Emerging Hyalomma lusitanicum: From identification to vectorial role and integrated control.

In the Mediterranean basin, the tick species Hyalomma lusitanicum Koch stands out among other species of the Hyalomma genus due to its wide distribution, and there is great concern about its potential role as a vector and/or reservoir and its continuous expansion to new areas because of climate warming and human and other animal movements. This review aims to consolidate all the information on H. lusitanicum, including taxonomy and evolution, morphological and molecular identification, life cycle, sampling methods, rearing under laboratory conditions, ecology, hosts, geographical distribution, seasonality, vector role and control methods. The availability of adequate data is extremely relevant to the development of appropriate control strategies in areas where this tick is currently distributed as well as in new areas where it could become established in the near future.

Are horse age and incision length associated with surgical site infection following equine colic surgery?

It is our clinical impression that age and incision length are more strongly associated with surgical site infection (SSI) following colic surgery than skin closure or wound protection method. Therefore, the objective of this observational clinical cohort study was to identify the risks for SSI in horses undergoing colic surgery. Data collection included pre-, intra-, and postoperative variables. Variables with P < 0.2 following univariable analysis were used in a logistic regression multivariable model. Variables with P <0.05 were included in the final model. Odds ratios (OR; 95% confidence intervals, 95% CI) were determined. The area under the curve (AUC) for the receiver-operator characteristic was calculated. The final multivariable model included breed (P = 0.008), incision length (P = 0.004), surgical procedure classification (P <0.001), and postoperative (PO) colic (P = 0.037; overall model P <0.001, and AUC was 0.81 [excellent discrimination between SSI vs. no SSI]). Warmbloods (OR 12.0; 95% CI 2.7-74.8), American breeds (OR 6.4; 95% CI 1.2-43.0), and Thoroughbreds (4.5; 95% CI 1.1-25.5) more commonly had SSI than other breeds (ponies/miniature horses, Draft breeds, Standardbreds, Arabians, and Crossbreeds [referent]). A higher SSI rate was associated with incision lengths >27 cm (3.7; 95% CI 1.5-9.9), heavily contaminated procedures (12.0; 95% CI 3.3-49.9), and horses with PO colic (2.7; 95% CI 1.1-6.8). SSI appeared to be more common after heavily contaminated procedures and in horses with PO colic, which probably resulted in more incisional contamination and trauma. Some breeds appeared to have higher odds of SSI. Age was not associated with SSI. The risk of developing SSI was higher for horses with an incision >27 cm; therefore, surgeons are encouraged to use the minimum incision length required to accomplish the necessary abdominal exploration and bowel manipulation in the safest manner possible.

Detection of Rickettsia aeschlimannii and Rickettsia africae in ixodid ticks from Burkina Faso and Somali Region of Ethiopia by new real-time PCR assays.

In the framework of cooperation for development projects in Burkina Faso and Ethiopia, we collected ixodid ticks from cattle, small ruminants and camels. We optimized new TaqMan Probe real-time PCR assays to detect Rickettsia aeschlimannii and Rickettsia africae OmpA gene in the collected samples. Rickettsia africae was identified in 75.0% Amblyomma variegatum (95%CI: 56.6-88.5), while R. aeschlimannii in 24.0% Hyalomma truncatum (95%CI: 9.4-45.1) and 50.0% H. rufipes (95%CI: 29.9-70.0) collected from cattle in different provinces throughout Burkina Faso. Ticks from the Libaan zone, Somali Region of Ethiopia, were also infected by R. africae (28.5% prevalence in Amblyomma gemma, 95%CI: 14.7-46.0) and R. aeschlimannii (27.0% H. truncatum, 95%CI: 5.0-62.9; 88.3% H. rufipes, 95%CI: 60.5-99.3). All tested ticks were adults. The developed diagnostic tools were highly sensitive and enabled us to rapidly classify R. aeschlimannii and R. africae, which were identified in Burkina Faso and in the Somali Region of Ethiopia for the first time. Further studies are needed to assess the zoonotic risk and prevalence of infection in local human populations, who have high contact rates with ticks and their animal hosts.

Detection of Invasive Borrelia burgdorferi Strains in North-Eastern Piedmont, Italy.

