Activation of several key components of the epidermal differentiation pathway in cattle following infestation with the cattle tick, Rhipicephalus (Boophilus) microplus.

The cattle tick, Rhipicephalus (Boophilus) microplus, and the diseases it transmits pose a persistent threat to tropical beef production. Genetic selection of host resistance has become the method of choice for non-chemical control of cattle tick. Previous studies have suggested that larval stages are most susceptible to host resistance mechanisms. To gain insights into the molecular basis of host resistance that occurs during R. microplus attachment, we assessed the abundance of proteins (by isobaric tag for relative and absolute quantitation (iTRAQ) and Western blot analyses) and mRNAs (by quantitative reverse transcription PCR (qRT-PCR)) in skin adjacent to tick bite sites from high tick-resistant (HR) and low tick-resistant (LR) Belmont Red cattle following challenge with cattle tick. We showed substantially higher expression of the basal epidermal keratins KRT5 and KRT14, the lipid processing protein, lipocalin 9 (LCN9), the epidermal barrier catalysing enzyme transglutaminase 1 (TGM1), and the transcriptional regulator B lymphocyte-induced maturation protein 1 (Blimp1) in HR skin. Our data reveals the essential role of the epidermal permeability barrier in conferring greater resistance of cattle to tick infestation, and suggest that the physical structure of the epidermal layers of the skin may represent the first line of defence against ectoparasite invasion. Crown Copyright. © Australian Society for Parasitology Inc.

Differential recognition by tick-resistant cattle of the recombinantly expressed Rhipicephalus microplus serine protease inhibitor-3 (RMS-3)

Rhipicephalus micro plus is an important bovine ectoparasite, widely distributed in tropical and subtropical regions of the world causing large economic losses to the cattle industry. Its success as an ectoparasite is associated with its capacity to disarm the antihemostatic and anti-inflammatory reactions of the host. Serpins are protease inhibitors with an important role in the modulation of host-parasite interactions. The cDNA that encodes for a R. microplus serpin was isolated by RACE and subsequently cloned into the pPICZ alpha A vector. Sequence analysis of the cDNA and predicted amino acid showed that this cDNA has a conserved serpin domain. B- and T-cell epitopes were predicted using bioinformatics tools. The recombinant R. microplus serpin (rRMS-3) was secreted into the culture media of Pichia pastoris after methanol induction at 0.2 mg l(-1) qRT-PCR expression analysis of tissues and life cycle stages demonstrated that RMS-3 was mainly expressed in the salivary glands of female adult ticks. Immunological recognition of the rRMS-3 and predicted B-cell epitopes was tested using tick-resistant and susceptible cattle sera. Only sera from tick-resistant bovines recognized the B-cell epitope AHYNPPPPIEFT (Seq7). The recombinant RMS-3 was expressed in P. pastoris, and ELISA screening also showed higher recognition by tick-resistant bovine sera. The results obtained suggest that RMS-3 is highly and specifically secreted into the bite site of R. microplus feeding on tick-resistant bovines. Capillary feeding of semi-engorged ticks with anti-AHYNPPPPIEFT sheep sera led to an 81.16% reduction in the reproduction capacity of R. microplus. Therefore, it is possible to conclude that R. microplus serpin (RMS-3) has an important role in the host-parasite interaction to overcome the immune responses in resistant cattle. (C) 2012 Elsevier GmbH. All rights reserved.

Etograma e orçamento temporal de um grupo de machos não pareados de Phyllostomus hastatus (Pallas 1767) (Phyllostomidae: Phyllostominae)

