell culture from fecal specific cell line MA-104 (Simian origin), and direct de- samples is the most conventional way of confirmatory di- tection has been facilitated by electromicroscopy. Immu- agnosis of rotavirus infection and gives the ultimate proof of nofluorescence test (IFT), immunoperoxidase test (IPT), virus association with the disease but it is less sensitive and is and viral RNA-based PAGE have also been employed to a laborious process. Isolation of BRV is performed in ro- detect the infectious agent. Latex agglutination test (LAT) tavirus-specific primary cell cultures (calf kidney cells) and has also been used for the rapid detection of rotavirus an- cell lines (MA 104-Simian origin, MDBK, HT-29, and PK- tigens [96, 97]. ELISA, being a highly sensitive and specific 15). Presence of virus is suspected by occurrence of cyto- test, has been developed by many workers and used for the pathic effect (CPE) including rounding and detachment of identification of rotaviruses [18]. cells in cell culture system. Enhancement of CPE has been shown to be increased by incorporation of trypsin in the medium in minute quantities and by the pretreatment of 2.9.1. Antigen Capturing Enzyme-Linked Immunosorbent fecal samples with trypsin [87]. )e viability of target virus in Assay (Ag-ELISA). Ag-ELISA is an assay for rapidly a specimen is critical for the success of virus isolation [101]. detecting a pathogen in a clinical specimen based on anti- Specimens should be kept at a low temperature and in a body (e.g., monoclonal antibody) recognition of the target transport medium during shipping to a diagnostic labora- antigen [98]. It has antibody attached to a solid surface tory and delivered to the lab as soon as possible after col- which can be a glass, plastic material, or membrane filter. lection [101]. )is antibody captures the target antigen if present in the sample. )en, there will be a cascade of colorimetric reac- tions to verify capturing of the antigen and visualize the 2.9.4. Rotavirus dsRNA PAGE. )e rotavirus dsRNA can be antigen-antibody reaction. Antigen can be quantitatively detected in clinical specimens by extraction of viral RNA and estimated as optical density (OD) measured by a spec- analysis by electrophoresis on a polyacrylamide gel followed trometry positively correlates with the amount of antigen. In by silver staining. During electrophoresis, the 11 segments of some situations, the commercial kit may be expensive, the rotavirus dsRNA, which are negatively charged mole- particularly for veterinary medicine [5]. cules, separate according to size [102]. )e patterns of dsRNA can be visualized in the gel by staining with silver nitrate, because silver ions form a stable complex with 2.9.2. Electron Microscopy (EM). Electron microscopy (EM) nucleic acids. )e gel can be stored after staining. )e is used for virus detection and identification based on migration patterns of the segments of rotavirus dsRNA allow morphological characteristics. )ere are two types of EM the classification of rotavirus strains into the “short” and methods: direct EM and immune-electron microscopy “long” electropherotypes [6]. (IEM) [99]. Two different staining techniques (positive and negative staining) are used to visualize the presence of target. In the direct EM, virus particles in a fluid sample matrix are 2.9.5. Polymerase Chain Reaction (PCR). Polymerase chain applied directly to a solid support and then are visualized by reaction (PCR) is frequently used test method for detecting EM after a contrast stain is applied. It is commonly referred rotavirus. It is a thermocyclic enzymatic amplification of to as “negative straining EM,” whereas positive staining is specific sequence of the target genes using a pair of 10 Veterinary Medicine International oligonucleotide primers that hybridize on each cDNA strand 2.9.7. Real-Time PCR. Real-time PCR is a PCR method of interest region in the genomic sequence. )e detection of which amplifies the target sequence and also quantifies the rotavirus dsRNA in fecal specimens consists of 4 steps: (i) amount of the target with higher sensitivity. Real-time re- viral dsRNA extraction, (ii) denaturation of the rotavirus verse transcription-PCR is a high-throughput robust easy- dsRNA, (iii) reverse transcription of dsRNA, and (iv) am- to-perform, quantitative, sensitive, and specific assay to plification of cDNA by PCR; PCR consists of (a) heating the detect viral nucleic acids [108]. Multiplex real-time PCR DNA to be amplified to separate the template strands, (b) based on SYBR Green and TaqMan assay have been de- annealing of two primers that are complimentary to the veloped for detection of group A human rotavirus. Multiplex region to be amplified, (c) extension of the primers by a heat real-time PCR has also been described to detect rotavirus stable DNA polymerase enzyme that uses each DNA strand along with other enteric pathogens in bovine fecal samples as template, and (d) repeating the process 30–40 times with [109]. Compared to conventional RT-PCR, real-time RT- the newly synthesized cDNA heat denatured and the en- PCR has been shown to be more rapid and more sensitive for zymes extending the primers attached to the separated single the detection and quantitation of rotavirus [106, 110]. For DNA strand. After completion of the reaction, the PCR rapid diagnosis of rotavirus in fecal samples, a SYBR Green products can be visualized on an agarose or acrylamide gel based real-time PCR assay was developed targeting the NSP4 by electrophoresis technique and special staining with gene [106]. ethidium bromide. Amplification of the target sequence is One of the primary advantages of real-time PCR is the determined based on molecular size and/or sequencing of ability to identify amplified fragments during the PCR the PCR product [5]. process. Real-time PCR measures the amount of the product. Standard PCR requires post-PCR analysis, possibly agarose gel electrophoresis. )e use of probe hybridization is often 2.9.6. Rotavirus Genotyping Using RT-PCR. With regard to used for characterization of the product by its sequence. reverse transcription-polymerase chain reaction (RT-PCR), )ough this method is more reliable and informative, it is ever since the initial report by Kary Mullis and coworkers in time-consuming and expensive. ELISA detections are also 1986 about in vitro enzymatic amplification of specific DNA time-consuming. Real-time PCR eliminates these needs. fragments from complex nucleic acid samples using PCR, a Amplicon recognition is achieved by monitoring the ac- number of different applications of the technique have grown cumulation of specific products during each cycle. Another exponentially. A novel G-typing was first reported by using advantage of real-time PCR over standard PCR is that the method based on RT-PCR amplification of the VP7 gene with entire process from amplification to analysis is performed in type-specific primers [103]. In addition to this, some study the same tube. )is differs from standard PCR where the used RT-PCR for serotyping of rotavirus virus and reported PCR product is moved and manipulated into other formats. that six VP7 serotypes or G types (G1-G4, G8, and G9) occur As a result, there is