Journal of Infection (2004) 48, 289302

www.elsevierhealth.com/journals/jinf

REVIEW

The zoonotic potential of rotavirus

Nigel Cooka,*, Janice Bridgerb, Kevin Kendallc, Miren Iturriza Gomarad,
Laila El-Attarb, Jim Grayd

a
Central Science Laboratory, Sand Hutton, York YO41 1LZ, UK
b
Royal Veterinary College, London, UK
c
Askham Bryan College, Askham Bryan, York, UK
d
Health Protection Agency, Colindale, London NW9 5HT, UK

Accepted 23 January 2004

KEYWORDS Summary Rotaviruses are generally species-specific, but cross-species transmission is

Zoonotic; Rotavirus;
Reassortants
possible, as has been demonstrated experimentally. Several case studies have
indicated infection of humans by animal rotaviruses. Comparison of genetic sequences
of human and animal rotaviruses often reveals close identity. Surveillance of
circulating rotaviruses in the human population has revealed the presence of several
uncommon genotypes. Many of these have been found in domestic animals, and it is
possible that they arose in the human population through zoonotic transmission. The
low incidence of uncommon strains would suggest that such transmission, or at least
the establishment of an animal rotavirus or a human/animal reassortant virus in the
human population, does not happen with any great frequency. However, many millions
of people will be exposed year on year to animal rotaviruses. This happens within
farming communities, and potentially to visitors to the countryside. There may be
some measure of environmental contamination through livestock excrement. This
exposure may not result in high levels of infection, but some infection could occur.
There may be a continual input of rotavirus strains or sequences into the human
population from the animal population albeit at a very low level.
Crown Copyright Q 2004 Published by Elsevier Ltd on behalf of The British Infection
Society. All rights reserved.

Introduction animals play a role as a source of rotavirus

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,1 speculation increased about whether
*Corresponding author. Tel.: 44-1904-462-623; fax: 44-
1904-462-111.
E-mail address: n.cook@csl.gov.uk
infection in humans. But it was observed, using
RNA RNA hybridization assays, that most of the
corresponding genes of animal and human
rotaviruses do not have a high degree of
homology whereas those of rotaviruses from
the same species do.2 4 These observations led
to the view that rotaviruses have a restricted
host range in nature due to lower fitness in non-
host tissues in terms of replication efficiency,
and under natural conditions animal rotaviruses

0163-4453/$30.00 Crown Copyright Q 2004 Published by Elsevier Ltd on behalf of The British Infection Society. All rights reserved.
doi:10.1016/j.jinf.2004.01.018
290 N. Cook et al.

do not infect humans or vice versa.1 There is after (e.g. a few days) another has initiated
however an alternative view that animal infection.8,12
rotaviruses can indeed infect humans and cause
disease whenever the chance exists.5 This is
based on the identification of unusual rotavirus Experimental evidence for zoonotic
types, with properties of strains more commonly transmission of rotavirus
found in animals, which were isolated from
various cases of human infection. These unusual
It has been demonstrated many times that animals
human rotavirus types may have arisen either as
of one species can be infected by rotaviruses which
whole virions or as genetic reassortants between
have been isolated from another species, including
human and animal strains during coinfection of a
humans. The early studies of this type were
single cell. The segmented nature of the genome
reviewed by Theil.13 Two more recent examples
suggests that, like other viruses with segmented
of experimental cross-species infectivity are
genomes such as influenza virus,6,7 rotaviruses
worthy of note. The first is that the human G1
are able to form new strains by a mechanism of
P[8] rotavirus strain Wa is pathogenic for exper-
reassortment. Reassortment can occur when two
imental pigs14 and is currently used as a pathogenic
rotaviruses of two different strains infect the
challenge inoculum in pigs to assess efficacy of
same cell, and during replication and packaging
potential human rotavirus vaccines.14,15 The rota-
they exchange genome segments. 8 The 11
virus NSP1 and NSP4 non-structural proteins show
genome segments of the parental virus strains
species specificity, but phylogenetic analyses of the
can theoretically reassort into 2048 (211) different
Wa NSP1 and NSP4 genes show that both genes
possible genome constellations, if reassortment is
cluster with those of porcine rotaviruses.16,17 In
random.9
addition, G1 rotaviruses are known to infect pigs.18
Gouvea and Brandtly9 hypothesized that rota-
It is possible that these relationships may explain
viruses exist as mixed populations of reassortants,
the molecular basis for the observed infectivity of
and that reassortment was the driving force
human rotavirus Wa for pigs. A second example of
behind diversity. A prerequisite of diversity is
experimental cross-species infectivity where the
co-circulation of many different rotavirus types
genetic composition of the infecting rotavirus has
in a population; and more diversity, and more
been examined is that of the bovine rotavirus
frequency of uncommon strains, is seen in years
isolate PP-1, which was highly pathogenic to
with the highest number of co-circulating
experimental pigs but replicated poorly in exper-
strains.10,11 Gouvea and Brandtly9 considered
imental cattle.18 Sequence analysis showed that
that mixed populations of rotaviruses are being
this G3 rotavirus had porcine NSP4 and VP4 genes
continually propagated in human and animal
but a bovine NSP1 gene. This strongly suggested
hosts, resulting in new and diverse progeny popu-
that PP-1 was a reassortant between bovine and
lations of rotavirus.
porcine rotaviruses but that a species-specific NSP1
There is a question as to whether transmission of
was not critical in cross-infection between cattle
rotaviruses from animals to humans are dead
and pigs. These two examples unequivocally
ends, i.e. do the viruses involved have any
demonstrate that cross-species rotavirus infection
capacity for secondary transmission to other
does occur, at least experimentally, and that this
humans and for initiating an outbreak? Viruses
infection can be pathogenic in the heterologous
which have arisen by reassortment between
species.
human and animal rotaviruses however, may have
greater fitness and become established in the
human population.