Embora exista uma enorme variação de histórias de vida e ecologia, a maioria das espécies de morcegos mostra-se, em algum nível, social. Os aspectos de ecologia comportamental de quirópteros tem sido estudados, embora a maior parte refira-se a colônias de maternidades em regiões de clima temperado. Repertórios comportamentais completos e orçamentos temporais são raros, especialmente para a região Neotropical. Para compreender melhor a sociabilidade neste grupo, mostra-se importante focar também em grupos neutros com relação a fatores associados à atividade reprodutiva. Neste sentido, o estudo de grupos de machos neotropicais pode apresentar respostas importantes. O comportamento de um grupo machos não pareados de Phyllostomus hastatus foi estudado dentro do seu abrigo por aproximadamente 100 horas entre os meses de janeiro e agosto de 2012. O grupo estava abrigado no forro de um telhado de uma casa em desuso da Vila Dois Rios, na Ilha Grande. Os comportamentos foram registrados com uma câmera sensível a infra-infravermelho Sony DCR-HC28 em modo night-vision. Quando necessário, utilizei uma luz de auxílio infravermelha. A partir dos vídeos eu elaborei primeiramente um etograma. Os comportamentos classificados como estados foram usados para fazer um orçamento temporal, usando metodologia de amostragem por varredura e amostragem instantânea. Adicionalmente, fiz algumas observações a respeito de horário de entrada e saída dos morcegos e do tamanho do grupo. Organizei os comportamentos em seis categorias, com um total de 24 comportamentos distintos. Os comportamentos descritos são consistentes com os publicados em outros etogramas de morcegos, inclusive de alguns megaquirópteros. Um comportamento mais notável foi ventilando, que parece raro entre os microquirópteros, mas provavelmente importante na termorregulação. Nos meses analisados os morcegos alocaram aproximadamente 50% do tempo ao estado dormindo; 14,6% ao estado parado; 15,3% ao estado ativo; 0,9% ao estado andando; 0,1% ao estado voando; 14,1% ao estado higiene; e 3,5% ao estado ventilando. O orçamento temporal foi semelhante aos descritos para outros microquirópteros no interior de abrigos, com uma maior prevalência do estado dormindo, e com picos de atividade (principalmente do estado higiene) antes e depois das saídas noturnas. A higiene parece ter um papel importante no controle de ectoparasitas, e talvez algum papel social, mas como a higiene de outros indivíduos só foi observada uma única vez, não pude concluir nada a respeito. O presente trabalho é o primeiro etograma para Phyllostomus hastatus e o primeiro etograma e orçamento temporal para um grupo de machos em Chiroptera. Observei algumas diferenças importantes do grupo estudado com trabalhos já publicados sobre essa espécie, e sugiro que essas diferenças sejam estudadas mais a fundo. Apesar desta dissertação trazer contribuições importantes, fica claro que ainda falta muito a ser examinado nesse campo.

Effects of land use and physicochemical water quality on grass shrimp, Palaemonetes pugio, and its parasitic isopod, Probopyrus pandalicola, in South Carolina, USA tidal creeks

Grass shrimp, Palaemonetes pugio, are a common inhabitant of US East and Gulf coast salt marshes and are a food source for recreationally and economically important fish and crustacean species. Due to the relationship of grass shrimp with their ecosystem, any significant changes in grass shrimp population may have the potential to affect the estuarine system. Land use is a crucial concern in coastal areas where increasing development impacts the surrounding estuaries and salt marshes and has made grass shrimp population studies a logical choice to investigate urbanization effects. Any impact on tidal creeks will be an impact on grass shrimp populations and their associated micro-environment whether predator, prey or parasitic symbiont. Anthropogenic stressors introduced into the grass shrimp ecosystem may even change the intensity of infections from parasitic symbionts. An ectoparasite found on P. pugio is the bopyrid isopod Probopyrus pandalicola. Little is known about factors that may affect the occurrence of this isopod in grass shrimp populations. The goal was to analyze the prevalence of P. pandalicola in grass shrimp in relation to land use classifications, water quality parameters, and grass shrimp population metrics. Eight tidal creeks in coastal South Carolina were sampled monthly over a three year period. The occurrence of P. pandalicola ranged from 1.2% to 5.7%. Analysis indicated that greater percent water and marsh coverage resulted in a higher incidence of bopyrid occurrence. Analysis also indicated that higher bopyrid incidence occurred in creeks with higher salinity, temperature, and pH but lower dissolved oxygen. The land use characteristics found to limit bopyrid incidence were limiting to grass shrimp (definitive host) populations and probably copepod (intermediate host) populations as well.

