Evaluation of selected growth factor expression in canine spirocercosis (Spirocerca lupi)-associated non-neoplastic nodules and sarcomas

The study aims to assess the expression of vascular endothelial growth factor (VEGF)-A, fibroblast growth factor (FGF) and platelet-derived growth factor (PDGF) in the progression of the spirocercosis-induced oesophageal nodule in the dog from early, non-neoplastic, inflammatory nodule to sarcomatous neoplasia. Triplicate 4 μm-thick sections from 62 paraffin blocks containing Spirocerca-induced nodules were treated according to the steptavidin-biotin (LSAB) immunohistochemical method using polyclonal goat-anti-canine VEGF and anti-human FGF and PDGF antibodies. The nodules were classified as early inflammatory (n=15), pre-neoplastic (n=27) and neoplastic (n=20). Additionally, 10 sections of normal distal third of the oesophagus and 21 non-spirocercosis-related sarcomas were evaluated and compared with the Spirocerca-induced nodules. Five non-overlapping high power fields per case were evaluated under the light microscope and the fibroblasts were evaluated for percentage of labelled cells. The intensity of labelling was further classified as weak (score 1) or strong (score 2). The intensity score was multiplied by the percentage of labelled fibroblasts to yield a field score and the final score was obtained by calculating an average of the five fields. Antigen labelling was compared between the different histological grades and the controls using the Kruskal-Wallis test followed by the Mann-Whitney test for comparison between specific groups. The level of significance was set at 0.05. There were significant differences between the groups' score in all the growth factors that were examined. The normal oesophagus showed no labelling for any of the growth factors. FGF scored highest in the non-spirocercosis-related sarcoma group (median 118, 3-194) followed by the spirocercosis-induced sarcoma (34.5, 0-138), pre-neoplastic nodule (8, 0-99) and early nodule (0, 0-30) groups. All the differences among the groups were significant. VEGF scored highest in the non-spirocercosis-related sarcoma group (median 47, 1-110) followed by the spirocercosis-induced sarcoma (26, 0-136), pre-neoplastic nodule (0, 0-62) and early nodule (0, 0-35) groups. PDGF scored highest in the non-spirocercosis-related sarcoma group (median 29.2, 0-70) followed by the pre-neoplastic nodule (23.4, 0-95), early nodule (13.6, 0-132) and spirocercosis-induced sarcoma (0, 0-47) groups. The expression of VEGF and FGF increased as the nodule progressed from early inflammation to sarcoma, but it was not limited to spirocercosis-induced sarcomas. The expression of PDGF in spirocercosis was restricted to the early stages of nodule progression. Further investigation is warranted to establish whether FGF, VEGF or PDGF play a role in the pathogenesis of the malignant transformation in canine spirocercosis or are they simply integral to angiogenesis induction?     

Idiopathic myelofibrosis accompanied by peritoneal extramedullary hematopoiesis presenting as refractory ascites in a dog

A 2.5‐year‐old spayed female American Pit Bull Terrier dog presented with a primary complaint of chronic refractory ascites. The dog's CBC displayed a moderate to severe macrocytic, hypochromic, nonregenerative anemia, and a moderate leukopenia as result of a moderate neutropenia and monocytopenia. Microscopic examination of the blood smear showed marked anisocytosis, mild polychromasia, mild acanthocytosis and ovalocytosis, moderate schistocytosis and poikilocytosis, and 4 metarubricytes/100 WBC. Abdominal ultrasonography revealed a homogenous, mild to moderately hyperechoic appearing liver as well as marked amounts of speckled anechoic to slightly hypoechoic peritoneal fluid. Cytology of the ascitic fluid demonstrated a sterile transudate, with evidence of a chronic inflammatory reaction as well as erythroid and myeloid precursor cells, and a few megakaryocytes with occasional micromegakaryocytes. Histologic sections of bone marrow, spleen, and liver were examined, using routine H&E stains, as well as a variety of immunohistochemistry and other special stains. Histopathology of the bone marrow and spleen revealed varying degrees of fibrosis, erythroid, and myeloid hyperplasia, as well as multiple small hyperplastic clusters of megakaryocytes. The megakaryocytes displayed many features of atypia such as increased cytoplasmic basophilia and occasional abnormal chromatin clumping with mitoses. Histopathologic examination of the liver disclosed evidence of mild extramedullary hematopoiesis. This case represents the first report of canine idiopathic myelofibrosis associated with peritoneal extramedullary hematopoiesis, resulting in refractory ascites. Although idiopathic myelofibrosis is a relatively rare condition in dogs, this case demonstrates that ascites caused by peritoneal implants of hematopoietic tissue may be the initial manifestation of myelofibrosis.     

