The decreased competition in expanding versus mature juniper woodlands is counteracted by adverse climatic effects on growth

World-wide deforestation is being reversed in Mediterranean continental areas, where abandonment of traditional practises favours the expansion of valuable habitats, like Juniperus thurifera woodlands. We hypothesised that pre-existing trees facilitate establishment in expanding woodlands, whereas in mature woodlands, competition leads to patch disaggregation. We compared the imprint of these processes on growth, demographic and spatial structure of expanding and mature J. thurifera woodlands. We selected plots where we geopositioned, aged and quantified the morphological characteristics of all trees. In the mature woodland, trees arranged in clumps and randomly in the expanding woodland. Competition negatively affected growth, was greater in the mature woodland and led to disaggregation of juvenile clumps. Differences in growth between the mature and the expanding woodland disappeared in climatically unfavourable years, suggesting that adverse climate constrains growth more in expanding than in mature woodlands. We suggest that change in the dispersal agents and a decrease of facilitation underlay differences in spatial patters between the expanding and the mature woodland. Observed effective recruitment in less than 30?years into the expanding woodland evidenced that propagule arrival and sapling survival do not constrain woodland expansion. Furthermore, growth of juveniles established in these new areas is favoured by reduced intra-specific competition. However, we expect growth in expanding woodlands to be negatively impacted by climate change. We conclude that under current global change scenario, conservation of J. thurifera woodlands is favoured by changes in land use, but greater frequency and severity of drier than usual episodes could hamper natural reforestation.     

Optimization of the protocols for double vaccination against Marek's disease by using commercially available vaccines: evaluation of protection, vaccine replication, and activation of T cells

Revaccination against Marek's disease is a widespread practice in some countries. The rationale of this practice is unknown, and there is no consensus in the protocols. Recently, we have demonstrated that administration of the first vaccine at 18 days of embryonation followed by a more protective second vaccine at hatch (18ED/1d) reproduced systematically the benefits of revaccination under laboratory conditions. Here, we have used the same model to optimize the revaccination protocols by using currently available vaccines and to determine whether two features associated with Marek's disease vaccine-induced protection (activation of T cells and replication of vaccine virus) are involved in the revaccination protocols. Protection conferred by three revaccination protocols (turkey herpesvirus [HVT] 18ED/HVT+SB-1 1d, HVT 18ED/CVI988 1d, and HVT+SB-1 18ED/ CVI988 1d) was evaluated. Revaccination protocols also were compared with single vaccination protocols (HVT 18ED, HVT+SB-1 18ED, HVT+SB-1 1d, CVI988 18ED, and CVI988 1d). Our results demonstrated that it is possible to improve efficacy of the currently available vaccines by using them in revaccination programs. Administration of HVT 18ED/CVI988 1d and HVT+SB-1 18ED/CVI988 1d were the two protocols that conferred the highest protection against a very early challenge (2 days of age) with very virulent plus Marek's disease virus strain 648A. In a separate experiment, we evaluated vaccine replication and activation of T cells in single and revaccination protocols. Our results demonstrated that replication of the second vaccine, although decreased compared with single vaccination, could be detected at 3 days (HVT, CVI988) or at 6 days (SB-1). Administration of the first vaccine (HVT) at 18ED resulted in a high percentage of activated T cells. Administration of a second vaccine (either HVT-SB-1 or CVI988) at 1d resulted in increased intensity of MHC-II stain in activated T cells.     

