Rotavirus excretion by village pigs in Papua New Guinea

Cohort studies were conducted on 29 pigs from 3 villages in the Highlands of Papua New Guinea. Animals ranged in age from 9 d to 5 m old. Three hundred and twenty nine faecal samples were collected from individual pigs followed over 3 to 6 w periods, and were examined for group A rotavirus antigen by ELISA, and rotaviral genomic RNA by polyacrylamide gel electrophoresis (PAGE). Electron microscopy was also conducted on selected samples. Group A rotavirus was detected in the faeces of 16 pigs with infected individuals coming from all villages. Non-group A rotavirus resembling group C was found in faeces from pigs from 2 villages. All of the group A rotaviruses examined had the same electrophoretype and this was distinct from that of the common type infecting humans in the area at the time of the study. None of the group A positive samples reacted with monoclonal antisera specific for human group A rotaviruses of serotypes 1, 2, 3, 4, or 8. The non-group A rotaviruses also all had identical electrophoretypes. In contrast to previous findings in intensive piggeries, rotavirus infection did not occur in all young pigs and was not limited to young animals under 2 m of age. Infected pigs varied in age from 12 days to 20 weeks of age. This pattern of infection was attributed to the non-intensive husbandry situations in the villages, with less opportunity for transmission to occur than in intensive piggeries.     

Group A rotavirus excretion patterns in naturally infected pigs

Cross-sectional and cohort studies were conducted in a piggery to investigate the excretion pattern of group A rotavirus in pigs. The cross-sectional survey revealed that 47 (9 per cent) of 521 pigs sampled were excreting rotavirus in the faeces. No rotavirus antigen was detected in the faeces of pigs either less than one week or over two months old. The prevalence of infection increased with age over the sucking period, and was greatest at five weeks old. Diarrhoea was observed in only eight (17 per cent) of the pigs excreting rotavirus. Sixteen piglets from four litters were selected and faecal samples collected daily from each animal from birth to two months old. All the piglets excreted group A rotavirus and the range of ages at which they first became infected was between 13 and 39 days. The average duration of excretion in individual piglets was 7.4 days. Ten of 13 sucking piglets which excreted rotavirus developed diarrhoea soon after it was first detected.     

Characterization of rotaviruses isolated from pigs with diarrhoea in Venezuela

The prevalence of porcine rotavirus infection was studied in 15 different herds located in the north-western region of Venezuela. The presence of rotavirus was studied by direct electron microscopy (EM) and by an enzyme-linked immunosorbent assay (ELISA). From 136 samples analyzed during the six months of the study (September 1983-February 1984), 38 (27.9%) were found to be positive for rotaviruses, with infection more common in animals that were 4-6 weeks old. Atypical rotaviruses were not detected in any of the samples examined. Most rotavirus positive specimens were subgrouped using specific monoclonal antibodies in an ELISA test. The majority of the samples (26 out of 38) were found to exhibit Subgroup I antigenicity. Only two specimens, collected from the same herd in two consecutive months, were found to belong to Subgroup II. To characterize further the circulating rotaviruses, electrophoretic analysis of the RNA genome was performed on samples selected from nine different herds. Great variability in the RNA electropherotypes was observed. No correlation was found between subgroup specificity and the migration of the two smaller segments (Genes 10 and 11), as has been described for human rotaviruses.     

Relative prevalence of typical and atypical strains among rotaviruses from diarrheic pigs in conventional swine herds

Polyacrylamide gel electrophoresis was conducted on genomic RNA extracted from rotaviruses detected in diarrheic pigs from conventional swine herds. Ninety samples contained sufficient virus for RNA band visualization and genome classification. Genome profiles were characteristic of typical group A rotaviruses in 67.8% of the 90 samples, of group B rotaviruses in 10.0%, and of group C rotaviruses in 11.1%. In 11.1% of the samples, the presence of more than 11 bands suggested concurrent infection with more than 1 strain of rotavirus. In infections among nursing pigs, 76.4% were group A rotaviruses, 7.4% were group B, 7.4% were group C, and 8.8% were coinfections. In infections among weaned pigs, 40.9% were group A, 18.2% were group B, 22.7% were group C, and 18.2% were coinfections. Coelectrophoresis with prototype OSU and Gottfried strains revealed a great diversity in electropherotype among field strains of rotavirus.     

Incidence of group A and atypical rotaviruses in Brazilian pig herds

The incidence of rotaviruses as a gastroenteritis causal agent in piglets was studied in 19 pig herds of Sao Paulo State, Brazil, during 1985. From 302 diarrhoea samples collected during January (summer), 65 were positive for rotavirus when analysed by enzyme-linked immunosorbent assay (ELISA) and polyacrylamide gel electrophoresis (PAGE). Sixty-two of these samples belonged to the classical group A rotavirus, three to atypical rotaviruses (ELISA negative and probably group B) and one elicited a mixed electropherotype of group A and atypical rotavirus and was ELISA positive. Atypical viruses appear to be very fragile and were rapidly degraded upon storage of samples at -20 degrees C. Three herds where atypical rotaviruses were present in January were sampled again in August (winter). Nine atypical isolates out of a total 21 positive samples (assayed by electron microscopy and PAGE) were detected again in two of them.     

