The leaf flavonoids of the orchidaceae

In a leaf survey of 142 species from 75 genera of the Orchidaceae, flavone C-glycosides (in 53%) and flavonols (in 37 %) were found to be the most common constituents. However, since these compounds are not found uniformly and their distribution shows a strong correlation with plant geography, it is not possible to represent the Orchidaceae by a single flavonoid profile. Thus, flavone C-glycosides are most common in tropical and subtropical species of the Epidendroid and Vandoid tribes (in 63%) and flavonol glycosides are more characteristic of temperate species of the Neottioid tribes (in 78%). By contrast 6-hydroxyflavones (in 6 species), luteolin (in 2 species) and tricin as the 5-glucoside (in 1 species) are all rare. Three new glycosides were characterised: scutellarein 6-methyl ether 7-rutinoside from Oncidium excavatum and O. sphacelatum, pectolinarigenin 7-glucoside from 0. excavatutn and Eria javanica, and luteolin 3′,4′-diglucoside from Listera ovata. The xanthones, mangiferin and isomangiferin were found in Mormolyca ringens, Maxillaria aff. luteo-alba and 5 Polystachya species and a mangiferin sulphate tentatively identified in P. nyanzensis. Other unusual phenolic constituents include 6,7-methylenedioxy- and 6,7-dimethoxycoumarins from Dendrobium densiflorum and D. farmeri, formed by the rearrangement during the extraction process from the corresponding O-glucosyloxycinnamic acids. The origin and relationship of the Orchidaceae to other monocot groups are discussed in the light of the flavonoid evidence.     

Sensitization of Children to Storage Mites in Kutahya, Turkey

Specific IgE against Acarus siro, Glycphagus domesticus, Tyrophagus putrescentiae, and Lepidoglyphus destructor have been investigated by ELISA in sera of 92 children. Of them, 41 were found to be specific IgE positive (≥ 0.35 IU/ml) against at least one of house dust mite species, Dermatophagoides pteronyssinus and Dermatophagoides farinae, by an immunoblot. In 65.9% of the dust mite-sensitized children, specific IgE against at least one of these mite species was found. Sensitization levels, including co-sensitization cases were found to be 35.7% against A. siro, 24.4% against T. putrescentiae, 31.7% against L. destructor, and 26.8% against G. domesticus. In non-sensitized children, dust mite sensitization level was found to be 25.5%. Breakdown of sensitization by individual species in this group was; against A. siro and T. putrescentiae at 7.8%, against L. destructor at 13.7%, and against G. domesticus at 9.8%. When all children were reckoned, 43.5% was found to be sensitized against at least one storage mite species, with sensitizations against A. siro at 18.5%, T. putrescentiae at 26.1%, L. destructor at 21.7%, and G. domesticus at 17.4%. In dust samples collected from the dwellings of children, distribution of species was found to be A. siro (17%), G. domesticus (23%), T. putrescentiae (29%), L. destructor (25%), and unidentified (6%). In Fisher''s chi-square test on SPSS program, there was a relationship between dust mite sensitization and storage mite sensitization (P < 0.05), but no meaningful relationship was found on the basis of individual mite species.     

Relative Abundance and Plasmodium Infection Rates of Malaria Vectors in and around Jabalpur,a Malaria Endemic Region in Madhya Pradesh State,Central India