Following reports of human cases of Lyme borreliosis from the Ossola Valley, a mountainous area of Piemonte, north-western Italy, the abundance and altitudinal distribution of ticks, and infection of these vectors with Borrelia burgdorferi sensu lato were evaluated. A total of 1662 host-seeking Ixodes ricinus were collected by dragging from April to September 2011 at locations between 400 and 1450 m above sea level. Additional 104 I. ricinus were collected from 35 hunted wild animals (4 chamois, 8 roe deer, 23 red deer). Tick density, expressed as the number of ticks per 100 m(2), resulted highly variable among different areas, ranging from 0 to 105 larvae and from 0 to 22 nymphs. A sample of 352 ticks (327 from dragging and 25 from wild animals) was screened by a PCR assay targeting a fragment of the 16S rRNA gene of B. burgdorferi s.l. Positive samples were confirmed with a PCR assay specific for the 5S-23S rRNA intergenic spacer region and sequenced. Four genospecies were found: B. afzelii (prevalence 4.0%), B. lusitaniae (4.0%), B. garinii (1.5%) and B. valaisiana (0.3%). Phylogenetic analysis based on the ospC gene showed that most of the Borrelia strains from pathogenic genospecies had the potential for human infection and for invasion of secondary body sites.

Wild ungulates as sentinel of BTV-8 infection in piedmont areas.

Bluetongue caused by the genotype 8 virus (BTV-8) appeared for the first time in BTV free areas in northern Italy in 2008. The presence of domestic animals outbreaks, abundant wild ungulates populations, and ongoing regional BTV control plans, made this area interesting to evaluate the role of wild ruminants in BTV-8 epidemiology. We analyzed spleen samples from hunted red deer (Cervus elaphus), roe deer (Capreolus capreolus) and Alpine chamois (Rupicapra rupicapra) by quantitative RT-PCR. Samples were collected from 2008 to 2011 in two provinces of Piedmont region. BTV-8 was detected in all ungulate species, confirming their receptivity to the infection. However, the viral load in the positive specimens was low, and decreased from 2008 to 2011. These results, together with the extinction of the epidemic following a regional livestock vaccination campaign, lead to hypothesize that wild ungulates were an epiphenomenon and they had not an important role in the domestic transmission cycle of BTV-8 in this area. In spite of this, wild ruminants appear to be good sentinels of BTV circulation and their monitoring could be useful for surveillance in piedmont areas.

Lyme borreliosis spirochetes and spotted fever group rickettsiae in ixodid ticks from Pianosa island, Tuscany Archipelago, Italy.

A study on tick fauna and tick-borne pathogens was undertaken in Pianosa, an island in the Tuscany Archipelago that constitutes an important stopping and nesting point for migratory birds. Ticks were removed from feral cats and a few terrestrial birds, and host-seeking ticks were collected by dragging. A total of 89 ticks were found on animals: 57 Ixodes ventalloi Gil Collado, 1936 and 32 Ixodes acuminatus Neumann, 1901. Host-seeking ticks were 354 Hyalomma spp. larvae and 18 Hyalomma spp. adults, identified as Hyalomma marginatum C.L. Koch, 1844 (n=11) and 7 Hyalomma detritum Schulze, 1919 (n=7). A sample of adult ticks was subjected to molecular analyses to look for Rickettsia spp. and Borrelia burgdorferi sensu lato (s.l.). Sequence analysis of the 5S-23S intergenic spacer region and OspA gene of B. burgdorferi s.l.-positive samples showed the presence of Borrelia spielmanii (n=3; 3.7%, 95% confidence interval [CI] 0.08-10.4) and Borrelia valaisiana (n=13; 13.6%, 95% CI 7.0-23.0) in Ixodes ticks from cats and terrestrial birds. Ixodes spp. were also infected by Rickettsia helvetica (n=19; 23.4%, 95% CI 14.7-34.2). Finally, we detected Rickettsia aeschlimannii in 3 out of 12 host-seeking Hyalomma spp. adults tested (25%, 95% CI 5.5-57.2). Our study shows the presence of several tick-borne pathogens in Pianosa. Hyalomma spp. and Ixodes ticks other than I. ricinus seem to be involved in their epidemiological cycle, and birds could contribute to the pathogen dispersal along their migration routes. This is the first finding of B. spielmanii in Italy. We hypothesize the involvement of peridomestic rodents or hedgehogs in its maintenance in Pianosa.

Prevalence of Campylobacter jejuni, Campylobacter coli and enteric Helicobacter in domestic and free living birds in North-Western Italy.

In order to investigate the prevalence of some thermophilic Campylobacter (C. jejuni and C. coli) and enteric Helicobacter (H. pullorum and H. canadensis) in domestic and wild birds, a total of 278 bird caecal samples were analyzed over a 2 year period in North-Western Italy. Samples were collected from poultry raised in intensive farming at the slaughterhouse (n=102, group A) and in small scale rural farms (n=60, group B) as well as from wild birds (n=116, group C). PCR amplifications were carried out on DNA extracted from caecal samples. Molecular assays targeted the hipO gene for C. jejuni, the asp gene for C. coli and the 16S rRNA gene of H. pullorum/H. canadensis. To differentiate H. pullorum from H. canadensis, PCR products were subjected to an ApaLI digestion assay. Prevalence of thermophilic Campylobacter and enteric Helicobacter was significantly different among groups (p<0.0001). Campylobacter infections were detected in all three bird groups (78.4% group A, 18.3% group B and 38.8% group C, respectively), Helicobacter infections were only detected in poultry, with H. pullorum infecting 68.6% of group A and 21.7% of group B birds. H. canadensis was detected in Guinea fowls (group A) and for the first time in pheasants (group B). Mixed infections by enteric Campylobacter and Helicobacter were shown in 53.9% of group A and in 5.0 % of group B. Our results show that both microorganisms commonly infect poultry, especially intensive farming animals. Only hooded crows among the wild bird group (group C), proved to be highly sensitive to Campylobacter infection.

