A new phoretic association: Bursaphelenchus minutus (Nematoda: Parasitaphelenchidae) and Orthotomicus erosus (Coleoptera: Scolitydae) recorded on Pinus pinea (L.)

A new phoretic association between Bursaphelenchus minutus and the bark beetle Orthotomicus erosus collected on Pinus pinea is described for the first time. Nematode identification was based on morphological observation and biometric measurements of males and females. Moreover, B. minutus was characterized by ITS sequences and RFLP profile.     

Determination of famoxadone, fenamidone, fenhexamid and iprodione residues in greenhouse tomatoes

BACKGROUND: Greenhouse tomato production is intended exclusively for fresh consumption. Typical greenhouse conditions provide a conducive environment for the development of different fungi, principally late blight and grey mould, that can destroy plants and fruits. For this reason, different fungicides with different sites of action are used in integrated pest management strategies. Famoxadone, fenamidone, fenhexamid and iprodione (fig. 1 ) are fungicides of difference classes with different actions that could be used to control tomato pests. RESULTS: The QuEChERS method showed good recoveries, and the analytical method allowed good separation of the fungicides selected. Good results were obtained in terms of repeatability and intermediate precision, with a coefficient of variation of < 9.8%. The limits of determination and quantification of the method were far below the maximum residual levels (MRLs) set by the EU for these fungicides in tomatoes. The analysis of fungicide residues showed that cv. Shiren accounted for higher residue levels than cv. Caramba. The dissipation curves were similar in the two cultivars, indicating that the decrease in pesticides was not related to the tomato type. CONCLUSIONS: Care should be taken when using pesticides in greenhouse conditions, because degradation mainly affects fruit growth. The size of the tomato, in particular its surface/weight ratio, is very important for defining pesticide residues. All fungicides used showed residue levels below the MRLs at the preharvest interval. Copyright © 2011 Society of Chemical Industry     

Widespread expression of SAA and Hp RNA in bovine tissues after evaluation of suitable reference genes

The serum amyloid A (SAA) and haptoglobin (Hp) are the most prominent acute phase proteins (APPs) in cow. Liver mainly produces APPs, but extra hepatic expression has also been demonstrated in some tissues. The major aim of the present study was to assess the constitutive SAA and Hp mRNA expression by quantitative PCR (qPCR) in a wide panel of 33 bovine tissues, including gastrointestinal tract, respiratory system, urogenital system, mammary gland, hematopoietic system, central nervous system, eye, thyroid and heart. Normalization of gene expression in different samples requires reference genes, which are stably expressed. Therefore, seven reference genes were investigated (ACTB, GAPDH, HMBS, SDHA, YWHAZ, SF3A1, EEF1A2) and three genes, namely SF3A1, HMBS and ACTB, were selected after assessing their stability with geNorm? and NormFinder^© softwares.The qPCR analysis confirmed liver as the principal source of SAA and Hp, but also identified both APPs’ mRNA in almost all tissues.The highest expression rate of SAA was found in thyroid, followed by pancreas and submandibulary gland. Hp mRNA expression was detected at high concentration in pancreas and submandibulary gland.The present data indicated a widespread expression of SAA and Hp also in non pathological conditions, thus envisaging a possible role as immunomodulatory and protective molecules. To understand where SAA and Hp come from is the prerequisite to their utilization as Acute Phase Reaction biomarkers.     

Insertion sequence IS256 in canine pyoderma isolates of Staphylococcus pseudintermedius associated with antibiotic resistance

Staphylococcus pseudintermedius is the most frequent staphylococcal species isolated from canine pyoderma. The control of S. pseudintermedius infection is often difficult due to the expanded antimicrobial resistance phenotypes. Antibiotic resistance in staphylococcal pathogens is often associated to mobile genetic elements such as the insertion sequence IS256 that was first described as a part of the transposon Tn4001, which confers aminoglycoside resistance in Staphylococcus aureus and in Staphylococcus epidermidis. In this study a collection of 70 S. pseudintermedius isolates from canine pyoderma was used to investigate antimicrobial susceptibility to 15 antibiotics and the presence of IS256, not revealed in S. pseudintermedius yet. Antibiotic resistance profiling demonstrated that all S. pseudintermedius isolates had a multi-drug resistance phenotype, exhibiting simultaneous resistance to at least five antibiotics; indeed methicillin resistant S. pseudintermedius isolates were simultaneously resistant to at least nine antibiotics and all were also gentamicin resistant. PCR analyses revealed the presence of IS256 in 43/70 S. pseudintemedius isolates. The association between the presence of IS256 and the resistance was particularly significant for certain antibiotics: cefovecin, amikacin, gentamicin and oxacillin (χ(2)p-value<0.05). However, there was a striking result in frequency of strains resistant to gentamicin and oxacillin, suggesting a specific association between the presence of the IS256 element and the determinants for the resistance to these antibiotics. To the best of our knowledge, this is the first report showing the detection of IS256 in S. pseudintermedius isolates and its association with antibiotic resistance. Our findings suggest that S. pseudintermedius may acquire antibiotic resistance genes through mobile genetic elements which may play a predominant role in the dissemination of multi-drug resistance.     