a decreased possibility of contaminating in group “A” human rotaviruses [104]. In their study, they the product with real-time PCR methods [106, 110]. could type about 89% of the samples [104]. Another report compared the tests and used PCR for identifying serotypes of 2.9.8. Restriction Fragment Length Polymorphism (RFLP). human and bovine rotaviruses, and PCR was shown to be Restriction Endonuclease (RE) analysis of field rotaviruses is a more sensitive (93%) than ELISA (82%) [105]. powerful tool to understand genomic diversity of rotaviruses RT-PCR is more sensitive (100%) and specific (99%) in circulating in environment. Apart from proving useful in comparison to ELISA and PAGE. As against RNA elec- monitoring the extent of genetic variation among rotavirus trophoresis and ELISA, it provides a more accurate detection strains within a population, RFLP may also prove valuable in of rotaviruses by 18.8% and 26.5%, respectively. In recent the examination of interspecies transmission and possible reports, it has been shown that increased detection and source of origin of rotavirus strain. Chang et al. [111] used quantification of group “A” rotavirus can be done by real- RFLF for P and G genotyping of bovine rotavirus A. Gouvea time RT-PCR. For easy screening of the fecal samples for et al. [103] analyzed 194 strains of rotavirus A representing all rotavirus A, a diagnostic RT-PCR assay was developed by known G types digestion with three restriction enzymes targeting the group specific VP6 gene [106]. (Sau96I, BstYI, and HaeIII) by direct digestion of amplified One researcher developed a one-step multiplex RT-PCR cDNA copies or by deduction of the restriction patterns from method for the simultaneous detection of five viruses causing known sequences. Digestion with Sau96I and HaeIII iden- diarrhoea in adult cattle, i.e., bovine group A rotavirus (ro- tified restriction sites commonly used for all, or mostly for all, tavirus A), bovine group B rotavirus (rotavirus B), bovine strains of rotavirus studied, whereas BstYl was the most group C rotavirus (rotavirus C/GCR), bovine coronavirus discriminating among rotavirus strains. (BCV), and bovine torovirus (BToV) [107]. In this report, the one-step multiplex RT-PCR was found to have higher sen- sitivity to detect rotavirus A than a single RT-PCR with 2.9.9. Reverse Transcription Loop-Mediated Isothermal conventional primers. )e results indicate that the one-step Amplification (RT-LAMP). Nemoto et al. [112] developed multiplex RT-PCR developed can be used for the detection of RT-LAMP for detection of equine rotavirus targeting P [12], rotavirus A, rotavirus B, rotavirus C, BCV, and BToV and can the most predominant P genotype worldwide. )e results be expected to be a useful tool for the rapid and cost-effective indicated that the RT-LAMP assay was specific for equine diagnosis and surveillance of viral diarrhea in adult cattle [94]. rotavirus and was found more sensitive than semi-nested Veterinary Medicine International 11

RT-PCR. Because RT-LAMP is easy to perform without the had antigenic similarities to some human strains, specula- need for a thermal cycler or gel electrophoresis, the RT- tion increased about whether animals play a role as a source LAMP assay should be applicable to diagnosis of equine of rotavirus infection in humans. )ere is however an al- rotavirus infections in diagnostic laboratories. ternative view that animal rotaviruses can indeed infect humans and cause disease whenever the chance exists. )is is based on the identification of unusual rotavirus types, with 2.9.10. Hybridization Assays. )e assessment of the genetic properties of strains more commonly found in animals, variability of rotavirus by hybridization assay, including blot which were isolated from various cases of human infection. techniques such as northern and southern blot and also )ese unusual human rotavirus types may have arisen either liquid assays, has been an alternative approach to PCR as- as whole virions or as genetic reassortants between human says. Most northern blot and liquid hybridization assays and animal strains during coinfection of a single cell [120]. have utilized cDNA or ssRNA probes synthesized from all )e segmented nature of the genome suggests that, like other segments in a single hybridization reaction and thus limit the viruses with segmented genomes such as influenza virus, amount of segment specific information available from the rotaviruses are able to form new strains by a mechanism of test [113]. Nonradiolabeled cDNA probes have been used for reassortment. Reassortment can occur when two rotaviruses G and P genotyping of bovine rotavirus A [114]. of two different strains infect the same cell, and during replication and packaging they exchange genome segments 2.9.11. Latex Agglutination Test (LAT). LAT is in principle [121]. )e 11 genome segments of the parental virus strains similar to ELISA test [115]. Antigen or antibody is coated on can theoretically reassort into 2048 [121, 122] different the surface of latex particles, which captures antibody and possible genome constellations, if reassortment is random. the target antigen, respectively. )e test has been applied for Gouvea and Brantly [123] hypothesized that rotaviruses the detection of a wide range of targets, such as bacteria, exist as mixed populations of reassortants and that reas- virus, hormones, drugs, and serum protein [116]. Latex sortment was the driving force behind diversity. A pre- particles are made of synthetic rubber and emulsified as requisite of diversity is cocirculation of many different billions of micelles of the same size of a desired diameter. rotavirus types in a population; and more diversity, as well as Usually, the size of particles ranges between 0.05 and 2 μm in more frequency of uncommon strains, is seen in years with diameter, and the presence of sulfate ions provides an in- the highest number of cocirculating strains [124]. Gouvea herent negative surface charge to the particles [117]. )is and Brandtly considered that mixed populations of rota- prepared latex particle can be further functionalized by viruses are being continually propagated in human and special processing, such as amidation, amination, carbox- animal hosts, resulting in new and diverse progeny pop- ylation, hydroxylation, or magnetization, to increase their ulations of rotavirus. With regard to new rotavirus strains binding stability and analytic attachment depending on the arising through reassortment, a concept of zoonotic genes purpose of test [117]. )e latex agglutination test is fre- may be developed. )ese can be defined as genes originating quently employed in diagnostic lab, because it can be a semi- in animal rotaviruses which can interact with genes of quantified test and is relatively cheap with rapid turnaround. human rotaviruses, to form infectious rotavirus particles Caution should be taken in interpreting marginal results as which are serially propagated in the human population [3]. false positive/negative results frequently occur due to Until recently, specific rotavirus types have been asso- nonspecific binding or interference [115]. ciated with specific animal species. For example, human rotaviruses most commonly