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Biomedical subjects

Claudio Genchi

Publications and source records attributed to Claudio Genchi.

15 recordsLinked to original sources

Genospecies of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks from the Autonomous Province of Trento, Italy.

Sequences of the variable intergenic spacer region 5S (rrfA) 23S (rrlB) rRNA were used to identify Borrelia genospecies present in Ixodes ricinus nymphs collected from the Lamar Lakes area of the Province of Trento, Italy (overall prevalence=6.3%). Four genospecies were identified, one for the first time in this Province (B. valaisiana), and three which have been noted previously (B. afzelii, B. garinii, and B. burgdorferi s.s.). In order to compare the genetic variability of these genospecies in Trento with that at a European level, our 21 sequences (15 new haplotypes) and all appropriate European Borrelia sequences registered in GenBank (up to the end of 2004) were subjected to a phylogenetic analysis (for a total of 73 sequences and 43 haplotypes). Clusters of sequences representing the five main European genospecies (afzelii, garinii, burgdorferi s.s., valaisiana, lusitaniae) are well-supported. At least two other groups of haplotypes (genospecies) are suggested by our analysis; moreover, divergent evolution may be occurring in several genospecies. The maximum uncorrected pairwise differences between sequences within genospecies ranges from 1.5% (B. burgdorferi s.s.), to 2.3% (B. garinii and B. valaisiana) to 4.7% (B. afzelii), and are not correlated with geographical distribution. Within the Province of Trento, these values for the same genospecies are 1.5%, 2.3%, 0.9%, 1.9%, respectively. These high mutation rates within genospecies suggest that the sequencing of haplotypes should continue if we are to fully understand and monitor the evolution and epidemiology of Borrelia.

Animals↗

A simple molecular method for discriminating common filarial nematodes of dogs (Canis familiaris).

Accurate diagnosis of canine filariosis is essential for choosing correct therapeutic approach. Therefore, reliable methods for discriminating among the different filarial infections in dogs are needed. The authors report simple and highly specific molecular methods that identify the three most common filarial nematodes of European dogs: Dirofilaria immitis, D. repens and Acanthocheilonema (syn. Dipetalonema) reconditum, based on (1) PCR amplifications of mitochondrial DNA (12S rDNA and coxI) with general filarial primers followed by digestion with restriction enzymes that generates band polymorphisms clearly discriminating the three species and (2) PCR amplifications with species-specific primers to support the restriction analysis, in particular in the case of multiple infections.

Animals↗

A dual anthelmintic treatment strategic scheme for the control of fasciolosis in dairy sheep farms.

A clinical longitudinal field trial was conducted on a dairy sheep farm in southern Italy to assess the effectiveness of a novel anthelmintic treatment strategic scheme against Fasciola hepatica. The scheme utilizes a dual anthelmintic treatment (DAT), i.e., the use of either one of two different anthelmintics on the flock, albendazole sulphoxide (SO) at 1-month intervals and rafoxanide at 2-month intervals, administered to the lactating and non-lactating animals, respectively. The DAT strategic scheme lasted 3 years. In Year 1 and Year 2, shotgun monthly DATs for 5 consecutive months (July, August, September, October, and November) were performed on the flock. In Year 3 there was only one monthly DAT, in July. Overall, the DAT scheme reduced the prevalence of F. hepatica infection by 94.4% (from an average prevalence of 71.1% during the pre-DAT period to an average prevalence of 4.0% during Year 3), and the eggs/gram of faeces (EPG) from 29.3 to 1.3. In conclusion, the DAT strategic scheme reported in the present study successfully reduced both the prevalence and EPGs of F. hepatica to a level at which there were no longer any clinical symptoms of the disease. This scheme did not influence the albendazole SO efficacy against GI nematodes and might be used for the treatment of fasciolosis in dairy sheep farms.

Albendazole↗

Is heartworm disease really spreading in Europe?