Epizootics of sudden death in tammar wallabies (Macropus eugenii) associated with an orbivirus infection

Epizootics of sudden death in tammar wallabies (Macropus eugenii) occurred at six research facilities and zoological gardens in New South Wales, Australia, in late 1998 and at one Queensland research facility in March 1999. There were 120 confirmed tammar wallaby deaths during this period; however, population censuses indicated that up to 230 tammar wallabies may have died. The majority of animals died without premonitory signs. A small proportion of wallabies exhibited increased respiratory rate, sat with a lowered head shortly before death or were discovered in lateral recumbency, moribund and with muscle fasciculations. Gross postmortem findings consistently included massive pulmonary congestion, mottled hepatic parenchyma and subcutaneous oedema throughout the hindlimbs and inguinal region. Approximately 30% of the animals examined also had extensive haemorrhage within the fascial planes and skeletal muscle of the hindlimb adductors, inguinal region, ventral thorax, dorsal cervical region and perirenal retroperitoneal area. The tissues of affected animals became autolytic within a short period after death. Bacteriological examination of tissues from 14 animals did not provide any significant findings. Toxicological examination of the gastric and colonic contents of four animals did not reveal evidence of brodifacoume or other rodenticides. Viruses from the Eubenangee serogroup of the Orbivirus genus were isolated from the cerebral cortex of nine, and the myocardium of two, tammar wallabies and the liver and intestine of another tammar wallaby. A similar orbivirus was also isolated from the cerebrospinal fluid of another tammar wallaby that died suddenly. The disease agent appears to be a previously unrecognised orbivirus in the Eubenangee serogroup. This is the first report of epizootics of sudden deaths in tammar wallabies apparently associated with an orbivirus infection.     

Bentonite as therapy for Lantana camara poisoning of cattle

Bentonite was compared with activated charcoal as therapy for lantana poisoning in calves dosed 5 d previously with leaf material of the common pink-edged red taxon of Lantana camara. Both therapies were given by stomach tube as a single dose at 5 g/kg. Five of 6 calves in each of the groups given bentonite and activated charcoal recovered while 5 of 6 calves in the control group died. Calves given bentonite took 3 d longer on average to recover fully than those given activated charcoal but the effects of the 2 therapies on plasma total bilirubin concentrations were statistically indistinguishable. Bentonite was judged to have promise as a cheap alternative to activated charcoal for therapy of lantana poisoning of cattle.     

Efficacies of low- to high-volume (960-10 700 litre ha− 1) citrus sprayers for applying petroleum spray oil to control chinese wax scale

Petroleum spray oil (2, 4 and 6% in water) was applied to Valencia orange, Citrus sinensis (L.) Osbeck, for the control of Chinese wax scale, Ceroplastes sinensis del Guercio, using a low-volume ( <2000 litre ha^?1)air-blast (LV AB) sprayer, a low- to high-volume (L-HV) (up to 7000 litre ha^?1) sprayer with four fan-assisted rotary atomiser (FARA) spray heads mounted on a vertical tower, and a high-volume (>7000 litre ha^?1) oscillating boom (HV OB) sprayer. The most effective sprayer was the L-HV FARA sprayer. The most cost-effective treatment was a 20 ml litre^?1 (60 litre oil ha^?1) spray applied at 3000 litre ha^?1 by the L-HV FARA sprayer. It gave mortality equivalent to a standard 20 ml litre^?1, 10 700 litre ha^?1 spray (214 litre oil ha^?1) applied by the HV OB sprayer but with 72% less spray and significantly less oil deposited per cm² of leaf area. Equivalent or significantly (P = 0·05) higher mortality than that given by the 10 700 litre ha^?1 HV OB spray was given by the 40 ml litre^?1, 3000 (120 litre oil ha^?1) and 60 ml litre^?1, 2180 and 3000 litre ha^?1 (130·8 and 180 litre oil ha^?1) L-HV FARA sprays, but the 60 ml litre^?1 sprays deposited more oil per cm² than the 20 ml litre^?1 HV OB spray and were considered to be potentially phytotoxic. The least effective sprayer was the LV AB sprayer, which applied a 60 ml litre^?1 spray (57·6 litre oil ha^?1) at 960 litre ha^?1. Linear relationships were established for Chinese wax scale mortality, transformed using an angular transformation (arcsin proportion), versus log₁₀ spray volume for the 20, 40 and 60 ml litre^?1 sprays applied by L-HV FARA at 1260,2180 and 3000 litre ha^?1, mortality versus log₁₀ μg oil cm^?2 and log₁₀ μg oil versus log₁₀ volume of oil sprayed.     