BackgroundThis study was undertaken in two Primary Health Centers (PHCs) of malaria endemic district Jabalpur in Madhya Pradesh (Central India).MethodsIn this study we had investigated the relative frequencies of the different anopheline species collected within the study areas by using indoor resting catches, CDC light trap and human landing methods. Sibling species of malaria vectors were identified by cytogenetic and molecular techniques. The role of each vector and its sibling species in the transmission of the different Plasmodium species was ascertained by using sporozoite ELISA.ResultsA total of 52,857 specimens comprising of 17 anopheline species were collected by three different methods (39,964 by indoor resting collections, 1059 by human landing and 11,834 by CDC light trap). Anopheles culicifacies was most predominant species in all collections (55, 71 and 32% in indoor resting, human landing and light trap collections respectively) followed by An. subpictus and An. annularis. All five sibling species of An. culicifacies viz. species A, B, C, D and E were found while only species T and S of An. fluviatilis were collected. The overall sporozoite rate in An. culicifacies and An. fluviatilis were 0.42% (0.25% for P. falciparum and 0.17% for P. vivax) and 0.90% (0.45% for P. falciparum and 0.45% for P. vivax) respectively. An. culicifacies and An. fluviatilis were found harbouring both P. vivax variants VK-210 and VK-247, and P. falciparum. An. culicifacies sibling species C and D were incriminated as vectors during most part of the year while sibling species T of An. fluviatilis was identified as potential vector in monsoon and post monsoon season.ConclusionsAn. culicifacies species C (59%) was the most abundant species followed by An. culicifacies D (24%), B (8.7%), E (6.7%) and A (1.5%). Among An. fluviatilis sibling species, species T was common (99%) and only few specimens of S were found. Our study provides crucial information on the prevalence of An. culicifacies and An. fluviatilis sibling species and their potential in malaria transmission which will assist in developing strategic control measures against these vectors.     

Prevalence of Malassezia species in pityriasis versicolor lesions in northeast Argentina

BackgroundMalassezia species normally colonize the skin but they can change their saprophytic state and invade the stratum corneum as pathogens.AimsTo determine the prevalence of Malassezia species isolated from patients with pityriasis versicolor (PV) and to analyse their distribution according to the location of the lesion on the body.MethodsThis study included 218 patients with PV and positive Malassezia cultures who resided in the city of Resistencia, a subtropical area located in northeast Argentina. Age, gender, and the body site of lesions were recorded. Strains were identified by PCR-RFLP.ResultsMalassezia sympodialis (37.7%) and Malassezia globosa (37.2%) were the most prevalent species isolated alone or in association with other Malassezia species in 82% of the patients. Malassezia furfur (21.3%) was the third most common species, followed by Malassezia slooffiae (1.7%), and Malassezia restricta (1.3%), which was found only in combination with M. globosa and M. sympodialis. Malassezia dermatis (0.4%) and Malassezia pachydermatis (0.4%) were each isolated once. None of the species affected a body site with statistical significance. Significant difference between genders according to age was found only in the 31–40-year-age group.ConclusionsThis study suggests that M. sympodialis and M. globosa represent the main species implicated in the pathogenicity of PV. M. furfur appears to be the third agent of importance in this geographical area. Statistical analyses showed none of the species was particularly associated with any one of the body sites.     

Survey of Zoonotic Trematode Metacercariae in Fish from Irrigation Canal of Togyo-jeosuji (Reservoir) in Cheorwon-gun,Gangwon-do,Republic of Korea

The infection status of zoonotic trematode metacercariae (ZTM) was investigated in total 568 freshwater fishes (19 species) from the irrigation canal of Togyo-jeosuji (Reservoir) in Cheorwon-gun, Gangwon-do, the Republic of Korea for 3 years (2018–2020). All fishes were examined using the artificial digestion method. The metacercariae of Clonorchis sinensis (CsMc) were detected in 180 (43.8%) out of 411 fish of positive species, and their infection intensity was 38 per fish infected (PFI). Especially, in 2 fish species, i.e., Pseudorasbora parva and Puntungia herzi, the prevalence was 82.1% and 31.3%, and the infection intensity with CsMc was 88 and 290 PFI, respectively. Metagonimus spp. metacercariae (MsMc) were found in 403 (74.1%) out of 544 fish of positive species, and their infection intensity was 62 PFI. In the pale chub, Zacco platypus, the prevalence of MsMc was 98.6%, and their infection intensity was 144 PFI. Centrocestus armatus metacercariae were detected in 171 (38.9%) out of 440 fish of positive species, and their infection intensity was 1,844 PFI. Echinostoma spp. metacercariae were found in 94 (19.6%) out of 479 fish of positive species, and their infection intensity was 3 PFI. Metorchis orientalis metacercariae were detected in 43 (29.3%) out of 147 fish of positive species, and their infection intensity was 4 PFI. By the present study, it has been confirmed that some species of ZTM, including CsMc and MsMc, are prevalent in fishes from the irrigation canal of Togyo-jeosuji in Cheorwon-gun, Gangwon-do, Korea.     

