Experimental reproduction of severe disease in CD/CD pigs concurrently infected with type 2 porcine circovirus and porcine reproductive and respiratory syndrome virus

Three-week-old cesarean-derived colostrum-deprived (CD/CD) pigs were inoculated with porcine circovirus type 2 (PCV2, n = 19), porcine reproductive and respiratory syndrome virus (PRRSV, n = 13), concurrent PCV2 and PRRSV (PCV2/PRRSV, n = 17), or a sham inoculum (n = 12) to compare the independent and combined effects of these agents. Necropsies were performed at 7, 10, 14, 21, 35, and 49 days postinoculation (dpi) or when pigs became moribund. By 10 dpi, PCV2/PRRSV-inoculated pigs had severe dyspnea, lethargy, and occasional icterus; after 10 dpi, mortality in this group was 10/11 (91%), and all PCV2/ PRRSV-inoculated pigs were dead by 20 dpi. PCV2-inoculated pigs developed lethargy and sporadic icterus, and 8/19 (42%) developed exudative epidermitis; mortality was 5/19 (26%). PRRSV-inoculated pigs developed dyspnea and mild lethargy that resolved by 28 dpi. Microscopic lesions consistent with postweaning multisystemic wasting syndrome (PMWS) were present in both PCV2- and PCV2/PRRSV-inoculated pigs and included lymphoid depletion, necrotizing hepatitis, mild necrotizing bronchiolitis, and infiltrates of macrophages that occasionally contained basophilic intracytoplasmic inclusion bodies in lymphoid and other tissues. PCV2/ PRRSV-inoculated pigs also had severe proliferative interstitial pneumonia and more consistent hepatic lesions. The most severe lesions contained the greatest number of PCV2 antigen-containing cells. PRRSV-inoculated pigs had moderate proliferative interstitial pneumonia but did not develop bronchiolar or hepatic lesions or lymphoid depletion. All groups remained seronegative to porcine parvovirus. The results indicate that 1) PCV2 coinfection increases the severity of PRRSV-induced interstitial pneumonia in CD/CD pigs and 2) PCV2 but not PRRSV induces the lymphoid depletion, granulomatous inflammation, and necrotizing hepatitis characteristic of PMWS.     

Prepuce and partial penile amputation for treatment of preputial gland neoplasia in two ferrets

Preputial tumours in ferrets are frequently malignant and therefore warrant prompt investigation. As many cases do not respond favourably to surgery, even in combination with radiation therapy, wide surgical resection has been recommended. Such a procedure may necessitate partial or total penile resection but outcomes have thus far not been well described. The current case series describes two ferrets in which surgical resection, including penile amputation, was performed using 10 and 5 mm margins, respectively. In the first case, no recurrence of preputial gland adenocarcinoma was noted for 32 months postsurgery, whereas multiple attempts at surgery and radiation therapy were unsuccessful in the second. These cases suggest that margins of at least 1 cm may help achieve a better outcome. Penile amputation for the treatment of preputial tumours appears to be well tolerated by ferrets, as demonstrated by these cases.     

IgE responses to bluetongue virus (BTV) serotype 11 after immunization with inactivated BTV and challenge infection

To test the hypothesis that development of a BTV-specific IgE response plays a role in clinical disease manifestation, the humoral immune response of cattle to inactivated and virulent BTV was studied. Three calves received three sensitizing immunizations of inactivated BTV, 3 weeks apart. The BTV-sensitized animals, two non-sensitized BTV-seropositive and 4 BTV-seronegative control cattle. were challenge-exposed with BTV-11, UC8 strain. All cattle inoculated with inactivated BTV developed group-specific non-neutralizing and serotype-specific neutralizing antibodies. The development of post-challenge-exposure neutralizing antibody titers was inversely correlated with protective immunity. None of the BTV-challenged animals showed clinical disease. The levels of IgE were greatest in the sensitized calves after virus challenge in comparison with control groups. The sequential development, specificity and intensity of virus protein-specific humoral responses were evaluated using immunostaining. After challenge exposure of BTV-sensitized and non-sensitized cattle, total and IgE antibodies reacted consistently within BTV structural proteins VP2, VP5 and VP7. Although no correlation was found between clinical disease and IgE, results add support to the hypothesis that IgE may be involved in the pathogenesis of clinical disease, since infection with BTV causes an increase in serum IgE levels. However, these results suggest that the levels of virus-specific reactivity may be an important factor in determining whether or not clinical disease manifestation occurs.     

