Species composition of Gasterophilus spp. (Diptera, Oestridae) causing equine gastric myiasis in southern Italy: parasite biodiversity and risks for extinction

Horse gastrointestinal myiasis caused by larvae of Gasterophilus spp. (Diptera, Oestridae) flies has a worldwide distribution and, where present, it is primarily caused by larvae of Gasterophilus intestinalis and Gasterophilus nasalis. Other species, i.e., Gasterophilus inermis, Gasterophilus pecorum and Gasterophilus haemorrhoidalis, present in different or in the same regions of the gastrointestinal tract, were only occasionally reported in very limited areas of eastern European Countries and in central Italy. With the aim to contribute data on the species composition of Gasterophilus and on the seasonal variation of the infection pattern in southern Italy, 152 native horses were necropsied from January to December 2003 and Gasterophilus larvae were collected from the stomach, the small intestine and the rectum of each of them. On the whole, 125 (82.2%) horses were infected by larvae of Gasterophilus spp. and 214 second stage larvae (L2) and 13,342 third stage larvae (L3) collected. Five species of Gasterophilus were identified with the following prevalence: G. intestinalis=95.2%, G. nasalis=44.8%, G. inermis=15.2%, G. pecorum=2.6% and G. haemorrhoidalis=0.8%. L3 were retrieved throughout the observation period with the highest mean burdens from January to August 2003 while the lowest mean was registered from September to November 2003. L2 were collected in February-March 2003 and from September to December 2003. The majority of the animals (n=66, 43.4%) were infected with a single Gasterophilus species, however, 45 animals (29.6%) harboured 2 species, 12 animals (7.9%) 3 species and 2 animals (1.3%) 4 species. The trend of abundance in the L3 of G. intestinalis and G. nasalis, the most represented species, was highly concordant (r=0.5, p<0.001). A retrospective comparison of our results and of other data from four seasons of observation (1983-1986) in central Italy showed that the number of G. inermis, G. pecorum and G. haemorrhoidalis have been decreasing relative to G. intestinalis and G. nasalis which may suggest tendency toward the extinction of the three former species of Gasterophilus.     

Note: A comparison of molecular diagnostic procedures for the detection of aster yellows phytoplasmas (16Sr-I) in leafhopper vectors

Different molecular procedures were compared for the detection of aster yellows phytoplasmas (AYP) in the leafhopper vectorsMacrosteles quadripunctulatus (Kirschbaum),Euscelidius variegatus (Kirschbaum) andEuscelis incisus (Kirschbaum). Polymerase chain reaction (PCR) with universal and group-specific primers designed on the 16S-rDNA sequence was most sensitive in nested assays. A dot-blot procedure with an oligoprobe designed on the 16S-rDNA was less sensitive and consistent to detect phytoplasmas in total insect DNA, but consistently detected amplicons from direct PCR. The dot-blot assay with a probe based on a phytoplasma plasmid sequence detected AYP in most vector specimens and did not react with DNAs from leafhoppers infected by flavescence dorée and psyllids infected by apple proliferation phytoplasmas. This last assay is almost devoid of contamination risks, faster and cheaper compared to PCR, therefore it has to be preferred for field-scale analysis of leafhopper populations. http://www.phytoparasitica.org posting Feb. 24, 2004.     

Cross-transmission studies with Hypoderma lineatum de Vill. (Diptera: Oestridae): attempted infestation of goats (Capra hircus)

The potential for cross-transmission of Hypoderma lineatum from cattle to domestic goats (Capra hircus) was examined using artificial infestation techniques. Two routes of infestation, subcutaneous injection and dermal penetration, were used to expose goats to newly hatched first instars. Presence of antibodies and appearance of circulating antigen (hypodermin C) were evaluated at selected intervals for up to 40 weeks post-infestation. In addition, immunoblots against H. lineatum first-instar proteins were conducted using sera taken at 10 weeks post-infestation. Goats were palpated for the presence of developing larvae at sub-dermal sites beginning at week 30 pi. No developing larvae were palpated at any time, regardless of the route of infestation nor was circulating antigen detected in any infested goats. Antibodies were present at weeks 6 and 10 and week 27 pi in both infested groups. Immunoblots indicated all infested goats produced antibodies to first instar H. lineatum antigens. H. lineatum appears to be incapable of completing development in domestic goats although the transient appearance of ELISA detectable antibodies and the presence of bands on immunoblots suggests that at least some larvae survive long-enough to engender a humoural response. The host specificity of H. lineatum is discussed in light of the general concepts of host-parasite relationships of oestrids.     

Early exposure to probiotics in a canine model of atopic dermatitis has long-term clinical and immunological effects

Probiotics modulate the immune response and may have protective effects against atopic dermatitis (AD). Clinical trials using dogs with spontaneous disease are limited by confounding factors such as different diets, environments and sensitizations while a more controlled evaluation is possible using experimental models. A validated model of canine AD showed that early exposure to Lactobacillus rhamnosus GG (LGG) significantly decreases allergen-specific IgE and partially prevents AD in the first 6 months of life. This study is a follow-up three years after discontinuation of LGG. Clinical signs were evaluated after allergen challenge with ragweed, timothy, Dermatophagoides farinae. Allergen-specific IgE, IL-10 and TGF-β were measured on the 1st day of challenge, before allergen exposure. Normal dogs were included as controls. Analyses included seven dogs in the non-probiotic and nine in the probiotic litter. For clinical scores, a 2-Group × 9-Time Analysis of Variance showed significant effects of group (p=0.0003, probiotic相似文献   

Towards a rapid molecular identification of the common phlebotomine sand flies in the Mediterranean region