Prevalence of ectoparasites in farmed Pangasius hypothalamus

Of the total 240 Pangasius hypothalamus (5 - 8.7cm) fry examined during September' O1 to February'02, 80 (33.33%) were found to be infested with one or more ecroparasites irrespective of genera or groups. Seven parasitic groups were identified with the highest average prevalence of Trichodinids (55%) followed by Dacrylogyrus spp. ( 42%), Episrylis spp. (8%), Apiosoma spp. (7%) Argulus spp. (5%), Gyrodacrylus spp. (4%) and Piscicola spp. (2%) the lowest prevalent group irrespective of months. Trichodinid and Dacrylogyms spp. were recorded to be the dominating parasitic groups among the seven both in terms of monthly prevalence and severity of infestation throughout the period of investigation. The highest prevalence (60%) of ectoparasite was recorded in December and the lowest (10%) in February irrespective of groups.

An immunoregulatory peptide from salivary glands of the horsefly, Hybomitra atriperoides

Horseflies are economically important blood-feeding arthropods and also a nuisance for humans, and vectors for filariasis. They rely heavily on the pharmacological propriety of their saliva to get blood meat and suppress immune reactions of hosts. Little information is available on horsefly immune suppressants. By high-performance liquid chromatography (HPLC) purification coupling with pharmacological testing, an immunoregulatory peptide named immunoregulin HA has been identified and characterized from salivary glands of the horsefly of Hybomitra atriperoides (Diptera, Tabanidae). Immunoregulin HA could inhibit the secretion of interferon-gamma (IFN-gamma) and monocyte chemoattractant protein (MCP-1) and increase the secretion of interteukin-10 (IL-10) induced by lipopolysaccharide (LIPS) in rat splenocytes. IL-10 is a suppressor cytokine of T-cell proliferative and cytokine responses. IL-10 can inhibit the elaboration of pro-inflammatory cytokines. Immunoregulin HA possibly unregulated the IL-10 production to inhibit IFN-gamma and MCP-1 secretion in the current experiments. This immunosuppression may facilitate the blood feeding of this horsefly. The current works will facilitate to understand the molecular mechanisms of the ectoparasite-host relationship. 2008 Elsevier Ltd. All rights reserved.

Toward an understanding of the molecular mechanism for successful blood feeding by coupling proteomics analysis with pharmacological testing of horsefly salivary glands

Horseflies are economically important blood-feeding arthropods and also a nuisance for humans and vectors for filariasis. They rely heavily on the pharmacological properties of their saliva to get a blood meal and suppress immune reactions of hosts. Little information is available on antihemostatic substances in horsefly salivary glands; especially no horsefly immune suppressants have been reported. By proteomics or peptidomics and coupling transcriptome analysis with pharmacological testing, several families of proteins or peptides, which act mainly on the hemostatic system or immune system of the host, were identified and characterized from 30,000 pairs salivary glands of the horsefly Tabanus yao (Diptera, Tabanidae). They are: (i) a novel family of inhibitors of platelet aggregation including two members, which possibly inhibit platelet aggregation by a novel mechanism and act on platelet membrane, (ii) a novel family of immunosuppressant peptides including 12 members, which can inhibit interferon-gamma production and increase interleukin-10 secretion, (iii) a serine protease inhibitor with 56 amino acid residues containing anticoagulant activity, (iv) a serine protease with anticoagulant activity, (v) a protease with fibrinogenolytic activity, (vi) three families of antimicrobial peptides including six members, (vii) a hyaluronidase, (viii) a vasodilator peptide, which is an isoform of vasotab identified from Hybomitra bimaculata, and interestingly (ix) two metallothioneins, which are the first metallothioneins reported from invertebrate salivary glands. The current work will facilitate the understanding of the molecular mechanisms of the ectoparasite-host relationship and help in identifying novel vaccine targets and novel leading pharmacological compounds.

