The tree that hides the forest: Cryptic diversity and phylogenetic relationships in the Palaearctic vector Obsoletus/Scoticus Complex (Diptera: Ceratopogonidae) at the European level

Background: Culicoides obsoletus is an abundant and widely distributed Holarctic biting midge species, involved in the transmission of bluetongue virus (BTV) and Schmallenberg virus (SBV) to wild and domestic ruminants. Females of this vector species are often reported jointly with two morphologically very close species, C. scoticus and C. montanus, forming the Obsoletus/Scoticus Complex. Recently, cryptic diversity within C. obsoletus was reported in geographically distant sites. Clear delineation of species and characterization of genetic variability is mandatory to revise their taxonomic status and assess the vector role of each taxonomic entity. Our objectives were to characterize and map the cryptic diversity within the Obsoletus/Scoticus Complex. Methods: Portion of the cox1 mitochondrial gene of 3763 individuals belonging to the Obsoletus/Scoticus Complex was sequenced. Populations from 20 countries along a Palaearctic Mediterranean transect covering Scandinavia to Canary islands (North to South) and Canary islands to Turkey (West to East) were included. Genetic diversity based on cox1 barcoding was supported by 16S rDNA mitochondrial gene sequences and a gene coding for ribosomal 28S rDNA. Species delimitation using a multi-marker methodology was used to revise the current taxonomic scheme of the Obsoletus/Scoticus Complex. Results: Our analysis showed the existence of three phylogenetic clades (C. obsoletus clade O2, C. obsoletus clade dark and one not yet named and identified) within C. obsoletus. These analyses also revealed two intra-specific clades within C. scoticus and raised questions about the taxonomic status of C. montanus. Conclusions: To our knowledge, our study provides the first genetic characterization of the Obsoletus/Scoticus Complex on a large geographical scale and allows a revision of the current taxonomic classification for an important group of vector species of livestock viruses in the Palaearctic region.[Figure not available: See fulltext.

Evaluation of an Indirect Immunofluorescence Assay for the Detection of Anaplasma phagocytophilum Antigen in Ovine Buffy Coat Smears

Diagnosis of anaplasmosis is challenging considering the great variation in clinical signs and the limitations of the available diagnostic assays, while the detection of carrier animals that play a significant role in disease epidemiology as reservoirs is of great significance. In this study, we evaluated the diagnostic accuracy of a newly developed indirect immunofluorescent assay (Ag-IFAT) for the detection of A. phagocytophilum antigens in buffy coat specimens, alone and in combination with cytology, using PCR as a reference. Blood samples were collected from 138 sheep of the Chios breed from six farms in Greece. A buffy coat was extruded from the centrifuged blood. Buffy coat smears were used for cytological examination and the Ag-IFAT assay. The Ag-IFAT assay presented excellent specificity (100%) and high sensitivity (85.4%) for the detection of A. phagocytophilum antigens in buffy coats, and it has an almost perfect agreement with PCR and cytology (κ value = 0.88 and 0.85, respectively). A. phagocytophilum antigens are likely to be detected using Ag-IFAT in a PCR-positive animal, as indicated by the good performance of the assay. Overall, this assay presents high diagnostic accuracy, and it could be used for the detection of animals during the early stage of infection. © 2022 by the authors. Licensee MDPI, Basel, Switzerland

Trichinella britovi as a risk factor for alternative pig production systems in Greece and Europe

Trichinellosis is an important zoonosis and the most common source of human infection is meat from pigs and wild boars. The European Union (EU) supports alternative pig production systems (organic and free-ranging) as sustainable farming systems. However, these systems that allow outdoor access for farm animals, may create new or reintroduce old risks to public health. During the last years, alternative pig production systems (free-ranging or organic pig) are growing in popularity in Greece due to the increasing interest of consumers for organic products. The majority of the trichinellosis outbreaks in the EU were associated with pork and meat products including wild boars. In Greece, from 2009 to 2012, 37 Trichinella spp. positive free-ranging pigs were reported in free-ranging pig farms of Northern-Eastern Greece (31 were identified as T. britovi). The recent re-emerging present of the Trichinella spp. infections in freeranging pigs and wild boars are a high risk for the consumers and should alarm the Public Health Authorities in Greece and the EU. During the last years, the organic or free-ranging pig production systems are growing in popularity in the EU. However, these systems increase the risk of Trichinella spp. infections, since pigs are possible to be infected by feeding on carcasses or the offal of hunted or dead wild animals. For this reason, it is important for Public Health Authorities to be focused on the training of hunters and farmers in order to avoid the transmission among free-ranging pigs and prevent the cases of human infection. © 2020 Urmia University. All rights reserved