Based on recently published surveys and newly acquired data, a study was conducted to verify the distribution of filarial worm (Filarioidea) infections in Europe, with particular emphasis on canine heartworm infection (Dirofilaria immitis). A Geographic Information System based on thermal regimen was constructed as a means to identify areas potentially suitable for heartworm transmission, taking into account that the development of D. immitis larvae in the mosquito does not occur below the threshold temperature of approximately 14 degrees C. Furthermore, a bionomic model of D. immitis in its mosquito vectors, which calculates the moving cumulative heartworm development unit parameter, was applied using the available temperature data to assess the theoretic transmission timing of heartworm in Europe. The results show that the earliest infection risk occurs in Spain on March 21 and the latest risk occurs in Spain on September 11. The longest risk period occurs in Spain (Murcia station: March 21-November 11), and the shortest risk period occurs in northeastern Europe. The study also provides the first risk assessment maps for Europe and suggests that if the actual climatic trend continues, filarial infection should spread into previously infection-free areas.

Animals↗

Heartworm disease in dogs: unusual clinical cases.

Two unusual cases of heartworm infection are described, including one of spontaneous rupture of cardiac chambers, most likely due to the progressive increase in pressure within the right ventricle caused by Dirofilaria immitis infection. Another case report describes persistent left cranial vena cava, which interfered with the surgical removal of heartworm via the left jugular vein.

Animals↗

Toxocara canis infection induces antigen-specific IL-10 and IFNgamma production in pregnant dogs and their puppies.

Toxocara canis (T. canis) is originally a parasite of canine bitches and their pups. The pathogenicity of T. canis infection is enhanced during pregnancy and puppyhood. The aim of this study was to investigate if modification of IFNgamma and IL-10 secretion occurs during infection in pregnant dogs and puppies. Analysis of cytokines secreted could let us hypothesize a role for IL-10 and/or IFNgamma in T. canis infection. We tested T. canis-specific production of IFNgamma and IL-10 by lymphocytes of pregnant dogs and their puppies after in vitro re-exposure to purified excretory/secretory antigen (ESAg) from T. canis. Blood mononuclear cells (BMC) isolated from pregnant dogs and their puppies were cultured in the presence of ESAg. Cultures' supernatants were tested for cytokine levels by ELISA. Results obtained showed that IL-10 concentrations increased during pregnancy in infected animals and in the meantime IFNgamma production decreased. In puppyhood, we observed that, IL-10 concentration decreased with the age of puppies mainly in infected animals while IFNgamma increased. In conclusion, our data suggests that BMC of infected dogs have a particular modification of IL-10 and IFNgamma synthesis. These data could be the basis to design immunotherapeutic approaches.

Animals↗

Expression and function of Toll-like receptor 2 in canine blood phagocytes.

Toll-like receptors (TLRs) are a family of highly conserved pattern recognition receptors (PRR) of mammals that participate in the activation of innate immune responses against microbial infections. Among these receptors, TLR2 is essential for the recognition of conserved structural components of bacteria, protozoa and fungi. Until now, expression of TLR2 in dogs has not been investigated. In this work we describe a partial sequence of the gene coding for canine TLR2 and show that TLR2 mRNA is constitutively expressed in canine blood PMNs. We also show that stimulation of purified PMNs with lipoteichoic acid (LTA), a ligand of TLR2, leads to the release of proinflammatory chemokine IL-8. Furthermore, TLR2 protein is easily detectable by flow cytometry on the canine peripheral blood granulocyte and monocyte cell surface, and slightly on lymphocytes. These findings suggest that, also in dogs as in humans the initial antibacterial response of PMNs could be elicited through engagement of TLR2.

Animals↗

Efficacy of a single milbemycin oxime administration in combination with praziquantel against experimentally induced heartworm (Dirofilaria immitis) infection in cats.

