Regeneration of Fitzroya cupressoides after indigenous and non-indigenous timber harvesting in southern Chilean forests

Fitzroya cupressoides (Cupressaceae) is an endemic and long-lived conifer of southern Chile and Argentina (40–43° S). This species has been subject to continuous exploitation since the 16th century, causing extensive population decline. Historically, the main labour force for the exploitation of F. cupressoides (alerce) was the indigenous Mapuche-Huilliche population, first under the command of the Spanish settlers and later, of non-indigenous Chileans. In coastal forests, timber of alerce has been harvested by Huilliche communities as well as by Chilean and international forestry companies. Records of the regeneration of this pioneer tree after exploitation in the Andean mountains have generally shown limited regeneration depending on the intensity of harvest. Because indigenous exploitation does not use machinery for timber harvesting, and is supposedly less utilitarian than commercial harvest, I propose that areas in the Coastal Range harvested by Huilliche communities should present higher regeneration of alerce than areas harvested by forestry companies. To test this hypothesis, I sampled 10 stands harvested and abandoned by forestry companies and 10 stands harvested by Huilliche communities in the coastal range of the Osorno Province (41° S, 400–800 m). In each stand, I estimated the density of regeneration (sapling stage), number of stumps, number of live adults, and the number of standing and fallen dead alerce. Each stand was characterized by elevation, forest-type, incidence of fire, and vegetation cover. Results showed that forests harvested by Huilliches had higher numbers of live, remnant adult trees with a dbh ≥ 60 cm than forests harvested by timber companies. The number of stumps with a dbh ≥60 cm was significantly higher in stands harvested by timber companies than in Huilliche stands. Despite large differences in sapling densities among stands, regeneration density of alerce was unrelated to the type of harvest used by indigenous people or forestry companies.     

Polymorphism of the PE domain of PE/PE_PGRS sequences in clinical isolates of Mycobacterium bovis in Mexico

Polymorphism of the PE domain of PE/PE_PGRS sequences was studied in Mycobacterium bovis isolates from different Mexican states. Samples were analyzed by spolygotyping and RFLP using IS6110 and a 235-bp fragment of the PE domain of PE/PE_PGRS as probes. With the PE probe, three different genotypes were observed, one being predominant in all states. These results confirm the high conservation of the PE domain and suggests a potential role for PE sequence as a stable genetic marker for bovine tuberculosis.     

Contrast‐enhanced ultrasound complements two‐dimensional ultrasonography in diagnosing gallbladder diseases in dogs

Gall‐bladder diseases are common in dogs and two‐dimensional ultrasonography is a current standard method for diagnosis and treatment planning. However, findings from this modality can be nonspecific. The aim of this retrospective, case series study was to describe conventional and contrast‐enhanced ultrasound (using SonoVue^®) findings in a group of dogs with histologically confirmed gall bladder disease. A total of 65 dogs were included. Branchlike, heterogeneous, and homogeneous contrast enhancement of echogenic intraluminal mass‐forming lesions was a contrast‐enhanced ultrasound characteristic of polypoid lesions due to cystic mucosal hyperplasia of the gallbladder and/or tumor, which had different wash‐in and washout characteristics. In dogs with mobile or immobile biliary sludge or mucocele, the echogenic intraluminal masses remained unenhanced. A double rim mark or enhancement defect in the gallbladder wall was a characteristic of edema or necrosis/rupture of the wall, respectively. Conventional ultrasonography correctly identified biliary sludge or mucocele in 36/37 dogs, cholecystitis/edema in 44/47 dogs, necrosis/rupture in 19/25 dogs, and gallbladder neoplasia in three of three dogs with these pathologies. It falsely identified biliary sludge or mucocele in eight of 28 dogs, cholecystitis/edema in three of 15 dogs, necrosis/rupture in 13/37 dogs, and gall‐bladder neoplasia in 20/59 dogs that did not have these pathologies. Contrast‐enhanced ultrasound correctly identified cholecystitis/edema in 42/47 dogs, but falsely identified cholecystitis/edema in three of 18 dogs. It correctly identified necrosis/rupture, benign polypoid lesions, and gallbladder neoplasia in all dogs with no false‐positive results. Findings supported contrast‐enhanced ultrasound as a complement to conventional ultrasonography for dogs with suspected gallbladder pathologies such as edema, necrosis, and rupture.     

