Porcine epidemic diarrhea virus reduces feed efficiency in nursery pigs.

Porcine epidemic diarrhea virus (PEDV) infects enterocytes and in nursery pigs, results in diarrhea, anorexia, and reduced performance. Therefore, the objective of this study was to determine how PEDV infection influenced growth performance and repartitioning of amino acids and energy in nursery pigs. A total of 32 barrows and gilts, approximately 1 wk post-wean (BW = 8.46 ± 0.50 kg), and naïve for PEDV were obtained, weighed, and allotted based on sex and BW to one of two treatments: 1) Control, PEDV naïve and 2) PEDV-inoculated (PEDV) with eight pens of two pigs each per treatment. On day post-inoculation (dpi) 0, PEDV pigs were inoculated via intragastric gavage with PEDV isolate (USA/Iowa/18984/2013). Pig and feeder weights were recorded at dpi -7, 0, 5, and 20 in order to calculate ADG, ADFI, and G:F. Eight pigs per treatment were euthanized on dpi 5 and 20, and tissues and blood were collected. At dpi 5, all PEDV pigs were PCR positive for PEDV in feces. Overall, PEDV pigs tended (P < 0.10) to increase ADFI, which resulted in reduced (P < 0.05) feed efficiency. At dpi 5, PEDV pigs had reduced (P < 0.05) villus height and increased (P < 0.05) stem cell proliferation in the jejunum compared with Control pigs. Pigs inoculated with PEDV had increased (P < 0.05) serum haptoglobin and increased insulin-to-glucose ratios compared with Control pigs at dpi 5. Markers of muscle proteolysis were not different (P > 0.05) between treatments within dpi; however, at dpi 5, 20S proteasome activity was increased (P < 0.05) in longissimus dorsi of PEDV pigs compared with Control pigs. Liver and jejunum gluconeogenic enzyme activities were not different (P > 0.05) between treatments within dpi. Overall, PEDV-inoculated pigs did recover the absorptive capacity that was reduced during PEDV infection by increasing proliferation of intestinal stem cells. However, the energy and nutrients needed to recover the epithelium may be originating from available luminal nutrients instead of muscle proteolysis and gluconeogenesis. This study provides insight into the effects of an enteric coronavirus on postabsorptive metabolism in nursery pigs.

Effects of porcine epidemic diarrhea virus infection on nursery pig intestinal function and barrier integrity.

The pig intestinal epithelium can be compromised by pathogens leading to reduced integrity and function. Porcine epidemic diarrhea virus (PEDV), recently detected in North America, exemplifies intestinal epithelial insult. Although several studies have investigated the molecular aspects and host immune response to PEDV, there are little data on the impact of PEDV on pig intestinal physiology. The objective of this study was to investigate the longitudinal impact of PEDV on nursery pig intestinal function and integrity. Fifty recently-weaned, 5-week-old barrows and gilts (BW=9.92±0.49kg) were sorted based on body weight (BW) and sex into two treatments: 1) Control or 2) PEDV inoculated. At 2, 5, 7, and 14days post inoculation (dpi), 4 pigs per treatment were euthanized and jejunum sections collected. PEDV antigen was detected in inoculated pigs by immunohistochemistry in 50% (2/4) at dpi 2, 100% (4/4) at dpi 5, and none at later time points. PEDV-infected pigs had reduced (P<0.05) villus height and decreased transepithelial resistance compared with controls. Total acidic mucins, particularly sialomucin, were reduced in PEDV pigs at dpi 2 and then increased compared with controls at dpi 7 and 14. In addition, PEDV pigs had increased stem cell proliferation (P<0.05) and a numerical increase in DNA fragmentation compared with controls through dpi 7 which coincided with an observed return of digestive function to that of controls. Collectively, these data reveal that PEDV infection results in time-dependent changes not only in intestinal morphology but also barrier integrity and function.

Nursery pig growth performance and tissue accretion modulation due to porcine epidemic diarrhea virus or porcine deltacoronavirus challenge.

