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Biomedical subjects

Domenico Otranto

Publications and source records attributed to Domenico Otranto.

36 records · Page 2Linked to original sources

Frequency, body distribution, and population size of Malassezia species in healthy dogs and in dogs with localized cutaneous lesions.

Malassezia species are commensal organisms of human and animal skin that occasionally act as opportunistic pathogens. The lipid-dependent species are associated with human skin disorders, whereas the non-lipid-dependent species (Malassezia pachydermatis) is considered as an opportunistic secondary pathogen affecting the canine skin surface and ear canal. This study evaluated the relationship between Malassezia yeasts, their population size, and the occurrence of skin lesions from healthy and skin-diseased dogs. The efficiency of cytological examination and fungal culture for Malassezia detection was also evaluated. From March 2002 to July 2003, 33 healthy dogs and 54 dogs with pruritic localized skin diseases were examined; skin swabs (1218) were collected from 7 anatomical sites for culture and cytological examination. Malassezia prevalence according to anatomical site and the agreement between cytological results and fungal cultures were statistically analyzed. Differences in mean colony forming unit counts between positive healthy and diseased dogs were evaluated using the Bonferroni test for post hoc pair-wise comparisons. In healthy dogs, Malassezia yeasts were most frequently isolated in the perianal and perioral areas. The frequency of isolation and population size of Malassezia species were higher in dogs with localized dermatitis, especially in affected areas, indicating a role for Malassezia in the occurrence of skin lesions. Malassezia pachydermatis was the species most commonly cultured from the skin and external ear canal of healthy and diseased dogs; isolation of lipid-dependent yeasts from healthy dogs was less frequent. Using fungal culture as the gold standard, cytological examination showed good relative specificity (95%) but very low relative sensitivity (30%).

Animals↗

Cattle grub infestation by Hypoderma sp. in Albania and risks for European countries.

In order to estimate the presence and the seroprevalence of hypodermosis in Albania, 625 head of cattle were bled during two sampling seasons (i.e. from February to March 2003 and from November to December 2003). The cattle came from three collection sites in Northern (site A), Central (site B) and Southern (site C) Albania. Milk samples were collected monthly from four animals from October 2002 to May 2003 during the lactating period. The animals were also clinically examined for the presence of warbles by manual palpation from April to July 2003 and 2004 and third instar larvae were collected and morphologically identified. Serum and milk samples were processed by ELISA. One hundred and thirty-three (38.6%) out of 344 and 116 (41.3%) out of 281 animals were found to be seropositive for Hypoderma during the first and the second sampling season. In particular, the animals from site C presented the highest percentage of seropositive results (i.e. 72.8% and 97.8% in the first and in second year, respectively) followed by the animals from sites A (i.e. 35.8% and 23.8% in the first and in second year, respectively) and B (i.e. 17.8% and 3.4% in the first and in second year, respectively). The kinetics of anti-Hypoderma antibodies in milk samples showed the highest antibody titres from October to February 2003. All the seropositive animals in both the sampling periods showed the presence of one or more warbles under the skin during April and May 2003 and 2004 and the third-stage larvae collected were morphologically identified as Hypoderma bovis. The results of this survey indicate that hypodermosis is widespread in Albania and that early prophylactic treatments must be carried out accordingly. The hope is not only to reduce the parasitic intensity with obvious benefits for livestock production, but also to avoid the risk of spreading this parasitic disease to neighboring countries.

Albania↗

Report in Europe of nasal myiasis by Rhinoestrus spp. in horses and donkeys: seasonal patterns and taxonomical considerations.

