Case Report of Transmissible Gastroenteritis Coronavirus Infection Associated with Small Intestine and Brain Lesions in Piglets.

This case study report describes a transmissible gastroenteritis coronavirus (TGEV) infection presented in a commercial pig herd. The clinical signs of infection appeared in newborn piglets, including medium morbidity and low mortality rates. Rectal swabs were collected from five different affected litters for laboratory examinations. Samples from two dead piglets and two euthanized affected piglets were collected for gross and histopathological examinations. All fecal samples were tested TGEV positive by real-time polymerase chain reaction (RT-PCR). Necropsy revealed nonspecific gross lesions. The histopathological examinations revealed villi fused with denuded tips and severe villus atrophy, leading to extensive epithelial flattening in middle and lower small intestine. The architecture pattern of villi presented columnar and cuboidal poorly differentiated enterocytes with mild subepithelial edema. In some enterocytes, pycnotic nuclei were detected. Microscopic examination of brain tissue revealed diffuse gliosis in the area of pia matter with mild congestion of the meningeal and parenchymal vessels and neuronal degeneration. In conclusion, this case study reported an epidemic TGEV infection in piglets, characterized by low mortality and medium morbidity rates accompanied by typical histopathological lesions in small intestine, as well as by coexisting brain lesions, that are described for the first time.

Coronavirus gene 7 counteracts host defenses and modulates virus virulence.

Transmissible gastroenteritis virus (TGEV) genome contains three accessory genes: 3a, 3b and 7. Gene 7 is only present in members of coronavirus genus a1, and encodes a hydrophobic protein of 78 aa. To study gene 7 function, a recombinant TGEV virus lacking gene 7 was engineered (rTGEV-Δ7). Both the mutant and the parental (rTGEV-wt) viruses showed the same growth and viral RNA accumulation kinetics in tissue cultures. Nevertheless, cells infected with rTGEV-Δ7 virus showed an increased cytopathic effect caused by an enhanced apoptosis mediated by caspase activation. Macromolecular synthesis analysis showed that rTGEV-Δ7 virus infection led to host translational shut-off and increased cellular RNA degradation compared with rTGEV-wt infection. An increase of eukaryotic translation initiation factor 2 (eIF2α) phosphorylation and an enhanced nuclease, most likely RNase L, activity were observed in rTGEV-Δ7 virus infected cells. These results suggested that the removal of gene 7 promoted an intensified dsRNA-activated host antiviral response. In protein 7 a conserved sequence motif that potentially mediates binding to protein phosphatase 1 catalytic subunit (PP1c), a key regulator of the cell antiviral defenses, was identified. We postulated that TGEV protein 7 may counteract host antiviral response by its association with PP1c. In fact, pull-down assays demonstrated the interaction between TGEV protein 7, but not a protein 7 mutant lacking PP1c binding motif, with PP1. Moreover, the interaction between protein 7 and PP1 was required, during the infection, for eIF2α dephosphorylation and inhibition of cell RNA degradation. Inoculation of newborn piglets with rTGEV-Δ7 and rTGEV-wt viruses showed that rTGEV-Δ7 virus presented accelerated growth kinetics and pathology compared with the parental virus. Overall, the results indicated that gene 7 counteracted host cell defenses, and modified TGEV persistence increasing TGEV survival. Therefore, the acquisition of gene 7 by the TGEV genome most likely has provided a selective advantage to the virus.

An antibody derivative expressed from viral vectors passively immunizes pigs against transmissible gastroenteritis virus infection when supplied orally in crude plant extracts.

