Hidden Genetic Variability, Can the Olive Moth Prays oleae (Lepidoptera: Yponomeutidae or Praydidae?) be a Species’ Complex?

Prays oleae is the second most important pest in Mediterranean olive groves, causing substantial damage on olive production. We used mitochondrial [cytochrome c oxidase subunit I (COI), and NADH dehydrogenase subunit 5 (nad5)] and nuclear [ribosomal protein S5 (RpS5)] amplicons to assess the population variability in five main olive producing regions from Tunisia, to support or dismiss the existence of two non-monophyletic groups within the species, as found within Portugal. Our phylogenetic analysis with cytochrome c oxidase subunit I (COI) indeed displayed two distinct and well-supported clades of P. oleae, which were corroborated by the haplotype network reconstructed with both mitochondrial and nuclear amplicons. We were also able to dismiss the hypothesis that one of the clades would not develop on olive fruits. No correlation was observed between clades differentiation and geographic distribution. The existence of cryptic species can impact on the management of agroecosystems and on the perception of how these moths responds to environmental changes

Bactrocera oleae (Diptera:Tephritidae) in Iran: An invasion from the middle west

Despite an age-old tradition of olive growing and its geographical location, Iran was apparently free of the olive fly, Bactrocera oleae (Rossi, 1790) (Diptera: Tephritidae), the major worldwide olive tree pest, until the last decade. However, this situation has changed radically since B. oleae was first reported in 2004, and the olive fly is now Iran’s most important pest of olive trees, and a very serious threat to olive production in the Northern and more humid parts of the country. Genetic analyses of mitochondrial markers of B. oleae collected in the traditional olive growing area in Northwestern Iran were used to determine the origin of this pest. The results indicate it was most likely imported on olive trees from the Central Mediterranean area.Project PEst-C/AGR/ UI0115/2011 and project ALENT-07-0324-FEDER-001747, by FEDER funds through the latter project and Operational Programme for Competitiveness Factors – COMPETE, and by L. Teixeira da Costa’s personal funds. LTC was partially supported by an FCT grant Ciência 2008-ICAAM

The mitochondrial genome of Prays oleae (Insecta: Lepidoptera: Praydidae).

Prays oleae is one of the most important olive tree pests and a species of interest in evolutionary studies, as it belongs to one of the oldest extant superfamilies of Ditrysian Lepidoptera. We determined its mitogenome sequence, and found it has common features for Lepidoptera, e.g. an 480% A + T content, an apparent CGA start codon for COX1 and an ATAGA(T)n motif in the control region, which also contains several copies of a 163-164 bp repeat. Importantly, the mitogenome displays the Met-Ile-Gln tRNA gene order typical of Ditrysia, consistent with the hypothesis that this is a synapomorphy of that clade

Isolation of entomopathogenic Bacillus from a biodynamic olive farm and their pathogenicity to lepidopteran and coleopteran insect pests

The occurrence of Bacillus entomopathogenic bacteria on a Tunisian biodynamic farm was determined by examining 75 samples from olive tree (Olea europaea L.) habitats. A total of 40 Bacillus isolates were characterized according to their phenotypic, physiological and biochemical parameters. Isolates of the species Bacillus subtilis, Bacillus mycoides, Brevibacillus brevis, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus sp. (1), Bacillus sp. (2) and a standard strain Btk HD-1 were used separately in feeding bioassays on fresh artificial diet against larvae of lepidopterans Prays oleae (Bernard) and Palpita unionalis (Hübner) and coleopterans Hylesinus oleiperda (F.) and Phloeotribus scarabaeoides (Bernard), which are olive tree pests. Larvae were successfully reared on an artificial diet with 25 g powdered olive tree leaves. Compared to the control data, only Btk and the isolates of B. licheniformis, P. polymyxa and B. brevis were entomopathogenic. Larval mortality assessed 7 days post-treatment showed high mortality rates with Btk to lepidopteran larvae (86.6% for P. oleae and 80.9% for P. unionalis) and low mortality against coleopteran pests. B. brevis isolates showed high mortality rates against P. oleae (up to 67.9%). B. licheniformis isolates caused up to 59.2% larval mortality for P. oleae and 43.6% for P. unionalis. Highest coleopteran mortality was achieved by P. polymyxa isolates (up to 55%). According to the 16S rDNA results, isolates of each of the three entomopathogenic strains were similar. Proteins in the strain supernatants were toxic to P. oleae larvae with LC50 values of 10.0 (B. brevis), 12.5 (B. licheniformis) and 37.6 μg/ml (P. polymyxa). Also, P. polymyxa showed an LC50 of 12.4 mg/l against P. scarabaeoides. Our results suggest that entomopathogenic Bacillus present locally in the biodynamic farm could be used in biological control programmes of olive tree pests

A rare coexistence of sarcoidosis with overlap syndrome and sarcoidosis with primary biliary cholangitis and Sjogren's syndrome: Two distinct case reports