Two species of Rhinoestrus (i.e. Rhinoestrus purpureus (Brauer) and Rhinoestrus usbekistanicus Gan) cause nasal myiasis in horses, donkeys and zebras. In the past 15 years myiasis caused by R. purpureus has been reported in Egypt and by R. usbekistanicus in Senegal and Niger, both in horses and in donkeys. With the aim to investigate the presence of this myiasis in autochthonous horses and donkeys from southern Italy and to study the seasonal trend of larval infection, 212 native horses were necropsied in two slaughterhouses in the Apulia region (site A) from January to November 2003, and 120 native horses and two donkeys in one slaughterhouse in Sicily (site B) from January to October 2003. Thirteen of 212 and 5 of 120 horses examined from sites A and B, respectively, were infected by nasal bot fly larvae. Both donkeys examined were positive. Two hundred and thirteen larvae, representing all stages, were collected from the throat region, the turbinates and beyond the lamina cribrosa of the ethmoid bone, in the cerebral cavity. Third larval stages were retrieved from April to September with the highest mean burden in site A in May and in July in site B. The simultaneous presence all three larval stages at site B suggests the existence of two or more overlapping generations. Larvae were identified on the basis of peritreme structures and arrangement of the spines on the dorsal surface of the third segment. Some L3 collected from sites A and B presented morphological characteristics of both R. usbekistanicus and R. purpureus. The presence of mixed features on the L3 of Rhinoestrus collected from the same animals may be explained by the existence of a single species of Rhinoestrus presenting different morphotypes or of two species of Rhinoestrus which are both present in southern Italy since this area is midway between eastern and African countries where these two species have been reported.

Age Factors↗

Specific identification of Habronema microstoma and Habronema muscae (Spirurida, Habronematidae) by PCR using markers in ribosomal DNA.

Gastric or cutaneous habronemosis caused by Habronema microstoma Creplin, 1849 and Habronema muscae Carter, 1865 is a parasitic disease of equids transmitted by muscid flies. There is a paucity of information on the epidemiology of this disease, which is mainly due to limitations with diagnosis in the live animal and with the identification of the parasites in the intermediate hosts. To overcome such limitations, a molecular approach, based on the use of genetic markers in the second internal transcribed spacer (ITS-2) of ribosomal DNA, was established for the two species of Habronema. Characterisation of the ITS-2 revealed sequence lengths and G+C contents of 296 bp and 29.5% for H. microstoma, and of 334 bp and 35.9% for H. muscae, respectively. Exploiting the sequence difference (approximately 40%) between the two species of nematode, primers were designed and tested by the polymerase chain reaction (PCR) for their specificity using a panel of control DNA samples from common equid endoparasites, and from host tissues, faeces or muscid flies. Effective amplification from each of the two species of Habronema was achieved from as little as 10 pg of genomic DNA. Hence, this molecular approach allows the specific identification and differentiation of the DNA from H. microstoma and H. muscae, and could thus provide a molecular tool for the specific detection of Habronema DNA (irrespective of developmental stage) from faeces, skin and muscid fly samples. The establishment of this tool has important implications for the specific diagnosis of clinical cases of gastric and cutaneous habronemosis in equids, and for studying the ecology and epidemiology of the two species of Habronema.

Animals↗

Otodectes cynotis (Acari: Psoroptidae): examination of survival off-the-host under natural and laboratory conditions.

The biological and environmental factors affecting survival off-the-host of Otodectes cynotis (Acari: Psoroptidae) ear mites were investigated under natural and laboratory conditions. From November 2000 to November 2002 mites were collected monthly from cats and divided into four groups according to sex and stage. In laboratory conditions, the mites were placed in an incubator with a steady 95% relative humidity (r.h.), a 10 degrees C. All the plates were examined by stereomicroscopy every 24 h until all the mites had died. The data were analysed statistically by multiple linear regression and survival analysis. At 10 degrees C, the maximum survival time of mites was between 15 and 17 days, while at 34 degrees C, it was between 5 and 6 days. The maximum survival time of adult females was significantly longer than that of other stages. No differences were observed in maximum survival times of mites that had been offered food and those that had not, or in the time (in days) to reach 50% mortality (LT50). When exposed to environmental conditions, the maximum survival time (12 days) was observed at temperatures ranging from 12.3 to 14.2 degrees C and r.h.s between 57.6 and 82.9%. Multiple regression analysis showed that temperature alone influenced the maximum survival time and LT50 of mites, and that the rate of survival declined linearly with increasing mean temperature. This basic understanding of off-host survival suggests that, places which have been inhabited by infected animals may need to be disinfected or remain vacated for at least 12 days before occupancy by clean cats or dogs.