Porcine epidemic diarrhea virus (PEDV) and porcine deltacoronavirus (PDCoV) are both members of the family which induce clinical signs of diarrhea, dehydration, and in some circumstances, mortality. Most research has been focused on isolation, genome sequencing, pathogenicity, and virulence of these viruses, but there is little information on long-term growth performance and tissue accretion of pigs inoculated with PEDV or PDCoV. Therefore, our objective was to determine the effect of PEDV or PDCoV infection on growth performance and tissue accretion over 42 d following inoculation. A total of 75 Choice Genetics Large White Pureline barrows and gilts (BW = 10.81 ± 0.81 kg) at approximately 2 wk post-wean and naïve for PEDV and PDCoV were selected. Pigs were allotted based on BW and sex, stratified across 3 treatments with 8 pens per treatment. Treatments were: 1) Control ( = 8); 2) PEDV inoculated ( = 8); and 3) PDCoV inoculated ( = 8). On day post inoculation (dpi) 2, 5, 7, and 14 pigs were euthanized for tissue collection and analyses from these tissues are discussed elsewhere. Pen feed intake and BW were recorded on dpi 2, 5, 7, and weekly thereafter until dpi 42. On 1 designated pig per pen, initial and final body composition was determined using dual-energy X-ray absorptiometry (DXA) and tissue accretion rates were calculated over 6 wk test period. Peak PEDV infection was noted at 3 dpi compared with 4 dpi for PDCoV pigs as determined by fecal swab quantitative real-time PCR (RT-PCR). Control pigs remained negative for PEDV and PDCoV throughout the experiment. Overall, Control and PDCoV pigs did not differ in ADG, ADFI or G:F ( > 0.05). Compared to Control and PDCoV pigs, the overall 42 d ADFI was reduced in the challenged PEDV pigs ( < 0.05) by 19 and 27%, respectively. PEDV did not significantly reduce the overall ADG or G:F compared with Control and PDCoV pigs; however, the biggest reduction in ADG and ADFI for PEDV pigs was within 14 dpi compared to the Control pigs ( < 0.05). Whole body tissue accretion was altered due to PED, with fat, lean, protein, and bone mineral accretion reductions by 24, 20, 21, and 42%, respectively ( < 0.05) compared with Control pigs. Overall, nursery pig performance was greatly impacted by PEDV challenge. Surprisingly, the PDCoV challenge did not negatively influence nursery pig performance. This study provides further insight into the longitudinal impact swine enteric coronaviruses have on growing pigs.

Swine Dysentery.

Swine dysentery is a severe enteric disease in pigs, which is characterized by bloody to mucoid diarrhea and associated with reduced growth performance and variable mortality. This disease is most often observed in grower-finisher pigs, wherein susceptible pigs develop a significant mucohemorrhagic typhlocolitis following infection with strongly hemolytic spirochetes of the genus Brachyspira. While swine dysentery is endemic in many parts of the world, the disease had essentially disappeared in much of the United States by the mid-1990s as a result of industry consolidation and effective treatment, control, and elimination methods. However, since 2007, there has been a reported increase in laboratory diagnosis of swine dysentery in parts of North America along with the detection of novel pathogenic Brachyspira spp worldwide. Accordingly, there has been a renewed interest in swine dysentery and Brachyspira spp infections in pigs, particularly in areas where the disease was previously eliminated. This review provides an overview of knowledge on the etiology, pathogenesis, and diagnosis of swine dysentery, with insights into risk factors and control.

The effect of porcine reproductive and respiratory syndrome virus and porcine epidemic diarrhea virus challenge on growing pigs I: Growth performance and digestibility.

