Molecular characterization of unusual bovine rotavirus A strains having high genetic relatedness with human rotavirus: evidence for zooanthroponotic transmission

We report here the genomic characterization of two rare rotavirus A (RVAs) G1P[11] and G9P[X] strains detected in cattle calves from two different geographical locations in India during routine rotavirus surveillance. These strains possessed unusual G types (VP7 gene) on a bovine/artiodactyl genotype constellation, G1‐P[11]‐I2‐Rx‐Cx‐Mx‐Ax‐N2‐T6‐E2‐H3 (HR‐B91) and G9‐P[X]‐I2‐Rx‐Cx‐Mx‐Ax‐N2‐T6‐E2‐H3 (WB‐H2). This is the first report on molecular characterization of G9 in cattle, and second report on G1 in cattle. The VP7 gene of HR‐B91 occupied lineage IIc within G1 while that of WB‐H2 occupied IIIb within G9 genotype. The latter was found to be very diverse from other RVA strains of G9 genotype, and this may emerge as a new genotype in due course. The study provides evidence of zooanthroponotic transmission of human G1 and G9 RVA genes to calves. Of note, the G9 genotype was found to serve as the ancestral genotype for G1. Phylogenetic analysis of remaining gene segments revealed close relatedness to artiodactyl or artiodactyl‐like human RVA strains. The findings of this study highlight the potential role of interspecies transmission and reassortment events in generating the rare rotavirus strains.     

Mycoplasma gallisepticum vaccination: further studies on egg transmission and egg production

Leghorn hens vaccinated twice with an inactivated Mycoplasma gallisepticum (MG) bacterin before egg production and subsequently challenged with virulent MG were protected against transmission of MG through the egg. Unvaccinated control hens transmitted MG through the egg at a high rate. When unvaccinated hens were vaccinated with MG bacterin 2 weeks after challenge with MG, there was no significant decrease in egg transmission. Hens vaccinated twice before laying did not suffer as severe egg-production drops as unvaccinated hens did when challenged with virulent MG. In a natural MG outbreak in a leghorn breeder flock, 50 hens were separated from the remainder of the flock and vaccinated with MG bacterin approximately 3 weeks after initial exposure. The unvaccinated hens transmitted MG through the egg at a rate three times higher than the rate of transmission of the vaccinated hens.     

Studies on neonatal calf diarrhoea caused by rotavirus: transmission of the disease and attempted vaccination of colostrum-deprived calves

Mild to severe scouring could be produced in colostrum-deprived calves with tissue culture-adapted rotavirus and feacal material from field cases of calf diarrhoea. The feaces of experimentally infected calves contained rotavirus for at least 3 days. Pathogenic bacteria were presented in one calf only and this calf also showed the most severe gastroenteritis. Eight calves were vaccinated with a live rotaviral calf diarrhoea vaccine and subsequently challenged with infective rotavirus. Mild scouring was observed after vaccination, but the calves remained normal after challenge. Rotavirus particles were detectable in the faeces for a few days after vaccination and challenge.     

Electrical stimulation on skin wound healing in the horse: preliminary studies

The effect of low-level direct-current stimulation on skin wound healing in the horse was assessed. Self-sustaining electrical circuits with electrodes were implanted subcutaneously in or near the wound. Stimulation by direct current (10 or 20 microA) was used to determine the effect on equine skin healing. The efficacy of electrotherapy was evaluated by sequentially comparing the clinical appearance of the wound and measuring the size of the granulating wound bed. The histologic appearance of the healing stimulated wounds was compared with that in nonstimulated control wounds created on 9 horses. Seemingly, electrical stimulation had no discernible effect on experimentally created skin wounds. Clinical observation and histologic examination of the wounds indicated that severe tissue reaction from the implanted electrodes and concurrent local infection produced local detrimental effects to wound healing.     

Familial cutaneous vasculopathy of German shepherds: clinical, genetic and preliminary pathological and immunological studies.