With regard to new rotavirus strains arising Epidemiological evidence for zoonotic
through reassortment, a concept of zoonotic transmission of rotavirus
genes may be developed. These can be defined as
genes originating in animal rotaviruses which can Until recently, specific rotavirus types have been
interact with genes of human rotaviruses, to form associated with specific animal species. For example,
infectious rotavirus particles which are serially human rotaviruses most commonly belong to G types
propagated in the human population. 1 4 and P types [4] and [8],19 whereas bovine
It is possible that reassortant rotaviruses can rotaviruses most commonly belong to G types 6, 8
arise not only through simultaneous infection by and 10 and P types [1], [5] or [11].20 Rotavirus
two different strains, but also after asynchro- subgroup II is strongly associated with human strains
nous infection where one strain infects a host, (L. Svensson: personal communication). As more
Zoonotic rotavirus
rotaviruses have been characterized, the host
species specificity of P and G types has become
less distinct. Human group A rotavirus strains that
posses genes commonly found in animal rotaviruses
have been isolated from infected children in both
developed and developing countries. Strains such as
G3 (found commonly in species such as cats, dogs,
monkeys pigs, mice, rabbits and horses), G5 (pigs
and horses), G6 and G8 (cattle), G9 (pigs and
lambs), and G10 (cattle) have been isolated from
the human population throughout the world.21 G
and P type combinations which are found in man
have also been found in animal species. For
example, G10P[11] was found in American and
Canadian cattle by Lucchelli et al.22 and in Indian
cows and buffaloes by Gulati et al.23 G3P[6] and
G4P[6] were found in pigs in Poland and the USA by
Winiarczyk et al.24 and G1P[8] and G5P[8] were
found in pigs in Brazil by Santos et al.25 The
emerging G9 strains26 28 may have arisen in humans
through transfer from animals. They have been
found in lambs and pigs.25,29 In humans, they appear
to cause more severe symptoms than the common
rotavirus strains,26,30 which might be due to less
immunity to these emerging strains, or to greater
virulence being conferred by their genetic makeup.
Several studies have indicated symptomatic
infection of humans by animal viruses. Nakagomi
and Nakagomi31 reported that almost all gene
segments of a rotavirus G3 strain (AU228) isolated
from a child with a pet cat were identical to those
of a feline rotavirus strain (FRV-1). Strains very
similar to this may have become established in
humans.5 A three week-old baby in an Israeli
household which had a young dog (, 6 months old)
was infected with an animal rotavirus G3 strain.32
An outbreak of group B rotavirus gastroenteritis
occurred in Maryland, USA in 1985.33 Six out of 16
people (adults and children) with gastroenteritis
excreted a virus similar to a group B rotavirus strain
in rats.
Das et al.34 reported that a G8 rotavirus which
had widely circulated in newborn infants in India,
causing asymptomatic infection, had VP7 and VP4
gene sequences which were identical to those of a
bovine rotavirus strain. It was not certain whether
this strain was a pure bovine virus, or whether some
of is genes were derived from human rotaviruses via
reassortment. Its NSP1 gene was highly homologous
to that of a G3 strain which was later isolated from
cattle, suggesting reassortment had occurred.35
Cooney et al.36 suggested that, since G8 strains
possess diverse P-types, the exchange of serotype
G8 VP7 genes between humans and cattle may have
occurred on more than one occasion, or may be
continuing. Nakagomi and Nakagomi5 considered
291
that available evidence suggested that whereas
some feline and canine rotavirus strains have
spread into human populations as whole virions,
bovine rotaviruses were involved in reassortment
with human rotaviruses, leading to the emergence
of unusual strains in various parts of the world.
Apparent dual infection with human and animal
rotaviruses has been observed. Nakagomi et al.37
recovered G1P[5] and G1P[8] strains from an infant
with severe diarrhoea. The G1P[5] rotavirus was
genotypically similar to bovine strains. It was not
isolated from the infant in high titre, and possibly
had little if any effect on the childs disease.
Nonetheless it would have had the potential to
reassort with the coinfecting strain.
It is possible that some human rotaviruses have
multiple origins. Holmes et al.38 speculated that an
unusual G8 P[14] type was derived from reassort-
ment between human, bovine and lapine strains.
This review will examine the potential for
zoonotic transmission of rotaviruses from a United
Kingdom perspective, describing the incidence of
uncommon rotavirus strains, detailing potential
routes of transmission of animal rotaviruses to
humans, and discussing risks of exposure, which
should be illustrative of the situation in a wider
geographical context.
Evidence for zoonotic group A
rotaviruses in the United Kingdom
During a survey of circulating rotavirus strains
carried out in the UK between 1995 and 1998,39
several uncommon genotypes were identified.
Reassortment between common human strains
could explain the presence of some of these
uncommon strains, but not all. Some of the strains
with unusual G and/or P types, i.e. G1P[9], G3P[6],
G3P[9], G8P[8], G9P[6] and G9P[8] could be the
result of zoonotic transmission or of gene transfer
by reassortment. Many of these virus types are
found circulating in domestic animals and pets.21
mara et al.40 considered that the G9
Iturriza-Go
strains resulted from the recent introduction into
the UK of G9P[6] rotavirus, followed by reassort-
ment with the common G1P[8], G3P[8] or G4P[8]
viruses. They observed that the G9P[8] strains
increased in incidence between 1995 and 1998,
whereas that of G9P[6] declined: this could suggest
that G9P[6] has a replicative disadvantage in
humans, which could be consistent with an animal
origin. Cubitt et al.26 observed that children
infected with G9 strains were older, and had more
severe symptoms than those infected with other
292
strains; this would support the recent introduction
hypothesis, since this population would thus lack
immunity to these strains. Furthermore, G9 strains
were not detected in archived rotaviruses collected
before 1995 in the UK.