Lysosomal storage disease caused by Sida carpinifolia poisoning in goats

A neurologic disease characterized by ataxia, hypermetria, hyperesthesia, and muscle tremors of the head and neck was observed for 2 years in a flock of 28 Anglo-Nubian and Saanen goats on a farm with 5 ha of pasture. Six newborns died during the first week of life, and five abortions were recorded. The predominant plant in the pasture was Sida carpinifolia. The disease was reproduced experimentally in two goats by administration of this plant. Three goats with spontaneous disease and the two experimental animals were euthanatized and necropsied. No significant gross lesions were observed. Fragments of several organs, including the central nervous system, were processed for histopathology. Small fragments of the cerebellar cortex, liver, and pancreas of two spontaneously poisoned goats and two experimentally poisoned goats were processed for electron microscopy. Multiple cytoplasm vacuoles in hepatocytes, acinar pancreatic cells, and neurons, especially Purkinje cells, were the most striking microscopic lesions in the five animals. Ultrastructural changes included membrane-bound vacuoles in hepatocytes, Kupffer cells, acinar pancreatic cells, Purkinje cells, and the small neurons of the granular cell layer of the cerebellum. Paraffin-embedded sections of the cerebellum and pancreas were submitted for lectin histochemical analysis. The vacuoles in different cerebellar and acinar pancreatic cells reacted strongly to the following lectins: Concanavalia ensiformis, Triticum vulgaris, and succinylated Triticum vulgaris. The pattern of staining, analyzed in Purkinje cells and acinar pancreatic cells coincides with results reported for both swainsonine toxicosis and inherited mannosidosis.     

Argentine strain of equine herpesvirus 1 isolated from an aborted foetus shows low virulence in mouse respiratory and abortion models

The equine herpesvirus 1 (EHV-1) was isolated in Argentina from an aborted equine foetus in 1979. This virus (SPv) has special restriction patterns (RP) in comparison with other Argentine isolates. In addition, SPv could be distinguished on the basis of its pathogenicity in baby mice inoculated intracerebrally. We studied the growth properties of the SPv in cell culture and its effects in a mouse respiratory and abortion model. We observed that SPv did not modify its capacity to grow in cell culture with respect to reference HH1 strain. Nevertheless, we found significant differences between the titres of the two strains at 8-14 h post-infection (PI). In this work we demonstrated that SPv showed low virulence in female at different stages of gestation, consistently, with results found in the mouse respiratory model. We considered that this low virulence of SPv could be related to its RP because the RP of HH1 strain are similar to those of the HVS25A strain and both showed effect on pregnant mice. More specific studies about genomic alterations to the SPv are necessary for identifying, more clearly, if the intra-strain variations have relation with the low virulence in the mouse respiratory and abortion model.     

Papillary adenocarcinoma of the extrahepatic bile duct in a Holstein cow

A 15-year-old female Holstein cow was presented with a history of anorexia and weight loss. Abnormal physical examination findings included poor body condition, dehydration, icterus, and photodermatitis on nonpigmented areas of the skin of the dorsum. At necropsy, the common bile duct was greatly enlarged, with thickened walls, and tightly adhered to the hepatic capsular surface and serosa of adjacent loops of the small intestine. Two sessile, yellowish, friable, well-circumscribed, cauliflower-like, bulging masses (14 and 8 cm in diameter) were attached to the inner surface of the common bile duct blocking its lumen. Microscopically, the tumor was composed of well-differentiated, columnar epithelial cells with oval or round, vesicular, basal nuclei arranged in papillary projections. Periodic acid-Schiff-positive material accumulated in the apical cytoplasm of the neoplastic cells. Immunostaining of the tumor cells was positive for cytokeratin (CK) of wide-spectrum screening and for CK 7. The diagnosis of papillary adenocarcinoma of the extrahepatic bile duct was based on the morphologic features of the neoplasm and evidence of local invasion. To the best of our knowledge, extrahepatic bile duct carcinomas have not been described in cattle.     

Paratuberculosis in red deer (Cervus elaphus): an immunohistochemical study

In the present study, we compared the utility of immunohistochemistry with serological and histological results for the characterization of Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) in tissues of affected red deer. Bacterial isolation was considered the standard reference. Samples were taken from seven clinically affected animals with typical macroscopic lesions. The enzyme linked immunosorbent assay (ELISA) and the gel diffusion tests (GD) were used for serological determinations. Samples from intestine and mesenteric lymph nodes were processed for bacterial isolation and histology. M. paratuberculosis was isolated from all the animals. Histologically, lymph nodes displayed necrosis and mineralization at the cortical and medullar areas. Ziehl-Neelsen stained bacteria were numerous inside macrophages and Langhans-type giant cells. Giant and epithelioid cells and lymphocytes were prominent at the ileal mucous membrane. The immunostaining of M. paratuberculosis was very clear inside epithelioid and giant cells. Image analysis was carried out to determine the immunostained area. There was total agreement among the methods employed. Immunohistochemistry can be very useful when the microorganism cannot be recovered from tissues or faeces.     