Adenocarcinoma of the parotid salivary gland in a cow

A 6‐year‐old Girolando dairy cow was presented for evaluation of a large subcutaneous facial mass. Fine‐needle aspirates of the mass contained many neoplastic cells with high nuclear:cytoplasmic ratios arranged in sheets and loosely cohesive clusters with streaming erythrocytes and neutrophils in the background. Neoplastic cells were 13–25 μm in diameter and were round to cuboidal with variably distinct borders. Based on the signalment, anatomic location, and cytologic findings, differential diagnoses included salivary adenocarcinoma, squamous cell carcinoma, and mucoepidermoid carcinoma. The cow was euthanized and a necropsy was performed. The primary neoplasm arose from the left parotid salivary gland and meastatic tumor was found in the regional lymph nodes and lung. Histologically, the tumor was composed of anastomosing and irregular solid islets surrounded by scant stroma. Cells were negative for periodic acid‐Schiff (PAS), PAS‐diastase, and Alcian blue pH 2.5 stains, used to detect mucin. On immunohistochemical analysis, neoplastic luminal salivary gland cells expressed cytokeratin, but not S100, α‐smooth muscle actin, or vimentin. Peripheral cells of neoplastic islets were immunoreactive for p63. The final diagnosis was nonsecretory adenocarcinoma of the parotid salivary gland.     

3-D ultrastructural distribution of collagen in human placental villi at term in relation to vascular tree

In order to understand the 3-D distribution of collagen in relation to vascularization, chorionic villi of human placentae, belonging to normal pregnancies at term, were studied by scanning electron microscopy (SEM) after alkali maceration techniques, and by transmission electron microscopy (TEM). The villous tree appeared made of an uninterrupted structure of collagen fibres. The collagen fibres connected the chorionic villi axis with their basal plates and organised differently according to the various levels of villous branching. The collagen of stem villi showed copious fibres. The external fibres (facing the villous surface) were arranged mainly longitudinally. The central core of the villi (inner fibres) were arranged concentrically around the wall of the fetal vessels. Both external and internal fibres formed stratified lamellae or small parallel bundles. The inner core of stem villi showed small holes housing capillary spaces. Mature intermediate and terminal villi showed a scarce amount of collagen arranged in thin concentric layer within the villous core, surrounding numerous dilated capillary and sinusoid spaces.These observations demonstrated that the extracellular matrix of human chorionic villi is highly compartmentalised and shows a variable structural 3-D distribution depending on the branching level of the villous tree, such a distribution ensures the most favourable microenvironment for feto-maternal exchanges and it is likely able to provide a modulated support to the developing chorionic fetal vessels and trophoblastic layer as well.     

Feline cutaneous toxoplasmosis: a case report

A 9-year-old female, domestic short hair cat was presented with sudden onset of polyuria/polydipsia, and hundreds of cutaneous nodules. Prior to referral, the cat had had four skin nodules that were treated with steroids. The four skin nodules then multiplied to form more than 100 ulcerated and nonulcerated nodules located all over the trunk. Clinical evaluation revealed hypothermia and respiratory distress. Cytology from both skin nodules and bronchoalveolar lavage showed macrophages and small organisms whose shape and size were indicative of Toxoplasma spp., or similar organisms. Feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) serology results were negative. The cat was seropositive for Toxoplasma (IgG 1 : 640) and Neospora (1 : 80) infections. The cat died soon after referral. Necropsy revealed pyothorax, necrotic/purulent pneumonia, haemorrhagic spots on kidneys and mesentery. Histopathology from skin nodules showed diffuse, deep necrotic dermatitis/panniculitis, vasculitis and disseminated free and grouped protozoa. The parasites were found in lungs, spleen, kidneys and liver. Immunohistochemistry on skin tissue with anti-Toxoplasma gondii and Neospora caninum antibodies gave positive results with both. Electron microscopy showed single and grouped tachyzoites with morphological features of T. gondii, often within macrophages. Samples of cutaneous nodules and bronchoalveolar fluid were examined by a polymerase chain reaction (PCR) assay for detecting apicomplexa coccidia. PCR results were consistent only with T. gondii infection. Therefore, immunohistochemistry positivity for N. caninum was considered a cross-reaction and a diagnosis of cutaneous and visceral toxoplasmosis was made.