The uncommon rotavirus strains in the UK study
made up 2.3% of the total identified.39 From this
data, it can be estimated that a minimum of 850 of
the reported 17,000 cases of rotavirus infection in
England and Wales annually were caused by strains
of unusual types. If this extrapolated to the whole
population, a total of 25,000 infections with
rotaviruses of unusual types might be expected
annually.39
The following sections will focus on the routes
whereby animal rotaviruses may be transmitted to
humans, to demonstrate the risk of exposure to
these agents in the UK population.
Direct contact with animals
In many developing countries, there is close contact
between humans and domestic livestock. In areas
prone to flooding, or with a monsoon climate, this
can increase the chances of contact with animal
faeces. In the UK and other developed countries,
contact with farm animals is relatively limited, in
terms of the proportion of the population working
on farms, but this contact could be sufficient to
allow ingress of animal rotavirus strains or genes
into the human population. Exposure of humans to
animal rotaviruses may also be promoted through
contact with domestic pets, particularly cats and
dogs.
Contact with farm animals
All farm workers handling livestock, especially the
young animals that are frequently handled, get
contaminated continuously with livestock faeces. It
is common for piglets, lambs and calves to be
handled frequently during the first few weeks of
life. Not many farm workers wear gloves, and they
may eat using soiled hands. Smoking also may be
another way to acquire the viruses. Clothing and
footwear can be heavily contaminated, then taken
into residential accommodation. Because farm
workers are predominantly engaged in manual
work, their hands can get quite rough and provide
cracks where viruses can become located, evading
removal by light washing. Aerosolised viruses
produced from cleaning practices, e.g. hosing of
pens, could also contaminate hair and skin. Often,
it is not farm workers practice to shower after
N. Cook et al.
work, but they can leave the farm and straightaway
participate in a recreational activity, for instance
meeting friends in a public house, where others can
get exposed to any pathogenic microorganisms
which they may harbour on their clothing or hands.
As well as faeces, dust and effluent may be
potential vehicles of transmission of rotaviruses
between animals and farm workers, and also to
others with access to farms. Within piggeries, there
is a continuous cycle of rotavirus infection, with
transmission of rotavirus from piglet to piglet, and
from litter to litter.41,42 Rotaviruses have been
detected in samples of dust, faeces and effluent
collected from a piggery.43 They were detected in
samples taken from farrowing and weaner rooms
but not from fattener and sow houses. The
following house had previously been cleaned and
disinfected, and the weaner house had actually
been free of piglets for 3 months. Porcine rota-
viruses can survive in faeces for at least 4 months
with only a 2 log10 reduction in the titre of
infectious virus.43
In the UK, there are large numbers of farm
animals. The latest DEFRA statistics show that in
2000 there was a total agricultural workforce of
550,000, of which approximately 50% will come into
direct contact with farm animals. This is just under
0.5% of the UK population.
As an illustrative example of a potential con-
sequence of farm workers exposure to rotavirus, a
hypothetical estimate of the number of infections
with bovine rotaviruses through contact with calves
on dairy farms can be made. In England, there are
approximately 18,000 dairy farms. It can be
assumed that, since young cattle need extensive
care, a minimum of 1 person per day per farm is in
direct contact with them. On dairy farms, the
calving season lasts all year. Therefore the number
of calf contact days per person per year in England
is 6,570,000. The average number of dairy cattle
per herd in approximately 80, each of which will
calve once per year. All the calves become infected
with rotavirus, for a 5-day period in their second or
third week of life, and shed virus in their faeces at a
concentration of at least 1 106 infectious rota-
virus particles per ml. If it is assumed that on
average each worker ingests 1 ml faecal material
per day, e.g. when eating lunch with soiled hands,
them they will ingest at least 1 103 infectious
rotavirus particles each day of contact with
infected calves. From the results of the study of
Ward et al.,44 ingestion of 1000 infectious human
rotavirus particles will cause infection in approxi-
mately 85% of people. Therefore, approximately
5.5 million infections per year could occur through
farm workers exposure to animal rotaviruses. Of
Zoonotic rotavirus
course, this exposure would not in every instance
result in infection, as animal rotaviruses will have
limited specificity for humans, and also farm
workers would have a degree of immunity against
the animal rotaviruses through previous contact.
But if there is, say, a one in a million chance of a
farm worker becoming infected through ingestion
of 1000 bovine rotavirus particles, then each year
on average at least five human infections will arise
this way in the UK. If the chance is one in a
thousand, then the number of human infections will
be 5000 per year. Many of these infections will be
asymptomatic. But secondary spread to family or
social contacts could occur. The opportunity for
reassortment between the bovine rotavirus strains
and any human rotavirus strains which may be
infecting the farm workers or their secondary
contacts also exists.
Exposure to animal rotaviruses will also occur
with farm workers handling suckler (beef) calves,
piglets and lambs, and similar calculations could
also be performed for these situations.
Non-farm workers can also have direct contact
with rotavirus-infected livestock. Opportunities are
afforded by access of children to farms, for
example (and perhaps significantly) during the
lambing season, and the proliferation of open
farms rearing exotic species of domesticated
animals, which may also harbour and excrete
rotaviruses. The protozoan parasite Cryptospori-
dium parvum may provide an analogous example. It
is zoonotic, being found in a range of animals
including cattle.45 The transmissive stage of the
parasite, the oocyst, cannot multiply outside a
host, but like a rotavirus particle, it can persist in an
infectious state in various environments.46 Also like
rotaviruses, C. parvum oocysts are shed in large
numbers in the faeces of infected animals.45
Cryptosporidiosis can be acquired through direct
contact with farm animals.47 50 There have been
outbreaks in non-farm workers after visits to open
farms, where faecally contaminated calves were
handled, or raw milk or animal feed consumed.51,52
There are approximately 300 farms in England
which actively allow school visits (data from www.
farmsforteachers.org.uk). Guidelines do, however,
exist to help avoid infection during such visits.50
They recommend amongst other things fresh bed-
ding for livestock to minimize the risk of children
coming into contact with faeces, and the removal
of animals with scour (diarrhoea).