Novel Clade of Rickettsia spp. from Leeches

Intracellular rickettsia-like structures were found in the tissues of a glossiphoniid leech, Torix tagoi, by transmission electron microscopy. Diagnostic PCR analysis using specific primers suggested that of the nine glossiphoniid species examined, two species, T. tagoi and Hemicrepsis marginata, harbored bacteria of the genus Rickettsia. A 1.5-kb eubacterial 16S rRNA gene segment obtained from each of these species was amplified by PCR, cloned, and sequenced. Phylogenetic analysis of the 16S rRNA gene demonstrated that the Rickettsia species found in the leeches constituted a novel clade that is distinct from the clade of arthropod-associated Rickettsia species. In natural populations, 97.7% (43 of 44) of T. tagoi leeches and 100% (9 of 9) of H. marginata leeches carried Rickettsia, suggesting that infection with Rickettsia is prevalent in these leeches. This is the first report of Rickettsia found in annelids.

     

Phylogeny and Diagnostic Markers of Six Tetranychus Species (Acarina: Tetranychidae) in Korea Based on the Mitochondrial Cytochrome Oxidase Subunit I

To understand the evolutionary relationship among six Tetranychus species in Korea, 598 nucleotides of cytochrome oxidase subunit I (COI) were sequenced and analyzed. The sequences were extremely rich in A+T (77% on average). The nucleotide differences in pairwise comparison among species ranged from 6.7% to 14.9%. Nucleotide substitutions based on all pairwise comparison showed 55% of transversions. The largest genetic distance was found between species T. viennensis and T. piercei. The inferred phylogenetic tree indicates that T. urticae, T. cinnabarinus and T. kanzawai are clustered together and showing sister-group relationship with T. truncatus at the highest bootstrap support. T. viennensis showed the largest genetic distance from all other species. T. piercei, a species recently found in Korea, was also distinct from the group composed of four species and also from T. viennensis. The restriction sites of Alu I, Dde I and Sau 3A were searched in the sequences of COI. The restriction fragment length polymorphisms (RFLPs) in COI genes and intraspecific variabilities of three restriction sites in diverse colonies of each species collected from different locations and various host plants were shown.     

Preliminary study of fish community structure in Amanos Mountain streams (Hatay-Turkey)

Fish diversity and distributions in 33 streams of the Amanos Mountains were investigated between December 2002 and November 2003. Some physical and chemical parameters, such as water temperature, dissolved oxygen, electrical conductivity, pH, and velocity of the streams were measured periodically at each site. The study area was grouped into three regions: AS (Asi Basin), IS (?skenderun Gulf) and CE (Çevlik region). Fish abundance and Shannon Diversity Index were computed for each region. CE region was the poorest compared to the others in terms of fish abundance and diversity. A total of nine species in four families (Cyprinidae, Balitoridae, Anguillidae, Blenniidae) were recorded and all species collected were found to be indigenous. Capoeta barroisi (42.2%) and Garra rufa (33.3%) were the most common and abundant species and Salaria fluviatilis (0.4%), Carasobarbus luteus (0.9%), Anguilla anguilla (1.0%) and Alburnus sellal (1.2%) were the most rarely encountered species in the Amanos Mts. The endangered species Alburnus orontis is also rarely found. Fish diversity was greater in perennial streams (0.62) than in intermittent streams (0.27). There are many adverse human impacts on fish fauna in the Amanos Mountain streams; however, numbers of intermittent streams impose the greatest threat to fish in the Amanos Mts.     