belong to G types 1–4 and P types [4] and [8] [125], whereas bovine rotaviruses most 2.10. Treatment. )ere is no specific treatment for rotaviral commonly belong to G types 6, 8, and 10 and P types [1], [5], infections. Treatment is based in providing supportive care or [11] [126]. )e rotaviruses have been characterized, and and managing clinical signs and potential complications. In the host species specificity of P and G types has become less livestock and companion animals, fluid administration is distinct. Human group A rotavirus strains that possess genes essential to replace losses from diarrhoea or vomiting, to commonly found in animal rotaviruses have been isolated correct acidosis and to restore electrolytes imbalance. Ad- from infected children in both developed and developing equate sodium concentration and appropriate glucose to countries. Strains such as G3 (found commonly in species sodium ratios are the most important components of an such as cats, dogs, monkeys, pigs, mice, rabbits, and horses), efficient rehydration solution [118]. In young animals, ad- G5 (pigs and horses), G6 and G8 (cattle), G9 (pigs and ministration of fluids can be performed by means of lambs), and G10 (cattle) have been isolated from the human oesophageal catheter; in older animals, intravenous ad- population throughout the world [127]. ministration is preferable. In affected piglets, administration Groups A to C have been shown to infect both humans of a plasma protein mixture, consisting of immunoglobulins, and animals [128]. Members of Rotavirus Group A are growth factors, and other biologically active peptides, has classified according to their glycoprotein (G) structures, been advocated to enhance small intestine recovery [119]. namely, G (G1, G2, G3, ..., Gn) genotypes, and their protein cleavage (P), namely, P (P [1], P [2], P [3], ..., P [n]) ge- 2.11. 2e Zoonotic Potential of Rotavirus. Rotaviruses have a notypes [129]. Currently, 36 G genotypes and 51 P genotypes wide host range, infecting many animal species as well as have been identified in humans and animals worldwide humans. As it was found that certain animal rotavirus strains [130]. G and P type combinations which are found in man 12 Veterinary Medicine International have also been found in animal species. For example, G10P immunity induced by vaccination is elicited to last during the [11] was found in American and Canadian cattle by Lucchelli first few years of children lives when the risk of severe in- et al. [131]. and in Indian cows and buffaloes by Gulati et al. fections is the greatest. In order to decrease the incidence of [132] G3P[6] and G4P[6] were found in pigs in Poland and disease in the herd, a good producer should maximize co- the USA and G1P [8] and G5P [8] were found in pigs in lostrum transfer, increase environmental sanitation, reduce Brazil by Santos et al. [133]. )e emerging G9 strains 26–28 stressors such as overcrowding or poor nutrition, and vaccinate may have arisen in humans through transfer from animals. bred cows for rotavirus at 60 and 30 days before calving [62]. )ey have been found in lambs and pigs [133, 134]. First-milking colostrum is a source of nutrients and of Epidemiologically, there exists evidence for zoonotic passively absorbed maternal antibodies, critical to protect transmission of rotaviruses. Human Group A rotavirus the newborn calf against infectious disease in the first weeks strains possessing genes commonly found in animal rotavi- and months of life. )e calf is born without most antibodies, ruses have been isolated from infected children in both de- including those that fight the infectious agents which cause veloped and developing countries. Strains such as G3 (found diarrhea. )e calf will acquire these antibodies only from commonly in species such as cats, dogs, monkeys, pigs, mice, colostrum [139]. Because of this, any effort to prevent di- rabbits and horse), G5 (pigs and horses), G6 and G8 (cattle), arrhea by vaccinating cows is wasted unless the calf actually G9 (pigs and lambs), and G10 (cattle) have been isolated from receives colostrum, preferably before it is two to four hours the human population through the world [127]. old. As the calf grows older, it rapidly loses its ability to In humans, they appear to cause more severe symptoms absorb colostral antibodies. Colostrum given to calves that than the common rotavirus strains [135], which might be due are more than 24 to 36 hours old are practically useless; to less immunity to these emerging strains, or to greater antibodies are seldom absorbed this late in life. )e neonatal virulence being conferred by their genetic makeup. Several calf should ideally receive 2 to 3L (for beef calves) or 3 to 4L studies have indicated symptomatic infection of humans by (in dairy calves) of colostrum within the first 6 hours after animal viruses. Nakagomi and Nakagomi, [136] reported that birth. )e colostrum contains antibodies, immune cells almost all gene segments of the rotavirus G3 strain (AU228) (neutrophils, macrophages, and T and B cells), comple- isolated from a child with a pet cat were identical to those of a ments, lactoferrin, insulin-like growth factor-1, trans- feline rotavirus strain (FRV-1). Strains very similar to this forming growth factor, interferon, and nutrients [140]. may have become established in humans [137]. A three-week- To improve the passive immunization of calves against old baby in an Israeli household which had a young dog (<6 rotavirus and coronavirus as well as against different strains months old) was infected with an animal rotavirus G3 strain of E. coli vaccination of the pregnant dam can be proposed. [113]. Das et al. [138] reported that a G8 rotavirus which had Usually cows are vaccinated twice (6 to 8 and 2 to 3 weeks) widely circulated in newborn infants in India, causing before parturition to stimulate the production of specific asymptomatic infection, had VP7 and VP4 gene sequences antibodies. )e primary function of colostrum is to enhance which were identical to those of a bovine rotavirus strain. the calf’s immune system through the passive transfer of Some feline and canine rotavirus strains have spread into both antibody and cell-mediated immunity. Ideally, calves human populations as whole virions, bovine rotaviruses were should receive colostrum from their dams although colos- involved in reassortment with human rotaviruses, leading to trum from several cows is often mixed and administration of the emergence of unusual strains in various parts of the world. colostrum feeding results in the transmission of BVDV, Apparent dual infection with human and animal rotaviruses bovine leukemia virus, and John’s disease that can be spread has been observed recovered G1P [5] and G1P [8] strains by infected or purchased colostrum [141]. from an infant with severe diarrhoea. )e G1P [5] rotavirus Specific IgG present in colostrum may protect against was genotypically similar to bovine strains. It was not isolated the more common enteropathogens causing calf diarrhea, from the infant in high titer and possibly had little, if any, such as rotavirus, coronavirus, and E. coli. Although vac- effect on the child’s disease. Nonetheless, it would have had cination of