Distribution, prevalence and intensity of protozoan and monogenean parasites of carp fingerlings in selected nursery ponds

An experiment was carried out on the distribution, prevalence and intensity of infestation of protozoan and monogenean parasites of carp fingerlings in two selected areas of Bangladesh. Six hundred and forty fingerlings of seven species viz. Catla calla, Hypophthalmicthys molitrix, Labeo rohita, Ctenophalyngodon idella, Cyprinus carpio, Barbodes gonionotus and Cirrhinus cirrhosus from different nursery ponds of Shambhuganj, Mymensingh, and Santaher, Bogra were examined during this investigation. This study revealed that carp fingerlings carried a large number of protozoan and monogenean parasites. Different protozoan parasites viz Trichodina domerguei, Trichodina reticulata, Myxobolus koi, Chilodonella cyprini and monogenean parasites such as Dactylogyrus extensus, Dactylogyrus catlarius and Dactylogyroides tripathi were identified in the experimental fishes throughout the study period. It has been observed that distribution, prevalence and intensity of parasite in carp fingerlings are species and zone specific. In case of C. cirrhosus, the highest prevalence of protozoan parasite viz Trichodina domerguei were 93.75% and 75.92% in Santhahar and Shambhuganj area respectively. Irrespective of host, the most prevalent ectoparasite was Dactylogyrus extensus, followed by Myxobolus koi, Chilodonella cyprini and Dactylogyroides tripathi. Relatively higher mortality of carp fingerlings was found at younger stage. The highest prevalent parasitic species was identified as Trichodina domerguei followed by Trichodina reticulata and Dactylogyrus catlarius were found 93.75% in Cirrhinus cirrhosus, 68.75% in Cyprinus carpio and 39.44% in Hypophthalmicthys molitrix respectively.

A note on some parasites of Merluccius capensis (Pisces) and their zoogeographical significance

During the Southeastern Atlantic Expedition of the German fishery research vessel "Walther Herwig" in 1967 the main emphasis lay on selective fishing of the South African hake Merluccius capensis (von BRANDT 1967). Some of the fish were found to be infested by ecto-and endoparasites both of which were collected whenever possible. Large plerocercoids of Dibothriorhynchus grossum whose adult stage lives in the South Atlantic Ocean in Lamna cornubica (L.SZIDAT, personal communication) were quite common as were cysticercoids of a Tetrarhynchus sp., which had also been reported in Cynoscion striatus off the Argentinian coast (MACDONAGH 1927, cited in Szidat, personal communication). Brownish nematodes were infesting the ovaries of several fish, but could not be identified. The most common ectoparasite to be observed was the parasitic isopod Livoneca raynaudii (fam. Cymothoidae) whose early larval stages were also found.

No evidence of host specialization in a parasitic pea-crab exploiting two echinoid hosts

The pinnotherid crab Dissodactylus primitivus lives parasitically on 2 burrowingechinoid species, Meoma ventricosa and Plagiobrissus grandis. The fecundity of female crabsvaries between hosts, and is higher when parasitizing P. grandis than M. ventricosa. Moreover, thehosts present great variations in morphology (size and density of spines). These characteristicssuggest the potential to differentiate crabs according to host species. We investigated the genetic(microsatellites) and morphometric (outline analysis) differentiation of this parasitic crab between2 host species at 1 Jamaican site (Western Lagoon, Discovery Bay), and compared it with geographicdifferentiation among 4 sites along the north coast of Jamaica. Greater genetic differencesbetween parasites of the 2 sympatric hosts than between parasites of a single host at different geographiclocations would indicate host differentiation. Genetic analyses (microsatellites) did notdetect spatial differentiation (probably due to local hydrography) or differentiation according tohost species. This lack of host differentiation could be explained by mobility of adult crabsbetween hosts. However, there was weak but significant morphological differentiation betweenfemale crabs from the 2 hosts. This morphological difference may reflect constraints due to hostmorphology.

Diel variation in egg-laying by the freshwater fish louse Argulus foliaceus (Crustacea : Branchiura)

Removal of deposited eggs could be a useful control strategy for the damaging fish ectoparasite Argulus foliaceus, but focused control requires knowledge of egg-laying patterns. Here, we investigated diel changes in the egg-laying behaviour of a natural population of A. foliaceus. Data were collected from 17-28 May 2004. Days were divided into 3 time periods: 06:00-14:00, 14:00-22:00 and 22:00-06:00 h. Significantly more egg clutches were laid from 06:00-14:00 h than during the other 2 time periods, which were not significantly different from each other. Significantly more egg clutches per hour were laid during hours of daylight as compared to hours of darkness. Significantly more egg clutches were laid in the top 1 m of the water column than at the bottom, and this was consistent throughout all 3 time periods. It is suggested that the increase in egg-laying activity during daylight hours may be due to a higher motivation to search for hosts during the night and an increased ability to locate silhouetted egg-laying sites during the day. These data can provide information useful for egg removal and control strategies.