Prevalence of non-strongyle gastrointestinal parasites of horses in Riyadh region of Saudi Arabia

This study aimed to provide recent data on the occurrence of non-strongyle intestinal parasite infestation in horses in the Riyadh region of Saudi Arabia as a basis for developing parasite control strategies. We conducted necropsy for 45 horses from September 2006 to November 2007 in the Riyadh region, Saudi Arabia. 39 out of 45 horses were infected with intestinal parasites with an infestation rate of 86.6%. Infestations with seven nematode species and two species of Gasterophilus larva were found. The most prevalent parasites were Strongyloides westeri (64.4%) and Parascaris equorum (28.8%) followed by Habronema muscae (22.2%). Trichostrongylus axei and Oxyuris equi were less common at (11.1%) and (8.8%), respectively. Habronema megastoma and Setaria equine were found in two horses only (4.4%). Gasterophilus intestinalis larvae were recovered from 39 horses (86.6%) and Gasterophilus nasalis larvae were found in 17 horses (37.7%). Season had a significant effect on the prevalence of P. equorum and G. nasalis, while age of horses had a significant effect only on the prevalence of P. equorum. The husbandry in Saudi Arabia appears to be conductive to parasites transmitted in stables or by insects rather than in pasture.     

The leaf flavonoids of the Zingiberales

A survey of flavonoids in the leaves of 81 species of the Zingiberales showed that, while most of the major classes of flavonoid are represented in the order, only two families, the Zingiberaceae and Marantaceae are rich in these constituents. In the Musaceae (in 9 species), Strelitziaceae (in 8 species) and Cannaceae (1 of 2 species) flavonol glycosides were detected in small amount and in the Lowiaceae no flavonoids were fully identified. In the Zingiberaceae kaempferol (in 22%), quercetin (72%) and proanthocyanidins (71%) are distributed throughout the family. The two subfamilies of the Zingiberaceae may be distinguished by the presence of myricetin (in 26%), isorhamnetin (10%) and syringetin (3%) in the Zingiberoideae and of flavone $\text{C-}\text{glycosides}$ (in 86% of taxa) in the Costoideae. A number of genera have distinctive flavonol profiles: e.g. Hedychium species have myricetin and quercetin. Roscoea species isorhamnetin and quercetin and Alpinia species kaempferol and quercetin glycosides. A new glycoside, syringetin 3-rhamnoside was identified in Hedychium stenopetalum. In the Zingiberoideae flavonols were found in glycosidic combination with glucuronic acid, rhamnose and glucose but glucuronides were not detected in the Costoideae or elsewhere in the Zingiberales. The Marantaceae is chemically the most diverse group and may be distinguished from other members of the Zingiberales by the occurrence of both flavone $\text{O-}$ and $\text{C-}\text{glycosides}$ and the absence of kaempferol and isorhamnetin glycosides. The distribution of flavonoid constituents within the Marantaceae does not closely follow the existing tribai or generic limits. Flavonols (in 50% of species). flavones (20%) and flavone $\text{C-}\text{glycosides}$ (40%) are found with similar frequency in the two tribes and in the genera Calathea and Maranta both flavone and flavonol glycosides occur. Apigenin- and luteolin-7-sulphates and luteolin-7,3′-disulphate were identified in Maranta bicolor and M. leuconeura var. kerchoveana and several flavone $\text{C-}\text{glycosides}$ sulphates in Stromanthe sanguinea. Anthocyanins were identified in those species with pigmented leaves or stems and a common pattern based on cyanidin-and delphinidin-3-rutinosides was observed throughout the group. Finally the possible relationship of the Zingiberales to the Commelinales, Liliales, Bromeliales and Fluviales is discussed.     

Prevalence of ectoparasitic arthropods on wild animals and cattle in the Las Merindades area (Burgos, Spain)

This paper reports the prevalence of ectoparasitic arthropods in sampled groups of wild (n = 128; 16 species) and domestic (n = 69; 3 species) animals in the Las Merindades area of the Province of Burgos, Spain. The study revealed that wild animals were more infested and with a wider variety of ectoparasites than domestic animals. The parasitic prevalence was 67% for wild animals and 48% for livestock. In this way, 39% of animals were infected by ticks. Ixodes ricinus and Ixodes hexagonus were the most prevalent species whereas Dermacentor reticulatus showed affinity for the fox and wolf. The overall prevalence of parasitisation by fleas was 27%. Ctenophthalmus spp. showed the wider range host in wild animals, while Pulex irritans was the most frequent specie found. The parasitic prevalences by lice (Trichodectes melis, Trichodectes canis and Trichodectes mustelae) and by mite (Neotrombicula spp., Laelaps agilis and Sarcoptes scabiei) were 4% and 12%, respectively. In both cases only wild animals were found parasited.     