To investigate the potential of antibody derivatives to provide passive protection against enteric infections when supplied orally in crude plant extracts, we have expressed a small immune protein (SIP) in plants using two different plant virus vectors based on potato virus X (PVX) and cowpea mosaic virus (CPMV). The epsilonSIP molecule consisted of a single-chain antibody (scFv) specific for the porcine coronavirus transmissible gastroenteritis virus (TGEV) linked to the epsilon-CH4 domain from human immunoglobulin E (IgE). In some constructs, the sequence encoding the epsilonSIP molecule was flanked by the leader peptide from the original murine antibody at its N-terminus and an endoplasmic reticulum retention signal (HDEL) at its C-terminus to allow the expressed protein to be directed to, and retained within, the endoplasmic reticulum. Western blot analysis of samples from Nicotiana clevelandii or cowpea tissue infected with constructs revealed the presence of SIP molecules which retained their ability to dimerize. The analysis of crude plant extracts revealed that the plant-expressed epsilonSIP molecules could bind to and neutralize TGEV in tissue culture, the levels of binding and neutralization reflecting the level of expression. Oral administration of crude extracts from SIP-expressing plant tissue to 2-day-old piglets demonstrated that the extracts which showed the highest levels of in vitro neutralization could also provide in vivo protection against challenge with TGEV.

Induction of protective immunity against transmissible gastroenteritis virus after exposure of neonatal pigs to porcine respiratory coronavirus.

OBJECTIVE: To test the ability of porcine respiratory coronavirus (PRCV) to induce protective immunity to antigenically related transmissible gastroenteritis virus (TGEV) in neonatal pigs. DESIGN: Neonatal pigs were exposed to PRCV when they were 2, 4, or 6 days old and challenge-exposed to virulent TGEV at 10 days of age. ANIMALS: 34 hysterectomy-derived, colostrum-deprived pigs. PROCEDURE: After challenge exposure, clinical signs were observed, body weight, antibody response, and virus shedding were measured, and mortality was determined. RESULTS: After exposure to PRCV, principals had a slightly slower rate of weight gain than did controls; with 1 exception (a PRCV-exposed pig that was dyspneic for 1 day), principals and controls remained clinically normal until shortly after challenge exposure, when all pigs became listless and anorectic and developed watery diarrhea. However, by day 3, most of the pigs that had been exposed to PRCV when they were either 2 or 4 days old began to recover and most (15/18) survived. Conversely, the clinical condition of most of the other pigs worsened and most (14/16) died. Pigs exposed to PRCV when they were 2 or 4 days old also differed from all other pigs in that they had serum virus-neutralizing antibodies for PRCV and TGEV at the time of challenge exposure. CONCLUSIONS: The PRCV can induce protective immunity to TGEV in neonatal pigs and such immunity develops at or about 6 days after exposure to PRCV. Moreover, protective immunity may be coincident with the appearance of virus-neutralizing antibody. CLINICAL RELEVANCE: Exposure to PRCV should enhance a TGE herd vaccination program.

A mathematical model of detection and dynamics of porcine transmissible gastroenteritis.

Transmissible gastroenteritis (TGE) is a viral disease causing dehydration, diarrhoea and death in pigs. The disease is widespread in pig-producing areas of the world but does not occur in Australia. A mathematical model of TGE spread within a pig herd is proposed and calibrated by reference to published data. The model is then applied to two situations of special interest; first to estimate the delay before detection of TGE (6 to over 30 days) when infection is first introduced into a herd of domestic or feral pigs, and second the effect of the disease in a population of feral pigs (could become endemic if transmission is high).

Simulation of the economic impact of transmissible gastroenteritis on commercial pig production in Australia.

A simulation model (AUSPIG) was used to predict the effect of an increase in piglet deaths, and a reduction in growth rate and an increase in feed conversion ratio of grower pigs on the profitability of two herds representative of the Australian pig industry caused by the introduction of transmissible gastroenteritis virus (TGE) into those herds. For each herd, mortality rates for piglets under 1 week of age of 50% and 95% were assumed to represent a 'moderate' and a 'severe' outbreak, respectively. A reduction in net revenue of 70% was predicted to occur in the 6 months after a 'moderate' outbreak of TGE (100% for a 'severe' outbreak). This represents a total loss of between $260 and $330 per breeding sow in the 12 months after infection with the TGE virus. The likely financial impact of an outbreak of TGE on an Australian piggery is substantial and should be considered when addressing quarantine issues.

Immunization of pregnant gilts with PRCV induces lactogenic immunity for protection of nursing piglets from challenge with TGEV.

