Outbreak of ocular disease associated with naturally-acquired canine herpesvirus-1 infection in a closed domestic dog colony

Objective  To describe clinical and virological findings of an outbreak of ocular disease attributed to naturally-acquired primary canine herpesvirus-1 (CHV-1) infection in a closed domestic dog colony.
Animals studied  Twenty-seven 10- to 16-week-old laboratory Beagles.
Procedure  Complete ophthalmic examinations were performed and ocular samples collected for CHV-1 polymerase chain reaction and virus isolation.
Results  The prevalence of ocular morbidity was 100% in examined dogs. Lesions were restricted to the ocular surface and included bilateral conjunctivitis (100% of dogs); punctate, dendritic, or geographic ulcerative keratitis (26% of dogs); and non-ulcerative keratitis (19% of dogs). Conjunctival petechiae were detected in 22% of dogs. Punctate and dendritic corneal ulcers were frequently organized into discrete groups or linear arrangements. Non-ulcerative keratitis appeared clinically as a perilimbal ring of superficial corneal vascularization and leukocyte infiltration. CHV-1 was detected in ocular samples by polymerase chain reaction or virus isolation in all dogs sampled.
Conclusions  In susceptible populations of domestic dogs, CHV-1 may be associated with outbreaks of highly contagious ocular infection in the absence of concurrent overt systemic disease. This naturally-acquired outbreak of CHV-1 infection provides an opportunity to report the spectrum and prevalence of ocular lesions associated with primary ocular CHV-1 infection in dogs. Conjunctivitis was the most frequent ocular lesion detected. Ulcerative and non-ulcerative keratitis were less prevalent and of variable clinical appearance. Dendritic ulcerative keratitis, a classic and relatively specific ocular lesion associated with alphaherpesvirus infection, was detected in < 20% of dogs.     

Protein digestibility of porcine colostrum by neonatal pigs

Colostrum plays an important role in neonatal growth and development. However, little is known about the digestion of macronutrients in colostrum of any species. This study was conducted with the neonatal piglet model to determine the digestibility of proteins in porcine colostrum. Twelve, 1-d-old, male piglets were selected from 3 litters (4 pigs/litter) and housed individually in metabolism crates with heating lamps to maintain a temperature of 35 °C. Colostrum (13 L) was collected from 400 sows (30 to 40 mL/sow) within 12 h postpartum after injection of oxytocin. All piglets were fed colostrum containing 0.25% (DM basis) chromium oxide as an external marker based on the following feeding program: 6 meals/d for an entire 3-d period; with 40 mL/meal for d 1 (240 mL/d), 55 mL/meal for d 2 (330 mL/d), and 70 mL/meal for d 3 (420 mL/d). Colostrum was hand-fed using baby milk bottles. Entire fecal samples with the chromium green color were collected each day after colostrum feeding. Fecal collection was terminated before the fading of the green color. Fecal samples were weighed (10.3 ± 1.0 g/pig), stored in − 20 °C, freeze-dried, and thoroughly ground for chemical analysis. Blood samples were collected at 0900 h of d 3 to obtain plasma samples for amino acid and immunoglobulin (Ig) G analysis. Digestibilities of crude protein and DM in colostrum, defined as the percentage of ingested colostral crude proteins and DM that disappeared in the gut, averaged 96.9 ± 0.4% and 98.3 ± 0.2%, respectively. Digestibility of total amino acids (protein-bound plus free amino acids) in the colostrum was 98.3 ± 0.1%, with the values being 98.5 ± 0.3, 98.2 ± 0.4, and 98.3 ± 0.3%, respectively, for Lys, Thr, and Arg. Plasma and colostral IgG content were 3.4 ± 0.3 and 3.8 ± 0.7 g/L, respectively. In conclusion, protein-bound and free amino acids in porcine colostrum were highly digestible and available to neonatal pigs.     

