Rhipicephalus microplus serine protease inhibitor family: annotation, expression and functional characterisation assessment

Background: Rhipicephalus (Boophilus) microplus evades the host's haemostatic system through a complex protein array secreted into tick saliva. Serine protease inhibitors (serpins) conform an important component of saliva which are represented by a large protease inhibitor family in Ixodidae. These secreted and non-secreted inhibitors modulate diverse and essential proteases involved in different physiological processes. Methods: The identification of R. microplus serpin sequences was performed through a web-based bioinformatics environment called Yabi. The database search was conducted on BmiGi V1, BmiGi V2.1, five SSH libraries, Australian tick transcriptome libraries and RmiTR V1 using bioinformatics methods. Semi quantitative PCR was carried out using different adult tissues and tick development stages. The cDNA of four identified R. microplus serpins were cloned and expressed in Pichia pastoris in order to determine biological targets of these serpins utilising protease inhibition assays. Results: A total of four out of twenty-two serpins identified in our analysis are new R. microplus serpins which were named as RmS-19 to RmS-22. The analyses of DNA and predicted amino acid sequences showed high conservation of the R. microplus serpin sequences. The expression data suggested ubiquitous expression of RmS except for RmS-6 and RmS-14 that were expressed only in nymphs and adult female ovaries, respectively. RmS-19, and -20 were expressed in all tissues samples analysed showing their important role in both parasitic and non-parasitic stages of R. microplus development. RmS-21 was not detected in ovaries and RmS-22 was not identified in ovary and nymph samples but were expressed in the rest of the samples analysed. A total of four expressed recombinant serpins showed protease specific inhibition for Chymotrypsin (RmS-1 and RmS-6), Chymotrypsin / Elastase (RmS-3) and Thrombin (RmS-15). Conclusion: This study constitutes an important contribution and improvement to the knowledge about the physiologic role of R. microplus serpins during the host-tick interaction.

Recombinant mid gut antigen (Bm95) as a vaccine against Indian Rhiphicephalus haemaphysaloides in Bos indicus cattle

In the present study, we report for the first time the efficacy of recombinant Bm95 mid gut antigen isolated from an Argentinean strain of Rhipicephalus microplus strain A in controlling the tick infestations in India. The synthetic gene for Bm95 optimized for expression in yeast was obtained and used to generate yeast transformants expressing Bm95 which was purified to apparent homogeneity. Liquid chromatography-mass spectrometry analysis of the purified protein confirmed its identity as Bm95. Vaccine was prepared by blending various concentrations of purified Bm95 with aluminium hydroxide as an adjuvant. lmmunogenicity studies of the vaccine in rabbits and cattle indicated that the vaccine was highly immunogenic. The efficacy studies of the vaccine was done in cattle. Naive Bos indicus cattle were vaccinated with the recombinant vaccine and were challenged with the larval, nymphal and adult forms of Rhiphicephalus haemaphysaloides. The vaccine protected the animals from larval, nymph and adult tick challenges with an efficacy of 98.7%, 84.6% and 78.9% respectively. The results obtained from the above studies clearly demonstrated the advantage and possibilities of the use of Bm95 in controlling R. haemaphysaloides infestations in the field. (C) 2010 Elsevier Ltd. All rights reserved.

Preliminary studies on food and feeding habits of Polypterus endlicheri and Polypterus senegalus in Lake Chad

The food and feeding habits of Polyprerus cncllicheri and Polypterus senegalus was carried out in the months of September to October. The food of 33 Polypierus endlicheri as observed include Tilapia species (89.3%), Eutropius niloticus (28.6%), Mayfly nymph (39.3%), Dragon fly larva (56.6%) fish remains (21.4%) and detritus (7.1%). The food of27 Polypterus senegalus as observed include Tilapia sp (88.4%), Eutropius niloticus (27.9%), may fly nymph (23.3%), Dragonfly nymph (34.9%) remains (21.1%) detritus (23.3%). (9 page document) The percentage occurrence of food item found in the stomach of Polypterus endlieheri is 93.3% while that of Polyprerus senegalus is 67.4%. The dominance of Tilapia sp was establish in the study, and there is no significant difference between the feeding habit of Polypterus endlicheri and Polyprerus senegalus.

The functional role of Gammarus (Crustacea, Amphipoda): shredders, predators, or both?