The level of passive protection against transmissible gastroenteritis virus (TGEV) was evaluated by experimentally infecting 12 pregnant gilts with different doses of porcine respiratory coronavirus (PRCV) and challenging their litters at 4 days of age. An overall survival rate of 70% was found for piglets nursing the 12 PRCV-infected gilts, compared to a 16% survival rate for piglets of nine uninfected control gilts. Six of the PRCV-infected gilts had adequate levels of immunity to resist infection with TGEV following the challenge of their litters. These six completely immuned gilts also solidly protected their litters from TGEV as shown by a 96% piglet survival rate through weaning at 3 weeks of age. The results suggest that respiratory infection with PRCV induces a substantial degree of protective lactogenic immunity against TGEV.

Experience with a planned exposure program for the control of enzootic transmissible gastroenteritis in swine.

Oral inoculation of pregnant sows and gilts with a homogenate of pig intestines containing live, virulent transmissible gastroenteritis (TGE) virus was associated with significant (P < 0.01) reduction of mortality in nursery pigs in a herd affected with enzootic TGE. The mortality of weaned pigs from April through June 1981, when sows were not vaccinated or inoculated, was 9.3%. Mortality of weanling pigs from July through December 1981 was 5.7% (P < 0.01), and stayed consistently between 2.6 and 4.8% from 1982 through 1990. After beginning the exposure program, there were no clinical signs attributable to TGE in the farrowing house or nursery until 1986, when recrudescence of TGE was evident in the nursery. Although the source of the virus could not be substantiated, recrudescence suggested that TGE virus would not be eradicated as long as exposure was continued. The most plausible explanation for the decrease in the incidence of diarrhea and mortality in the nursery pigs after initiation of the planned exposure program is that a higher, longer lasting amount of lactogenic immunity was provided to suckling pigs. Presumably over time, this resulted in less TGE virus carryover into the nursery by weanling pigs, thereby reducing and confining the environmental challenge to the nursery. Strict all-in, all-out pig movement in the nursery with thorough cleaning and disinfecting presumably further reduced the environmental challenge.

Risk factors associated with transmissible gastroenteritis in swine.

Commercial production data base records from 2 Illinois farms, on which epizootic or enzootic transmissible gastroenteritis (TGE) was experienced, were accessed for an epidemiologic study. Risk factors investigated were sow parity, source of sows, location of farrowing crates, and breeding practices. At farm 1, an epizootic was experienced; at farm 2, an epizootic of TGE followed by enzootic TGE was experienced. Initially, crude risk ratios were calculated for these risk factors, and the crude risk ratios were subsequently adjusted for confounders and interactions, using multiple logistic regression techniques. After adjustment, parity-3 sows were 2.3 times more likely to have litters with TGE than were sows of all other parities on farm 1, and parity-1 sows were 2.6 times more likely to have litters that experienced TGE than were sows of all other parities on farm 2. A single boar on each farm was linked to increased likelihood of a sow's litter contracting epizootic TGE on each farm. Enzootic TGE was maintained by the periodic influx of outside-source gilts on farm 2; these gilts were 2.2 times more likely to have litters with TGE than were sows derived from farm 2. Sows housed in farrowing crates located under the cold air inlet of farm 2 were 1.7 times as likely as sows located in other rows to have litters with enzootic TGE.

A double-protease-resistant variant of transmissible gastroenteritis virus and its ability to induce lactogenic immunity.

A virus resistant to 2 major intestinal proteases (trypsin and alpha-chymotrypsin) was derived from the attenuated Purdue strain of transmissible gastroenteritis virus. Its enzymatic stability was confirmed, in vitro, by exposure to proteolytic enzymes and to porcine intestinal fluids. Vaccination of 5 seronegative pregnant sows with the variant virus by a series of 2 oral and 1 IM inoculations resulted in high titers of neutralizing antibody in serum and colostrum. The mean antibody titer in milk whey decreased 44-fold within 1 week after parturition. At 3 days of age, the 40 pigs delivered by these sows were challenge exposed orally with virulent transmissible gastroenteritis virus. Pigs nursing the 5 vaccinated sows underwent a relatively mild clinical course of illness. The average mortality of these 40 pigs was 33%. Thirty-six pigs which had been raised by 4 nonvaccinated sows had a more severe illness, greater daily weight loss, and higher mortality (92%).