A genodermatosis affecting the German shepherd breed has been recognized in 26 dogs in Ontario since 1991. Clinical signs, first noted in young puppies, are manifested as pyrexia and lethargy. The main cutaneous lesions are footpad swelling and depigmentation, but there is also crusting and ulceration of ear tips and tail tips, and focal depigmentation of the nasal planum. Affected puppies show no consistent abnormalities in hematological or biochemical parameters, and immunological tests (antinuclear antibody and rheumatoid factor titer, immunoglobulin levels, and CD4+ and CD8+ T-lymphocyte percentages) are normal. Bone marrow analysis has shown myeloid hyperplasia in 5 of 7 cases and myelofibrosis has been detected in 1 case. All but 3 of the 19 clinical cases have been strongly positive for platelet factor-3; however, normal puppies routinely develop positive platelet factor-3 tests. Furthermore, affected pups all had normal numbers of platelets on repeat complete blood counts. Light microscopic examination of footpad biopsies reveals a multifocal nodular dermatitis in which neutrophils and mononuclear inflammatory cells surround foci of dermal collagenolysis, and degenerative and inflammatory vessel lesions. Depigmented lesions have a mild, cell-poor, interface dermatitis, characterized by single cell necrosis of the basal cells, in addition to the nodular dermatitis. Similarities and differences between this disease, a condition known as collagen disorder of the footpads of German shepherds and other forms of cutaneous vasculitis in the dog are discussed. The cause and the pathogenesis of the disease are yet to be elucidated; however, pedigree analysis indicates an autosomal recessive inheritance pattern.(ABSTRACT TRUNCATED AT 250 WORDS)     

Border disease: virus persistence, antibody response and transmission studies

Three groups of five and one group of four oestrus-synchronised sheep were inoculated with Border disease (BD) virus at 52 +/- 2 days after their first service. Transmission of virus to offspring as demonstrated by virus isolation, detection of viral antigen and, or antibody response occurred in 12 of 19 sheep and probably in four others which aborted or produced stillborn lambs. Both apparently normal and clinically affected animals excreted virus in saliva, urine and faeces, and excretion and contact transmission to sheep and pigs persisted for up to two and a half years. Most of the tissues of infected sheep contained virus titres between 10(3.5) and 10(5.5) TCID50 per g. The immune response in the lambs varied, in some it began before birth, in others a transient or low level response was observed in the first or second year, while others remained serologically negative for two and a half years.     

Studies on rotavirus infection and diarrhoea in young calves.

The occurrence of diarrhoea in calves was monitored during the first three weeks of life. Calves fed amounts of colostrum sufficient to produce serum Ig levels in excess of 30 mg per ml did not develop diarrhoea, whereas calves fed less colostrum did. Rotaviruses and mycoplasma-like particles were observed in the faeces of calves with and without diarrhoea. The epidemiology of rotavirus infection in calves is discussed.     

Epizootiological and virological studies of foal diarrhea associated with serotype 3 rotavirus

Epizootiological and virological studies were conducted on foal diarrhea occurring in 3 foal-raising locations in a light horse farm from March to July, 1987. At the first location, although 27 (75%) of 36 foals had developed diarrhea, the isolation rate of rotavirus (RV) was low (5/14 feces, 36%). Many of the foals had the disease as early as 23 days after birth. At the second and third locations, 21 (27%) of 78 foals and 41 (76%) of 54 foals were affected with diarrhea. Isolation rates of RV were 90% (20/22 feces) and 100% (26/26 feces), respectively. The diseased cases were observed throughout the short period from June to early July and on foals aged from 63 to 65 days. These findings suggested the importance of RV as a causal agent of foal diarrhea at the latter location, RV and/or other agents at the former location. All the 3 strains of RV represented from each location were identified as serotype 3 by plaque reduction neutralization test against the antiserums of serotypes 1 to 6 of RV. However, the 3 strains showed low cross-reactivity with antiserum of the serotype 3 of the equine RV.     