the dam prior to calving may boost colostrum the potential to reassort with the coinfecting strain [136]. IgG concentrations [118, 142], vaccinate the cows and pregnant heifers with any necessary calf diarrhea vaccines well prior to calving. Vaccines that contain rotavirus, 2.12. Control and Prevention of Rotavirus Infections. coronavirus, and the K99 E. coli antigens can be helpful in Rotaviruses are infectious and comparatively resistant to preventing calf diarrhea. )ese are best given to the cow inactivation by chemical disinfectants and antiseptics. prior to calving so it can make antibodies and secrete them Control and prevention measures against rotavirus infection into the colostrum. When the calf ingests this enriched are not so easy for its mass distribution and tendency to colostrum, it will be protected against these major agents stability in different climate situation and are shed in high [143]. In animals, the concept of passive immunization is concentrations in feces of infected animals. )e primary based on maternal antibodies that are transferable through strategy to reduce the burden of rotavirus infections is the placenta or are secreted in the colostrum providing vaccination. Vaccination protocol differs from the ap- transient protective immunity to offspring against clinically proaches implemented to protect infants and children manifest RVA infection. Rotavirus vaccines have been de- against rotavirus disease [79]. veloped to control the neonatal calf diarrhea associated with In humans, the primary objective is the reduction of rotavirus infection. Most of the commercial vaccines are maternal antibody level by the age of 4–6 months; active combined with more than one agent [144]. Veterinary Medicine International 13

Commercial RVA vaccines are administered parenter- BCoV: Bovine coronavirus ally to cows and sows during the late stage of gestation, in BRoV: Bovine rotavirus order to elicit a strong maternal immunity that is readily cDNA: Complementary DNA conferred to newborn animals. Some studies have demon- CPE: Cytopathic effect strated vaccine failure or breakthroughs that have been DMEM: Dulbecco’s modified eagle medium related to a number of factors, including inadequate man- DNA: Dioxy nucleotide ribonucleic acid aging conditions of animals or antigenic differences between DNTPs: Dioxy nucleotide triphosphates vaccine and field RVA strains, even if vaccine and field dsRNA: Double stranded RNA strains shared partially their surface antigen specificities. ELISA: Enzyme-linked immunosorbent assay Moreover, optimum management and hygienic practices MDBK: Madin-Darby bovine kidney cell can minimize the incidence of rotaviral diarrhea in farm mRNA: Messenger RNA animals. To control secondary bacterial infection, antibiotics NSP: Nonstructural proteins and fluid and electrolyte therapy to restore the fluid reserve PBS: Phosphate-buffered saline have to be given due importance so that the mortality rate in PCR: Polymerase chain reaction calves could be minimized [87]. pH: Power of hydrogen RNA: Ribonucleic acid 3. Conclusion and Recommendations rpm: Revolution per minute RT-PCR: Reverse transcriptase polymerase chain reaction Diarrheal disease caused by rotavirus poses a great health UK: United Kingdom problem in calves that interrupts production benefits with VP: Structural proteins. reduced weight gain and increased mortality, and its po- tential for zoonotic spread [68]. Rotavirus is a major pathogen responsible for diarrheal disease in calves resulting Data Availability in loss of productivity and economy of farmers. However, )e datasets used and/or analyzed during the current study various facets of diarrheal disease caused by rotavirus in are available from the corresponding author on reasonable calves in world are inadequately understood. Awareness of request. the advantage of colostrum feeding is not enough, but also times of colostrum administration to neonate calves are crucial for the ultimate development of immune status Conflicts of Interest against pathogens including rotavirus infection. Calving )e authors declare that they have no conflicts of interest. areas should have well-drained grass lots or pastures visible from the barn area and calving areas should be selected or landscaped to allow for adequate drainage. Enteric disease Authors’ Contributions like rotavirus infection is a vital health problem in calves that )is manuscript was written entirely by the authors. interrupts production benefits with reduced weight gain and increased mortality, and the virus potential for its zoonotic spread; it is imperative to determine the disease burden and Acknowledgments responsible risk factors. )is is very useful to execute ef- )e authors acknowledge Oda Bultum University for pro- fective preventive measures such as practicing early colos- viding different facility and reading materials used to pre- trum feeding in newborn calves, vaccination in dams, and pare this manuscript. improving livestock management. Rearing healthy dairy calves to weaning time requires maximizing the calf’s level of immunity against disease while minimizing its exposure to References infectious agents. Based on the above conclusion, the fol- [1] A. A. Alfieri, M. E. Parazzi, E. Takiuchi, K. C. Medici,´ and lowing recommendations were forwarded: A. F. Alfieri, “Frequency of group a rotavirus in diarrhoeic (i) Awareness creation for researcher and government calves in Brazilian cattle herds, 1998–2002,” Tropical Animal Health and Production, vol. 38, p. 521, 2006. regarding the effect of rotavirus infection in calf’s [2] G. M. Barrington, J. M. Gay, and J. F. Evermann, “Biosecurity health and growth performance and national for neonatal gastrointestinal diseases,” Veterinary Clinics of economy is very important North America: Food Animal Practice, vol. 18, no. 1, pp. 7–34, (ii) Further studies of rotavirus infection in calves 2002. covering larger areas of the country need to be [3] N. Cook, J. Bridger, K. Kendall, M. I. Gomara, L. El-Attar, conducted and J. Gray, “)e zoonotic potential of rotavirus,” Journal of Infection, vol. 48, no. 4, pp. 289–302, 2004. [4] D. M. Foster and G. W. Smith, “Pathophysiology of diarrhea Abbreviations in calves,” Veterinary Clinics of North America: Food Animal Practice, vol. 25, no. 1, pp. 13–36, 2009. Ag: Antigen [5] S. R. Barua, “Clinico-pathology and molecular character- Ag- Antigen capturing enzyme-linked ization of bovine rotavirus infection in calves in South- ELISA: immunosorbent assay Eastern part of Bangladesh,” 2019. 14 Veterinary Medicine International