Evaluation of novel adjuvant Eimeria profilin complex on intestinal host immune responses against live E. acervulina challenge infection

The effects against avian coccidiosis of two novel adjuvants, Quil A/cholesterol/dimethyl dioctadecyl ammonium bromide/Carbopol (QCDC) and QCDC/Bay R1005 (R)/cytosine-phosphate-guanosine (CpG) oligodeoxynucleotides (CpG ODN [T]) (QCDCRT) emulsified with profilin, a conserved Eimeria recombinant protein, were determined in broiler chickens. Chickens were subcutaneously immunized with isotonic saline (control group), profilin (P), profilin emulsified with QCDC (P-Q), or profilin with QCDCRT (P-QR) at 2 and 9 days post-hatch and orally challenged with 1.0 x 10(4) sporulated oocysts of Eimeria acervulina (EA) at 7 days postimmunization. All profilin-immunized groups showed increased body weight gain when compared to the control group, and the P-QR group had significantly higher body weight gain than did those of the P and P-Q groups following EA challenge infection. All groups immunized with profilin showed significantly decreased intestinal lesions compared with the control group, with the P-QR group showing the lowest intestinal lesions among the profilin-treated groups. Finally, the P-QR group showed greater CD4+/CD8+ and TCR1+/TCR2+ splenocytes and higher antiprofilin serum antibody titers compared with the P and P-Q (or both) groups following EA challenge infection. These results further suggest that vaccination of chickens with profilin, in combination with the QCDCRT adjuvant, may provide a novel control strategy against EA infection in commercial flocks.     

Differential glutathione S-transferase isozyme activities in rice and early watergrass seedlings

Glutathione S -transferase (GST) isozymes were investigated in one-leaf-stage rice ( Oryza sativa L. cv. Nipponbare) and early watergrass ( Echinochloa oryzicola Vasing) shoots after being induced by treatment with a combination of pretilachlor [2-chloro-2',6'-diethyl-N-(2-propoxyethyl)acetanilide] and fenclorim (4,6-dichloro-2-phenylpyrimidine) using DEAE-Sephacel anion exchange chromatography. Non-treated plants contained GST isozymes that had activity to the following substrates: three isozymes for l-chloro-2,4-dinitrobenzene (CDNB), six isozymes for pretilachlor and three isozymes for fenclorim in rice shoots; and four isozymes for CDNB, three or four isozymes for pretilachlor and two or three isozymes for fenclorim in early watergrass shoots. Glutathione S -transferase isozyme activities of non-treated plants were higher in rice than in early watergrass, especially in the case of GST activity with fenclorim as a substrate. Pretreatment of rice roots with a combination of pretilachlor and fenclorim increased the activity of the constitutively expressed isozymes that exhibited activity with CDNB, pretilachlor and fenclorim. This pretreatment also caused the appearance of one new GST(fenclorim) isozyme. Pretreatment of early watergrass roots with a combination of pretilachlor and fenclorim produced almost no increase in activity of some constitutively expressed isozymes that exhibited activity to CDNB and fenclorim, although it partly increased the peaks to corresponding to pretilachlor. The induction of GST was higher in rice than that in early watergrass. These results indicated that the isozyme pattern and substrate specificity of GST isozymes in rice were different from those in early watergrass. Furthermore, the selectivity of pretilachlor between rice and early watergrass may be related to different constitutively expressed GST(pretilachlor) isozyme activities and the induction of GST(pretilachlor) isozyme activities in the combination treatment.     

An outbreak of highly pathogenic avian influenza subtype H5N1 in broiler breeders, Korea

Highly pathogenic avian influenza (HPAI) was diagnosed in broiler breeders, submitted to the National Veterinary Research and Quarantine Service in South Korea. Grossly, the dead breeders had lesions consistent with HPAI, including pancreatic mottling, splenomegaly, pulmonary edema and congestion, and hemorrhages in the mucosa of the proventriculus, gizzard and small intestine, and on the serosal surface. Microscopically, there were necrotized hepatitis and pancreatitis, lymphocytic meningoencephalitis, myocarditis, and interstitial pneumonia. Influenza viral antigen was demonstrated in areas closely associated with histopathologic lesions. The AI virus was isolated from cecal tonsils, feces, trachea, and kidney of the chickens. The isolated virus was identified as the highly pathogenic H5N1, with a hemagglutinin proteolytic cleavage site deduced amino acid sequences of QREKRKKR/GLFGAGLFGAIAG. In order to determine the pathogenicity of the isolate, eight 6-week-old specific pathogen free chickens were inoculated intravenously with the virus, and all the birds died within 24 hr after inoculation. This is the first report of an outbreak of HPAI in the chickens in South Korea.     

