Identification of Equine Herpesvirus 5 in Horses with Lymphoma

Equine multinodular pulmonary fibrosis, equine herpesvirus 5 (EHV-5), and multicentric lymphoma were discovered in one patient. Review of gamma herpesvirus activity in humans revealed a propensity for lymphoproliferative disorders associated with infection. The objective was to determine the frequency of EHV-5 in lymphoma tissues and compare with the frequency found in the lymph nodes of clinically normal horses. Case control investigation of lymphoma-positive tissues and analysis via polymerase chain reaction (PCR) for EHV-5 was performed on 12 horses. Prospective collection and PCR analysis of lymph nodes (mesenteric or submandibular) for EHV-5 was performed on 21 control horses. Thirteen samples of lymphoma-positive tissues and fluid were submitted for PCR analysis for EHV-5. Of these, 67% was positive. In the control horse population, 14% was positive for EHV-5 (P = .004). Neoplastic samples positive for EHV-5 were classified as T-cell rich B-cell lymphoma (three), T-cell lymphoma (one), one was nondifferentiated, and two were not stained. Gamma herpesviruses in humans have been associated with lymphoproliferative diseases such as Kaposi sarcoma and Burkitt lymphoma. This study reveals an increased frequency of EHV-5 (gamma herpesvirus) in horses diagnosed with lymphoma compared with healthy control horses. Although the exact role this virus plays in the initiation or perpetuation of lymphoproliferative neoplasia is unknown, EHV-5 may be an etiologic agent associated with the development of some types of equine lymphoma.     

Equine Multinodular Pulmonary Fibrosis and Lymphoma in a Horse Associated with Equine Herpesvirus-5

A 9-year-old mare presented with a 2-week history of partial anorexia, multiple swellings in the area of the throatlatch, and purulent nasal discharge. On initial presentation, the horse had submandibular and retropharyngeal lymphadenopathy, mild ventral edema, and weight loss. Thoracic radiographs revealed a pulmonary interstitial pattern. Necropsy revealed enlarged lymph nodes throughout the body. The lung parenchyma contained multiple random, well-circumscribed nodules, which, on cut section, were pale, tan colored, and very firm with a distinct line of demarcation from the surrounding normal parenchyma. The subendocardium of the left ventricle, left atrium, and, multifocally, the right ventricle contained white, gritty areas of mineralization. There was marked subintimal mineralization of the aorta and pulmonary artery. Histopathology of the lymph nodes revealed effacement of the parenchyma by a neoplasm composed of large numbers of small mature lymphocytes, fewer large lymphocytes, and scattered moderate numbers of histiocytes. Immunohistochemistry tests for CD3, CD79a, and CD20, confirmed the lymphoma was T-cell-rich B-cell lymphoma. The lungs contained marked interstitial fibrosis with alveolar histiocytosis. Polymerase chain reaction test results of lymph node and lungs were positive for equine herpesvirus-5 (EHV-5), a gammaherpesvirus. Gammaherpesvirus infection has been associated with lymphoma and pulmonary fibrosis in other species. This report describes the association between EHV-5 and both pulmonary fibrosis and lymphoma.     

Treatment of Metastatic Equine Melanoma with a Plasmid DNA Vaccine Encoding Streptococcus Pyogenes EMM55 Protein

A 19-year-old castrated male Arab/Quarter horse presented with an extensive history of cutaneous metastatic melanoma. Over a period of 8 months, a total of 8 doses of plasmid DNA vaccine expressing the Streptococcus pyogenes emm55 gene (pAc/emm55) were administered intratumorally at 300 μg/dose via a needless injector. Upon completion of the vaccination protocol, the size of the injected lesions, on average, were reduced by 40.3% from the initial size measurements. Lesions that were not injected were reduced by 47.6%. The overall reduction in total tumor burden was 42.3%. Tumor regression was also associated with the augmentation of antimelanoma IgG antibody response, thus implying that an induction of an effective antimelanoma response would be of great advantage in the management of equine melanoma.     