Evidence against transmission of rotavirus
from livestock
In a study undertaken in Panama between August
293
1979 and March 1980, Ryder et al.53 surveyed the
prevalence of antibodies to rotavirus in, and the
incidence of rotavirus diarrhoea among cattle
ranchers, their families, and their livestock. One
hundred and eighty farm workers, and 161 family
contacts were studies. The animals comprised 512
cattle, 35 pigs and 19 sheep. At the start of the
study, less than 30% of the people and animals on all
the farms had detectable antibody (IgG) to rota-
virus, and there was no difference between the
workers who had daily exposure to the animals and
the women and children who rarely had contact
with the animals. No antibodies to rotavirus were
detected in pigs and sheep. By March, approxi-
mately 70% of the humans and nearly 40% of the
cattle had developed detectable antibodies to
rotavirus. No correlation could be observed
between the rates of rotavirus infection in humans
and animals, either overall or on individual farms.
Rates of rotavirus infections among humans were
the same on farms with cattle as on those with pigs
and sheep. Also, infection rates were similar in all
workers, regardless of how long they had been
working with animals. Looking within the workers
families, however, it was observed that there was a
high correlation between infection of children and
infection of adults. Infection rates were higher
amongst parents with four or more children than
amongst those with fewer numbers or no children.
The authors of the study concluded that for
rotavirus infections in humans, children are the
reservoir.
While the antibody prevalences were low in this
study, its results suggested that repeated episodes
of rotavirus infection occurred in individuals,
perhaps at intervals as short as 1 year. No typing
of rotavirus isolated was performed. So it cannot be
ruled out that some of the human infections were
related to the animal infections. As more of the
farms studied had animal containment structures
contiguous with the workers housing, close contact
between children and animals may have occurred
with some regularity and transmission from children
to adults may have involved animal rotavirus
strains.
Contact with pets
Several case studies have indicated infection of
humans by direct contact with household pets.
Nakagomi and Nakagomi31 reported that a rotavirus
group A G3 strain isolated from a child with a pet
cat was identical to a feline rotavirus strain. A 3-
week old baby in an Israeli household which had a
young dog (, 6 months old) was infected with a
canine rotavirus group A G3 strain.32
294
Rotavirus infections in dogs are commonly sub-
clinical.54 Human rotavirus has been shown, exper-
imentally, to infect dogs.55 Dogs are also
susceptible to infection with porcine56 and bovine
rotaviruses.57 Sequence analysis of VP7 genes from
some canine and feline rotavirus strains revealed a
high degree of homology between them and may
reflect interspecies transmission.58
In a study of outpatients with diarrhoea on a
Native American reservation, Engleberg et al.59
found that dog ownership was significantly P
0:05 associated with rotavirus infection. Ownership
of cats, cattle, horses, or any other animal was not
associated with illness or seropositivity to rotavirus.
The most significant factor was actually contact
with children under 2 years. Dogs were not thought
to be the reservoir of infection, but to act as
transmitters of the virus by rooting around on the
refuse piles within the reservation (which contained
soiled nappies) and carrying viruses on their paws or
snouts.
In a survey of healthy cats in an animal shelter in
Melbourne, Australia between March and October
1984, Birch et al.60 found that 5% shed G3 type
group A rotavirus. They noted that the most recent
human infections (March to May 1984) were also due
to G3 rotavirus, and would not rule out the
possibility of interspecies transfer.
Of the 24.5 million UK households, just under
around 45% own a pet. In 2001 the number of
households owning pets was: dogs: 4.8 million;
cats: 4.8 million. The number of pet dogs was 6.1
million and pet cats 7.5 million (data from the Pet
Food Manufacturers Association; www.pfma.com).
Also in the UK, there are around 1 million horses and
3 million horse riders (data from the British Horse
Society www.bhs.org.uk). Handling and care of
infected pets could directly expose humans to
rotaviruses. Also, household contamination of
objects and surfaces with faeces from infected
animals may also allow transmission of rotaviruses
to humans.
Contamination of the environment and
food
Rotaviruses in livestock excreta
The excreta from infected cattle, pigs and sheep
contains large numbers of infectious rotavirus
particles. This is a potential source of contami-
nation in various ways. Viruses in excreta deposited
in fields could pass via run-off water into fresh
waters such as rivers or lakes. Aerosolised virus
N. Cook et al.
could be produced through disturbance of excreta,
e.g. during cleaning practices. Also, excreta from
cattle and pigs can be stored, then spread onto
land. All the excreta are stored, including that from
diarrhoeic animals. It is likely that rotaviruses
could, via manure or slurry, contaminate arable
land, water sources, and possibly also crops. As an
example, the potential extent of rotavirus con-
tamination of land via cattle slurry from an average
dairy farm in the UK can be considered.
The average number of dairy cattle per herd is
approximately 80, each of which will calve once per
year. One dairy cow produces 53 l excreta per day
(data from Anonymous61); assuming it is all stored,
the cattle slurry produced will be 1.5 106 l. There
are 80 calves produced per year, all of which
become infected with rotavirus for a 5-day period in
their second or third week of life. Assuming each
calf produces 2 l of rotavirus-contaminated faeces
per day during infection, the total volume of
excreta the calves produce over the year is 800 l,
containing at least 106 infectious rotavirus particles
per ml. After mixing with the slurry produced by the
older cattle, and assuming homogeneity, the
concentration will be at least 5 105 infectious
rotavirus per litre of slurry. A recent ADAS
recommendation62 is that slurries should be pre-
ferentially stored for 3 months prior to land
application. Bovine rotaviruses have a decay rate
in stored slurry of less than 2 logs in 12 weeks;63
therefore, in the current example, the concen-
tration of rotavirus after 3 months would be around
5 103 per litre. If the slurry is spread onto land,
the contamination per hectare can be estimated.