A new species and record of branchial parasitic isopods (Crustacea: Isopoda: Bopyridae: Pseudioninae) of porcellanid crabs from the Philippines

Branchial bopyrids infesting porcellanid crabs from the Philippines were investigated based on intertidal collections made in 1999-2000. Crabs of the genus Petrolisthes collected from sites in the northern Philippines were examined and two parasite species were found. One new pseudionine species found infesting Petrolisthes sp. [cf. Petrolisthes asiaticus (Leach)] is described as Aporobopyrus galleonus (prevalence 6.1%); this species is distinguished from other members of the genus by a setose palp on the maxilliped of the females, barbula morphology, and male characters including the possession of pleopods. This represents the second described species of Aporobopyrus from the Philippines, and the first from porcellanid crabs. In addition, Pleurocrypta macrocephalaNierstrasz and Brender à Brandis, 1923 (originally described from Indonesia) was found infesting the same unidentified Petrolisthes sp. (prevalence 2.6%); this is the first report of the species from the Philippines.     

The Distribution and Diversity of Bartonella Species in Rodents and Their Ectoparasites across Thailand

Our study highlights the surveillance of Bartonella species among rodents and their associated ectoparasites (ticks, fleas, lice, and mites) in several regions across Thailand. A total of 619 rodents and 554 pooled ectoparasites (287 mite pools, 62 flea pools, 35 louse pools, and 170 tick pools) were collected from 8 provinces within 4 regions of Thailand. Bandicota indica (279), Rattus rattus (163), and R. exulans (96) were the most prevalent species of rats collected in this study. Real-time PCR assay targeting Bartonella-specific ssrA gene was used for screening and each positive sample was confirmed by PCR using nuoG gene. The prevalence of Bartonella DNA in rodent (around 17%) was recorded in all regions. The highest prevalence of Bartonella species was found in B. savilei and R. rattus with the rate of 35.7% (5/14) and 32.5% (53/163), respectively. High prevalence of Bartonella-positive rodent was also found in B. indica (15.1%, 42/279), and R. norvegicus (12.5%, 5/40). In contrast, the prevalence of Bartonella species in ectoparasites collected from the rats varied significantly according to types of ectoparasites. A high prevalence of Bartonella DNA was found in louse pools (Polyplax spp. and Hoplopleura spp., 57.1%) and flea pools (Xenopsylla cheopis, 25.8%), while a low prevalence was found in pools of mites (Leptotrombidium spp. and Ascoschoengastia spp., 1.7%) and ticks (Haemaphysalis spp., 3.5%). Prevalence of Bartonella DNA in ectoparasites collected from Bartonella-positive rodents (19.4%) was significantly higher comparing to ectoparasites from Bartonella-negative rodents (8.7%). The phylogenetic analysis of 41 gltA sequences of 16 Bartonella isolates from rodent blood and 25 Bartonella-positive ectoparasites revealed a wide range of diversity among Bartonella species with a majority of sequences (61.0%) belonging to Bartonella elizabethae complex (11 rodents, 1 mite pool, and 5 louse pools), while the remaining sequences were identical to B. phoceensis (17.1%, 1 mite pool, 5 louse pools, and 1 tick pool), B. coopersplainensis (19.5%, 5 rodents, 1 louse pool, and 2 tick pools), and one previously unidentified Bartonella species (2.4%, 1 louse pool).     