Animals↗

Genotyping of Cryptosporidium isolates from Chamelea gallina clams in Italy.

Chamelea gallina clams collected from the mouths of rivers along the Adriatic Sea (central Italy) were found to harbor Cryptosporidium parvum (genotype 2), which is the lineage involved in zoonotic transmission. The clams were collected from the mouths of rivers near whose banks ruminants are brought to graze. This paper reports the environmental spread of C. parvum in Italy and highlights the fact that genotyping of seaborne Cryptosporidium isolates is a powerful tool with which to investigate the transmission patterns and epidemiology of this microorganism.

Animals↗

Molecular characterization of the first internal transcribed spacer of ribosomal DNA of the most common species of eyeworms (Thelazioidea: Thelazia).

Spirurid nematodes of the genus Thelazia cause ocular infection in several mammals and are transmitted by dipteran flies. Of the 10 described species, T. gulosa, T. rhodesi, T. skrjabini (affecting cattle), T. lacrymalis (affecting horses), and T. callipaeda (affecting carnivores and humans) are the most common. The aim of this study was to characterize the first internal transcribed spacer (ITS1) ribosomal DNA sequences of these 5 species. The length of the ITS1 sequences ranged from 357 bp (T. lacrymalis) to 905 bp (T. callipaeda). Interspecific differences ranged from 35 to 77%. Polymorphic sites were detected in each species. Intraspecific variation varied from 0.3 to 2.5%. The characteristics of the ITS1 of Thelazia spp. show similarities to those of other Spirurida. The findings of this present study show that the ITS1 represent a powerful genetic marker for the molecular identification of eyeworms and a useful tool for developing molecular epidemiological techniques to study Thelazia spp. transmission patterns and prevalence in definitive and intermediate hosts.

Animals↗

Seroprevalence and associated risk factors of neosporosis in beef and dairy cattle in Italy.

A cross-sectional serological survey for Neospora caninum was carried out on beef and dairy cattle in southern and northern Italy. A total of 111 herds and 1140 animals were tested using an ELISA assay (CHEKIT-Neospora) to detect anti-N. caninum antibodies. Management and individual data were collected and analysed both by linear and logistic multiple-regression models in order to find good predictors of the cattle seroprevalence and anti-N. caninum antibody level. At least one seropositive animal was found in 49 herds (44.1%), of which 31 (39.7%) from southern Italy and 18 (54.5%) from northern Italy. A total of 126 head of cattle (11%) were found to be seropositive and the seroprevalence was lower in southern (8.7%) than in northern Italy (16%). One of the best predictors of neosporosis seroprevalence in this study was the practice of self-rearing replacement heifers. Further risk factors were linked to higher stocking density, i.e. animals farmed in large herds and with no summer or permanent grazing practices were more likely to be seropositive than others. Farms with two or more dogs had higher herd seropositivity than farms with one or no dogs and this factor interacted significantly with the farm size and presence of poultry. Among individual characteristics, seropositivity was higher in animals sampled in mid- or late-pregnancy compared to animals either in early pregnancy or not pregnant. There was a significant interaction between the factors for pregnancy status and grazing practices. None of the epidemiological data recorded was a good predictor of the anti-N. caninum antibody level.

Animal Husbandry↗

Efficacy of injectable and pour-on microdose ivermectin in the treatment of goat warble fly infestation by Przhevalskiana silenus (Diptera, Oestridae).

The prophylactic efficacy of microdoses of injectable and pour-on ivermectin formulations against larval stages of Przhevalskiana silenus was assessed in naturally infected goats in the region of Calabria (southern Italy).Sixty-eight goats from two goat farms were divided into five groups: one group remained untreated, while the other four groups were treated with microdoses of ivermectin (5 and 10 microg/kg injectable formulation and 10 and 20 microg/kg pour-on formulation). The microdoses of ivermectin were fully effective in the treatment of goat warble fly infestation (GWFI) as no larvae emerged from the warbles in the treated groups, while all the larvae emerged in the control groups. Irrespective of the type of formulation used, the difference between the treated groups and the control group was statistically significant (P< 0.001). By contrast, no statistical differences were found between the goats treated with the injectable formulation and those receiving the pour-on applications, and between the two doses of the injectable and pour-on formulations used. Given the plasma concentrations it attains at its lowest dose (0.052 - 0.042 ng/ml for the injectable formulation and 0.030 ng/ml for the pour-on) the injectable formulation seems to offer the most reliable route for the administration of ivermectin microdoses and it is acceptable for milk consumption. The introduction of ivermectin in the early eighties and the use of microdoses in some cases have made it possible to control cattle hypodermosis in large areas of Europe. As with cattle hypodermosis, the administration of ivermectin microdoses in goats is particularly interesting because of the low costs involved and the low levels of residues found in goat milk; it may thus constitute the basis for GWFI control campaigns in areas where the disease is prevalent.