The area of land required for spreading waste from
one dairy cow housed for 6 months is 0.19 ha.61 So
53 182.5 9.6 103 l of cattle slurry is spread
onto 0.19 ha. Therefore 1 ha will be spread with
approximately 5 104 l of cattle slurry. This area
could then be contaminated with 2.5 108 infec-
tious rotavirus particles. In other words, 1 m2 could
be contaminated with 2.5 104 infectious rotavirus
particles.
There is a round-the-year application of cattle
slurry onto land in the UK.64 This will mediate a
continuous introduction of animal rotaviruses into
the environment. Besides cattle slurry, cattle
manure, pig waste, and sheep excrement will also
provide vehicles for environmental contamination
with animal rotaviruses. Consequent human
exposure to animal rotaviruses through agricultural
work is likely. With regard to farm visits, or
recreational activities such as rambling, the Cryp-
tosporidium analogy may again be illustrative. The
number of cases of cryptosporidiosis in England and
Wales65 and Southern Scotland66 declined in 2001
Zoonotic rotavirus
after the outbreak of foot and mouth disease.
Hunter et al.67 considered that this was probably
due to the removal of access to the countryside,
one of the control measures imposed, which
prevented people from coming into contact with
farm and wild animals and their excrement. It can
be reasonably expected that exposure to animal
rotaviruses also occurs through similar contact, and
it would be interesting to examine rotavirus
incidence likewise. Furthermore, the potential
also exists for contamination of watrcourses and
food crops with animal rotaviruses, via excrement.
Contaminated water
There are several enteric viruses which have been
proven to be transmissible by water,68 and water
has been the vehicle in a number of outbreaks of
rotavirus gastroenteritis worldwide.
A risk of contamination of watercourses with
animal rotaviruses exists through animal excreta.
As is known to occur with Cryptosporidium,46,69
rotaviruses shed from infected animals on pasture-
land could be transported via run-off waters into
watercourses.
It has been shown that enteric virus excreted
from cattle can be present in river water. Ley
et al.70 demonstrated that bovine enterovirus, a
picornavirus which can cause diarrhoea and abor-
tions but which is generally asymptomatic, and can
be detected in the faeces of cattle could also be
found in pasture pools, run-off streams and an
adjacent river. In a study conducted in Switzerland,
Metzler et al.71 detected rotavirus by RT-PCR in
approximately 40% of samples (23/58), which
included sewage effluent, river water, lake water,
spring water and well water. Five isolates of
infectious rotaviruses were isolated from the
samples by cell culture. When typed, three isolates
were G8P[1], one was G8 but unidentified P type,
and another was unidentified. Rotaviruses of the
G8P[1] type have been exclusively detected in
cattle, but when Metzler et al.71 examined 18
faecal samples from diarrhoeic calves in the area of
their study, G8P[1] rotaviruses were not detected.
They did not state whether any human clinical
samples were also screened, and the origin of the
unusual rotaviruses was therefore not clear. Grata-
cap-Cavalier et al.72 detected bovine and porcine
(group A but type not reported) rotaviruses in
drinking water from homes in Isere, France, where
there were children with rotaviral gastroenteritis.
The rotavirus isolates were not the same as those
which were infecting the children, but the authors
considered that consumption of animal rotavirus-
295
contaminated water could facilitate the occurrence
of human-animal virus reassortants.
The MAFF Water Code61 recommends that farm-
yard manure or slurry should not be spread within
10 m of any ditches, streams, rivers or ponds. But a
recent survey of farms in England62 revealed that
this precaution against contamination of water-
courses was not being observed in all cases, with
22% of respondents with manure/slurry and water-
courses spreading less than the recommended
distance. There has been much experimental work
done to show that enteric viruses can be trans-
ported through soils,73 and a recent field study74
demonstrated that poliovirus seeded onto soil could
be transported 21.5 m to contaminate a well.
Rotaviruses may be able to persist in freshwater
for several days. In a study by Raphael et al.,75 it
was demonstrated that a 100-fold decline in
infectious particles in river water took approxi-
mately 10 days at 20 8C, and 32 days at 4 8C. Thus,
rotaviruses have the potential for wide dissemina-
tion through watercourses after a contamination
event. Humans could consequently become
exposed to animal viruses through drinking
untreated water, either directly or after failure of
a potable water treatment or distribution system.
Bathing or recreational water would be another
likely source of exposure.
In the UK, there have been no reported water-
borne outbreaks of rotavirus. But sporadic cases of
waterborne viral gastroenteritis are difficult to
recognise and are likely to go unnoticed.68 This
will be even more apposite of any animal rota-
viruses, given the probable low risk of symptomatic
infection. But the size of the recorded outbreaks in
other countries demonstrates the magnitude of
exposure which waterborne human rotavirus trans-
mission can cause, and gives some indication of the
potential exposure of a human population to animal
rotaviruses, which could be continually present in
some water sources. It would be informative to
verify, and ascertain the extent of, this presence by
a survey of freshwater sources for animal
rotaviruses.
Contaminated food
Animal rotaviruses could potentially be transmitted
by foods directly, i.e. through consumption of meat
from infected animals, or indirectly, through con-
sumption of foods contaminated with organic
waste. Another indirect route of transmission
might be handling of food, such as fresh produce,
sold on farms, by farm workers exposed to infected
animals.
The foods which are normally associated with
296
transmission of enteric viral disease are those which
are eaten raw, such as soft fruit and salad
vegetables, or only lightly cooked, such as shell-
fish.76,77 However, foods which have been contami-
nated by infected handlers post-cooking can also
cause outbreaks of viral disease.