Genetic diversity and novel combinations of G4P[19] and G9P[19] porcine rotavirus strains in Thailand

Several epidemiological studies reported the detection of rotavirus strains bearing unusual combinations of genetic background of human and porcine rotaviruses. This observation supports the hypothesis of interspecies transmission of rotaviruses in humans and pigs. The aims of this study were to investigate the genotypes and molecular characteristics of rotaviruses in piglets with diarrhea in several farms from two provinces in Thailand. A total of 207 fecal specimens collected from diarrheic piglets were screened for the presence of groups A, B, and C rotaviruses. Group A rotaviruses were detected in 41 out of 207 (19.8%) fecal specimens tested. A wide variety of G-P combination rotavirus strains were detected in this study. The G4P[6] was identified as the most prevalent genotype (39.0%), followed by G4P[23] (12.2%), G3P[23] (7.3%), G4P[19] (7.3%), G3P[6] (4.9%), G3P[13] (4.9%), G3P[19] (4.9%), G9P[13] (4.9%), G9P[19] (4.9%), G5P[6], and G5P[13] each of 2.4%. Furthermore, G5 and G9 in combinations with P-nontypeable strains were also found at each consisting of 2.4% (n = 1) of the collection. It was interesting to note that among diversified porcine rotavirus strains, novel combinations of G4P[19] and G9P[19] strains were detected for the first time in this study. Nucleotide sequences of VP4 and VP7 of these strains were closely related to human rotaviruses reported previously. The data implies that these porcine rotaviruses were probably generated in nature from the reassortment between the viruses of human and porcine origin. This study provides valuable epidemiological information and molecular characteristics of porcine rotaviruses circulating in piglets with diarrhea in northern Thailand.     

Development and evaluation of a nested-PCR assay for <Emphasis Type="Italic">Senecavirus A</Emphasis> diagnosis

Senecavirus A (SVA) has been associated with vesicular disease in weaned and adult pigs and with high mortality of newborn piglets. This study aimed to establish a nested-PCR assay for the routine diagnosis of SVA infection. Tissue samples (n = 177) were collected from 37 piglets of 18 pig farms located in four different Brazilian states. For the nested-PCR, a primer set was defined to amplify an internal VP1 fragment of 316 bp of SVA genome. Of the 37 piglets, 15 (40.5%) and 23 (62.2%) were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. The SVA RNA was detected in 61/177 (34.5%) samples with the RT-PCR, while the nested-PCR assay showed 84/177 (47.5%) samples with the virus (p < 0.05). According to the herds, 11 (61.1%) and 16 (88.9%) of the 18 pig herds were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. Nucleotide sequencing analysis revealed similarities of 98.7–100% among SVA Brazilian strains and of 86.6–98% with SVA strains from other countries. The nested-PCR assay in this study was suitable to recover the SVA RNA in biological specimens, piglets, and/or herds that were considered as negative in the RT-PCR assay, and is proposed for the routine investigation of the SVA infection in piglets, especially when other techniques are not available or when a great number of samples has to be examined.     

Identification of atypical rotaviruses in outbreaks of preweaning and postweaning diarrhea in Québec swine herds.

Intestinal contents of diarrheic pigs from 120 outbreaks of diarrhea were examined for the presence of atypical rotaviruses. Pigs involved in these outbreaks were aged two days to five weeks and samples collected over a period of one year originated from different regions of the province of Québec. Samples were analyzed by polyacrylamide gel electrophoresis (PAGE) for the presence of viral RNA and genome profiles were compared to those of porcine rotavirus A/OSU, B/Ohio and C/Cowden. Based on electropherotypes both typical (group A) and atypical (groups B and C) rotaviruses were identified. Rotaviruses could be demonstrated in 25.8% of outbreaks and together atypical rotaviruses accounted for 46.7% of rotavirus-positive outbreaks. Other common enteropathogens were often present in conjunction with rotaviruses in the preweaning and postweaning outbreaks studied.     

