Anaplasma Species in Ticks Infesting Mammals of Sardinia, Italy.

Ticks are hematophagous ectoparasites that are recognized for their ability to vector a wide variety of pathogens of viral, bacterial, protozoal, and helminthic nature to vertebrate hosts. Among the different diseases transmitted by ticks, also called "Tick-Borne Diseases" (TBD), many are zoonotic. Pathogens of the genus Anaplasma refer to obligate intracellular bacteria within the Rickettsiales order transmitted mainly through tick bites and considered as well-established threats to domestic animals, livestock, and humans, worldwide. In this retrospective study, 156 ticks collected from twenty goats, one marten, and one cattle from several Sardinian sites, were examined by molecular analyses to detect the presence of Anaplasma species. A total of 10 (10/156; 6.4%) ticks were shown to be Anaplasma-positive by PCR screening. After sequence analyses, A. phagocytophilum was detected in four Rhipicephalus sanguineus s.l. (3.3%) and four Rh. bursa (11%) ticks from goats, while one Rh. sanguineus s.l. (0.8%) and one Rh. bursa (2.8%) collected from the marten and cattle, respectively, exhibited 100% of identity with A. marginale strains. In this study, we provide the first description and molecular detection of A. marginale and A. phagocytophilum in ticks of the Rhiphicephalus genus in Sardinia. Considering the growing impact of tick-borne Anaplasma pathogens on human health, further studies are necessary to monitor the prevalence of these pathogens in Sardinia.

Molecular Investigation of Theileria and Babesia Species in Domestic Mammals from Sardinia, Italy.

Piroplasmoses are tick-borne diseases caused by hemoprotozoan parasites of veterinary and public health significance. This study focuses on the molecular identification and characterization of species belonging to the Theileria/Babesia genera in 152 blood samples, collected from 80 horses and 72 cattle from several farms in Sardinia, by targeting the 18S rRNA gene. The PCR results highlighted that 72% of the samples were positive for Theileria/Babesia spp., with a rate of infection of 68% and 75% for the horses and cattle, respectively. Sequencing and the BLASTn analysis showed that the 18S rRNA generated in this study has 99-100% homology with the B. bigemina, T. orientalis/sergenti/buffeli, T. equi and T. annulata strains isolated from different hosts worldwide. These findings improve the knowledge on Babesia and Theileria infections in domestic mammals and confirm the significant prevalence of piroplasmosis among subclinical and carrier animals throughout the island. Furthermore, the presence of T. annulata, reported for the first time in the study area, expands the repertoire of pathogens already detected in Sardinia. Our results gather updates on the diversity and distribution of piroplasms in Sardinia and suggest the need to develop procedures to improve animal and public health safety.

Molecular detection of Theileria sergentii/orientalis/buffeli and Ehrlichia canis from aborted ovine and caprine products in Sardinia, Italy.

The economic impact and losses caused by abortion of small ruminants represent an important threat to livestock industry worldwide. Infectious agents are the most commonly causes of small ruminant abortion and many of which pose a serious threat to human health. The management of abortion outbreaks is essential to understand the transmission, prevention and control of the zoonotic diseases. This study aimed to increase the knowledge about the common known zoonotic pathogens causing abortion (Coxiella burnetii, Chlamydia abortus and Toxoplasma gondii) circulating in Sardinia. In addition, the occurrence of other infectious agents that, until now, had never been identified in abortion samples and which might be cocirculating during the abortion outbreaks were also considered. In this study, 125 abortion samples collected from 91 small ruminant farms were screened for the presence of Babesia/Theileria spp., Ehrlichia canis, Anaplasma spp., Chlamydia spp., C. burnetii and T. gondii by PCR analyses and sequencing. This is the first evidence on the presence of Theileria sergenti/orientalis/buffeli group and Eh. canis in 22 (22/125; 18%) and 26 (26/125; 21%) abortion products from small ruminants, respectively. Chlamydia abortus, C. burnetii and T. gondii were also detected in brain, liver, spleen and placentae at 46% (58/125), 34% (42/125) and 2% (2/125), respectively. This study highlights that pathogens with epizootic and zoonotic potential are circulating in the island and could be involved directly or in association with other pathogens as possible cause of ruminant abortion. Further studies are needed to fully assess the impact of Theileria sergenti/orientalis/buffeli group and Eh. canis on ruminant abortion and their real zoonotic risk in the island.