Although rotaviruses should be as easily trans-
mitted by foods as most other enteric viral
pathogens, foodborne disease outbreaks are seldom
reported. In England and Wales, the most common
identified cause of foodborne viral disease are
noroviruses,78 which between 1992 and 1995,
accounted for 6% of reported outbreaks of food-
borne gastroenteritis, and 95% of foodborne gastro-
enteritis outbreaks where a viral agent was
positively identified.78 The reason for this dispro-
portionate figure may be that most of the cases
occurred in large outbreaks, and the food vehicle
was relatively easily identified. It is unlikely that
this would occur with animal rotaviruses, again
given the probable low risk of symptomatic infec-
tion. However, the possibility of sporadic food-
borne infection cannot be ruled out. Also, the
correlates of protection from rotaviruses and
noroviruses are poorly understood although symp-
tomatic reinfection with noroviruses is not uncom-
mon. Repeated rotavirus infections are usually
asymptomatic or very mild.
Group C rotaviruses were the cause of a large
foodborne outbreak affecting schoolchildren and
teachers in seven elementary schools in Fukui City,
Japan:79 this affected more than 3000 individuals.
One particular preparation center supplied lunch to
all the affected schools, and this was the basis for
attributing the outbreak to infected food, although
no particular food item was incriminated through
questionnaire responses and the virus could not be
detected in food samples. The rotavirus isolated
from the patients was immunologically similar to
porcine group c rotaviruses, and Matsumoto et al.79
speculated whether it was actually of porcine
origin. As pork is usually thoroughly cooked it is
unlikely that it was the vehicle in this outbreak;
post-cooking contamination of foods by improper
hygienic practice (e.g. contact with raw pork on
preparation surfaces) cannot however be dismissed
as a possibility.
There is a potential risk of transmission of animal
rotaviruses if crops, particularly those which are
eaten raw, are exposed to farmyard waste. There is
no published information on detection or survival of
rotaviruses in crops or soil amended with farmyard
waste, although information accruing from a
European research project on Cryptosporidium in
food and water may be useful with regard to
analogous situations involving protozoan para-
N. Cook et al.
sites.80 Group A rotaviruses have been detected
by EM and ELISA in lettuces sold in Costa Rican
markets.81 The detection by EM implied that the
viruses were present on the lettuces in high
numbers (at least 105 106 particles per gram of
lettuce), but their source was undetermined.
The Advisory Committee for the Microbiological
Safety of Foods78 considered that there was a
possible role of sludge derived from human sewage
as a potential source of viral contamination of
crops. This concern could also be appropriate with
regard to animal rotaviruses in manure or slurry. In
a report to the Department for the Environment,
Transport and the Regions, Carrington et al.82,83
reviewed various experiments on virus survival in
sewage sludge-amended soils. They produced a
quantitative interpretation of viral decay rates
from the data produced by these studies, and
calculated decimal reduction times for the viruses
involved (enteroviruses including poliovirus). These
studies under review were mostly performed with
North American soil types and indigenous climatic
conditions, and information is lacking on persist-
ence of viruses in soils and conditions pertaining to
other countries. Carrington et al.83 pointed out that
in countries like the United Kingdom, mean soil
temperatures seldom exceed 15 8C at 10 cm depth
is summer, and are about 5 8C in winter. They
suggested that viral decay rates would be slow
under such conditions, with decimal reduction
times from 24 days to over 100 days. This estimate
is supported by the work of Damgaard-Larsen
et al.,84 who studied the survival of enteroviruses
in sludge-amended soil under Danish winter con-
ditions. In December 1975 they added Coxsack-
ievirus B3 to municipal sludges, which were then
placed on soil lysimeters, and dug in by spade.
Samples were taken each month until May 1976.
The viral titre fell by 0.5 1 log10 per month, but
some viruses could still be detected at the end of
the experiment. Carrington et al.83 considered that
cultivation of soil after sludge application would
encourage viral decay by encouraging evaporation,
but this practice could also result in aerosolisation
of virus particles. Irrigation of crops with contami-
nated water or organic waste is a potential means
of contaminating foofdstuffs with enteric viruses,
and studies have demonstrated that rotaviruses can
be transferred to the surfaces of vegetables and
persist there for several days, following the
application of sewage sludge to effluent. Once on
foodstuffs such as vegetables, viruses may persist
under normal storage conditions over the times
usual between purchase and consumption.85 Using
simian rotavirus SA-11 as a model, Badawy et al.86
showed that rotavirus could survive on lettuce,
Zoonotic rotavirus
radishes and carrots for up to 30, 30, and 25 days,
respectively, at refrigeration temperature; at room
temperature, however, the survival periods were
25, 4, and 15 days, respectively.
It is well known that zoonotic agents can be
transmitted via meat contaminated at slaughter.
Petersen et al.87 devised a rating scheme whereby
the public health significance of the potential
transmission of zoonotic agents by beef could be
prioritized. The list of 25 pathogens included
rotavirus, which was ranked 15th (the first three
were E. coli O157:H7, Salmonella, and Listeria, in
that order). Young animals, which are more likely to
be infected with rotavirus and display symptoms,
are not likely to be slaughtered, but older animals
could be infected subclinically, and rotaviruses in
faecal material could contaminate their meat at
slaughter. Abattoirs in the UK grade livestock on
their cleanliness, using a 1 5 scale (1 clean).
Only clean animals get killed. This reduces the
chances that carcasses are contaminated with
zoonotic agents from faecal material from the skin.