Gammarus spp. are traditionally viewed under the functional feeding group (FFG) concept as herbivorous 'shredders'. Although recent studies suggest that Gammarus should also be viewed as predators, this latter role remains contentious. Here, in a laboratory experiment, we objectively examine the balance between shredder and predator roles in a common freshwater species. Gammarus pulex preyed significantly on mayfly nymph, Baetis rhodani, in both the presence and absence of excess leaf material. There was no significant difference in predation where the alternative food, that is, leaf material, was present as compared to absent. Also, G. pulex shredded leaf material in the presence and absence of B. rhodani. However, shredding was significantly reduced where alternative food, that is, B. rhodani prey, was present as compared to absent. Further, G. pulex had a clear leaf species preference. Our results suggest that Gammarus function as both predators and shredders, with the balance of the two roles perhaps depending on food availability and quality. We discuss implications for the use of the FFG concept in assessing freshwater processes, and the role that Gammarus predation may play in structuring macroinvertebrate communities.

Sexual behavior and factors affecting female reproduction in house and field crickets

The objective of this investigation was to clarify the adaptive significance of female sexual behaviours in the house cricket, Acheta domesticus, and the Texas field cricket, Gryllus integer. Experiments were focussed primarily on: nutritional factors affecting female reproductive success; the ontogeny of female sexual behaviours; female mating frequency and progeny production; and the pattern of sperm competition. Reproduction of singly mated female A. domesticus assigned to 3 nutritional regimes was compared . Females fed a vitamin and protein-enriched mouse chow, cannibalistic females, and starved females produced on the average, 513 , 200 and 68 offspring, respectively. Cannibals probably could not obtain the same amounts of essential nutrients as females fed mouse chow. Reabsorption of oocytes was likely the major factor contributing to the decreased reproduction of starved females. In addition, female !. domesticus fed mouse chow, but allowed constant access to males produced 11 times as many offspring than did females fed corn meal. Females fed corn meal probably could not absorb or synthesize enough dietary lipids, thus resulting in poor ovariole growth. Female !. domesticus first mate at an average adult age of 7 days, closely corresponding to when they first exhibit positive phonotaxis. Females mate repeatedly and often consume the externally attached spermatophore. In ~. domesticus, females allowed constant access to males produced significantly more offspring than did single maters. Similarly, doubly mated G. integer females produced more offspring than did single maters. This difference resulted largely from the failure of many single maters to reproduce. Remating by female crickets partly functions in offsetting the possibility of a failed initial mating. Nymph production increased significantly with the time the spermatophore was attached in singly mated ~. domesticus. Spermatophore consumption by the female was not affected by male guarding behaviour, and the interval between mating and eating of the spermatophore may often be shorter than the time required for maximum insemination. Some degree of sperm depletion in singly mated !. domesticus and G. integer may have occurred. The patterns of daily offspring production of singly and multiplymated females suggests that a factor provided by a male during mating stimulates female oviposition and/or egg production. Female crickets also might acquire nutrition from spermatophore consumption, a benefit that is augmented by female multiple mating. The electrophoretic examination of various allozymes in ~. integer did not permit determination of a pattern of sperm competition. However, the possibility of last male sperm predominance is related to male guarding behaviour.

Independent Chronicle Vol. XLVI No. 3470- July 14, 1814

A semi-weekly paper that was published from 1801 to 1817. It was previously called the Independent chronicle and the universal advertiser, and was later the Independent chronicle and Boston patriot (Semiweekly). Publishers were Abijah Adams and Ebenezer Rhoades. Topics of interest in this issue include: Page 1: report of Independence Day celebration including multiple toasts to the military; Page 2: report of South Carolina militia marching home; report of U.S. schooner Alligator being lost to a tornado in Port Royal; report of 10,000 militia and 2,000 regular Army troops to be used in defense of the District of Columbia and vicinity; report U.S. is to try to raise 100,000 militia for defense; news of British deserters providing information to the U.S.; report of skirmishes along the U.S. coast; statement from U.S. Navy Capt. David Porter stating he had taken possession of Sir Henry Martin's Island; report of British troops burning a militia barracks in Lewiston, N.Y.; 4 accounts of the battle of Chippewa and the taking of Fort Erie; statement from Maj. Gen. Brown in praise of his troops in the battle of Chippewa; copy of General Brown's orders to attack at Fort Erie; essay arguing against U.S. citizens boarding enemy ships; report of peace treaty rumours; Page 3: account of Capt. Porter's capture and subsequent escape from the British; list of American prisoners held on the British ship Nymph; report of British cargo ship captured by U.S. war ship; Proclamation from James Madison pardoning military deserters if they return to military service; announcement to officers and soldiers in the military offering reward for the return of military deserters; Page 4: 2 reward advertisements for return of military deserters;

Maid of the Mist, Niagara Falls, U.S.A.