Administration, Topical↗

Current status and epidemiological observation of Thelazia callipaeda (Spirurida, Thelaziidae) in dogs, cats and foxes in Italy: a "coincidence" or a parasitic disease of the Old Continent?

Thelazia callipaeda is a spirurid nematode which causes ocular infections in dogs and man and, occasionally, in cats, foxes and rabbits. The intermediate host and vector of T. callipaeda is unknown. For a long time T. callipaeda incidence was reported only from the Russian Federation and the Far East, but recently it has also been found in Italy. In order to investigate the spread of T. callipaeda in Italy, a survey was carried out in two sites, site A in the Piedmont region (North West Italy), and site B in the Basilicata region (Southern Italy). Dogs, cats and foxes in site A and dogs in site B were examined for eyeworms, using different procedures and timing. From January 1995 to August 2002, 91 dogs, 4 cats and 903 fox carcasses were examined in site A, and from October 1999 to January 2003, 443 dogs were examined in site B, and the eyeworms collected were identified using morphological keys. Twenty-one (23.07%) and 185 (41.76%) of the dogs from sites A and B, respectively, were found to be infected by eyeworms; furthermore, all the cats examined and 46 fox carcasses (5.1%) were positive for eyeworms. All the nematodes collected were identified as T. callipaeda. These results indicate that T. callipaeda is not confined to Eastern Europe and Asia, but that it has spread to the Old Continent, and to both Northern and Southern Italy. Considering the high prevalence of infected dogs reported in some municipalities (e.g. 60.14% of 138 dogs examined in a municipality from site B), it is assumed that one or more vectors are significantly present in the areas under investigation. Furthermore, there is good reason to believe that T. callipaeda is also present in other European countries. Speculation as to the origins of this parasitic infestation in Europe and the biology of T. callipaeda and its vector/s is also discussed.

Animals↗

Molecular differentiation of Hypoderma bovis and Hypoderma lineatum (Diptera, Oestridae) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

The most variable region of the cytochrome oxidase I (COI) gene of Hypoderma bovis(1) and Hypoderma lineatum(2) (Diptera, Oestridae) was amplified by PCR and the amplicons were sequenced and analysed. PCR products were digested with three restriction enzymes, namely BfaI, HinfI and TaqI, providing informative profiles. H. bovis and H. lineatum sequences revealed an inter-specific variation rate of 8.5%, and an intra-specific variation rate of 0.87 and 0.29%, respectively. The results showed that the COI gene region examined was useful for the differentiation of H. bovis and H. lineatum and that a PCR-RFLP assay is a practical tool for their identification, offering additional diagnostic and epidemiological instruments for the study of cattle grub infestation.

Animals↗

Assessment of cattle grub (Hypoderma spp.) prevalence in northeastern Italy: an immunoepidemiological survey on bulk milk samples using ELISA.