Although rotaviruses have been detected in
shellfish,88 shellfish have not been associated with
transmission of rotaviral disease. Metcalf et al.89
speculated that this may reflect some property of
the stability of rotaviruses, e.g. loss of infectivity
upon uptake by shellfish through possible removal
of surface spikes. However, Ansari et al.90 believed
that shellfish harvested from sewage-polluted areas
may be a likely vehicle for rotavirus, and, as
children seldom eat raw shellfish, and adults
seldom display symptoms of infection, shellfish-
associated rotavirus gastroenteritis may go unno-
ticed. Ley et al.70 found bovine enterovirus in
oysters in a river which received run-off from
pasture where cattle grazed. The virus could also
be detected in the cattle faeces, and the run-off
streams.
Contaminated air
Many viral infections can be spread through air91
and it has been suggested that rotaviruses may be
airborne.92 An airborne route of rotavirus trans-
mission has been demonstrated in experimental
animals responsible for outbreaks of epizootic
diarrhoea in infant mice.93,94 Although possibly
borne by aerosol droplets into the respiratory
tract, rotaviruses may not replicate there, but be
translocated by mucocilliary activity into the
gastrointestinal tract and ingested.95 Airborne
droplet spread of rotavirus has been suggested to
explain the universal occurrence of rotavirus
disease in children during the first 4 years of life,
regardless of the level of hygiene that prevails or of
297
the quality of food and water available to them.
Cook et al.92 drew some similarities with the
epidemiology of measles, such as the marked
seasonality of disease, the ubiquity of infection,
and the high attack rate, to support the airborne
route hypothesis for spread of rotavirus.
In a study of rotavirus in pigs in Venezuela,
Utrera et al.96 noted an increase in the number of
infections during periods of low rainfall and
humidity. They suggested that the low relative
humidity promoted an increase in aerosol for-
mation, particularly in relation to virus-laden dust
from faecally contaminated surfaces, which could
result in an increased spread of infection. A dry
atmosphere, especially in combination with lower
air temperature, appears to be conducive to the
start of outbreaks of rotavirus gastroenteritis.97,98
The dry air indoors when it is heated during cold
weather has been postulated99 as one of the causes
of the seasonality of rotavirus infection, as it could
encourage aerosol transmission of rotavirus. The
handling of rotavirus-contaminated material can
create aerosols of various sizes. The larger of these
settle out rapidly and can contaminate the immedi-
ate surroundings, while the smaller (usually , 5 mm
diameter) become droplet nuclei which may remain
airborne for prolonged intervals.99,100
Ansari et al.90 reviewed information derived
from experimental study of the survival of rotavirus
in air. Although there are differences in results
which may be due to differences in the rotavirus
strains and the experimental protocols which have
been employed, overall the information indicates
that rotavirus could survive in air long enough to
pose a risk of infection to persons or animals in the
same environment.
Potential routes of airborne exposure to animal
rotaviruses exist. For instance, intensive pig units
are force-ventilated, as a form of temperature
control and also to prevent airborne disease such as
pneumonia in the pigs. The air is sucked out either
through the side or through the roof. It is
conceivable that this could result in dissemination
of aerosolised rotavirus, to which not only workers
but also people living in the vicinity could be
exposed. Also, spreading of animal wastes onto
land, in particular by slurry spreading mechanisms,
which generate aerosol-sized droplets, could result
in such exposure. In the UK, over 90% of slurry is
applied to land by broadcast spreading,101 and it has
been shown that the technique can disseminate
bacteria over distances of up to 650 m, especially
under windy conditions.102 A current DEFRA project
(WA0804) is conducting extensive studies on bac-
terial pathogen dissemination by land spreading of
solid manure or slurry, and has identified the
298
highest risks to occur for instance from the use of
splashplate tankers, spreading of fresh slurry (no
pathogen die-off), and during high windspeeds (N.
Nicholson: personal communiation).
Contaminated surfaces
Viral contamination of various objects and surfaces
can occur either directly by contact with faeces or
indirectly through virus-contaminated aerosols.91,
103,104
Hands frequently contact environmental
surfaces, and the potential for transfer of rotavirus
between surfaces and hands was demonstrated by
Ansari et al.:105 their findings showed that rotavirus
could retain infectivity for several hours on skin,
and could transfer in an infectious state to other
surfaces. Abad et al.106 examined the survival of
rotavirus on surfaces composed of several
materials, aluminum, china, glazed, tile, latex
and polystyrene, commonly found in households,
and found that infectious virus could persist for at
least 60 days. Moe and Shirley107 found that
rotavirus infectivity could persist for up to a week
under conditions similar to those found in the home
in temperate countries, namely 30% relative
humidity and 20 8C. This could permit the survival
of infectious virus on household objects long
enough to infect persons living in the same
environment with an individual or animal which is
shedding rotavirus. There is some inferential
evidence to indicate that virus-contaminated car-
pets may be a vehicle for transmission of gastroin-
testinal infection.108
Occupational exposure to animal pathogens may
be a source of household contamination. Rice
et al.109 showed that Salmonella enterica could
be cultured from the contents of vacuum cleaners
used in houses where one or more occupant was in
contact with infected cattle, or involved in a
veterinary clinic outbreak of feline salmonellosis,
or engaged in research on the bacterium. Thirty
percent (8/26) of the samples taken from the
households whose occupant(s) had contact with
infected cattle were S. enterica positive, whereas
no positive samples n 12 were identified from
houses with no known occupational exposure. It is
quite possible that occupational exposure to rota-
virus-infected animals could similarly be a source of
household contamination.
Conclusions
There is evidence that zoonotic transmission of
rotaviruses, or at least rotavirus genes, can occur.