The efficacy of a combination of milbemycin oxime and praziquantel in preventing the establishment of experimentally induced heartworm (Dirofilaria immitis) infection was investigated in a study involving 24 young domestic short-hair cats. The animals were inoculated with 50 infective larvae on day 0. Subsequently they were divided into two groups of 12 animals each. The animals in group 1 were treated once with medicated tablets containing 4 mg milbemycin (minimum dose 2 mg/kg body weight) and 10 mg praziquantel (MILBEMAX) on day 30 after infection. Cats in group 2 received placebo tablets on the same day. On day 183 post-infection a blood sample was taken from each animal before euthanasia and necropsy. The blood samples were tested for the presence of microfilariae and the necropsied animals were examined for the presence of adult worms. Microfilariae were not found in any of the investigated cats. No heartworms were found in the animals in group 1 (treated with medicated tablets). Out of the 12 placebo-treated cats 1 was heartworm-free, whereas all the others were found to be infected with 1-3 adult heartworms.

Animals↗

A novel alpha-Proteobacterium resides in the mitochondria of ovarian cells of the tick Ixodes ricinus.

An intracellular bacterium from Ixodes ricinus ticks collected in Italy was characterized by electron microscopy (EM), PCR sequencing of the 16S rRNA gene, molecular phylogenetic analysis, and in situ hybridization (ISH). This bacterium was shown by EM to be present in the cytoplasm, as well as in the mitochondria of ovarian cells. When universal 16S rRNA bacterial primers were used, PCR amplification of ovarian DNA followed by cloning and sequencing resulted in the same sequence being found in each sample. Phylogenetic analysis of this sequence showed that the bacterium from which it was derived, tentatively designated IricES1, is part of a novel clade in the alpha subdivision of the Proteobacterium: ISH and PCR assays of various tissues performed with oligonucleotides specific for the IricES1 16S rRNA showed that IricES1 is restricted to ovarian cells. Based on the results obtained, we inferred that the bacteria seen by EM in ovarian cells are a single type of bacteria, corresponding to IricES1. PCR screening of 166 ticks from various parts of Italy and one site in England showed that IricES1 was present in 96% of adult females and 44% of nymphs (unsexed). No adult males were found to be infected. Despite the apparent parasitism of host mitochondria by IricES1, the available information suggests that the bacterium has an obligate relationship with its host, although this must be confirmed.

Alphaproteobacteria↗

Prevalence of Bartonella henselae in Italian stray cats: evaluation of serology to assess the risk of transmission of Bartonella to humans.

Bartonella henselae is the major etiological agent of cat scratch disease in humans. Cats act as the natural reservoir of B. henselae and can transmit the infection to humans by a bite or scratch. The prevalence of B. henselae in cat populations was evaluated by serological and bacteriological tests. A total of 769 stray cats from three urban and three rural areas in northern Italy were sampled between January 1999 and December 2000. The positive and the negative predictive values of serological tests with respect to bacteremic status were evaluated. Tests of a total of 140 cats (18%) resulted in detection of bacteremia. A total of 540 cats were tested by serology; 207 (38%) were seropositive. Of the 531 cats tested by both methods, the results for 65 (12.2%) showed both bacteremia detection and seropositivity. The molecular typing of the isolates showed that 20.6% of bacteremic cats were infected with B. henselae type I strain, 61.1% were infected with B. henselae type II, and 18.3% were coinfected with both. A statistically significant difference in antibody and bacteremia prevalences among geographical areas was detected. Statistical analysis showed no association between characteristics such as seroprevalence-bacteremic status, sex, general health status, and the presence of ectoparasites. The negative predictive value of serological test was 84.7%, and the positive predictive value was 31.8%. Receiving operator characteristic analysis of the data showed that serological tests had a low predictive value in relation to the bacteremic status of a cat; in surveys aimed at assessing the real risk of B. henselae infection in a human population, therefore, we suggest the use of blood culture as the reference test. Nevertheless, both blood culture assays and serological tests for Bartonella infection should be performed for a complete evaluation of the health status of cats.

Animals↗

Full season efficacy of moxidectin microsphere sustained release formulation for the prevention of heartworm (Dirofilaria immitis) infection in dogs.