Intestinal Helminthic Infections in Striped Field Mice,Apodemus agrarius,from Two Southern Regions of Korea

The present study was performed to know the infection status of intestinal helminths in a most common species of field mice, Apodemus agrarius, from 2 southern regions of Korea. Total 133 and 103 mice were collected by the mouse trap in Hapcheon-gun, Gyeongsangnam-do and Gurye-gun, Jeollanam-do, respectively, from July 2005 to June 2006. The small intestine of each mouse was resected and longitudinally opened with a pair of scissors. The intestinal contents were washed with 0.85% saline until the supernatant became clear. Helminths were collected with naked eyes or under a stereomicroscope from the sediment of the intestinal content. More than 11 species of helminths (4 nematode spp., 5 trematode spp., and 2 cestode spp.) were recovered. Among these, heligmosomoid nematodes (97.5%) was the most highly and heavily infected species. As the members of trematodes, Plagiorchis muris, Brachylaima sp., Echinostoma hortense, Echinostoma cinetorchis, and unidentified echinostome larvae were found in the small intestines of 35 (14.8%), 12 (5.1%), 6 (2.5%), 1 (0.4%), and 1 (0.4%) mice respectively. Two species of tapeworms, Hymenolepis nana and Hymenolepis diminuta were also detected in 79 (33.5%) and 21 (8.9%) mice, respectively. Conclusively, heligmosomoid nematodes were the most prevalent (dominant) species among more than 11 helminth species detected, and Brachylaima sp. fluke is newly added in the list of intestinal trematodes in Korea.     

Arthropods (Acarina, Coleoptera, Siphonaptera) in nests of hoopoe (Upupa epops) in Central Europe

In 1993–1995 and 2009–2011, during investigation of reproduction biology of hoopoe, 63 nests of this species were collected after fledging of the chicks in Slovakia and Austria and their arthropod fauna was analyzed. Altogether 5,481 individuals and 34 species of mesostigmatic mites were found in 82.5% of the nests examined. The nidicolous mite Androlaelaps casalis was most abundant and frequent, representing 91.7% of all individuals. The richest in species were saprophilous mites (64.7% of all recorded species), while ectoparasites of the genera Dermanyssus and Ornithonyssus represented only 0.4% of all mites. Only 8 beetle species represented by 65 individuals were found in 18 nests. The dominant trophical group were carnivores (mainly nidicolous Gnathoncus buyssoni) with almost identical representation in (86.2%), followed by a similar representation of necrophags (10.8%). Unlike nests of other birds, the typical nidicolous species Haploglossa puncticollis and the fungivors were absent due to the dry character of studied countryside and placement of the nests in boxes situated in vineyard cottages. Only one species of fleas, Ceratophyllus gallinae — parasitizing first of all in the passeriform birds and being particularly abundant in the cavity nesting birds was recorded in hoopoe nests.     

Prevalence and Fingerprinting of Listeria monocytogenes Strains Isolated from Raw Whole Milk in Farm Bulk Tanks and in Dairy Plant Receiving Tanks

The incidence of Listeria species in raw whole milk from farm bulk tanks and from raw milk in storage at a Swedish dairy plant was studied. Listeria monocytogenes was found in 1.0% and Listeria innocua was found in 2.3% of the 294 farm bulk tank (farm tank) milk specimens. One farm tank specimen contained 60 CFU of L. monocytogenes ml⁻¹. L. monocytogenes was detected in 19.6% and L. innocua was detected in 8.5% of the milk specimens from the silo receiving tanks at the dairy (dairy silos). More dairy silo specimens were positive for both Listeria species during winter than during summer. Restriction enzyme analysis and pulsed-field gel electrophoresis were applied to 65 isolates of L. monocytogenes, resulting in 16 different clonal types. Two clonal types were shared by the farm tank milk and the dairy silo milk. All except one clonal type belonged to serovar 1/2a. In the dairy silo milk five clonal types were found more frequently and for a longer period than the others. No Listeria species were found in any other samples from the plant.     

Species of Root-knot Nematodes and Fungal Egg Parasites Recovered from Vegetables in Almería and Barcelona,Spain

Intensive vegetable production areas were surveyed in the provinces of Almería (35 sites) and Barcelona (22 sites), Spain, to determine the incidence and identity of Meloidogyne spp. and of fungal parasites of nematode eggs. Two species of Meloidogyne were found in Almería—M. javanica (63% of the samples) and M. incognita (31%). Three species were found in Barcelona, including M. incognita (50%), M. javanica (36%), and M. arenaria (14%). Solanaceous crops supported larger (P < 0.05) nematode numbers than cucurbit crops in Almería but not in Barcelona. Fungal parasites were found in 37% and 45% of the sites in Almería and Barcelona, respectively, but percent parasitism was never greater than 5%. Nine fungal species were isolated from single eggs of the nematode. The fungi included Verticillium chlamydosporium, V. catenulatum, Fusarium oxysporum, F. solani, Fusarium spp., Acremonium strictum, Gliocladium roseum, Cylindrocarpon spp., Engiodontium album, and Dactylella oviparasitica. Two sterile fungi and five unidentified fungi also were isolated from Meloidogyne spp. eggs.     