Epidemiological studies of piglet diarrhoea in intensively managed Danish sow herds. IV. Pathogenicity of porcine rotavirus

Colostrum-deprived piglets inoculated with rotavirus 24 h after birth developed a profuse diarrhoea that spread to non-inoculated, colostrum-deprived litter mates and, occassionally, to colostrum-fed piglets. Case fatality rates in these 3 categories of piglets were 63.2%, 35.7% and 8.3%, respectively. Surviving piglets recovered in 1-2 weeks, but shedded virus via the faeces for up to 3 weeks p.i. The D-xylose test revealed severe malabsorption, with extremely flat absorption curves for up to 3-4 weeks p.i. Malabsorption was more marked in piglets with a long-lasting faecal virus excretion than in piglets where virus disappeared from the faeces within 10 days p.i. Infected piglets (colostrum-fed and colostrum-deprived) had decreased weight gains and were 5 days older at a bodyweight of 25 kg than non-inoculated controls. It is concluded that rotavirus is probably of significance in diarrhoeal syndromes in suckling piglets, alone or in combination with E. coli or other pathogens.     

Phylogenetic analysis of porcine rotavirus in Argentina: increasing diversity of G4 strains and evidence of interspecies transmission

Group A rotaviruses are one of the most frequently detected viral agents associated with neonatal diarrhea in piglets. In order to characterize rotavirus (RV) strains circulating in Argentinean swine, four porcine production farms located in Buenos Aires were studied. RV strains genotyped as P[6]G4, P[6]G8 and P[1]G6 were found in piglets under 30 days of age, without diarrhea. Phylogenetic and sequence analysis of the VP7 gene from G4 strains available in databases, reveals five porcine new lineages (III-VII) and three sublineages (VIIa-VIIc). The G4 porcine Argentinean strains were grouped with a porcine RV strain isolated in Brazil and another RV strain isolated from a child with diarrhea in Mexico, constituting an American lineage (VII). On the other hand, porcine G6 and G8 were closely related to RV's circulating in Argentinean cattle and South-American camelids, respectively. The fact that G4 porcine lineages were epidemiologically related to human strains, and G6 and G8 Argentinean porcine strains were found related to bovine and South-American camelids, respectively, suggests that pigs might play a crucial role as reservoir and generator of newly adapted emerging RV strains for human and other species.     

Aujeszky's disease of sheep: experimental studies on the excretion and horizontal transmission of the virus

Five 5-month-old merino lambs were nasally inoculated with 10(5.0) TCID50 of Aujeszky's disease virus (ADV). The dynamics of virus excretion in the nasal discharges--in agreement with the histologic findings--indicated that ADV also replicates in extraneural sites, in the upper and lower parts of the respiratory tract. The virus was excreted continuously in the nasal discharges, even during the incubation period. The titres, with certain fluctuations, increased gradually up to the final stage of the fatal disease. Following the onset of the clinical disease, the titre of excreted virus (ranging from 10(4.0) to 10(6.0) TCID50/0.1 ml) was comparable with the ADV content found in the nasal discharge of naturally infected piglets. However, the horizontal transmission of ADV to contact lambs failed.     

Epizootiology of bovine rotavirus infection.

Published information on rotaviruses as pathogens, the source of virus infection and the method of transmission of infection under normal conditions are reviewed. The antigenic differences between rotavirus isolates from children, calves, pigs, foals and mice are discussed. Bovine rotaviruses isolated in the USA and the UK were shown to be closely related antigenically and the US vaccine strain protected calves from challenge with the UK rotavirus. Nineteen normally reared calves, with 20 or more ZnSO4 units of serum delta globulin, were susceptible to rotavirus inoculation at two days of age. They developed diarrhoea, showed body weight loss but recovered. Three calves with less than 10 ZnSO4 units of serum delta globulin developed diarrhoea and died. In a serological survey of 654 adult cows and calves from three herds, between 2 per cent and 37 per cent of individuals in a group had low rotavirus antibody titres and were probably susceptible to rotavirus infection. These were found in all age groups of animals studied, whether or not the group had suffered a recent rotavirus epizootic. It was not possible to predict whether an epizootic would develop on the basis of a serological survey.