Paraganglioma,pituitary adenoma,and osteosarcoma in a dog

An 11‐year‐old neutered male Alaskan Malamute mixed‐breed dog was presented with a complaint of polyuria/polydipsia (PU/PD), weight loss, tachypnea, regurgitation, and a previous history of nontreated osteosarcoma of the right distal radius, diagnosed 21 months prior. On physical examination, an abdominal mass was palpated. The abdominal mass was aspirated and cytologically diagnosed as a neuroendocrine tumor, suspected to be a pheochromocytoma. Laboratory examination revealed a mild anemia and thrombocytopenia, markedly elevated ATP and ALP activities, and moderate hypercalcemia. A low‐dose dexamethasone suppression test and endogenous adrenocorticotropic hormone (ACTH) concentration were compatible with pituitary hyperadrenocorticism. On urinalysis, proteinuria was noted as well as a high urine metanephrine/creatinine ratio, consistent with a diagnosis of pheochromocytoma. The dog was treated with supportive care and euthanized 6 months later due to decreasing quality of life. On necropsy, an extra‐adrenal pheochromocytoma (paraganglioma) was diagnosed in the caudal abdomen, and a pituitary adenoma and an osteosarcoma of the right distal radius were confirmed.     

Molecular epidemiology of Mycobacterium bovis: usefulness in international trade

Tuberculosis (TB) represents a barrier for free trade of livestock between Mexico and the United States of America (US). In spite of efforts from Mexico to export TB-free animals, some of those found with TB lesions in slaughterhouses in the US are traced back to that country. Therefore, the purpose of this study was to determine, through molecular epidemiology, the most probable source of infection for cattle found with TB lesions in the US. Ninety M. bovis isolates, 50 from Mexico obtained from cattle in 8 different states, and 40 from the US from cattle, deer, elk and feral pigs from 7 different states were included in the study. All samples were analyzed in both laboratories, Mexico and the US, following the same protocol for molecular analysis by spoligotyping. Twenty-seven clusters, ranging from 1 to 18 genetically similar strains were found. Some clustering by country was observed, strains from cattle and deer in Michigan in the US fell into the same cluster, suggesting transmission between species. These results, combined with epidemiological information suggest that despite of the possibility that some animals with lesions in the US come from Mexico as false negatives, the US has its own source of infection, must probably in dairy cattle and wildlife. Genetic diversity of isolates from Mexico was larger than that in the US, which could be a consequence of the endemic status of the disease and the indiscriminate movement of animals between regions.     

Morphological and acrosomal changes of canine spermatozoa during epididymal transit

Background

During epididymal transit, functional and structural modifications leading to full maturation enable male gametes to reach, recognize and fertilize the oocytes. In dogs, little is known on the modifications of spermatozoa during the passage in the epididymis. The aim of this study was to describe the motility, morphology and acrosomal patterns of canine spermatozoa retrieved from the epididymis caput, corpus and cauda.

Results

After the dilution required for the collection of epididymal content, sperm motility was significantly higher (P <0.0001) in the cauda compared to corpus and caput.Proportions of spermatozoa with normal morphology were significantly higher in corpus (P =0.02) and cauda (P <0.0001) compared to caput. Overall morphological abnormalities of the head and neck/midpiece were similar in the three different epididymal regions. A significantly increased prevalence of tail defects, mainly represented by single bent tails, was observed in the corpus compared to caput (P <0.0001) and cauda (P =0.006).Numbers of immature sperm with cytoplasmic droplets decreased from the proximal to the distal region of the epididymis. Particularly, proximal cytoplasmic droplets were more frequently found in spermatozoa collected from the caput epididymis than in the corpus (P <0.0001) and in the cauda (P <0.0001), whereas the occurrence of distal cytoplasmic droplets was higher in the corpus than in the caput (P =0.0003) and in the cauda (P <0.05).Significantly higher proportions of spermatozoa with intact acrosomes were retrieved from the cauda epididymis than from the caput (P =0.03) and the corpus (P =0.008). This difference was mainly due to a lower proportion of spermatozoa with abnormal acrosomes (mainly swollen acrosomes) rather than with absent acrosomes.

Conclusions

Canine spermatozoa undergo several modifications in the epididymis. The acquisition of progressive motility, migration of the cytoplasmic droplet and acrosomal reshaping lead to mature spermatozoa which are then stored in the cauda epididymis. From this site, spermatozoa can be retrieved and used in assisted reproductive techniques as a valuable tool for propagating genetic traits of high value individuals that dies accidentally or undergoes orchiectomy for medical purposes. Further investigations should be also focused on the potential use of spermatozoa recovered from other epididymal regions.     