Use of conventional and real-time polymerase chain reaction for confirmation of Mycobacterium avium subsp. paratuberculosis in a broth-based culture system ESP II.

The ESP II Culture System (ESP II), a broth-based culture system, has been modified and optimized for culturing Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) in animal feces since 2000. Conventional and real-time polymerase chain reaction (PCR) assays based on the IS900 sequence were performed as confirmatory tests for M. paratuberculosis in ESP II liquid culture medium. There were no differences between test results of conventional and real-time PCR assays. During the 5-week incubation period, if acid-fast bacilli (AFB) were detected in ESP culture-positive samples, IS900 PCR assays were performed to confirm whether those AFB were M. paratuberculosis. At the end of the 5-week incubation, AF staining was performed on all ESP II-negative cultures to screen any false-negative cultures; IS900 PCR assays were performed on AFB-positive cultures. During a period of 1 year, of a total of 18,499 ESP II cultures, 2,814 (15.2%) PCR confirmation assays were performed. Of those, 2,259 (80%) were both ESP and PCR positive; 104 (4%) were ESP positive and PCR negative; 423 (15%) were ESP negative and PCR positive; 28 (1%) were both ESP and PCR negative. The AF-staining step after the 5-week incubation produced 423 (15%) more PCR-positive cultures. Of a total of 2,814 AFB-positive cultures, 132 (5%) were not confirmed as M. paratuberculosis. Further studies are needed for speciation of non-M. paratuberculosis isolates.     

Both anti-oxidation and lipid-carbohydrate conversion enhancements are involved in priming-improved emergence of Echinacea purpurea seeds that differ in size

This study evaluated the effects of priming on emergence responses of purple coneflower (Echinacea purpurea L. Moench) seeds. The seeds that differ in seed size were either primed with moistened vermiculite (solid matrix priming) or primed in non-aerated −0.5 MPa polyethylene glycol 6000 solution at 25 °C for 6 days (osmopriming), followed by air-drying to their initial moisture level. The tetrazolium staining tests indicated that both large and small seeds were biochemically viable. No notable difference in germination percentage was found between large and small seeds. However, extensive cavity was visible in portions of small seeds in comparison with large seeds. Large seeds accumulated more antioxidants and had greater activities of anti-oxidative enzymes than small seeds. They also had greater isocitrate lyase and malate synthase activities than small seeds. As a result, large seeds had higher emergence percentage and faster emergence speed as compared to that of small seeds. Both solid matrix priming and osmopriming increased emergence percentage and shortened mean emergence time of purple coneflower seeds by increasing the activities of anti-oxidative enzymes and decreasing the levels of malondialdehyde and total peroxide accumulation. Moreover, priming also enhanced the anti-oxidative activities of treated seeds. The activities of isocitrate lyase and malate synthase were also increased in primed seeds. The enhanced anti-oxidation and lipid-carbohydrate conversion activities might explain in part why primed purple coneflower seeds emerged better than non-primed seeds.     

Doublecortin-immunoreactive neuronal precursors in the dentate gyrus of spontaneously hypertensive rats at various age stages: comparison with Sprague-Dawley rats

Spontaneously hypertensive rats (SHRs) are widely accepted in medical research because this model has been used for studies in neurodegenerative diseases such as vascular dementia and stroke. In the present study, we observed newly generated neuronal precursors using doublecortin (DCX, a marker of neural proliferation and differentiation) in the subgranular zone of the dentate gyrus in SHRs compared to Sprague-Dawley rats (SDRs) at various age stages. DCX immunoreactivity, immunoreactive cell numbers and its protein level in the dentate gyrus of the SHRs were higher than those in the SDRs at postnatal month 1 (PM 1). At PM 8, DCX immunoreactivity, immunoreactive cell numbers and protein levels in both groups were markedly decreased compared to those at PM 1; however, they were higher than those in the SDRs. They were decreased in the both groups with age: DCX immunoreactive cells in the SDRs were few at PM 12. Our results indicate that newly generated neuronal precursors are more abundant in SHRs than in SDRs during their life.