Lectin histochemical study of lipopigments present in the cerebellum of Solanum fastigiatum var. fastigiatum intoxicated cattle

This study was carried out to investigate the pattern of lectin binding in the cerebellum of calves poisoned with Solanum fastigiatum var. fastigiatum. For the experimental reproduction of the illness, S. fastigiatum var. fastigiatum was collected from farms where the intoxication occurs. The dried ground plant was administered to two 1-year-old cattle by a ruminal cannula. The animals received 5 g/kg b.w. daily, 5 days a week, during periods of 107 and 140 days. After these periods the animals were bled to death. For the histological study, transverse sections of the cerebellum were used. Paraffin-embedded sections were incubated with the following biotinylated lectins with different specificity: Concanavalia ensiformis (Con-A). Glycine max (SBA). Dolichos hiflorus (DBA), Ulex europeus-I (UEA-I). Triticum vulgaris (WGA), succynyl-WGA (sWGA). Arachis hypogaea (PNA), Ricinus communis-I (RCA-I) and Bandeirea simplicifolia-I (BS-I). Avidin-biotin-peroxidase complex was applied as a detection system. Purkinje cells showed vacuolation in the pericaryon. The stored material present in the cells reacted strongly with the following lectins: Con-A. sWGA, WGA and RCA-I. An irregular affinity was observed with PNA and DBA. The lectin-binding pattern was compatible with a glycolipid storage disease.     

Measurement of Mechanical Forces Acting on the Area Pellucida of the Early Chick Embryo

We have designed a method for measuring mechanical forces acting on the area pellucida of the chick embryo between stages 1 to 5 of HAMBURGER and HAMILTON (1951). The coordinates of several points on the outline of the area pellucida were used as input, from which a computer program measured the changes in the position of similar points to produce a table of values which, we argue, reflect mechanical forces in the embryo. The results obtained are shown to be reproducible between embryos. The computer program and the measurements obtained were then used to predict the behaviour of surgical incisions made in the area pellucida, using the tables obtained from direct measurements of embryos and the coordinates of various points on the surface of the operated embryos. This procedure was applied to embryos cultured by the method of NEW (1955) and in-ovo. It was found that in those embryos cultured by EWS technique the computer predictions correspond very closely to shape measurements made directly from embryos. However, the correspondence is less close when applied to embryos in-ovo.     

Blood calcium,glucose and haematology profiles of parturient bitches diagnosed with uterine inertia or obstructive dystocia

Bitches with dystocia most often present with clinical signs of uterine inertia (UI). The aetiology of myometrial dysfunction in most of these cases is still not elucidated. We compared blood ionized calcium (iCa) and glucose concentrations in bitches diagnosed with primary UI (PUI, n = 14), secondary UI (SUI, n = 6) or obstructive dystocia (OD, n = 6), and we described their haematology profiles. Bitches diagnosed with UI had a patent birth canal and delivered no puppies yet (PUI) or only part of the whole litter (SUI). The OD group had no UI and showed strong abdominal contractions. Blood iCa did not differ between the PUI, SUI and OD groups and was not influenced by litter size. There was a significant positive relationship (R² = .241, p = .013) between iCa concentrations and the dam's body weight. Glucose concentrations were also not significantly different between dystocia groups or influenced by body weight and litter size. Hypocalcaemia was detected in 11 bitches, and hypoglycaemia in two bitches. Pregnancy‐associated anaemia was seen in about one‐third of the bitches. Eight of 12 dogs had increased platelet counts, and ten had leukocytosis with mature neutrophilia. Although iCa did not differ between dystocia groups, low concentrations may have contributed to the development of UI in some of the small size bitches. Hypoglycaemia was uncommon, and therefore, we consider low glucose concentrations not to have played an important role in the pathogenesis of UI in our study population. Pregnancy‐associated anaemia, thrombocytosis, leukocytosis and mature neutrophilia were common findings in otherwise healthy bitches diagnosed with different forms of dystocia.