Cultivation affected the level of inorganic phosphorus more than the organic pool of the Pampas soils in Argentina

The effects of land use on soil organic and inorganic phosphorus (P) stocks were assessed in the Pampas, Argentina. Three hundred and eighty-six paired sites widely distributed over an area of ca. 50 Mha were sampled. Land use types included soils under trees, uncropped soils, cropped soils at the pasture phase of a mixed rotation, cropped soils at the crop phase of a mixed rotation, and flooded soils. Slight differences in organic P stocks were found among land uses. Organic P was 21–35% lower in flooded soil than in the other treatments in the 0–100 cm depth. Inorganic P was significantly lower (ca. 27%) in pasture and cropped soils than in the uncropped controls at 0–25 cm depth. The ratios of organic P/inorganic P and organic C/organic P decreased with depth and did not significantly differ among the sites. The influence of cultivation on inorganic P to a depth of 100 cm depended on the initial phosphorus content of the soil. Soils rich in phosphorus lost substantial amounts of their phosphorus stocks, in some cases losses were as high as 70%, whereas phosphorus-poor soils presented only small changes in their inorganic P levels.     

Rebaling of silage and haylage and its effects on forage microbial and chemical composition—A pilot study

Use of big bale silage and haylage can be difficult on farms where daily forage consumption is comparatively low as speed of deterioration of forage after bale opening may be faster than feed-out rate. Production of smaller bales at harvest is possible, but expensive and work-intensive. Therefore, a pilot study of rebaling forage stored in big bales to smaller bales was conducted. Three separate experiments were included, where microbial and chemical composition of silage and haylage was studied before and after rebaling. In Experiment III, residual big bale forage stored and opened together with rebaled forage was included. Results showed that rebaled haylage and silage had higher yeast counts compared to initial forage; however, residual bales in Experiment III had yeast counts similar to rebaled forage, indicating an effect of storage time rather than of rebaling. In Experiment II, mould counts were higher in rebaled compared to initial silage, but not in haylage. Chemical composition was similar in initial and rebaled forage except for ammonia-N. In Experiment III, ammonia-N was higher in rebaled compared to initial and residual forage and was the only chemical variable affected by rebaling. Bale temperature during aerobic storage followed ambient temperature until day 6–8 in Experiment I and until day 14 in Experiment III where ambient temperature was lower. In conclusion, rebaling can be done without large changes in chemical composition of the forage, but yeast and mould counts may be higher in rebaled forage, and this risk should be considered when using this procedure.     

Partial chemical composition,free soluble sugars and unavailable carbohydrates in the embryonic axis and seed coat ofPhaseolus vulgaris L. (Canario group)

Four varieties of common bean were examined, but there were no significant differences among them. Crude protein content (46.7% and 49.0%) was the main constituent in embryonic axes. The average values of ash, crude fiber and ether extract were 4.2%, 3.7% and 3.0% respectively. Starch was approximately three times more abundant (7.5%) than cellulose (2.7%). Hemicellulose B values were lower (5.5%) than hemicellulose A (9.2%). The mean value of the pectic substances in embryonic axes was 12.3%. The free ethanol-soluble sugars ranged from 2.9% to 4.9%. Verbascose, stachyose, raffinose, sucrose, and galactose were identified and quantified in embryonic axes. Seed coat flour contained cellulose (30.6%), ether extract (0.4%), ash (2.8%), and protein (5.4%). Free ribose, mannose, arabinose, and galactose were present in seed coats in amounts ranging from 0.009% to 0.031%.     

In vitro and in vivo effects of toceranib phosphate on canine osteosarcoma cell lines and xenograft orthotopic models

Canine osteosarcoma (OSA) is the most common primary malignant bone tumour in dogs, and it has a high metastatic rate and poor prognosis. Toceranib phosphate (TOC; Palladia, Zoetis) is a veterinary tyrosine kinase inhibitor that selectively inhibits VEGFR‐2, PDGFRs and c‐Kit, but its efficacy is not yet fully understood in the treatment of canine OSA. Here, we evaluated the functional effects of TOC on six OSA cell lines by transwell, wound healing and colony formation assays. Subsequently, two cell lines (Wall and Penny) were selected and were inoculated in mice by intrafemoral injection to develop an orthotopic xenograft model of canine OSA. For each cell line, 30 mice were xenografted; half of them were used as controls, and the other half were treated with TOC at 40 mg/kg body weight for 20 days. TOC inhibited cell growth of all cell lines, but reduced invasion and migration was only observed in Penny and Wall cell lines. In mice engrafted with Penny cells and subjected to TOC treatment, decreased tumour growth was observed, and PDGFRs and c‐Kit mRNA were downregulated. Immunohistochemical analyses demonstrated a significant reduction of Ki67 staining in treated mice when compared to controls. The results obtained here demonstrate that TOC is able to slightly inhibit cell growth in vitro, while its effect is evident only in a Penny cell xenograft model, in which TOC significantly reduced tumour size and the Ki67 index without modifying apoptosis markers.