Coexistence of Tick-Borne Pathogens in Ticks Collected from their Hosts in Sardinia: an Update.

PURPOSE: In recent decades, the incidence and distribution of tick-borne diseases have increased worldwide, attracting the attention of both clinicians and veterinarians. In Sardinia, notifiable tick-borne diseases are spreading and Mediterranean spotted fever (MSF) rickettsiosis continues to be endemic with an incidence of 10/10,000 inhabitants per year. Furthermore, ticks can transfer more than one pathogen after a single blood meal from a coinfected host or after multiple feeding on different infected hosts. The aim of this study was to update information on ticks and tick-borne diseases, focusing also on the presence of coinfection in Sardinian ticks. METHODS: The presence of protozoan (Theileria and Babesia species) and bacterial pathogens (Rickettsia spp., Anaplasma spp., Ehrlichia canis, Chlamydia spp., Bartonella spp., and Coxiella burnetii) was evaluated in 230 ticks collected from different hosts in Sardinia. RESULTS: PCR and sequencing analyses highlighted that the 59% of ticks were infected with at least one pathogen while the 15% resulted in coinfection by double and triple pathogens. Among the double co-infections, those of E. canis/C. burnetii, Babesia sp. Anglona/Ch. psittaci and Babesia sp. Anglona/C. burnetii revealed a statistically significant index of coinfection. CONCLUSION: This study identifies new pathogens in Sardinian ticks and updates the information about tick-borne diseases in the island. We also provide new results on the presence of coinfections in collected ticks. The knowledge about the diversity of ticks and tick-borne diseases circulating in Sardinia is a necessary step toward implementing effective tick-borne disease prevention and control programs.

Molecular characterization of Toxoplasma gondii Type II in sheep abortion in Sardinia, Italy.

During 2009-2010, 161 tissue samples (142 placentas, 16 brains, and 3 livers) from aborted ovine fetuses on Sardinia Island, Italy, were tested for toxoplasmosis. Organs that showed a positive result by nested polymerase chain reaction (PCR) targeting the ITS1 region for Toxoplasma gondii were also amplified with 11 genetic markers (SAG1, 5'-SAG2, 3'-SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) and then subjected to PCR/RFLP for genetic typing. T. gondii DNA was found in 5 placentas, 14 brains, and 2 livers by PCR analysis and all isolates displayed Type II alleles at all 11 loci with all 11 markers. The results indicate that the Type II T. gondii is associated with ovine abortion.

Occurrence, distribution, and role in abortion of Coxiella burnetii in sheep and goats in Sardinia, Italy.

Between 1999 and 2002, 9349 sera and 517 aborted samples (422 foetuses and 95 placenta) were analysed from 675 sheep and 82 goat farms distributed all over the island of Sardinia. After abortion notification, sera collected at random from adult animals were examined to detect antibodies specific to Coxiella burnetii by ELISA, whereas foetuses and placenta were analysed by PCR assay. Specific IgG antibodies were detected in 255 (38%) sheep farms and in 39 (47%) goat herds whereas 40 ovine (10%) and 3 (6%) caprine foetuses were C. burnetii PCR-positive. Although C. burnetii DNA was amplified from different types of tissues, placenta was the tissue with the highest detection rate. Seroprevalence analysis indicates that C. burnetii distribution in sheep and goats is very high, but PCR results demonstrate that C. burnetii has a relatively low role in abortion, especially in goats.