Porcine reproductive and respiratory syndrome (PRRS) and porcine epidemic diarrhea (PED) are two diseases costly to the U.S. swine industry. The objective of this study was to determine the impact of PRRS virus and PED virus, alone or in combination, on growth performance, feed efficiency, and digestibility in grower pigs. Forty-two gilts (16 ± 0.98 kg BW) naïve for PRRS and PED were selected and allocated to 1 of 4 treatments. Treatments included 1) a control, 2) PRRS virus infected, 3) PED virus infected, and 4) PRRS+PED coinfection (PRP). Pigs in treatments 2 and 4 were inoculated with a live field strain of PRRS virus via intramuscular and intranasal routes at 0 d after inoculation (dpi). Treatments 3 and 4 were orally inoculated with a cloned PED virus at 15 dpi. Infection with PRRS virus was confirmed by quantitative PCR and seroconversion. Infection with PED virus was confirmed with PCR. Control pigs remained PRRS and PED virus negative throughout the study. All pigs were offered, ad libitum, a standard diet with free access to water. During the test period, PRRS reduced ADG and ADFI by 30 and 26%, respectively ( < 0.05), compared with control pigs, whereas PRP decreased ADG, ADFI, and G:F by 45, 30, and 23%, respectively ( < 0.05). Additional reductions in ADG and G:F were detected in PRP pigs compared with singular PED or PRRS treatments (33 and 16%, respectively). The impact of PED, alone or in combination, on performance (15-21 dpi) reduced ADG (0.66 vs. 0.35 vs. 0.20 kg/d; < 0.01), ADFI (1.22 vs. 0.88 vs. 0.67 kg/d; = 0.003), and G:F (0.54 vs. 0.39 vs. 0.31; = 0.001) compared with control pigs. Compared with control pigs, PRRS infection did not reduce apparent total tract digestibility (ATTD) of nutrients and energy. However, PED infection, alone or in combination, decreased ATTD of DM and energy by 8 and 12%, respectively ( < 0.05). Compared with control pigs, PRP reduced N and OM ATTD by 13 and 3%, respectively ( < 0.05). No significant differences in apparent ileal digestibility (AID) were detected between virus challenges. However, Lys AID tended to be reduced in both PED treatments compared with the control (10 and 12%; = 0.095). Altogether, PRRS reduced growth but did not alter digestibility. Pigs challenged with PED and, to a greater extent, the coinfection of PED and PRRS viruses had reduced ADG, ADFI, G:F, and ATTD of nutrients and energy.

The effect of porcine reproductive and respiratory syndrome virus and porcine epidemic diarrhea virus challenge on growing pigs II: Intestinal integrity and function.

The objective of this study was to determine if intestinal function and integrity is altered due to porcine reproductive and respiratory syndrome (PRRS) virus and porcine epidemic diarrhea (PED) virus infection in growing pigs. Forty-two gilts (16.8 ± 0.6 kg BW), naïve for PRRS and PED, were selected and randomly assigned to 1 of 4 treatments: 1) a control (CON; = 6), 2) PRRS virus challenge only (PRRS; = 12), 3) PED virus challenge only (; = 12), or 4) coinfection of PRRS + PED viruses (PRP; = 12). Treatments 2 and 4 were inoculated with a live field strain of PRRS virus on d 0 after inoculation. Treatments 3 and 4 were inoculated with PED virus on 14 d after inoculation (dpi) and all pigs were euthanized 7 d later (21 dpi). Infection with PRRS virus was determined by viremia and seroconversion. Fecal quantitative PCR was used to confirm PED virus infection. Control pigs remained PRRS and PED virus negative throughout the study. Compared with the CON, intestinal morphology was unaffected by PRRS. As expected, PED and PRP treatments resulted in duodenum, jejunum, and ileum villus atrophy compared with the CON treatment ( < 0.01). Ex vivo transepithelial electrical resistance (TER) did not differ between CON and PRRS pigs (P < 0.05) but was reduced by 40% in PED alone ( < 0.01). Interestingly, TER was increased ( < 0.01) in the PRP pigs. Active transport of glucose was increased in PRRS pigs over CON pigs ( < 0.01), whereas PED had pigs increased ( < 0.01) active glutamine transport over the CON pigs. Jejunum GLUT2 mRNA abundance and sucrase, maltase, and Na+/K+ adenosine triphosphatase activities tended to be increased in PRRS pigs compared with CON pigs ( < 0.06). The jejunum AA transporter, SLC6A14, and mucin 2 mRNA abundance tended to be increased in PED-only pigs ( < 0.10). These data suggest that PRRS infection supports a higher affinity for glucose uptake, whereas PED favors glutamine uptake. Interestingly, digestive machinery during PED challenge remained intact. Altogether, PED but not PRRS challenges alter intestinal morphology and integrity in growing pigs.