N. Cook et al.
The low incidence of uncommon strains in the UK,
however, would suggest that such transmission, or
at least the establishment of an animal rotavirus or
a human/animal reassortant virus in the human
population does not happen with any great fre-
quency. However, the example of the G9 strains
may be instructive. These viruses appeared
recently, possibly from animals: even if such
transmission took place through a single event,
they are now found worldwide. It cannot be
completely dismissed that zoonotic transmission
of rotaviruses could occur in the UK, nor can it be
predicted when such an occurrence could take
place. The rapid movement of people of diverse
populations throughout the world allows the intro-
duction and spread of rotaviruses generated in
developing countries through zoonotic transmission
and reassortment. This could have implications for
a vaccine strategy: it may be that if symptomatic
infections arising from the common human rota-
virus types were reduced or eliminated, then other
infectious strains may still exist within the animal
population.
In the UK, contact with animals, and the input
into the environment of animal wastes, must
frequently expose many thousands of people to
animal rotaviruses. This may not result in high
levels of infection, but some infection could occur.
There may be a continual, albeit very low level, of
input of rotavirus strains or sequences into the
human population from the animal population.
Because of the nature of the exposure, this input
will occur continually, year on year.
It is clear that there is much information lacking,
which would allow a full assessment of any risk
which animal rotaviruses pose to the UK population
through zoonotic transmission. In particular, there
is no data on rotaviruses circulating in animals in
the UK. Whilst detailed surveys of the G and P types
have been undertaken recently in man in the UK, no
comparable survey has been conducted in animals
and there are no published surveys of the G and P
types circulating in cattle/pigs/lambs/cats/dogs/
rodents in the UK. There is a lack of sequence data
from animal rotaviruses, which prevents close
comparison between human and animal strains,
and little is known about the long-term molecular
epidemiology of rotavirus infections in animals. The
replicative or pathogenic potential of animal
rotaviruses in human hosts is not optimally
assessed. There is no information on survival of
rotaviruses in slurry- or manure-amended soil, and
there is no information on whether rotaviruses may
be detected in UK watercourses, or on foodstuffs in
the UK.
Several questions remain to be answered. For
Zoonotic rotavirus
example: is genotyping correct? Some published
G10 primers cross-react with G3 types.20 Do
common human strains have a zoonotic back-
ground, i.e. are gene segments other than those
encoding VP7 and VP4 derived from animal strains?
Examining only G and P sequences from clinical
isolated does not reveal whether other segments,
such as segment 11 (NSP4) may have a zoonotic
background. Are there species-specific clusters of
genotypes, e.g. G types? Species-specific genetic
clusters of NSP4 have been described.17
There are several recommendations which can
be made. In order of priority, these are:
1. To establish whether there is any relationship
between rotavirus strains infecting humans and
animals, a survey should be performed of
concomitantly circulating strains. The survey
should compare rural and urban areas, with
perhaps two examples of each. The study should
involve GP practices for collection of human
samples, and veterinary practices for collection
of animal samples (livestock and pets). The rural
areas should be chosen for high prevalence of
different livestock types, e.g. North Humberside
for pigs, Cheshire for cattle. The study should
focus on group A rotaviruses, as the typing and
classification scheme for these is more fully
developed than for other groups (although
information on group B and C rotaviruses would
also be useful), and examine the G and P types of
the isolates. Genotyping should be performed
with fully validated primer sets. To provide a
fuller picture, the NSP1 and NSP4 genes of each
isolate should also be typed. Sequencing of RT-
PCR amplicons should be performed as far as
possible, to facilitate identification of lineages.
2. A database of animal rotavirus sequences should
be established. Attributes such as (1) Species of
origin (2) Strain name, (3) Isolation date, (4)
Geographical origin of isolate and (5) Sampling
method, i.e. whether it was sub-cultured or
sampled directly, should be included. This
information could be used to order the compli-
cated rotavirus classification system as well as
determine phylogenetic relationships more effi-
ciently. These attributes would be equally as
useful for human rotavirus isolates, and this
information should be included in future
accessions.
3. In order to further develop the phylogenetic
analysis it would be advantageous to find
historical rotaviral sequences to include in any
phylogenetic analysis. If a human sequence was
segregating into a human clade in the past but
was segregating into an animal clade at a later
299
date, then this would be a further proof that
segments were being transferred from animal to
human rotaviruses.
4. The potential for infection of humans by animal
rotaviruses through direct contact with animals
could be studied by monitoring a group, e.g.
incoming veterinary students, likely to handle
infected livestock or pets.
5. It would be beneficial to acquire dose response
information to directly determine the infectivity
of animal rotaviruses to humans. Examination of
the data from vaccine trails should give reason-
able information in this regard.
6. The evidence for cross-species infection raises
interesting questions about the genetic and
cellular basis of species specificity, and this
should be established. This would provide an
element of prediction of replication in heter-
ologous species.
7. Studies to determine how long animal rotavirus
can survive in soil amended with manure or slurry
would provide useful exposure assessment data.
Such studies should model UK agricultural prac-
tice, and involve in situ fields settings using
lysimeters over several seasons, as well as
laboratory microcosm experimentation, to
thoroughly determine survival potential and
examine the factors which may influence it.
The potential for transfer of animal rotaviruses
to crops in soil which has been amended with
manure or sewage sludge could also be studied in
these tests.
8. Examination of watercourses for animal rota-
viruses would facilitate assessment of the extent
of exposure to viruses by this means.
If these recommendations are followed, the
information obtained will allow a fuller under-
standing of the zoonotic potential of rotavirus.
Acknowledgements
We acknowledge the support of the United Kingdom
Department for the Environment, Food and Rural
Affairs (Defra). The views expressed in this paper
are the authors and not necessarily those held by
Defra. We would like to thank the following for
consultation and advice: Dr Ulrich Desselberger; Dr
David Cubitt, Great Ormond Street Hospital,
London; Mr Andrew Schofield, Minster Veterinary
Practice, York; Professor Lennart Svensson, Uni-
versity of Linko
ping, Sweden. Many thanks also to Dr
Nick Nicholson, of ADAS, Boxworth, Cambridge and
to Professor Huw Smith of the Scottish Parasite
Diagnostic Laboratory, Glasgow.
300
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