Hypodermosis is a myiasis of cattle mainly found in the Northern Hemisphere. In Italy it has been reported primarily in southern regions with a farm prevalence reaching a peak of 85%, while few data are available in the north of the country. This paper reports the results of a serological survey carried out on hypodermosis in Vicenza province (northeastern Italy). During January and February 1999, bulk milk samples were collected from 224 dairy farms and tested, using an hypodermosis ELISA kit. Epidemiological data, such as farm location, herd size, number of cows, grazing management, treatments against Hypoderma larvae, presence of imported animals, were gathered and statistically analysed in relation to the serological results. In 15 farms the presence of warbles was monitored by manual palpation and larvae collected for identification.Ninety-seven (43.3%) samples were detected as positive, whereas 12 (5.4%) and 115 (51.3%) were inconclusive and negative, respectively. Thirteen farms were positive upon parasitological examination and the larvae collected were identified as Hypoderma bovis.Seroprevalence was highly correlated with the practice of free grazing and slightly, but significantly, correlated with the number of cows on the farm. The mean antibody level of positive herds was significantly higher on farms without imported animals. On farms with warbled animals there was a high inverse correlation between antibody level and quantity of milk in the tanks and the number of lactating cows. The ELISA test on bulk milk samples has been shown, under field conditions, to be a useful, cost-effective and easy test for the diagnosis of bovine hypodermosis.

Animal Husbandry↗

Molecular detection of Anaplasma platys in dogs using polymerase chain reaction and reverse line blot hybridization.

Several polymerase chain reactions (PCRs) and a reverse line blot hybridization (RLB) method were used to identify Anaplasma platys in dogs held in a kennel in Italy. Whereas PCR techniques confirmed the presence of A. platys, the RLB method not only correlated the results obtained by PCR but also ruled out the presence of other species such as Ehrlichia canis or E. chaffeensis. There was no correlation between infection status and age or breed of the dogs. Polymerase chain reaction performed on the Rhipicephalus sanguineus ticks collected from those dogs showed that they were also infected with A. platys. Sequences obtained from some samples and compared with those within the GenBank also confirmed the presence of A. platys.

Anaplasma↗

Molecular approaches to the study of myiasis-causing larvae.

Among arthropod diseases affecting animals, larval infections - myiases - of domestic and wild animals have been considered important since ancient times. Besides the significant economic losses to livestock worldwide, myiasis-causing larvae have attracted the attention of scientists because some parasitise humans and are of interest in forensic entomology. In the past two decades, the biology, epidemiology, immunology, immunodiagnosis and control methods of myiasis-causing larvae have been focused on and more recently the number of molecular studies have also begun to increase. The 'new technologies' (i.e. molecular biology) are being used to study taxonomy, phylogenesis, molecular identification, diagnosis (recombinant antigens) and vaccination strategies. In particular, more in depth molecular studies have now been performed on Sarcophagidae, Calliphoridae and flies of the Oestridae sister group. This review discusses the most topical issues and recent studies on myiasis-causing larvae using molecular approaches. In the first part, PCR-based techniques and the genes that have already been analysed, or are potentially useful for the molecular phylogenesis and identification of myiasis-causing larvae, are described. The second section deals with the more recent advances concerning taxonomy, phylogenetics, population studies, molecular identification, diagnosis and vaccination.

Animals↗

Epidemiology of bovine tick-borne diseases in southern Italy.

This investigation was carried out in an area covering part of three southern Italian regions: Campania, Basilicata and Apulia. Eighty-one farms were involved using the formula suggested by Thrusfield; they were equally distributed over the area which was subdivided into 81 geo-referenced sub-areas. In May and June 1999 from a total of 506 cattle, older than 18 months, blood-samples were taken and ticks were collected and identified. Serum samples were tested for antibodies of Bahesia bigemina, Babesia bovis and Anaplasma marginale with an ELISA technique. Eight farms (9.8%) out of the 81 examined were positive for B. bigemina only, 3 (3.7%) for A. marginale only, and 70 (86.4%) for both. None of the animals of any farm was found to be positive for B. bovis. Out of the 506 sera tested, 117 (23.1 %) were positive for B. bigemina only, 58 (11.5%) forA. marginale only and 250 (49.4%) for both species; 81 (16.0%) were negative for all of them. Ticks were collected on animals on 62 (76.5%) out of the 81 farms. Adult ticks (1 410) were collected and identified; the highest number belonged to the Rhipicephalus bursa species (65.5%), followed by Rhipicephalus turanicus (8.6) and Haemaphysalis punctata (8.4). The results showed that B. bigemina, A. marginale and their potential vectors are common in the area examined and indicated that there is a risk for animals imported from tick-borne disease-free areas.

Anaplasma↗