Detection and Zoonotic Potential of Trichinella spp. from Free-Range Pig Farming in Greece

Trichinellosis is a serious parasitic zoonosis, which is widely distributed around the world. Pork meat is still the predominant source of outbreaks of human trichinellosis in many countries. The aim of this study is to examine the impact of Trichinella spp. as an important risk factor on the free-range pig farming sector in Greece. In 2009, during routine testing for the detection of Trichinella larvae at slaughterhouses and the National Reference Laboratory for Parasites (NRL), a total of 826,426 pigs were tested with the magnetic stirrer method for Trichinella spp. at slaughterhouses, including 2,892 samples from free-range pigs. Two positive samples were detected: one positive for Trichinella britovi and one positive for Trichinella spp. (unspecified) in the samples from wild farmed free-range pigs. It is alarming that one of these cases was connected with clinical signs of trichinellosis in five persons of the same family in northeastern Greece, who consumed undercooked pork meat from a free-range pig farm. During 2010, a total number of 1,295,034 pigs were tested with same method, including 4,159 samples from free-range pig farms. Five positive samples for Trichinella spp. (unspecified) were detected from 4,159 free-range pigs tested by the Greek NRL. Moreover, 363 serum samples from free-range pigs were serologically tested with enzyme-linked immunosorbent assay (ELISA). Moreover, 363 serum samples from farmed free-range pigs were serologically tested with ELISA, and 15 samples were found positive. Finally, the present study is the first report of detection of T. britovi in Greece. In conclusion, based on the results of the present study, Trichinella spp. is a high-risk factor for the free-range pig farming in Greece

Health and Production of Greek Organic Pig Farming: Current situation and perspectives

For many years, outdoor pig farming has been one of the most important livestock production sectors in Greece. Since the 1960s, the introduction of high-yielding pig genotypes, under intensive production systems, has replaced almost to disappearance the traditional -based on the native pig breed- outdoor pig farms. Organic pig production systems in Greece are growing in popularity over the last years due to the increasing interest of consumers for organic products. The National Greek projects for organic pig farming started in 2002 and since then it has increased, representing the 15% of total organic livestock production in 2007. According to the Directorate of Organic Agriculture in the Ministry of Rural Development and Food, the development of organic pig farming industry in West Greece, Thessaly and North Greece was impressive from 2002 to 2007. A significant but more gradual development of organic pig farming was seen in the island of Crete, while insignificant development was observed in the Central Greece. The development of organic pig farming especially in the Northern part of the country started from 2003-2004 onwards. In 2002, the total number of organic pigs was just 1.288, while in 2007 it had reached 175.000 organic pigs in the country. However, a significant reduction has been noticed since 2008, mainly due to the national funding cuts for organic pig farming, as well as the increase of feeding expenses. Moreover, training and specialization of organic pig farmers and their investments on modernization and equipment / housing facilities were insufficient, resulting in animal health problems, poor growth performance, poor carcass quality and high-cost production. The most common health problems in Greek organic pig farming are respiratory problems, gastrointestinal problems, claw and skin problems, parasitic infections and high piglet mortality. Increasing health problems were attributed mainly to poor housing conditions (predispose to various infectious micro-organisms), and the contact of organic pigs with rodents that act as tank of several pathogens (Trichinella spp and Toxoplasma gondii). The housing condition of organic farming may predispose animals to various infectious micro-organisms, normally no longer present indoors because of the strict hygienic measures that are taken. An important risk factor in organic pig production is the more frequent, compared with conventional swine industry, in contact with rodents. Rodents are actors - tank of several pathogens, some of which are hazardous to public health, such as Trichinella spp and Toxoplasma gondii. This report aims to present updated information about the health status, production and development of organic pig farming in Greece during the last decade, as well as the potential of this particular productive activity for future development