Morphological and phylogenetic analysis of Raillietina spp. in indigenous chickens (Gallus gallus domesticus) in Bangladesh

Raillietina spp. (Cestoda: Davaineidae), the most common cestodes in indigenous chickens, cause a substantial production loss in poultry industry in Bangladesh. Here, we estimated the prevalence, confirmed the species and determined the genetic pattern of species of Raillietina using molecular tools. We collected and examined 375 chickens randomly from household of different villages of Mymensingh sadar and Gouripur upazila, Mymensingh district and adult parasites were isolated and identified. Genomic DNA was extracted from collected parasites, amplified ITS-2 and ND-1 genes, sequenced and analyzed. Out of 375 samples, 270 (72.0%) were found positive with Raillietina species and mean worm burden was 10.46 ± 0.56. Microscopically, three species of Raillietina, such as R. cesticillus (37.9%), R. echinobothrida (41.1%) and R. tetragona (52.8%) were detected on the basis of their morphological features. The total length, length and width of scolex, sucker and rostellum were also measured. Among different factors, age, farming nature and flock size of chickens were significantly (p < 0.05) influenced Raillietina infections. For further validation, the sequences of ITS-2 gene generated in this study were matched with reference sequences of R. cesticillus, R. echinobothrida and R. tetragona and found 99.63% − 100% similarity. The phylogenetic analyses of ITS-2 and ND-1 sequences were clustered together with the reference sequences of R. cesticillus, R. echinobothrida and R. tetragona confirming microscopic identification. This is the first confirmation of species of Raillietina along with the prevalence of the species, which will be helpful for the formulation of a control strategy and provide basic information for further molecular study.     

Longitudinal Analysis of Tick Densities and Borrelia, Anaplasma, and Ehrlichia Infections of Ixodes ricinus Ticks in Different Habitat Areas in The Netherlands

From 2000 to 2004, ticks were collected by dragging a blanket in four habitat areas in The Netherlands: dunes, heather, forest, and a city park. Tick densities were calculated, and infection with Borrelia burgdorferi and Anaplasma and Ehrlichia species was investigated by reverse line blot analysis. The lowest tick density was observed in the heather area (1 to 8/100 m²). In the oak forest and city park, the tick densities ranged from 26 to 45/100 m². The highest tick density was found in the dune area (139 to 551/100 m²). The infection rates varied significantly for the four study areas and years, ranging from 0.8 to 11. 5% for Borrelia spp. and 1 to 16% for Ehrlichia or Anaplasma (Ehrlichia/Anaplasma) spp. Borrelia infection rates were highest in the dunes, followed by the forest, the city park, and heather area. In contrast, Ehrlichia/Anaplasma was found most often in the forest and less often in the city park. The following Borrelia species were found: Borrelia sensu lato strains not identified to the species level (2.5%), B. afzelii (2.5%), B. valaisiana (0.9%), B. burgdorferi sensu stricto (0.13%), and B. garinii (0.13%). For Ehrlichia/Anaplasma species, Ehrlichia and Anaplasma spp. not identified to the species level (2.5%), Anaplasma schotti variant (3.5%), Anaplasma phagocytophilum variant (0.3%), and Ehrlichia canis (0.19%) were found. E. canis is reported for the first time in ticks in The Netherlands in this study. Borrelia lusitaniae, Ehrlichia chaffeensis, and the human granylocytic anaplasmosis agent were not detected. About 1.6% of the ticks were infected with both Borrelia and Ehrlichia/Anaplasma, which was higher than the frequency predicted from the individual infection rates, suggesting hosts with multiple infections or a possible selective advantage of coinfection.