in october 2004 , a 3-month - old kitten ( felis domesticus ) was adopted 
                 from the society of the prevention of cruelty to animals ( east london , eastern cape 
                 province , south africa ) and lived with its owner on a farm 23 km outside the city . 
                
it had been neutered and had been vaccinated at 10 months of age with an adjuvanted 
                 inactivated vaccine against rabies ( rabisin ; merial , lyon , france ) , but no 
                 subsequent vaccinations were given .
the cat spent most of the day indoors , but went 
                 out at night and returned in the morning .
it appeared dull and physically unbalanced and its pupils were dilated but it 
                 was not aggressive .
the cat was humanely killed , and its brain was sent to the 
                 onderstepoort veterinary institute for rabies testing . on june 17 , 2005 , a 6-month - old puppy ( canis familiaris ) was brought 
                 by its owner to a veterinarian in the rural town of nkomazi ( mpumalanga province , 
                 south africa ) .
after 
                 symptoms were treated , the dog was discharged , but it was brought back 11 days later 
                 because it was paralyzed , dehydrated , and had a fixed stare .
the dog was humanely killed , and its brain 
                 was sent to the onderstepoort veterinary institute for rabies testing . direct immunofluorescent antibody test with an anti - rabies conjugate cross - reactive 
                 with african lyssaviruses showed numerous and strongly stained inclusion bodies in 
                 every field of impression smears of both brain samples .
isolation of virus was 
                 attempted by suckling mouse brain passage and cell culture ( neuroblastoma cells ; 
                 diagnostic hybrids , athens , oh , usa ) ; both methods were successful for the cat 
                 sample . however , neither method yielded an isolate from the dog sample , despite a 
                 lyssavirus - specific reaction in the original brain sample by direct 
                 immunofluorescent antibody test .
subsequently , antigenic characterization was performed with a panel of 16 monoclonal 
                 antibodies to the nucleocapsid protein of rabies virus ( canadian food inspection 
                 agency , nepean , ontario , canada ) .
gt , genotype ; + , positive reactivity ; var , reactivity with some regional 
                         variants ;  , negative reactivity .
final confirmation of mokv in both case samples was obtained by reverse 
                 transcription  pcr , nucleotide sequencing , and phylogenetic analysis as 
                 described ( 12 ) .
phylogenetic analysis ( figure ) showed 
                 that the virus isolated from the cat sample ( designated mokv173/06 ) belonged to the 
                 same lineage of mokv isolates that were recovered from cats in the same region of 
                 south africa ( 12 ) .
however , the virus detected in the dog sample ( designated mokv404/05 ) 
                 appeared to represent a different south african mokv lineage that was 
                 phylogenetically positioned between known south african and zimbabwean lineages . 
                
this mokv had nucleotide similarities of 88.1%90.4% and 
                 85.3%88.5% with viruses from zimbabwe and south africa , respectively .
phylogenetic tree based on 267 nt of partial nucleoprotein gene sequences of 
                         moloka virus ( mokv ) identified with the n1-n2 primer set as described ( 12 ) .
the tree shows phylogenetic positions of 2 recently identified 
                         cases of mokv infection from south africa ( mokv173/06 from a cat and 
                         mokv404/05 from a dog ) ( in boldface ) relative to previously characterized 
                         mokv isolates from south africa ( sa ) and zimbabwe ( zim ) and lagos bat virus 
                         ( lbv ) as the outgroup .
infections with mokv are rare ; only 23 isolates are known . during the past 2 decades , 
                
because these viruses are not 
                 exclusive to south africa ( 212 ) , lack of isolates from other regions of africa indicates a lack of active 
                 surveillance and limited diagnostic capabilities in many african laboratories . to 
                 our knowledge ,
the 2 cases of infection with mokv we report are the first in 8 years 
                 from south africa .
clinical signs in the dog and cat , including general neurologic 
                 manifestations with a lack of aggression , are often signs that warrant submitting 
                 samples for rabies testing .
whether 
                 these variations are caused by neutral genetic drift or reflect different 
                 epidemiologic features is not known .
phylogenetically , divergence of these viruses 
                 into different lineages indicates active cycles and evolutionary changes that occur 
                 independently , but in close proximity ( a few hundred kilometers apart ) . although the epidemiology of mokv is incomplete , the case for a reservoir host(s ) 
                 among small terrestrial animals of limited range
together with recent isolations of rabies - related lyssaviruses in a human ( 13 ) and wild animals ( 14,15 ) , these reports emphasize the endemicity of these lyssaviruses in south 
                 africa .
public health implications of african rabies - related lyssaviruses should be 
                 recognized by laboratory workers , researchers , veterinarians , wildlife personnel , 
                 gamekeepers , and pet owners
. a better understanding of the epidemiology of these 
                 viruses is vital and can only be achieved by improved surveillance and 
             awareness .
 we recently identified 2 mokola viruses from domestic mammals ( a dog and a cat ) 
                     in south africa . 
 these cases occurred 8 years after the last reported case of 
                     infection with this virus . 
 our findings emphasize the endemicity of 
                     rabies - related lyssaviruses in south africa and the need to better understand 
                     the epidemiology of mokola viruses . 