Characterization of Porcine Epidemic Diarrhea Virus Isolate US/Iowa/18984/2013 Infection in 1-Day-Old Cesarean-Derived Colostrum-Deprived Piglets.

Porcine epidemic diarrhea virus (PEDV) was first recognized in North America in April 2013 and has since caused devastating disease. The objective of this study was to characterize disease and viral detection associated with an original North American PEDV isolate inoculated in neonatal piglets. Thirty-six 1-day-old cesarean-derived and colostrum-deprived piglets were randomly assigned to the control (n = 16) or challenged group (n = 20); the latter were orogastrically inoculated with 1 ml of US/Iowa/18984/2013 PEDV isolate titered at 1 × 10(3) plaque-forming units per milliliter. Rectal swabs were collected from all piglets prior to inoculation and every 12 hours postinoculation (hpi) thereafter, with 4 control and 5 challenged piglets euthanized at 12, 24, 48, and 72 hpi. One piglet had a positive real-time quantitative polymerase chain reaction test on rectal swab at 12 hpi, and all remaining piglets were positive thereafter, with highest viral quantities detected at 24 and 36 hpi. Diarrhea was evident in 30% and 100% of challenged piglets at 18 and 24 hpi, respectively. Viral antigen was detected in enterocytes by immunohistochemistry in the duodenum and ileum of piglets euthanized at 12 hpi and was apparent throughout the small intestine of all piglets thereafter, with villus height:crypt depth ratios consistently below 4:1. Viremia was confirmed in 18 of 20 pigs at euthanasia. Clinical disease was severe and developed rapidly following infection with an original North American PEDV isolate, with lesions, viremia, and antigen detection possible by 12 hpi.

Evidence of Oxidative Injury in Pigs With Melanosis Coli.

Melanosis coli is a dark discoloration of the colon due to accumulation of pigment-laden macrophages in the lamina propria. Three case submissions were received where rectal discoloration was reported at slaughter in pigs from separate production systems and melanosis coli was confirmed microscopically. Tissues from affected and unaffected cohort pigs were evaluated for evidence of oxidative damage using immunohistochemical staining for 3-nitrotyrosine, 4-hyroxynonenol, and malondialdehyde. Affected colons had significantly greater immunolabeling for all 3 target compounds than unaffected colons (P ≤ .001, all analyses). Hepatic vitamin E levels were low in both affected and unaffected pigs, and there was a trend toward lower values in affected pigs. Given the limited number of slaughter-collected samples available for this investigation, further study is warranted to elucidate the possible association between low vitamin E concentrations and oxidative damage in cases of melanosis coli in pigs.

Pathogenesis of porcine epidemic diarrhea virus isolate (US/Iowa/18984/2013) in 3-week-old weaned pigs.