Evaluation of killed and modified live porcine rotavirus vaccines in cesarean derived colostrum deprived pigs

Twenty-eight cesarean derived, colostrum deprived (CDCD) piglets were used to evaluate the efficacy of killed and modified live rotavirus (MLV) vaccines against challenge with virulent A-1 and A-2 rotaviruses. Two killed rotavirus vaccines were evaluated: an experimental vaccine and a commercially available vaccine. Efficacy parameters included: average daily weight gains, rotavirus shedding in feces, morbidity incidence and duration, and rotavirus serum antibody conversion post-vaccination and post-challenge. Piglets vaccinated orally/intramuscularly with the modified live vaccine were completely protected from A-1 and A-2 virulent rotavirus challenge. Nonvaccinated control piglets and piglets receiving killed rotavirus vaccines developed diarrhea, shed virus and exhibited reduced weight gains post-challenge. Only the MLV rotavirus vaccine was able to prevent virus shedding in feces after virulent challenge. Both controls and pigs which received killed vaccines intraperitoneally, orally or intramuscularly shed virus in the feces for 7 days post-challenge and virus peak titers approached 10(7) fluorescent antibody infectious dose (FAID)50/g feces. These studies clearly reflected the inability of killed rotavirus vaccines to induce active local immunity to rotaviral diarrhea in piglets.     

Prevalence and risk factors associated with Cryptosporidium spp. infection in young domestic livestock in India

A total of 938 faecal samples (461 cattle calves, 264 buffalo calves, 55 lambs, 116 kids and 42 piglets) from different livestock farms and individual small holdings in six targeted states of India were collected and screened by modified Ziehl–Neelsen staining technique to determine the prevalence of Cryptosporidium spp. and its association with age, sex, season and faecal consistency in domesticated animals. Overall, 16.2 % of the animals were positive for Cryptosporidium infection with prevalence of 16.3, 24.2, 1.8, 3.5 and 19.1 % in cattle calves, buffalo calves, lambs, kids and piglets, respectively. The prevalence of infection was significantly higher (p?<?0.05) in bovines (19.3 % cattle and 33.7 % buffalo) below 1 month of age than in animals between 1 and 3 months of age. But in piglets, it was higher in the age group of 1 to 3 months (22.6 %) than in younger animals (9.1 %). Also, higher prevalence (p?>?0.05) was recorded in females than in males. Seasons had a significant effect (p?<?0.05) on the prevalence of infection in large ruminants, with the highest prevalence in monsoon (cattle 28.8 % and buffalo 36.6 %) followed by pre-monsoon and post-monsoon season. However, in case of sheep and goats, the prevalence was higher (p?>?0.05) in post-monsoon than in monsoon season. A high degree of association was noticed between Cryptosporidium infection and diarrhoea in ruminants screened during the present study. But, in case of pigs, the prevalence was higher in non-diarrhoeic than in diarrhoeic animals. Genotyping of Cryptosporidium spp. based on nested PCR amplification of partial 18S rRNA and its subsequent digestion with SspI, VspI and MboII restriction enzymes revealed prevalence of Cryptosporidium parvum in representative number of positive samples of cattle, buffalo and goats.     

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.     

The shedding of group A rotavirus antigen in a newly established closed specific pathogen-free swine herd.

A longitudinal study was undertaken in a newly established specific pathogen-free (SPF) swine herd to determine the dynamics of rotavirus antigen shedding in a closed swine facility. Pregnant SPF gilts which populated the herd, and their offspring, were monitored weekly for three consecutive lactations. Fecal samples were assayed for the presence of group-specific viral antigen by a solid phase immunoassay (ELISA). Results indicate that in the week prior to farrow, 35% of samples from gilts/sows contained rotavirus antigen. During nursing, 37% of the gilts'/sows' fecal samples also contained virus antigen. Over the course of three farrowings, every gilt/sow in the herd excreted virus antigen. Virus antigen was present in 25% of the samples tested from nursing pigs and in 70% of the samples tested from pigs in the postnursing period; 95% of the litters excreted virus antigen either while nursing or postweaning. Seasonal incidence in virus antigen excretion was noted with proportionally more suckling pigs virus antigen-positive in summer and proportionally more sows/gilts positive during winter. Diarrhea occurred only rarely in the sampled population. Although piglets shed rotavirus subclinically, ELISA positive feces from piglets of each lactation caused severe disease when fed to neonatal gnotobiotic pigs. Electropherotyping of these passaged viruses indicated minor variation in RNA banding patterns over time.     