The present study reports two simple molecular approaches allowing a rapid identification of the most prevalent species of phlebotomine sand flies in the Mediterranean region. A PCR protocol for the amplification of ITS2 ribosomal region and a PCR-RFLP on a mitochondrial DNA fragment (cytb-nd1) were settled in order to identify and discriminate among Phlebotomus perniciosus, Phlebotomus neglectus, Phlebotomus perfiliewi, Phlebotomus papatasi and Sergentomyia minuta. The ITS2 regions showed a certain degree of interspecific variability, which led to PCR amplicons of different sizes, i.e., 450, 490, 460, 480 and 530 bp for P. perniciosus, P. neglectus, P. perfiliewi, P. papatasi, and S. minuta, respectively. Analogously, the digestion of a mitochondrial DNA amplicon with Ase I enzyme showed five different restriction profiles, which allowed the unequivocal differentiation of the sand fly species examined. These methods might represent useful tools for a molecular large scale screening of phlebotomine sand fly species caught in areas where leishmaniasis is endemic, in order to plan appropriate epidemiological surveillance programs for both Leishmania spp. and their vectors.     

A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilaria immitis and Dirofilaria repens in dogs and mosquitoes

This study describes a duplex real-time polymerase chain reaction (PCR) assay for the detection and differentiation between Dirofilaria immitis and Dirofilaria repens in dog blood and mosquitoes. Regions of a cytochrome oxidase 1 (cox1) mitochondrial DNA fragment and the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA were amplified from microfilariae and adult worm samples, using a sensitive SsoFast? EvaGreen(?) based real-time PCR method coupled with melting-curve analysis. The limit of the real-time PCR in detecting microfilaria and adult worm DNA was also tested both in dog blood and in artificially infected microfilarial. Two peaks at different melting temperatures (T(m)) for D. immitis (mean ± SD=75.7 ± 0.3°C) and D. repens (mean ± SD=70 ± 0.7°C), respectively, were obtained for microfilarial and adult positive controls of both species when examined separately and together. The real-time PCR protocol was also efficient in detecting microfilarial and adult DNA of both species when tested in samples spiked with DNA from Aedes albopictus, in Aedes aegypti experimentally infected by D. repens and in Culex pipiens naturally infected by D. repens and D. immitis. The high sensitivity of real-time PCR confirmed its reliability in detecting small amounts of genomic DNA either in dog blood or mosquitoes (2.5 pg/μl and 3 × 10(-1)pg/μl for D. immitis and D. repens, respectively). This assay is proposed as a tool for the epidemiological surveillance of the two most important Dirofilaria species in areas where they are endemic and sympatric.     

Digital Agenesia in Martina Franca Donkey Foal: A Case Report

A 4-day-old male Martina Franca donkey foal was evaluated for a forelimb alteration. Clinical examination and radiographs revealed the agenesia of the distal digit. Biochemical parameters were normal, and ultrasonographic evaluation did not identify any relievable organ alteration. Karyotype study revealed an abnormality on chromosome 1. The foal was discharged with a distal limb bandage in which a palmar splint was applied. A poor prognosis for the functionality of the limb was given. In endangered species, such as the Martina Franca donkey, the excessive inbreeding could result in an increase in genetic disorders. These findings shed new light on the possible pathogenesis of the digital dysgenesia. The study of the karyotype could be a useful approach to detect genetic alterations that could or could not be expressed in the animal, especially in endangered species in which a risk of an excessive inbreeding is considerable. These defects should be considered in the choice and selection of the breeders.     

In vitro antifungal susceptibility of Malassezia pachydermatis from dogs with and without skin lesions

Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.     

Effects of milk feeding, frequency and concentration on weaning and buffalo (Bubalus bubalis) calf growth, health and behaviour

Growth, weight at birth and daily weight gain (DWG) on 12 water buffalo calves, starting from 6 days of age until completion of weaning, was investigated in this study. Different feeding regimens were given to two groups of animals with regard to daily milk replacer: (1) group 1 (G1) received a double concentration in single administration; whereas (2) group 2 (G2) received the same amount of milk replacer split twice daily. Blood samples were collected from each calf on days 6, 30, 60 and 90 to evaluate acute phase proteins (haptoglobin), bactericide activity, lysozime, total protein content and biochemical parameters. No differences were observed between the two groups in terms of dry matter intake, feed efficiency and live body weight at the end of the study. Interestingly, a significantly (P?<?0.05) reduced DWG was observed earlier in G1 (day 45) than in G2 (day 60). Gastrointestinal disorders were not recorded throughout the experimental period, and no significant differences were recorded between the two groups for all considered parameters. This study confirms the possibility of utilising one daily administration of milk replacer in water buffalo calf during weaning. This new approach facilitates calves management, without interfering with calves growing performances.     

Cross-sectional study on acrylamide hemoglobin adducts in subpopulations from the European Prospective Investigation into Cancer and Nutrition (EPIC) Study

Acrylamide exposure was investigated in subgroups of the EPIC study population (510 subjects from 9 European countries, randomly selected and stratified by age, gender, and smoking status) using hemoglobin adducts of acrylamide (HbAA) and its primary metabolite glycidamide (HbGA). Blood samples were analyzed for HbAA and HbGA by HPLC/MS/MS. Statistical models for HbAA and HbGA were developed including body mass index (BMI), educational level, and physical activity. A large variability in acrylamide exposure and metabolism between individuals and country groups was observed with HbAA and HbGA values ranging between 15-623 and 8-377 pmol/g of Hb, respectively. Both adducts differed significantly by country, sex, and smoking status. HbGA values were significantly lower in high alcohol consumers than in moderate consumers. With increasing BMI, HbGA in nonsmokers and HbAA in smokers decreased significantly. In the assessment of potential health effects related to acrylamide exposure, country of origin, BMI, alcohol consumption, sex, and smoking status should be considered.