Porcine epidemic diarrhea virus (PEDV) is associated with clinical diarrhea in naïve swine of all ages. This report describes timing of antibody generation and disease progression following infection with a US PEDV isolate by assessing fecal viral shedding, morphometric analysis of intestinal lesions, and magnitude of immunohistochemical staining. Sixty-three, 3-week-old pigs were randomly allocated into control (n=27) and challenged (n=36) groups. Challenged pigs were administered 1 mL of 1 × 10(3) PFU/mL of US/Iowa/18984/2013 PEDV isolate by oro-gastric gavage. Three control and four challenged pigs were necropsied on days post-inoculation (dpi) 1, 2, 3, 4, 7, and weekly thereafter, until study termination on dpi 35. Clinical disease, fecal shedding, body weight, and temperature were monitored during the study period. Diarrhea was observed in challenged pigs beginning for some on dpi 2, affecting a majority of pigs by dpi 6 and subsiding by dpi 10. Average daily gain was significantly lower (P<0.001) for one week post-infection in challenged pigs. PEDV was detected in feces by PCR on dpi 1 and continued in a subset of pigs until dpi 24. PEDV-specific antigen was detected in villous enterocytes of challenged pigs by immunohistochemistry (IHC) on dpi 1, 2, 3, 4, 7, and 14. Microscopic lesions included severe diffuse atrophic enteritis with significantly reduced (P<0.001) villous length observed on dpi 3, 4, and 7. Under the conditions of this study, fecal shedding of PEDV and IHC staining can precede and continue beyond the observation of clinical signs, thus increasing the risk of viral transmission.

Cessation of clinical disease and spirochete shedding after tiamulin treatment in pigs experimentally infected with "Brachyspira hampsonii".

With the emergence of "Brachyspira hampsonii" associated with swine dysentery in North America, identification of effective treatments and interventions is a pressing need. Denagard® (tiamulin hydrogen fumarate) Liquid Concentrate 12.5% is approved in the United States for treatment of dysentery caused by Brachyspira hyodysenteriae at 0.006% in the water. In this study, the effectiveness of tiamulin in resolving clinical disease, eliminating viable spirochete shedding, and reducing neutrophilic colitis following infection with either "B. hampsonii" or B. hyodysenteriae was evaluated. Seventy-eight 7-week-old crossbred pigs were divided into three groups [sham-inoculated (n = 18), "B. hampsonii"-inoculated (n = 30), and B. hyodysenteriae-inoculated (n = 30)]. Each inoculum group was divided into three subgroups which received either 0.006% tiamulin, 0.018% tiamulin, or no medication. Both levels of tiamulin resolved clinical disease within 24 h of treatment initiation, eliminated spirochete shedding within 72 h of treatment initiation, and resolved and/or prevented histologic lesions in pigs infected with either Brachyspira spp.

Comparison of sesion severity, distribution, and colonic mucin expression in pigs with acute swine dysentery following oral inoculation with "Brachyspira hampsonii" or Brachyspira hyodysenteriae.

Swine dysentery is classically associated with infection by Brachyspira hyodysenteriae, the only current officially recognized Brachyspira sp. that consistently imparts strong beta-hemolysis on blood agar. Recently, several strongly beta-hemolytic Brachyspira have been isolated from swine with clinical dysentery that are not identified as B. hyodysenteriae by PCR including the recently proposed species "Brachyspira hampsonii." In this study, 6-week-old pigs were inoculated with either a clinical isolate of "B. hampsonii" (EB107; n = 10) clade II or a classic strain of B. hyodysenteriae (B204; n = 10) to compare gross and microscopic lesions and alterations in colonic mucin expression in pigs with clinical disease versus controls (n = 6). Gross lesions were similar between infected groups. No histologic difference was observed between infected groups with regard to neutrophilic inflammation, colonic crypt depth, mucosal ulceration, or hemorrhage. Histochemical and immunohistochemical evaluation of the apex of the spiral colon revealed decreased expression of sulphated mucins, decreased expression of MUC4, and increased expression of MUC5AC in diseased pigs compared to controls. No difference was observed between diseased pigs in inoculated groups. This study reveals significant alterations in colonic mucin expression in pigs with acute swine dysentery and further reveals that these and other microscopic changes are similar following infection with "B. hampsonii" clade II or B. hyodysenteriae.

Amyloid deposition in 2 feline thymomas.