Sarcoidosis and the overlap syndrome of autoimmune hepatitis and primary biliary cholangitis (PBC) share common clinical, biological, and histological features. The simultaneous occurrence of these diseases have been reported in few cases and suggests that a common pathway which may contribute to granuloma formation in both conditions.We report the cases of two female patients having an association of sarcoidosis and inflammatory liver diseases.The first case is of a 61-year-old woman had been monitored for an overlap syndrome of PBC and autoimmune hepatitis (AIH). Therefore, treatment with azathiprine has been initiated associated with ursodeoxycholic acid (UDCA). Azathioprine had to be discontinued due to digestive intolerance, specifically chronic diarrhea and abdominal pain. The patient remained clinically stable on UDCA and her liver function tests were stable for years, until she developed symptoms of progressive dyspnea without any other associated signs. Chest computed tomography (CT) revealed mediastinal enlargement, bilateral pulmonary nodules, and symmetrical adenomegalies in the mediastinum. The bronchoalveolar lavage (BAL) revealed increased cellularity, with a notable elevation in lymphocyte count (48 %) and a CD4/CD8 ratio of 4. The patient underwent mediastinoscopy; a biopsy of the right laterotracheal (4R) adenomegaly was performed. Histological examination of the lymph node showed epithelioid and giant-cell tuberculoid lymphadenitis without necrosis, compatible with sarcoidosis. Ophthalmological and cardiac assessments were normal. Plethysmography test was normal and there was no need for corticosteroid treatment; a surveillance was planned. Treatment with UDCA was pursued. The second case is of a 50-year-old woman with no medical history presented symptoms including dry eyes and mouth, inflammatory-type polyarthralgia affecting knees and wrists, bilateral Raynaud's phenomenon, right hypochondrium pain, and worsening dyspnea over six months. Liver analysis showed elevation of alkaline phosphatase (ALP) to three times upper limit of normal (ULN) and gamma-glutamyltransferase (GGT) to 5 times ULN. This cholestasis was associated with an increase in transaminase activity to 5 times ULN for over six months. Immunological tests revealed positive anti-nuclear antibodies (ANA), anti-Ro52, anti-M23E, and anti-centromere antibodies. Chest-CT showed multiple bilateral bronchiolar parenchymal micronodules mostly in the upper and posterior regions without any mediastinal adenomegaly. Bronchial endoscopy was normal, and biopsies indicated chronic inflammation. The BAL revealed increased cellularity, characterized by a high lymphocyte count (51.7 %) and a CD4/CD8 ratio of 2.8. Biopsy of minor salivary gland revealed grade 4 lymphocytic sialadenitis. Skin biopsy revealed an epithelioid granuloma without caseous necrosis. Liver biopsy performed in the presence of cytolysis and moderate hepatic insufficiency, revealed granulomatous hepatitis and cholangitis lesions along with septal fibrosis suggestive of PBC. The diagnosis of cutaneous and pulmonary sarcoidosis with PBC and Sjögren's syndrome was retained. The spirometry and diffusing capacity for carbon monoxide value were normal. Treatment involved UDCA, corticosteroids, and azathioprine, leading to clinical and biological improvement.Sarcoidosis shares some clinical manifestations with autoimmune liver diseases, primarily PBC. A hepatic granuloma with a different appearance and location can guide the diagnosis. Early diagnosis and appropriate management can avoid serious complications and improve prognosis

Hyper-proteolytic mutant of Beauveria bassiana, a new biological control agent against the tomato borer

The world tomato production is threatened by the invasive tomato borer Tuta absoluta. Difficulties in managing this pest were imposed mainly by the development of resistance in strains treated with conventional chemical insecticides. Resistance problems were even reported to insecticides of natural origin, leading to search for other control alternatives. P2 is a spontaneous mutant of the entomopathogenic fungus Beauveria bassiana. It was previously selected from a local strain (P1) and was characterized as hyper-producer of extracellular proteases. Here, the insecticidal potential of P1 and P2 strains was evaluated against T. absoluta larvae under laboratory conditions. Both strains were effective but P2 showed stronger effect than P1; median lethal concentration of P2 is tenfold lower than that of P1. Enzymatic assay analysis showed that extracellular enzymes are differently expressed by the two strains, especially proteases and chitinases which are known as cuticle degrading enzymes. The major expressed subtilisin-like protease (SBP) was upregulated at the transcriptional level in P2 strain. Proteomic analysis revealed four SBP isoforms which are highly overexpressed in this strain compared to P1. Post-translational regulation, most probably phosphorylation, was further suggested to control the SBP protease expression in B. bassiana P1 and P2 strains. The enzymatic profile in the two strains might explain their different insecticidal potential against the tomato borer. This is the first report showing such efficiency of Beauveria strains against this dangerous pest. Particularly, P2 strain showed high virulence reaching almost total larval mortality within 5 days post-application. It thus should be recommended as a new tool for the biocontrol of T. absoluta