Feeding regime and management of local Lao pigs in Central Lao PDR

A survey was carried out in smallholder farming systems in the districts of Borrikun, Khamkeuth, and Pakkading, in central Lao People's Democratic Republic, to assess the feeding regime and health status of local Lao pigs. In total, 216 farmers, 72 in each district, were interviewed using a questionnaire. Feed samples were collected for nutrient analysis and pen soil samples for mineral content determination. Farmers kept sows of native breeds such as Moo Lath, Moo Chid, and Moo Hmong. The number of sows per household was low (1.1?±?0.1, 1.1?±?0.1, and 1.2?±?0.1 in Borrikun, Khamkeuth, and Pakkading, respectively) and pigs were mainly fed with rice bran and fresh cassava roots. Daily protein and mineral (Calcium, phosphorus, potassium, and sodium) allowances were lower than requirements. In all districts, the number of piglets at birth was around seven and pre-weaning piglet mortality was around 20 %. The most common causes of piglet death, before weaning, were diarrhea, runts, and squashed piglets. Most piglets were weaned after 4 months of age, but in the Borrikun and Pakkading districts, 11–16 % of households were weaned at <4 months. In all districts, pig management was mainly women's responsibility (around 90 %) and most farmers spent less than 1 h per day on pig management. A feasible and cheap way to increase nutrient supply to local pigs in central Lao People's Democratic Republic would be to include more fresh plant material (e.g., thick head, taro) and salt in the diet. Information and education on improving pig management should be targeted towards women.     

First Isolation of Rotavirus Associated with Neonatal Diarrhoea in Guanacos (Lama guanicoe) in the Argentinean Patagonia Region

Group A rotavirus (RV) and coronavirus (CV) are common viral pathogens associated with neonatal diarrhoea in numerous animal species. The purpose of this work was to investigate the presence of these viral agents in two farm populations of captured guanacos (Lama guanicoe) in the Argentinean Patagonia region, that developed severe diarrhoea outbreaks. Stool and serum samples were analysed for RV and bovine CV antigen and antibody by enzyme‐linked immunosorbent assay. Rotavirus was detected in faeces from two new‐born guanacos with acute diarrhoea, one in each farm. After electrophoretic analysis, each isolated strain, showed a distinctive long dsRNA electropherotype characteristic of group A rotaviruses (4:2:3:2). In addition, 95% (38 of 40) of the sampled animals were positive for RV antibodies, suggesting a high prevalence of RV infection in the populations tested. No evidence of CV circulation by antigen or antibody analysis was observed. To our knowledge, this is the first report of the detection and isolation of RV associated with neonatal diarrhoea in Lama guanicoe.     

Transfer of maternal antibody against group A rotavirus from sows to piglets and serological responses following natural infection

An enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of antirotaviral antibody in sera and faeces from pigs and used to study the dynamics of antirotaviral antibody responses in three cohorts of pigs. Piglets acquired antirotaviral antibody by sucking their dams soon after birth. Antirotaviral antibodies of IgA and IgG classes were detected in both colostrum and milk of all sows tested but IgM class antibodies were not. The antibody levels in colostrum were eight to 32 times higher than those in milk which was collected 18 days post partum. The levels of antibody in piglets' sera were comparable to those in colostrum but declined quickly to low levels by one month old. Maternal antibody was also detected in the faeces of piglets up to 18 days old. Natural rotavirus infection occurred in each of these cohorts when the geometric mean ELISA titres of maternal antibody in their sera declined to 1/1600 (by days 21, 25 and 30 for cohorts 1, 2 and 3, respectively). However, a positive correlation was not obtained between the levels of antirotaviral antibody and protection in individual litters within each of the cohort groups. In each of the cohorts, rotavirus infection usually occurred in one or two piglets first and then spread to other piglets in the same cohort. It is therefore suggested that maternally derived antibody is protective against rotavirus infection in piglets only for the first one or two weeks. Following natural infection with rotavirus, increases in serum antibodies were detected in two of the three cohorts by 20 to 30 days after the average time of onset of faecal shedding of virus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epidemiology of typical and atypical rotavirus infections in New Zealand pigs

Polyacrylamide gel electrophoresis (PAGE) and enzyme-linked immunosorbent assay (ELISA) were employed to investigate the epidemiology of typical and atypical rotavirus infections in five piggeries. Of 152 faecal samples examined, 46 (30 per cent) were positive by ELISA for group A rotavirus. Rotaviruses with electrophoretic patterns resembling groups A, B and C were detected. At least two and up to five different rotavirus electrophoretypes (typical and/or atypical) were detected in each of the five piggeries. Out of 152 faecal samples examined, 28 (18 per cent) contained rotaviruses with group A electrophoretypes, 9 (6 per cent) with group C but only 1 with Group B. Six samples contained both group A and group C rotaviruses. No common electrophoretypes of group A or C rotaviruses were detected in these five piggeries. The PAGE technique was also used to analyze group A rotavirus isolated sequentially from another piggery over a three year period. A single electrophoretype was found during the first two years, but in the third year a different electrophoretype was detected.     

2014-2019年青藏高原牦牛轮状病毒血清学检测与分析

为明确青藏高原4个省区牦牛轮状病毒(RV)的流行情况,本研究采用双抗原一步夹心ELISA法,对2014-2019年采集自西藏、青海、甘肃、四川的630份牦牛血清样品进行了RV抗体检测.结果显示:630份青藏高原牦牛RV血清抗体总阳性率为18.57％(117/630),西藏地区牦牛RV抗体阳性率与其他地区相比较高;201...