Ultrasonographic Characteristics of Ovulatory Follicles and Associated Endocrine Changes in Cyclic Ewes Treated with Medroxyprogesterone Acetate (MAP)-releasing Intravaginal Sponges and Equine Chorionic Gonadotropin (eCG)

The aim of this study was to assess the ultrasonographic characteristics of ovulatory follicles in cyclic Western White Face ewes (December) that had received intravaginal sponges containing medroxyprogesterone acetate (MAP; 60 mg) for 12 days, with or without an injection of 500 IU of equine chorionic gonadotropin (eCG) at sponge removal. We hypothesized that quantitative echotextural attributes of the follicles in ewes treated only with MAP would differ from those in MAP/eCG-treated ewes, reflecting the increased antral follicular growth and secretory function under eCG influence. Digital images of ovulatory follicles obtained at 0 and 24 h after MAP sponge removal and at 24 h before ovulation in the eCG-treated (five ewes, 13 follicles) and control (six ewes, 9 follicles) animals, were subjected to computerized analyses. The mean diameter of ovulatory follicles increased (p < 0.001) 24 h after eCG treatment. The mean pixel intensity and heterogeneity of the follicular antrum (p < 0.001), as well as mean pixel intensity of the follicular wall and perifollicular ovarian stroma (p < 0.05), were greater in eCG-treated animals compared with control ewes 24 h after sponge removal and at 24 h before ovulation. Mean serum concentrations of oestradiol-17beta tended to increase (p = 0.06) 24 h after eCG treatment and the eCG-treated ewes exceeded (p < 0.05) control animals in progesterone concentrations from days 9-15 after ovulation. Our results support the hypothesis that large antral follicles in eCG-treated ewes exhibit distinctive echotextural characteristics. Follicular image attributes in eCG-treated ewes appear to be indicative of the changes in follicular morphology and secretory activity caused by the administration of the exogenous gonadotropin, which has both FSH- and LH-like activities.     

Effects of FGF10 on Bovine Oocyte Meiosis Progression,Apoptosis, Embryo Development and Relative Abundance of Developmentally Important Genes In Vitro

Fibroblast growth factor (FGF10) acts at the cumulus oocyte complex, increasing the expression of cumulus cell expansion‐related genes and oocyte competency genes. We tested the hypothesis that addition of FGF10 to the maturation medium improves oocyte maturation, decreases the percentage of apoptotic oocytes and increases development to the blastocyst stage while increasing the relative abundance of developmentally important genes (COX2, CDX2 and PLAC8). In all experiments, oocytes were matured for 22 h in TCM‐199 supplemented with 0, 2.5, 10 or 50 ng/ml FGF10. In Experiment 1, after maturation, oocytes were stained with Hoechst to evaluate meiosis progression (metaphase I, intermediary phases and extrusion of the first polar body) and submitted to the TUNEL assay to evaluate apoptosis. In Experiment 2, oocytes were fertilized and cultured to the blastocyst stage. Blastocysts were frozen for analysis of COX2, CDX2 and PLAC8 relative abundance. In Experiment 1, 2.5 ng/ml FGF10 increased (p < 0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50 ng/ml FGF10 (21, 14 and 12%, respectively) and FGF10 decreased the percentage of oocytes that were TUNEL positive in all doses studied. In Experiment 2, there was no difference in the percentage of oocytes becoming blastocysts between treatments and control. Real‐time RT‐PCR showed a tendency of 50 ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10 ng/ml FGF10 to increase CDX2. In conclusion, the addition of FGF10 to the oocyte maturation medium improves oocyte maturation in vitro, decreases the percentage of apoptotic oocytes and tends to increase the relative abundance of developmentally important genes.     

Effect of compost and casing treatments of insecticides against the sciarid Bradysia ocellaris (Diptera: Sciaridae) and on the total yield of cultivated mushrooms,Agaricus bisporus

BACKGROUND: Bradysia ocellaris (Comstock) is one of the major pests of cultivated mushrooms, Agaricus bisporus (Lange) Imbach. The feeding of sciarid fly larvae causes damage to the cultivation substrates, compost and casing. A comparative study of insecticidal treatments incorporated into compost and casing against B. ocellaris was conducted during 2003–2004. Treatments with an organophosphate, diazinon, a pyrazole, fipronil, and an insect growth regulator (IGR), triflumuron, were incorporated into casing. Another IGR, cyromazine, was incorporated into compost. Insecticidal incorporation into compost and casing has been associated with reductions in mushroom yield. Therefore, the effect of these insecticidal treatments on the overall yield was evaluated. RESULTS: A combined treatment, incorporation of cyromazine at 10 mg kg^?1 into compost and of triflumuron at 20 mg kg^?1 into casing, provided 100% control against B. ocellaris. There was no apparent reduction in yield as a result of incorporation of the tested chemicals into either compost or casing. CONCLUSION: The results obtained from this study suggest that incorporation of insecticides into both compost and casing is required to control a moderate to high pressure of insect pests. There were no significant yield reductions as a result of the incorporation of pesticides in the Australian mushroom‐growing system. Copyright © 2009 Society of Chemical Industry