[6] Y. Cho, “Ecology of calf diarrhea in cow-calf operations,” a worldwide perspective,” Vaccine, vol. 28, no. 31, Dissertations thesis, IOWA State University, Ames, IA, USA, pp. 5100–5108, 2010. 2012. [25] L. Johne and C.-H. Von Bonsdorff, “Frequent reassortments [7] J. F. Mee, “Newborn dairy calf management,” Veterinary may explain the genetic heterogeneity of rotaviruses: analysis Clinics of North America: Food Animal Practice, vol. 24, no. 1, of Finnish rotavirus strains,” Journal of Virology, vol. 76, pp. 1–17, 2008. no. 23, pp. 11793–11800, 2002. [8] J. B. Pesavento, A. M. Billingsley, E. J. Roberts, R. F. Ramig, [26] A. R. Laird, J. R. Gentsch, T. Nakagomi, O. Nakagomi, and and B. V. V. Prasad, “Structures of rotavirus reassortants R. I. Glass, “Characterization of serotype G9 rotavirus strains demonstrate correlation of altered conformation of the VP4 isolated in the United States and India from 1993 to 2001,” spike and expression of unexpected VP4-associated phe- Journal of Clinical Microbiology, vol. 41, no. 7, pp. 3100–3111, notypes,” Journal of Virology, vol. 77, no. 5, pp. 3291–3296, 2003. 2003. [27] E. W. Rainsford and M. A. McCrae, “Characterization of the [9] C. Bj¨orkman, C. Svensson, B. Christensson, and NSP6 protein product of rotavirus gene 11,” Virus Research, K. De Verdier, “Cryptosporidium parvum and Giardia vol. 130, no. 1-2, pp. 193–201, 2007. intestinalis in calf diarrhoea in Sweden,” Acta Veterinaria [28] D. Chen, C. Q. Zeng, M. J. Wentz, M. Gorziglia, M. K. Estes, Scandinavica, vol. 44, no. 3, p. 145, 2003. and R. F. Ramig, “Template-dependent, in vitro replication [10] Y. S. Malik, N. Kumar, K. Sharma et al., “Epidemiology and of rotavirus RNA,” Journal of Virology, vol. 68, no. 11, genetic diversity of rotavirus strains associated with acute pp. 7030–7039, 1994. gastroenteritis in bovine, porcine, poultry and human [29] J. T. Patton and C. O. Gallegos, “Rotavirus RNA replication: population of Madhya Pradesh, Central India, 2004–2008,” single-stranded RNA extends from the replicase particle,” Virus, vol. 20139 pages, 2013. Journal of General Virology, vol. 71, no. 5, pp. 1087–1094, [11] K. De Verd Er, “Infektionspanoramat vid diarr´eer hos 1990. svenska kalvar,” Sven. Vet, vol. 58, p. 29, 2006. [30] C. F. Arias, P. Romero, V. Alvarez, and S. L´opez,“Trypsin [12] D. Reynolds, J. Morgan, N. Chanter et al., “Microbiology of activation pathway of rotavirus infectivity,” Journal of Vi- calf diarrhoea in Southern Britain,” Veterinary Record, rology, vol. 70, no. 9, pp. 5832–5839, 1996. vol. 119, no. 2, pp. 34–39, 1986. [31] J. Matthijnssens, M. Ciarlet, S. M. McDonald et al., “of [13] F. Bendali, H. Bichet, F. Schelcher, and M. Sanaa, “Pattern of rotavirus strain nomenclature proposed by the rotavirus diarrhoea in newborn beef calves in South-West France,” classification working group (RCWG),” Archives of Virology, Veterinary Research, vol. 30, pp. 61–74, 1999. vol. 156, no. 8, pp. 1397–1413. [14] M. M. Alam, A. Huque, S. Nigar, M. Haque, [32] M. K. Estes and H. B. Greenberg, “Rotaviruses,” in Fields N. S. Choudhury, and M. U. Ahmed, “Rotavirus infection in Virology, pp. 1347–1401, Lippincott Williams Wilkins, children and calves in association with acute gastroenteritis.” Philadelphia, PA, USA, 6th edition, 2013. Bangladesh Veterinary Journal, vol. 28, pp. 35–40, 1994. [33] E. Mihalov-Kovacs,´ A.´ Gellert,´ S. Marton et al., “Candidate [15] G. Abraham, P. L. Roeder, and R. Zewdu, “Agents associated new rotavirus species in sheltered dogs, Hungary,” Emerging with neonatal diarrhoea in Ethiopian dairy calves,” Tropical Infectious Diseases, vol. 21, no. 4, pp. 660–663, 2015. Animal Health and Production, vol. 24, no. 2, pp. 74–80, [34] S. Ghosh and N. Kobayashi, “Exotic rotaviruses in animals 1992. and rotaviruses in exotic animals,” Virus Disease, vol. 25, [16] R. Bishop, G. P. Davidson, I. H. Holmes, and B. J. Ruck, no. 2, pp. 158–172, 2014. “Virus particles in epithelial cells of duodenal mucosa from [35] P. O. Tonietti, A. S. Hora, F. D. F. Silva, V. L. A. Ruiz, and children with acute non-bacterial gastroenteritis,” 2e F. Gregori, “Phylogenetic analyses of the VP4 and VP7 genes Lancet, vol. 302, no. 7841, pp. 1281–1283, 1973. of porcine group a rotaviruses in São Paulo State, Brazil: first [17] A. M. Paredes, D. T. Brown, R. Rothnagel et al., “)ree- identification of G5P [23] in piglets,” Journal of Clinical dimensional structure of a membrane-containing virus,” Microbiology, vol. 51, pp. 2750–2753, 2013. Proceedings of the National Academy of Sciences, vol. 90, [36] J. Matthijnssens, M. Ciarlet, E. Heiman et al., “Full genome- no. 19, pp. 9095–9099, 1993. based classification of rotaviruses reveals a common origin [18] F. A. Murphy, E. P. J. Gibbs, M. C. Horzinek, and between human Wa-Like and porcine rotavirus strains and M. J. Studdert, Veterinary Virology, Elsevier, Amsterdam, human DS-1-like and bovine rotavirus strains,” Journal of )e Netherlands, 1999. Virology, vol. 82, no. 7, pp. 3204–3219, 2008. [19] N. Ved, Modecular Characterisation and genotyping of [37] T. Nakagomi, O. Nakagomi, Y. Takahashi, M. Enoki, Human and Animal Rotaviruses Isolates from Northern In- T. Suzuki, and P. E. Kilgore, “Incidence and burden of ro- dia, Jiwaji University, Gwalior, India, 2014. tavirus gastroenteritis in Japan, as estimated from a pro- [20] M. K. Estes and A. Z. Kapikian, D. M. Knipe, P. M. Howley, spective sentinel hospital study,” 2e Journal of Infectious D. E. Griffin et al., Rotaviruses Fields Virology, Kluwer Health Diseases, vol. 192, no. s1, pp. S106–S110, 2005. Williams Wilkins, Philadelphia, PA, USA, 2007. [38] A. F. Dennis, S. M. McDonald, D. C. Payne et al., “Molecular [21] E. J. Anderson and S. G. Weber, “Rotavirus infection in epidemiology of contemporary G2P [4] human rotaviruses adults,” 2e Lancet Infectious Diseases, vol. 4, no. 2, cocirculating in a single U.S. community: footprints of a pp. 91–99, 2004. globally transitioning genotype,” Journal of Virology, vol. 88, [22] M. Hagbom, “Rotavirus disease mechanisms diarrhea, no. 7, pp. 3789–3801, 2014. vomiting and inflammation : how and why,” Medical Dis- [39] CDC, “Rotavirus surveillance–worldwide, 2001-2008,” sertation, Linkoping¨ University, Linkoping,¨ Sweden, 2015. MMWR. Morbidity and Mortality Weekly Report, vol. 57, [23] K. Pak, , Chapter two, pp. 8–13, 2011. p. 1255, 2008. [24] C. Rodrigo, N. Salman, V. Tatochenko, Z. Meszner,´ and [40] M. Wu, T. Dallman, K. B´anyaiet al., “Rotavirus surveillance C. Giaquinto, “Recommendations for rotavirus vaccination: in Europe, 2005–2008: web-enabled reporting and real-time Veterinary Medicine International 15

analysis of genotyping and epidemiological data,” Journal of [57] M. Zrelli, L. Messadi, L. Ben Miled, M. H. Jemli, and Infectious Diseases, vol. 200, pp. S215–S221, 2009. N. Haddad, “Infectious agents associated with calf diarrhea [41] S. Sharma, V. K. Paul, M. K. Bhan, and P. Ray, “Genomic in Tunisia [neonatal enteritis],” Revue de Medecine´ characterization of nontypeable rotaviruses and detection of Vet´ erinaire´ , vol. 11, 1990. a rare G8 strain in Delhi, India,” Journal of Clinical Mi- [58] H. Langoni, A. C. Linhares, F. A. De Avila, A. V. Da Silva, crobiology, vol. 47, no. 12, pp. 3998–4005, 2009. and A. O. Elias, “Contribution to the study of diarrhea [42] K.-P. Hwang, Y.