The authors report the efficacy of an injectable, moxidectin sustained release (SR) formulation for the prevention of canine heartworm infection in endemic areas in northern and central Italy. Three field trials were carried out on a total of 324 dogs. Two hundred forty-three dogs were treated with moxidectin SR 6 months apart and 81 dogs (positive controls) with moxidectin tablets given monthly for 5 consecutive months during the risk season each year throughout the study. Results of testing for microfilariae and circulating adult female antigens were negative for all the experimentally treated dogs at the 6, 7, 11 and 19 months after the last injection. No adverse reactions to moxidectin SR were observed but a moderate pain at palpation and swelling (5-6 cm) at the injection site after the first treatment. In the study areas, prevalence of Dirofilaria immitis infection calculated by testing dogs which had no preventive treatment in the previous transmission season ranged from 33 to 63%. This study confirms the efficacy and safety of injectable, moxidectin SR formulation in the prevention of heartworm infection in dogs and demonstrates that the prophylactic efficacy lasts for the full season and strongly suggests that the product gives 1-year protection.

Administration, Oral↗

Geographical information systems and bootstrap aggregation (bagging) of tree-based classifiers for Lyme disease risk prediction in Trentino, Italian Alps.

The risk of exposure to Lyme disease in the province of Trento, Italian Alps, was predicted through the analysis of the distribution of Ixodes ricinus (L.) nymphs infected with Borrelia burgdorferi s.l. with a model based on bootstrap aggregation (bagging) of tree-based classifiers within a geographical information system (GIS). Data on L ricinus density assessed by dragging the vegetation in 438 sites during 1996 were cross-correlated with the digital cartography of a GIS, which included the variables altitude, exposure and slope, substratum, vegetation type and roe deer density. Ticks were more abundant at altitudes below 1,300 m a.s.l., in the presence of limestone and vegetation cover with thermophile deciduous forests and high densities of roe deer. A bootstrap aggregation procedure (bagging) was used to produce a model for the prediction of tick occurrence, the accuracy of which was tested on actual tick counts assessed by a further dragging campaign carried out during 1997 to determine infection prevalence and resulted in average 77%. Other tests of the model were made on additional and independent data sets. The prevalence of infection with Borrelia burgdorferi s.l, determined by polymerase chain reaction on 2,208 nymphs collected by random dragging in 245 transects selected within eight areas where the model predicted the occurrence of I. ricinus during 1997, was 17.5% and was positively correlated to tick abundance and roe deer density. These findings were used to relate the output of the bagged model (probability of tick occurrence) to the density of infected nymphs through a stepwise model selection procedure and thus to produce a GIS digital map of the probability distribution of infected nymphs in the Province of Trento at high resolution scale (50 by 50-m cell resolution). The application of the bagging procedure increased the accuracy of the prediction made by a single classification tree, a well-known classification method for the analysis of epidemiological data.

Animals↗

First detection of spotted fever group rickettsiae in Ixodes ricinus from Italy.

Ixodes ricinus from Italy were examined for the first time to detect whether rickettsiae were present. Using molecular methods, we detected three different spotted fever group rickettsiae, including Rickettsia helvetica. Our results raise the possibility that bacteria other than R. conorii are involved in rickettsial diseases in Italy.

Animals↗

Ultrasonographic features of peritoneal cestodiasis caused by Mesocestoides sp. in a dog and in a cat.

Peritoneal infections caused by Mesocestoides spp. are rare in dogs and cats. Little data exist on the role of abdominal ultrasonography for diagnosis and therapy management of the disease. We describe the ultrasonographic features of peritoneal cestodiasis in a dog and in a cat. In the dog, abdominal ultrasound allowed both a presumptive diagnosis and the collection of tissue samples to confirm peritoneal larval infection. Ultrasound was also very useful for therapy management. In the second patient the ultrasonographic features of tetrathyridial infection in a cat in which the parasite was observed as an incidental finding during ovariohysterectomy are described.

Animals↗