Seasonal and vertical patterns of egg-laying by the freshwater fish louse Argulus foliaceus (Crustacea : Branchiura)

Argulus foliaceus is a damaging fish ectoparasite for which new control measures are being developed based on egg-removal, In an attempt to develop further understanding of seasonal and vertical egg-laying patterns in this parasite, egg-laying activity was monitored over the period 14 April to 17 November 2003 in 2 rainbow trout Oncorhynchus mykiss fisheries in Northern Ireland, UK. At Site 1, egg-laying was continuous from 21 April to 17 November, when water temperature was above 8 to 10 degrees C. At Site 2, egg-laying was continuous from 4 June to 29 October. In the early months of the season, egg-laying was recorded mainly within the top 1 m of the water column; however, a significant shift to deep water egg-laying was recorded between 7 July and 17 November at Site 1 and between 20 August and 29 October at Site 2. Egg clutches were preferentially laid at depths of up to 8.5 m during this time (Site 2), a feature of egg-laying hitherto unappreciated. Temperature and dissolved oxygen did not differ significantly among depths, but there was an increase in water clarity over time. However, the precise environmental triggers for deep water egg-laying are still unclear. These new insights into the reproductive behaviour of this species will be useful in developing control methods based on egg-removal.

Parasitic foraminifers on a deep-sea chiton (Mollusca, Polyplacophora, Leptochitonidae) from Iceland

Epibiotic foraminifers selectively settle on the most food-rich area of the host substrate, even when the species acts as a facultative ectoparasite in later life stages. In 398 specimens examined of the deep-sea chiton Leptochiton arcticus from Iceland, 46% show evidence of infestation by foraminifers, with many showing extensive shell damage from present and past bioeroding epibionts. Disturbances to the inner layer of the host shell are indicative of parasitism, as evidenced both by wound healing calcification and protrusions of the foraminiferan tubules. The epibionts employ different feeding strategies at different stages of their life cycle, taking advantage of nutrient availability from the posterior respiration currents and excrement of the chitons as juveniles, and feeding parasitically as adults. Epibiont persistence on individual hosts-through successive generations, or long-term continuous bioerosion by epibionts-allow larger adult parasitic foraminifers of Hyrrokkin sarcophaga to penetrate the thick tail valve of a chiton and feed parasitically on the host tissue. The proportion of chitons infested increases with host size, indicating that epibionts are accumulated through a chiton's life, seemingly without major detriment to host survivorship.

Reproductive success of parasitized males in a marine reef fish

Parasitism is hypothesized to reduce reproductive success in heavily parasitized males because females
may preferentially mate with less parasitized males (parasite-mediated sexual selection) or parasites may compromise
male competitiveness. In marine systems, this hypothesis is largely unexplored. This paper provides the first confirmed record of a copepod ectoparasite (Caligus buechlerae Hewitt 1964) on the common triplefin (Forsterygion lapillum) and evaluates the hypothesis that males parasitized with C. buechlerae experience lower reproductive success than unparasitized males (as determined
by the presence and area of eggs within male nests). We found that 38 % of males we surveyed were infected with
at least one C. buechlerae, with a median of two individuals per infected male. About 32 % of males were defending
eggs, with 62.5 % of those males infected with at least one parasite. Males of greater total length (TL) were both
more likely to be infected and more likely to be defending eggs. However, when statistically accounting for the effects
of TL, parasite infection had no effect on the probability of defending eggs, or the average surface area of eggs when
present. Positive covariation in fish length, the presence of eggs and parasite infection observed here potentially suggest
that the importance of parasitic infection on reproductive success may depend upon the strength of selection for larger male body size. Our study is one of the few studies to investigate the effects of ectoparasites on reproductive success in reef fish and also provides a quantitative measure of infection for a widespread species within New Zealand.