A simplified surgical technique for correction of orbital adipose prolapse in the horse

A Quarter Horse gelding underwent standing surgical correction of an orbital adipose prolapse through a subconjunctival approach removing the prolapsed fat without use of electrocautery or closure of the conjunctiva. This technique provided an acceptable long‐term cosmetic outcome. There was no recurrence of orbital adipose prolapse up to 15 months post operatively. The surgical procedure performed in this case is a simple technique, which could be performed in the field under sedation and local anaesthesia.     

Consequences for the bovine embryo of being derived from a spermatozoon subjected to post-ejaculatory aging and heat shock: development to the blastocyst stage and sex ratio

The objective was to determine whether aging of sperm caused by incubation at normothermic (38.5 C) or heat shock (40 C) temperatures for 4 h prior to oocyte insemination affects sperm motility, fertilizing ability, competence of the resultant embryo to develop to the blastocyst stage and blastocyst sex ratio. In the first experiment, the percent of sperm that were motile was reduced by aging (P<0.001) and the reduction in motility was greater for sperm at 40 C compared to sperm at 38.5 C (P<0.01). In the second experiment, oocytes were inseminated with aged sperm. A smaller percent of oocytes fertilized with sperm aged at either temperature cleaved by Day 3 after insemination than oocytes fertilized with fresh sperm (P<0.05). There was no effect of sperm aging on the percent of oocytes or cleaved embryos that developed to the blastocyst stage. Aging of sperm before fertilization at 38.5 C reduced the percent of blastocysts that were male (P=0.08). In the third experiment, incubation of sperm at 38.5 C or 40 C for 4 h did not reduce fertilizing ability of sperm as determined by pronuclear formation at 18 h post insemination. In conclusion, aging of sperm reduced cleavage rate and the percent of blastocysts that were males but had no effect on the developmental capacity of the embryo. The effect of aging on cleavage rate may represent reduced motility and errors occurring after fertilization and pronuclear formation. Aging at a temperature characteristic of maternal hyperthermia had little additional effect except that polyspermy was reduced. Results indicate that embryo competence for development to the blastocyst stage is independent of sperm damage as a result of aging for 4 h at normothermic or hyperthermic temperatures.     

Modulation of the immunogenicity of the Trypanosoma congolense cysteine protease, congopain, through complexation with ��2-macroglobulin

The protozoan parasite Trypanosoma congolense is the main causative agent of livestock trypanosomosis. Congopain, the major lysosomal cysteine proteinase of T. congolense, contributes to disease pathogenesis, and antibody-mediated inhibition of this enzyme may contribute to mechanisms of trypanotolerance. The potential of different adjuvants to facilitate the production of antibodies that would inhibit congopain activity was evaluated in the present study. Rabbits were immunised with the recombinant catalytic domain of congopain (C2), either without adjuvant, with Freund’s adjuvant or complexed with bovine or rabbit α₂-macroglobulin (α₂M). The antibodies were assessed for inhibition of congopain activity. Rabbits immunised with C2 alone produced barely detectable anti-C2 antibody levels and these antibodies had no effect on recombinant C2 or native congopain activity. Rabbits immunised with C2 and Freund’s adjuvant produced the highest levels of anti-C2 antibodies. These antibodies either inhibited C2 and native congopain activity to a small degree, or enhanced their activity, depending on time of production after initial immunisation. Rabbits receiving C2-α₂M complexes produced moderate levels of anti-C2 antibodies and these antibodies consistently showed the best inhibition of C2 and native congopain activity of all the antibodies, with maximum inhibition of 65%. Results of this study suggest that antibodies inhibiting congopain activity could be raised in livestock with a congopain catalytic domain-α₂M complex. This approach improves the effectiveness of the antigen as an anti-disease vaccine candidate for African trypanosomosis.     