-C. Huang, K. Banyai´ et al., “Severe gas- etiology in neonate dairy calves in São Paulo State, Brazil,” troenteritis associated with G3P [9] rotavirus in Taiwan,” Brazilian Journal of Veterinary Research and Animal Science, Infection, vol. 39, no. 3, pp. 271–275, 2011. vol. 41, pp. 313–319, 2004. [43] V. Martella, M. Ciarlet, K. Banyai et al., “Identification of [59] A. Kam, D. Khelef, R. Kaidi et al., “)e frequency of the group a porcine rotavirus strains bearing a novel VP4 (P) shedding of cryptosporidium parvum, F5 Escherichia coli, genotype in Italian Swine Herds,” Journal of Clinical Mi- rotavirus, coronavirus and Salmonella spp. in young dairy crobiology, vol. 45, no. 2, pp. 577–580. calves in Mitidja Area (Algeria),” Bulletin of the University of [44] J. T. Patton, “Rotavirus diversity and evolution in the post- Agricultural Sciences and Veterinary Medicine, vol. 68, 2011. vaccine world,” Discovery Medicine, vol. 13, no. 68, pp. 85– [60] D. Snodgrass, H. Terzolo, D. Sherwood, I. Campbell, 97, 2012. J. Menzies, and B. Synge, “Aetiology of diarrhoea in young [45] S. M. McDonald, J. Matthijnssens, J. K. McAllen et al., calves,” Veterinary Record, vol. 119, no. 2, pp. 31–34, 1986. “Evolutionary dynamics of human rotaviruses: balancing [61] R. De La Fuente, A. Garcıa,́ J. A. Ruiz-Santa-Quiteria et al., reassortment with preferred genome constellations,” PLoS “Proportional morbidity rates of enteropathogens among Pathogens, vol. 5, no. 10, p. e1000634, 2009. diarrheic dairy calves in Central Spain,” Preventive Veteri- [46] Y. Hoshino, S. Honma, R. W. Jones et al., “A porcine G9 nary Medicine, vol. 36, no. 2, pp. 145–152, 1998. rotavirus strain shares neutralization and VP7 phylogenetic [62] M. Izzo, P. Kirkland, V. Mohler, N. Perkins, A. Gunn, and sequence lineage 3 characteristics with contemporary human J. House, “Prevalence of major enteric pathogens in Aus- G9 rotavirus strains,” Virology, vol. 332, no. 1, pp. 177–188, tralian dairy calves with diarrhoea,” Australian Veterinary 2005. Journal, vol. 89, no. 5, pp. 167–173, 2011. [47] R. E. Randall and S. Goodbourn, “Interferons and viruses: an [63] J. E. Tate, A. H. Burton, C. Boschi-Pinto, A. D. Steele, interplay between induction, signalling, antiviral responses J. Duque, and U. D. Parashar, “2008 estimate of worldwide and virus countermeasures,” Journal of General Virology, rotavirus-associated mortality in children younger than 5 vol. 89, no. 1, pp. 1–47, 2008. years before the introduction of universal rotavirus vacci- [48] R. Lamb and R. M. Kurg, “Orthomyxoviridae: the viruses and nation programmes: a systematic review and meta-analysis,” their replication,” in Fields Virology, pp. 1487–1531, 2e Lancet Infectious Diseases, vol. 12, no. 2, pp. 136–141, Northwestern University, Chicago, IL, USA, 2001. 2012. [49] W. Diederichsen, V. Munford, A. M. Villaç, T. A. Ramos [64] WHO, Global Rotavirus Information and Surveillance Bul- Caruzo, and M. L. Racz,´ “Characterization of mixed infec- letin, WHO, Geneva, Switzerland, 2013. tions with different strains of bovine rotavirus in an outbreak [65] M. Hagbom, C. Istrate, D. Engblom et al., “Rotavirus of diarrhea in dairy herds in Goi´as,Brazil,” Brazilian Journal stimulates release of serotonin (5-HT) from human en- of Microbiology, vol. 31, pp. 140–145, 2000. [50] L. Svensson, S. Sharma, J. Nordgren et al., “Both lewis and terochromaffin cells and activates brain structures involved secretor status mediate susceptibility to rotavirus infections in nausea and vomiting,” PLoS Pathogens, vol. 7, no. 7, in a rotavirus genotype–dependent manner,” Clinical In- p. e1002115. fectious Diseases, vol. 59, pp. 1567–1573, 2014. [66] T. Bizuneh, A. Abebe, and E. Lema, “Rotavirus infection in [51] K. T. Kaljot, R. D. Shaw, D. H. Rubin, and H. B. Greenberg, under-five children in Jimma hospital, Southwest Ethiopia,” “Infectious rotavirus enters cells by direct cell membrane Ethiopian Journal of Health Development, vol. 18, pp. 19–24, penetration, not by endocytosis,” Journal of Virology, vol. 62, 2004. no. 4, pp. 1136–1144, 1988. [67] A. Gelaw, C. Pietsch, and U. G. Liebert, “Molecular epide- [52] M. S. Mcnulty, “)e etiology, pathology and epidemiology of miology of rotaviruses in Northwest Ethiopia after national viral gastroenteritis,” Annales De Recherches Veterinaires, vaccine introduction,” Infection, Genetics and Evolution, vol. 14, 1983. vol. 65, pp. 300–307, 2018. [53] S. S. M. Ammar, K. Mokhtaria, B. B. Tahar et al., “Prevalence [68] U. Seid, F. Dawo, A. Tesfaye, and M. Ahmednur, “Isolation of rotavirus (GARV) and coronavirus (BCoV) associated and characterization of coronavirus and rotavirus associated with neonatal diarrhea in calves in Western Algeria,” Asian with calves in central part of Oromia, Ethiopia,” Veterinary Pacific Journal of Tropical Biomedicine, vol. 4, pp. S318–S322, Medicine International, vol. 2020, Article ID 8869970, 2014. 10 pages, 2020. [54] S. R. Jindal, N. K. Maiti, and M. S. Oberoi, “Genomic di- [69] O. Lundgren and L. Svensson, “Pathogenesis of rotavirus versity and prevalence of Rotavirus in cow and buffalo calves diarrhea,” Microbes and Infection, vol. 3, no. 13, in Northern India,” Revue Scientifique et Technique de l’OIE, pp. 1145–1156, 2001. vol. 19, pp. 871–876, 2016. [70] J. M. Ball, P. Tian, C. Q.-Y. Zeng, A. P. Morris, and [55] R. B. Rai, A. Hansha, S. Rai et al., “Prevalence of rota and M. K. Estes, “Age-dependent diarrhea induced by a rotaviral coronavirus infections in calves of Barabanki and Raebareli nonstructural glycoprotein,” Science, vol. 272, no. 5258, districts of Uttar Pradesh,” Indian Journal of Veterinary pp. 101–104, 1996. Pathology, vol. 35, pp. 73-74, 2011. [71] Y. Ge, A. Mansell, J. E. Ussher et al., “Rotavirus NSP4 triggers [56] I. AlRobaiee and M. Farwachi, “Prevalence of rotaviral in- secretion of proinflammatory cytokines from macrophages fection in diarrheic neonatal calves in Mosul city, Iraq,” via toll-like receptor 2,” Journal of Virology, vol. 87, no. 20, Veterinary World, vol. 6, no. 8, p. 538, 2013. pp. 11160–11167, 2013. 16 Veterinary Medicine International

[72] A. P. Morris, J. K. Scott, J. M. Ball, C. Q.-Y. Zeng, [89] M. A. Soltan, Y.-L. Tsai, P.-Y. A. Lee et al., “Comparison of W. K. O’Neal, and M. K. Estes, “NSP4 elicits age-dependent electron microscopy, ELISA, real time RT-PCR and insulated diarrhea and Ca2+ mediated I−influx into intestinal crypts of isothermal RT-PCR for the detection of rotavirus group a CF mice,” American Journal of Physiology-Gastrointestinal (RVA) in feces of different animal species,” Journal of Vi- and Liver Physiology, vol. 277, no. 2, pp. G431–G444, 1999. rological Methods, vol. 235, pp. 99–104, 2016. [73] B. N. Fields, D. M. Knipe, P. M. Howley, K. D. Everiss, and [90] R. L. Ward, D. I. Bernstein, E. C. Young, J. R. Sherwood, H.