Two cases of feline thymoma with amyloid deposition were encountered between 1982 and 2010. Neoplastic cells were separated by abundant, pale eosinophilic, homogeneous material that was congophilic and birefringent. Ultrastructurally, the neoplastic cells were connected by desmosomes, and the extracellular deposits were composed of nonbranching, hollow-cored fibrils, 8-10 nm in diameter. In the case with sufficient archived tissue for additional sections, the amyloid remained congophilic following potassium permanganate incubation, and the neoplastic cells were immunoreactive for pancytokeratin. The histologic, histochemical, ultrastructural, and immunohistochemical features of both neoplasms are consistent with epithelial-predominant thymoma with the unusual feature of intratumoral amyloid deposition. The affinity of the amyloid for Congo red following potassium permanganate incubation is consistent with non-AA amyloid. The ultrastructural findings were consistent with amyloid production by the neoplastic epithelial cells.

Spatial distribution of putative growth factors in the guinea pig placenta and the effects of these factors, plasma, and bile on the growth and chemotaxis of Campylobacter jejuni.

The pregnant guinea pig is an effective model for studying abortifacient Campylobacter spp, and previous experiments have demonstrated that C. jejuni IA3902 has a marked predilection for the subplacenta while sparing the placental disc in this species. In the study described here, the growth and chemotaxis of IA3902 and a reference strain (NCTC 11168) are compared in the presence of subplacental and placental factors, as well as bile and plasma, from pregnant and nonpregnant guinea pigs. Both strains grew better in subplacental versus placental disc tissue extracts at 24 hours; however, only IA3902 maintained this enhancement at 48 hours. Histochemistry and lectin histochemistry were used to localize mucin, iron, and l-fucose within the placental unit. Mucin was most abundant in subplacental lacunae, the junctional zone, and visceral yolk sac placenta, while iron was most abundant in the placental disc, and L-fucose-containing surface glycans were limited to the visceral yolk sac placenta. These 3 individual factors, along with progesterone and estradiol, were evaluated for effects on growth and chemotaxis of C. jejuni. Mucin, iron, and L-fucose were growth promoting, while l-fucose was also chemoattractive for both strains. Progesterone, estradiol, and pregnant guinea pig plasma did not affect growth or chemotaxis, and no difference was observed when bile from pregnant and nonpregnant animals was compared. These findings demonstrate the presence of specific factors within the guinea pig placental unit that drive chemotaxis and enhance growth of C. jejuni, shedding light on potential mechanisms underlying the fetoplacental tropism observed with this strain.

Expression of toll-like receptors 2 and 4 in subplacental trophoblasts from guinea pigs (Cavia porcellus) following infection with Campylobacter jejuni.

Toll-like receptors 2 and 4 (TLR2 and TLR4) are well-characterized cell surface receptors that recognize specific pathogen-associated molecular patterns and play an important role in pathogen recognition and activation of the innate immune system. Variable expression of TLR2 and TLR4 has been described in trophoblasts from normal and diseased placentas; yet, there are limited data regarding trophoblast TLR expression in response to specific placental pathogens, and TLR expression in the guinea pig placenta has not been described. The guinea pig is an effective model for Campylobacter-induced abortion of small ruminants, and the authors have shown by immunohistochemistry that C jejuni localizes within syncytiotrophoblasts of the guinea pig subplacenta. The present study was designed to determine if the expression of either TLR2 or TLR4 would be affected in subplacental trophoblasts following infection with C jejuni. Immunohistochemistry for TLR2 and TLR4 was performed on placenta from guinea pigs that aborted following inoculation with C jejuni and from sham-inoculated controls. Quantitative assessment of TLR expression was performed, and mean immunoreactivity for TLR2 was significantly higher in subplacental trophoblasts from animals that aborted compared with uninfected controls (P = .0283), whereas TLR4 expression was not statistically different (P = .5909). These results suggest that abortion in guinea pigs following infection with C jejuni is associated with increased TLR2 expression in subplacental trophoblasts and may reveal a possible role for TLR2 in the pathogenesis of Campylobacter-induced abortion.