Detection of Antibodies Against Toxoplasma gondii in Filter Paper-Dried Blood Dot Spots Compared with Serum in Pigs and Assessment of Variation Associated with Packed Cell Volume

The aim of this study was to assess the agreement between anti-Toxoplasma gondii IgG antibody detection in serum and filter paper (FP) blood spots using the indirect immunofluorescence antibody assay (IFA) and to evaluate the potential impact of the packed cell volume (PCV) on antibody detection in FPs. A pair of a serum and an FP sample was collected from 96 sows at various farms in Greece, with previously identified high seropositivity and/or risk factors associated with high seropositivity against T. gondii. The PCV value was determined using the microhematocrit method. IFA was used for the detection of antibodies against T. gondii. T. gondii-specific antibodies were detected in 45.8% serum samples and 41.6% FP samples showing almost perfect agreement. Detection in FP samples presented high sensitivity (87.1-92.8%) and excellent specificity (100%) when compared with detection in serum, regardless of the PCV values. The findings of this study support the reliability of FPs for the evaluation of the serological status of swine against T. gondii. FPs could be a good alternative sample type compared with serum for large-scale epidemiological studies. © 2022, Mary Ann Liebert, Inc., publishers

Phlebotomine sandflies and factors associated with their abundance in the leishmaniasis endemic area of Attiki, Greece

Leishmaniasis is a parasitic disease of animals and humans caused by several Leishmania species and transmitted by phlebotomine sandflies. The aim of the present study was to identify the species of field collected phlebotomine sandflies in the endemic area of the Attiki during 4 consecutive years, to isolate the Leishmania parasites from the infected sandflies, and identify possible factors associated with sandfly abundance in the area. A total of 542 trappings were made in 46 collection sites, in purely urban areas, periurban areas, and purely rural areas in Attiki. Out of the 3254 sandflies trapped, 1448 (44.43%) were female and 241 (16.64%) of the females were blood fed while Leishmania infantum DNA was detected in the 0.41% of them. Regarding sandfly species, the most prevalent was Phlebotomus tobbi (41.52%) followed by Sergentomyia minuta (27.44%), P. neglectus (14.83%), P. simici (11.08%), P. papatasi (3.68%), P. similis (0.89%), and P. alexandri (0.56%). Periurban areas were found to have the highest density of sandfly populations. © 2017, Springer-Verlag GmbH Germany, part of Springer Nature

Ectoparasite infestations of urban stray dogs in Greece and their zoonotic potential

The presence of ectoparasites is very common in stray dogs worldwide. So far there are limited data on the spread of external parasites of dogs in Greece. Due to the potential risk of human infestation and/or transmission of zoonotic agents, knowledge of their abundance and distribution is essential in order to better define preventive measures. Therefore, the aim of this study was to assess the prevalence of external parasites of stray dogs from the area of Thessaloniki Greece. Out of a total of 242 stray dogs (85 males and 157 females) tested, the greatest percentages of infestation of 46.28% and 43.8% by fleas (Ctenocephalides felis and Ctenocephalides canis) and ticks (Rhipicephalus sanguineus) respectively were observed. Moreover, 15 dogs (6.20%) were found infested by Sarcoptes scabiei var canis, 8 (3.31%), by Otodectes cynotis while 4 (1.65%) dogs were infested by Cheyletiella sp and Demodex canis. Finally, three dogs (1.24%) were found infested by lice (Trichodectes canis). A statistically significant higher percentage of external parasites was detected in young dogs compared to adults (P=0.0001) and in males compared to females (P<0.0001), while this difference was not statistically significant in short haired compared to long haired and in small size compared to large size dogs (P= 0.6938 and P=0.9934, respectively). The results of this study support the assumption that stray dogs constitute a considerable source of ectoparasites and their infestation is indicative of the presence and abundance of ectoparasites in an area. © 2016, Malaysian Society for Parasitology. All rights reserved