Biochemical markers of bone metabolism in growing thoroughbreds: a longitudinal study

This study describes longitudinal changes in serum levels of biochemical markers of bone cell activity in a group of 24 thoroughbred foals from birth to 18 months of age. The markers of bone formation included the type I collagen carboxy-terminal propeptide (PICP), the bone-specific isoenzyme of alkaline phosphatase (BAP), and osteocalcin (OC). Levels of the cross-linked telopeptide of type I collagen (ICTP), a marker of bone resorption, and the N-terminal propeptide of type III collagen (PNIIIP), a marker of soft tissue turnover, were also measured. Levels of all markers fell significantly between birth and 18 months of age (70-80 per cent); this decrease being most marked between 0 and 6 months. However, a transient increase in levels of the markers then occurred between 6 and 14 months of age. The timing of this increase was specific for each parameter. ICTP and OC concentrations increased between October and December. PICP concentrations increased between December and April whereas the increase in PIIINP was coincident with the peak in weight gain between April and June. Changes in BAP concentration were less distinct at this time. Season was shown to have significant effects on the biochemical markers independent from the effect of age. Concentrations of all markers decreased with increasing body weight and at any given age heavier horses had lower marker levels. These results show that biochemical markers of bone cell activity and soft tissue turnover follow characteristic patterns of change in growing thoroughbreds influenced by age, season and bodyweight. The demonstration that the reference ranges for the biochemical markers change from month to month means that single samples from individuals are of little value for monitoring bone cell activity in growing thoroughbreds.     

Expression of receptor tyrosine kinases VEGFR-1 (FLT-1), VEGFR-2 (KDR), EGFR-1, PDGFRa and c-Met in canine primary brain tumours

Inhibition of tumour growth and angiogenesis by targeting key growth factor receptors is a promising therapeutic strategy for central nervous system tumours. Characterization of these growth factor receptors in canine primary brain tumours has not been done. Using quantitative real‐time TaqMan polymerase chain reaction (PCR), we evaluated the expression of messenger RNA (mRNA) for five tyrosine kinase growth factor receptors (vascular endothelial growth factor receptor [VEGFR]‐1, VEGFR‐2, endothelial growth factor receptor [EGFR]‐1, platelet‐derived growth factor receptor a [PDGFRa], and c‐Met) relative to normal cerebral cortex in 66 spontaneous canine primary brain tumours. Increased expression of VEGFR‐1 and VEGFR‐2 mRNA was greatest in grade IV astrocytomas (glioblastoma multiforme) and grade III (anaplastic) oligodendrogliomas. EGFR‐1 mRNA expression was more consistently increased than the other receptors in all tumour types, while increased PDGFRa mRNA expression was mostly restricted to oligodendrogliomas. The similarities in increased expression of these tyrosine kinase growth factor receptors in these canine tumours, as compared to data from their human counterparts, suggest that common molecular mechanisms may be present.     

Lectin histochemical investigations of the distal gut of chicks with special emphasis on the follicle-associated epithelium

Carbohydrates on epithelial cell surfaces play an important role as attachment sites for different microorganisms like bacteria, viruses and protozoa. To obtain more information about the distribution of carbohydrates on the luminal surface along the intestine, lectin histochemical studies on different gut segments of chicks of different age groups were carried out using a panel of 13 lectins with specificities for Man, Glc, Gal, GalNAc, GlcNAc or GlcNAc oligosaccharides and Sia. Furthermore, we tried to find out whether previously reported specificities of certain lectins for M cells (membranous or multifold cells) in the bursa of Fabricius (BF) can be observed also on M cells of the intestine. As a result we were able to demonstrate binding of all lectins employed in these studies in all investigated gut segments. In some cases, the application of the same lectin led to varying staining intensities of the same histological structures in different age-groups (e.g. staining of the brush border with WGA, LEA, MAA or Conarva) or different gut segments (e.g. staining of goblet cells with CMA II, LEA and MPA). Hence, terminal carbohydrate residues of glycoconjugates on the intestinal epithelium vary depending on age and organ site. As glycoconjugates can act as attachment sites for microorganisms, these differences in the distribution of sugar residues may be one explanation for the site-specificity of certain pathogens. Furthermore, the binding of lectins to the follicle-associated epithelium (FAE) of the BF differs from that to the FAE of the intestine again stressing the site specificity of lectin binding. Thus, up to now no universal M-cell marker along the chicken intestine exists.     

Experimental infection of weanling pigs with Salmonella typhisuis: effect of feeding low concentrations of chlortetracycline, penicillin, and sulfamethazine

Clinical and pathologic variables of Salmonella typhisuis infections were studied in weanling pigs. The influence of daily feeding of low concentrations of chlortetracycline, penicillin, and sulfamethazine on S typhisuis infection also was determined. Ten pigs (group 1) given feed containing low concentrations of chlortetracycline, penicillin, and sulfamethazine when orally inoculated with S typhisuis became pyretic, developed a mild neutrophilia, and had increased serum agglutinating antibody titers, but were clinically normal. Lesions were not present, and S typhisuis was not isolated from 2 group-1 pigs that were killed and necropsied on postinoculation day 8. Then, the antimicrobial agents were withdrawn from the feed of the remaining 8 pigs for 6 days. The pigs were reinoculated with S typhisuis on postinoculation day 16 and developed mild clinical disease with sustained high rectal temperatures. Severe necrotizing typhlocolitis and ulcerative proctitis were found at necropsy in all 8 of the pigs. Similar findings were induced in 7 additional pigs (group 2) that were concurrently inoculated with S typhisuis, but that had no previous exposure to S typhisuis. Successful isolation of S typhisuis required special care and media.