Functional characterization of bovine TIRAP and MyD88 in mediating bacterial lipopolysaccharide-induced endothelial NF-κB activation and apoptosis

Mastitis is a prevalent disease in dairy cows. Gram-negative bacteria, which express the pro-inflammatory molecule lipopolysaccharide (LPS), are responsible for the majority of acute clinical cases of mastitis. Previous studies have identified differential susceptibility of human and bovine endothelial cells (EC) to the pro-inflammatory and injury-inducing effects of LPS. The Toll-like receptor (TLR)-4 signaling pathway, which is activated by LPS, has been well studied in humans, but not in ruminants. Human myeloid differentiation-factor 88 (MyD88) and TIR-domain containing adaptor protein (TIRAP) are critical proteins in the LPS-induced NF-κB and apoptotic signaling pathways. To assess the role of the bovine orthologs of these proteins in bovine TLR-4 signaling, dominant-negative constructs were expressed in bovine EC, and LPS-induced NF-κB activation and apoptosis evaluated. The results from this study indicate that bovine MyD88 and TIRAP play functional roles in transducing LPS signaling from TLR-4 to downstream effector molecules involved in NF-κB activation, and that TIRAP promotes apoptotic signaling.     

Aluminum accumulation in nineteen Melastomataceae species from three contrasting plant formations in acid soils

This study describes the aluminum (Al) accumulation in relation to macronutrient and micronutrient elements in 19 Melastomataceae species in the Guayana Region in Venezuela. The purpose was to investigate the Al accumulation in four tribes and different life forms. Aluminum accumulation was predicted in the basal tribes Miconieae and Merianieae in contrast to the derived tribes and herbs from any tribe, which generally do not accumulate Al. The survey was done in a vegetation continuum, which includes a savanna shrubland, a palm‐swamp community, and an evergreen forest in the Guayana region in southeastern Venezuela. The highest value of soil Al concentration was found in the savanna shrubland, where ten lignified Miconiae and one Merianeae Al accumulators were present. At the forest, the site with highest soil acidity, four Al‐accumulator tree species from Miconiae were found. Miconia lepidota showed similar Al foliar concentrations in the savanna shrubland and forest, but foliar Ca was lower in the forest, even though it was the site with highest Ca in the soil. At the palm‐swamp community, the Melastomeae shrub Macairea pachyphylla was found with an Al concentration of 0.59 g kg^–1 in leaves and 0.16 g kg^–1 in bark. At the same site, Al accumulation occurred in one Microlicieae species, one Miconieae species, and in the Melastomeae herbs Pterogastra divaricata (13.25 g [kg dry mass]^–1) and Pterolepis trichotoma (17.83 g kg^–1). The report of Al hyperaccumulation in P. trichotoma is new for the genus, and Al accumulation in herbs is considered exceptional. The foliar Al concentration was positively correlated (p < 0.005) with Fe (r = 0.64, n = 20) and Zn (r = 0.63). The analysis of the relationships between soil Al, Fe, or Zn and the concentrations of these elements in leaves revealed they were not significantly correlated. The results indicate Al hyperaccumulation in two herbaceous Melastoamataceae species and suggest Al accumulation in this life form deserves future research. However, they also confirm the highest number of Al accumulators in lignified species of the ancient tribe Miconiae (14 out of 19 species studied).     

Distribution of Several Heavy Metals in Tidal Flats Sediments within Bahía Blanca Estuary (Argentina)

Sediment pollution by metals is of high interest considering that it can affect marine life. The estuaries' quality may be reflected by the environmental intertidal zone condition. Subsurface sediments collected at the nude tidal flats from three sampling stations in the Bahía Blanca Estuary were analyzed for total metals concentrations (Hg, Cd, Pb, and Cr), distribution, and geochemical partitioning. Most of the elements (Hg, Cd, and Cr) have shown highest concentration values in the industrial-influenced area. Maximum value of Pb was obtained where the main freshwater input discharges. Intertidal sediments have presented higher values of Cr than the subtidal ones. Cd and Pb contents near the industrial area were strongly higher in the subtidal zones. The distribution of Cd and Pb demonstrated the occurrence of a diffusion pattern from the land toward the sea, showing a dependence on both the metal itself and/or the source. Not all studied metals have shown the highest content in the fine fraction. The chemical partitioning in the fine fractions offered evidence that the tidal flats were an important source as well as sink of metals to the adjacent coastal area. The studies of intertidal sediments provide an integrative knowledge on the potential effects of different trace metals in the environment and they must be used in the contamination studies within coastal areas.