-J. Kung, Fundamental Virology, Lippincott-Raven Phil- D. R. Knowlton, and G. M. Schiff, “Human rotavirus studies adelphia, Philadelphia, PA, USA, 1996. in volunteers: determination of infectious dose and sero- [74] G. P. Davidson and G. L. Barnes, “Structural and functional logical response to infection,” Journal of Infectious Diseases, abnormalities of the small intestine in infants and young vol. 154, no. 5, pp. 871–880, 1986. children with rotavirus enteritis,” Acta Paediatrica, vol. 68, [91] G. L. Barnes, S. L. Callaghan, C. D. Kirkwood, no. 3, pp. 181–186, 1979. N. Bogdanovic-Sakran, L. J. Johnston, and R. F. Bishop, [75] M. C. Medici, L. A. Abelli, P. Guerra, I. Dodi, G. Dettori, and “Excretion of serotype G1 rotavirus strains by asymptomatic C. Chezzi, “Case report: detection of rotavirus RNA in the staff,” 2e Journal of Pediatrics, vol. 142, no. 6, pp. 722–725, cerebrospinal fluid of a child with rotavirus gastroenteritis 2003. and meningism,” Journal of Medical Virology, vol. 83, no. 9, [92] S. Ramani, R. Arumugam, N. Gopalarathinam et al., “In- pp. 1637–1640, 2011. vestigation of the environment and of mothers in trans- [76] T. Nakagomi and O. Nakagomi, “Rotavirus antigenemia in mission of rotavirus infections in the neonatal nursery,” children with encephalopathy accompanied by rotavirus Journal of Medical Virology, vol. 80, no. 6, pp. 1099–1105, gastroenteritis,” Archives of Virology, vol. 150, no. 9, 2008. pp. 1927–1931, 2005. [93] D. I. Bernstein, “Rotavirus overview,” Pediatric Infectious [77] K. Dhama, R. S. Chauhan, M. Mahendran, and S. V. S. Malik, Disease Journal, vol. 28, no. 3, pp. S50–S53, 2009. “Rotavirus diarrhea in bovines and other domestic animals,” [94] CDC, “Delayed onset and diminished magnitude of rotavirus Veterinary Research Communications, vol. 33, no. 1, pp. 1–23, activity–United States, November 2007-May 2008,” MMWR. 2009. Morbidity and Mortality Weekly Report, vol. 57, p. 697, 2008. [78] P. H. Dennehy, “Transmission of rotavirus and other enteric [95] J. J. Hull, E. N. Teel, T. K. Kerin, M. M. Freeman, pathogens in the home,” 2e Pediatric Infectious Disease M. D. Esona, and J. R. Gentsch, “United States rotavirus Journal, vol. 19, pp. S103–S105, 2000. strain surveillance from 2005 to 2008: genotype prevalence [79] V. Martella, K. Banyai,´ J. Matthijnssens, C. Buonavoglia, and before and after vaccine introduction,” Pediatric Infectious M. Ciarlet, “Zoonotic aspects of rotaviruses,” Veterinary Disease Journal, vol. 30, 2011. Microbiology, vol. 140, no. 3-4, pp. 246–255, 2010. [96] S. Svensson, A. E. Castro, and B. I. Osbum, “Comparison of [80] H. Jayaram, M. K. Estes, and B. V. V. Prasad, “Emerging polyacrylamide gel electrophoresis, an enzyme-linked-im- themes in rotavirus cell entry, genome organization, tran- munosorbent assay, and an agglutination test for the direct scription and replication,” Virus Research, vol. 101, no. 1, identification of bovine rotavirus from feces and coelec- pp. 67–81, 2004. trophoresis of viral RNA’s,” Journal of Veterinary Diagnostic [81] U. Desselberger and H.-I. Huppertz, “Immune responses to Investigation, vol. 2, no. 3, pp. 184–190, 1990. rotavirus infection and vaccination and associated correlates [97] N. Reidy, G. Lennon, S. Fanning, E. Power, and H. O’Shea, of protection,” Journal of Infectious Diseases, vol. 203, no. 2, “Molecular characterisation and analysis of bovine rotavirus pp. 188–195, 2011. strains circulating in Ireland 2002–2004,” Veterinary Mi- [82] F. R. Vel´azquez,D. O. Matson, M. L. Guerrero et al., “Serum crobiology, vol. 117, no. 2–4, pp. 242–247, 2006. antibody as a marker of protection against natural rotavirus [98] R. M. Lequin, “Enzyme immunoassay (EIA)/enzyme-linked infection and disease,” 2e Journal of Infectious Diseases, immunosorbent assay (ELISA),” Clinical Chemistry, vol. 51, vol. 182, no. 6, pp. 1602–1609, 2000. no. 12, pp. 2415–2418, 2005. [83] R. I. Glass, J. Bresee, B. Jiang, U. Parashar, E. Yee, and [99] C. D. Brandt, H. W. Kim, W. J. Rodriguez et al., “Com- J. Gentsch, “Rotavirus and rotavirus vaccines,” in Hot Topics parison of direct electron microscopy, immune electron in Infection and Immunity in Children III, Springer, Berlin, microscopy, and rotavirus enzyme-linked immunosorbent Germany, 2006. assay for detection of gastroenteritis viruses in children,” [84] R. F. Ramig, “Pathogenesis of intestinal and systemic rota- Journal of Clinical Microbiology, vol. 13, no. 5, pp. 976–981, virus infection,” Journal of Virology, vol. 78, no. 19, 1981. pp. 10213–10220, 2004. [100] J. A. Ribes, J. P. Seabolt, and S. B. Overman, “Performance [85] J. Chanock, H. Nirwati, R. Triasih et al., “Maternal antibodies characteristics of VIDAS and Directigen respiratory syn- to rotavirus: could they interfere with live rotavirus vaccines cytial virus (RSV) antigen detection assays and culture for in developing countries?” Vaccine, vol. 29, no. 6, the identification of RSV in respiratory specimens,” Journal pp. 1242–1247. of Clinical Microbiology, vol. 40, no. 5, pp. 1818–1820, 2002. [86] M. Patel, A. L. Shane, U. D. Parashar, B. Jiang, J. R. Gentsch, [101] A. Schielke, M. Filter, B. Appel, and R. Johne, “)ermal and R. I. Glass, “Oral rotavirus vaccines : how well will they stability of hepatitis E virus assessed by a molecular bio- work where they are needed most ?” 2e Journal of Infectious logical approach,” Virology Journal, vol. 8, no. 1, p. 487, 2011. Diseases, vol. 200, pp. 39–48, 2009. [102] WHO, Manual of Rotavirus Detection and Characterization [87] A. D. Steele, A. Geyer, and G. H. Gerdes, “Rotavirus in- Methods, World Health Organization, Geneva, Switzerland, fections,” in Infectious Diseases of Livestock, pp. 1256–1264, 2009. Oxford University Press, Oxford, UK, 2004. [103] V. Gouvea, R. I. Glass, P. Woods et al., “Polymerase chain [88] R. E. Holland, “Some infectious causes of diarrhea in young reaction amplification and typing of rotavirus nucleic acid farm animals,” Clinical Microbiology Reviews, vol. 3, no. 4, from stool specimens,” Journal of Clinical Microbiology, pp. 345–375, 1990. vol. 28, no. 2, pp. 276–282, 1990. Veterinary Medicine International 17

[104] O. Nakagomi and T. Nakagomi, “Molecular evidence for surface chemistry for enhancing a latex homogeneous im- naturally occurring single VP7 gene substitution reassortant munoassay of C-reactive protein,” Analytical Chemistry, between human rotaviruses belonging to two different vol. 73, no. 14, pp. 3417–3425, 2001. genogroups,” Archives of Virology, vol. 119, no. 1-2, [118] I. Lorenz, J. Fagan, and S. J. More, “Calf health from birth to pp. 67–81, 1991. weaning. II. management of diarrhoea in pre-weaned [105] K. Taniguchi, T. Urasawa, S. Urasawa, and T. Yasuhara, calves,” Irish Veterinary Journal, vol. 64, p. 9, 2011. “Production of subgroup-specific monoclonal antibodies [119] B. A. Corl, S. A. Mathews Oliver, X. Lin et al., “Conjugated against human rotaviruses and their application to an en- linoleic acid reduces body fat accretion and lipogenic gene zyme-linked immunosorbent assay for subgroup determi- expression in neonatal pigs fed low-or high-fat formulas,” nation,” Journal of Medical Virology, vol. 14, no. 2, 2e Journal of Nutrition, vol. 138, no. 3, pp. 449–454, 2008. pp. 115–125, 1984. [120] M. Gorziglia, G. Larralde, A. Z. Kapikian, and [106] G. Kang, M. Iturriza-Gomara, J. G. Wheeler et al., “Quan- R. M. Chanock, “Antigenic relationships among human titation of group a rotavirus by real-time reverse-tran- rotaviruses as determined by outer capsid protein VP4,” scription-polymerase chain reaction: correlation with Proceedings of the National Academy of Sciences, vol. 87, clinical severity in children in South India,” Journal of pp. 7155–7159, 2006. Medical Virology, vol. 73, no. 1, pp. 118–122, 2004. [121] R. F. Ramig, “Genetics of the rotaviruses,” Annual Review of [107] M. Fukuda, K. Kuga, A. Miyazaki et al., “Development and Microbiology, vol. 51, pp. 225–255, 2002. application of one-step multiplex reverse transcription PCR [122] J. Flores, M. Sereno, C. J. Lai et al., Use of Single-Stranded for simultaneous detection of five diarrheal viruses in adult Rotavirus RNA Transcripts for the Diagnosis of Rotavirus cattle,” Archives of Virology, vol. 157, no. 6, pp. 1063–1069, Infection, the Study of Genetic Diversity Among Rotavi- 2012. ruses, and the Molecular Cloning of Rotavirus Genes. [108] M. J. Espy, J. R. Uhl, L. M. Sloan et al., “Real-time PCR in Double-Stranded RNA Viruses, Elsevier, Amsterdam, )e clinical microbiology: applications for routine laboratory Netherlands, 1983. testing,” Clinical Microbiology Reviews, vol. 19, no. 1, [123] V. Gouvea and M. Brantly, “Is rotavirus a population of pp. 165–256, 2006. reassortants?” Trends in Microbiology, vol. 3, no. 4, [109] Y.