Detection of "Candidatus Rickettsia sp. strain Argentina"and Rickettsia bellii in Amblyomma ticks (Acari: Ixodidae) from Northern Argentina.

Ixodid ticks were collected from vegetation and from humans, wild and domestic mammals in a rural area in the semi-arid Argentine Chaco in late spring 2006 to evaluate their potential role as vectors of Spotted Fever Group (SFG) rickettsiae. A total of 233 adult ticks, identified as Amblyomma parvum, Amblyomma tigrinum and Amblyomma pseudoconcolor, was examined for Rickettsia spp. We identified an SFG rickettsia of unknown pathogenicity, "Candidatus Rickettsia sp. strain Argentina", in A. parvum and A. pseudoconcolor by PCR assays targeting gltA, ompA, ompB and 17-kDa outer membrane antigen rickettsial genes. Rickettsia bellii was detected in a host-seeking male of A. tigrinum. Amblyomma parvum is widespread in the study area and is a potential threat to human health.

Observations on Antricola ticks: small nymphs feed on mammalian hosts and have a salivary gland structure similar to ixodid ticks.

Ticks use bloodmeals as a source of nutrients and energy to molt and survive until the next meal and to oviposit, in the case of females. However, only the larvae of some tick species are known to feed upon bats; females are obligatorily autogenous, and nymphal stages are believed to not feed. We investigated the presence of blood in a natural population of nymphal Antricola delacruzi ticks collected from bat guano; their ability to feed upon laboratory hosts; and the microscopic structure of both salivary glands and gut. DNA amplification of gut contents of freshly collected material was positive for a mammal in 4 of 11 first instar nymphs, but we were unsuccessful in the amplification of host bloodmeal DNA from late instar nymphs. All early nymphal stages (n = 10) fed on rabbits, and host DNA was detected and sequenced from gut contents. However, all the large nymphs (n = 10) rejected feeding, and host DNA remained undetected in these ticks. All stages of A. delacruzi have salivary glands similar in morphology to the ixodid agranular Type I salivary gland acini and to granular Type II or Type B acini. All stages of A. delacruzi had a similar gut structure, consisting of digestive cells in the basal portion that contained hematin granules. Neither regenerative nor secretory cell traces were observed in the sections of gut.

Detection of Babesia caballi in Amblyomma variegatum ticks (Acari: Ixodidae) collected from cattle in the Republic of Guinea.

A reverse line blot hybridisation (RLB) assay was applied to screen Amblyomma variegatum adult ticks (n = 504) collected from N'Dama cattle in the Republic of Guinea. In a PCR, the V1 hypervariable region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for species of the genera Anaplasma and Ehrlichia, and the V4 hypervariable region of the 18S rRNA gene was amplified with primers specific for members of the genera Theileria and Babesia. Amplified PCR products from A. variegatum ticks were hybridised onto a membrane, to which oligonucleotide probes species-specific for Ehrlichia/Anaplasma and Theileria/Babesia parasites were covalently linked. No pathogens belonging to Ehrlichia/Anaplasma species were found, while 10 DNA samples resulted positive for Babesia caballi and 5 samples for Theileria velifera. This is the first report of B. caballi in A. variegatum ticks. One of the B. caballi positive samples was sequenced. This new strain (BcabGuinea) showed a 97% similarity to the Z15104 B. caballi GenBank sequence.

Detection of beak and feather disease virus (BFDV) and avian polyomavirus (APV) DNA in psittacine birds in Italy.

Beak and feather disease (psittacine circovirus) and Budgerigar fledgling disease (avian polyomavirus) are viral diseases that can frequently affect captive psittacine birds. We designed the first survey to investigate the presence of beak and feather disease virus (BFDV) and Avian polyomavirus (APV) inside the population of captive psittacine birds in Italy. Samples were collected in 18 Italian psittacine breeding centres and four trade centres over a 4-year period. A total of 1516 birds were tested for BFDV and 877 birds were tested for APV by means of a polymerase-chain-reaction (PCR) assay. BFDV was found in 122 (8.05%) and APV in 7 (0.79%) birds. No significant difference in infection rate was found between imported and locally raised parrots. We report the first BFDV DNA isolation in wild birds imported to Italy from Papua New Guinea.