-I. Cho, W.-I. Kim, S. Liu, J. M. Kinyon, and K. J. Yoon, pp. 159–162, 1995. “Development of a panel of multiplex real-time polymerase [124] V. Jain, B. K. Das, M. K. Bhan, R. I. Glass, J. R. Gentsch, and chain reaction assays for simultaneous detection of major I. S. S. C. Laboratories, “Great diversity of group a rotavirus agents causing calf diarrhea in feces,” Journal of Veterinary strains and high prevalence of mixed rotavirus infections in Diagnostic Investigation, vol. 22, no. 4, pp. 509–517, 2010. India,” Journal of Clinical Microbiology, vol. 39, no. 10, [110] X. L. Pang, B. Lee, N. Boroumand, B. Leblanc, pp. 3524–3529, 2001. J. K. Preiksaitis, and C. C. Yu Ip, “Increased detection of [125] J. R. Gentsch, P. A. Woods, M. Ramachandran et al., “Review rotavirus using a real time reverse transcription-polymerase of G and P typing results from a global collection of rotavirus chain reaction (RT-PCR) assay in stool specimens from strains: implications for vaccine development,” Journal of children with diarrhea,” Journal of Medical Virology, vol. 72, Infectious Diseases, vol. 174, pp. S30–S36, 2011. no. 3, pp. 496–501, 2004. [126] L. El-Attar, W. Dhaliwal, M. Iturriza-G´omara, and [111] K. O. Danchin, A. V. Parwani, and L. J. Saif, “)e charac- J. C. Bridger, “Identification and molecular characterization terization of VP7 (G type) and VP4 (P type) genes of bovine of a bovine G3 rotavirus which causes age-independent group A rotaviruses from field samples using RT-PCR and diarrhea in cattle,” Journal of Clinical Microbiology, vol. 40, RFLP analysis,” Archives of Virology, vol. 141, no. 9, no. 3, pp. 937–942, 2002. pp. 1727–1739, 1996. [127] U. Desselberger, M. Iturriza-Gomara, and J. J. Gray, “Ro- [112] M. Nemoto, Y. Morita, H. Niwa et al., “Rapid detection of tavirus epidemiology and surveillance,” Novartis Foundation equine coronavirus by reverse transcription loop-mediated Symposium, vol. 238, pp. 125–152, 2001. isothermal amplification,” Journal of Virological Methods, [128] M. Ciarlet, F. Reggeti, C. I. Piña, and F. Liprandi, “Equine vol. 215-216, pp. 13–16, 2015. rotaviruses with G14 serotype specificity circulate among [113] O. Nakagomi, M. Mochizuki, Y. Aboudy, I. Shif, Venezuelan horses,” Journal of Clinical Microbiology, vol. 32, I. Silberstein, and T. Nakagomi, “Hemagglutination by a no. 10, pp. 2609–2612, 1994. human rotavirus isolate as evidence for transmission of [129] M. K. Estes, E. L. Palmer, and J. F. Obijeski, “Rotaviruses: a animal rotaviruses to humans,” Journal of Clinical Micro- review,” Current Topics in Microbiology and Immunology, biology, vol. 30, no. 4, pp. 1011–1013, 1992. vol. 105, pp. 123–184, 1983. [114] G. Prasad, Y. Malik, and R. Pandey, “G and P genotyping of [130] RCWG, “List of accepted genotypes,” 2018, https://rega. bovine group a rotaviruses in faecal samples of diarrhoeic kuleuven.be/cev/viralmetagenomics/virusclassification/ calves by DIG-labelled probes,” 2005. rcwg. [115] D. Polpanich, P. Tangboriboonrat, A. Elaissari, and [131] A. Lucchelli, S. Y. Kang, M. K. Jayasekera, A. V. Parwani, R. Udomsangpetch, “Detection of malaria infection via latex D. H. Zeman, and L. J. Saif, “A survey of G6 and G10 se- agglutination assay,” Analytical Chemistry, vol. 79, no. 12, rotypes of group a bovine rotaviruses from diarrheic beef and pp. 4690–4695, 2007. dairy calves using monoclonal antibodies in ELISA,” Journal [116] J. Park, S. Kurosawa, J. Watanabe, and K. Ishihara, “Eval- of Veterinary Diagnostic Investigation, vol. 6, no. 2, uation of 2-methacryloyloxyethyl phosphorylcholine poly- pp. 175–181, 1994. meric nanoparticle for immunoassay of C-reactive protein [132] B. R. Gulati, O. Nakagomi, Y. Koshimura, T. Nakagomi, and detection,” Analytical Chemistry, vol. 76, no. 9, R. Pandey, “Relative frequencies of G and P types among pp. 2649–2655, 2004. rotaviruses from Indian diarrheic cow and buffalo calves,” [117] S. Perez-Amodio, P. Holownia, C. L. Davey, and C. P. Price, Journal of Clinical Microbiology, vol. 37, no. 6, pp. 2074– “Effects of the ionic environment, charge, and particle 2076, 1999. 18 Veterinary Medicine International

[133] N. Santos, R. C. C. Lima, C. M. Nozawa, R. E. Linhares, and V. Gouvea, “Detection of porcine rotavirus type G9 and of a mixture of types G1 and G5 associated with wa-like VP4 specificity: evidence for natural human-porcine genetic reassortment,” Journal of Clinical Microbiology, vol. 37, no. 8, pp. 2734–2736, 1999. [134] R. Gray, “Vi rology ©,” Archives of Virology, vol. 4, pp. 737–743, 1995. [135] W. D. Cubitt, A. D. Steele, and M. Iturriza, “Characterisation of rotaviruses from children treated at a London hospital during 1996: emergence of strains G9P2A [6] and G3P2A [6],” Journal of Medical Virology, vol. 61, no. 1, pp. 150–154, 2000. [136] T. Nakagomi and O. Nakagomi, “RNA-RNA hybridization identifies a human rotavirus that is genetically related to feline rotavirus,” Journal of Virology, vol. 63, no. 3, pp. 1431–1434, 1989. [137] K. Nishikawa, Y. Hoshino, K. Taniguchi et al., “Rotavirus VP7 neutralization epitopes of serotype 3 strains,” Virology, vol. 171, no. 2, pp. 503–515, 1989. [138] M. Das, S. J. Dunn, G. N. Woode, H. B. Greenberg, and C. D. Rao, “Both surface proteins (VP4 and VP7) of an asymptomatic neonatal rotavirus strain (1321) have high levels of sequence identity with the homologous proteins of a serotype 10 bovine rotavirus,” Virology, vol. 194, no. 1, pp. 374–379, 1993. [139] S. A. Edwards, D. M. Broom, and S. G. Collis, “Factors affecting levels of passive immunity in dairy calves,” British Veterinary Journal, vol. 138, no. 3, pp. 233–240, 1982. [140] R. L. Larson, J. W. Tyler, L. G. Schultz, R. K. Tessman, and D. E. Hostetler, “Management strategies to decrease calf death losses in beef herds,” Journal of the American Veter- inary Medical Association, vol. 224, no. 1, pp. 42–48, 2004. [141] A. C. B. Berge, D. A. Moore, and W. M. Sischo, “Field trial evaluating the influence of prophylactic and therapeutic antimicrobial administration on antimicrobial resistance of fecal Escherichia coli in dairy calves,” Applied and Envi- ronmental Microbiology, vol. 72, no. 6, pp. 3872–3878, 2006. [142] H. P. Heckert, I. Bardella, B. Brunner, and R. Brunner, “Effects of maternal immunization of cows-a field study,” Praktische Tierarzt, vol. 86, 2005. [143] P. Pithua, S. J. Wells, S. M. Godden, and E. A. Raizman, “Clinical trial on type of calving pen and the risk of disease in Holstein calves during the first 90d of life,” Preventive Veterinary Medicine, vol. 89, no. 1-2, pp. 8–15, 2009. [144] H. Papp, B. Laszl´ o,´ F. Jakab et al., “Review of group A ro- tavirus strains reported in swine and cattle,” Veterinary Microbiology, vol. 165, no. 3-4, pp. 190–199, 2013.