The image is described as "(5) 'Maid of the Mist' - Nymph of the mighty cataract, Niagara Falls, U.S.A.".

Comparison of two methods for collecting free-living ticks in the Amazonian forest

In the present study, we compared 2 methods for collecting ixodid ticks on the verges of animal trails in a primary Amazon forest area in northern Brazil. (i) Dragging: This method was based on passing a 1-m(2) white flannel over the vegetation and checking the flannel for the presence of caught ticks every 5-10 m. (ii) Visual search: This method consisted of looking for guesting ticks on the tips of leaves of the vegetation bordering animal trails in the forest. A total of 103 adult ticks belonging to 4 Amblyomma species were collected by the visual search method on 5 collecting dates, while only 44 adult ticks belonging to 3 Amblyomma species were collected by dragging on 5 other collecting dates. These values were statistically different (Mann-Whitney Test, P = 0.0472). On the other hand, dragging was more efficient for subadult ticks, since no larva or nymph was collected by visual search, whereas 18 nymphs and 7 larvae were collected by dragging. The visual search method proved to be suitable for collecting adult ticks in the Amazon forest: however, field studies should include a second method, such as dragging in order to maximize the collection of subadult ticks. Indeed, these 2 methods can be performed by a single investigator at the same time, while he/she walks on an animal trail in the forest. (C) 2010 Elsevier GmbH. All rights reserved.

Baltic winter (amped folk instruments-ethereal and haunting with female vocals)

Baltic Winter - a Slavic mermaid rests upon a rock on the edge of the Baltic Sea. It is night time during the depths of winter. She risks not being able to return to the sea as it could freeze in the time that she drapes the snow crested pine trees with her ethereal calls. Her voice travels through the Medieval forest in search of her nymph . . . will he hear her? Or will her calls only serve to sing her loneliness?

OK . . . now for the technical . . . I wrote this with a documentary in mind. I have used a sampled Finnish/Estonian Kantele, a touch of an acoustic guitar, female vocals, percussion, and a couple of synths.

Baltic winter (beautiful ethereal sweeping music with haunting female vocals)

Baltic Winter - a Slavic mermaid rests upon a rock on the edge of the Baltic Sea. It is night time during the depths of winter. She risks not being able to return to the sea as it could freeze in the time that she drapes the snow crested pine trees with her ethereal calls. Her voice travels through the Medieval forest in search of her nymph . . . will he hear her? Or will her calls only serve to sing her loneliness?

OK . . . now for the technical . . . I wrote this with a documentary in mind. I have used a sampled Finnish/Estonian Kantele, a touch of an acoustic guitar, female vocals, percussion, and a couple of synths.

Baltic winter (under 1 minute haunting female vocals and ethereal)

Baltic Winter Shorts 02 - a Slavic mermaid rests upon a rock on the edge of the Baltic Sea. It is night time during the depths of winter. She risks not being able to return to the sea as it could freeze in the time that she drapes the snow crested pine trees with her ethereal calls. Her voice travels through the Medieval forest in search of her nymph . . . will he hear her? Or will her calls only serve to sing her loneliness?

OK . . . now for the technical . . . I wrote this with a documentary in mind. I have used a sampled Finnish/Estonian Kantele, a touch of an acoustic guitar, female vocals, percussion, and a couple of synths.

Baltic winter (under 1 minute haunting female vocals and ethereal)

Baltic Winter Short 01 - a Slavic mermaid rests upon a rock on the edge of the Baltic Sea. It is night time during the depths of winter. She risks not being able to return to the sea as it could freeze in the time that she drapes the snow crested pine trees with her ethereal calls. Her voice travels through the Medieval forest in search of her nymph . . . will he hear her? Or will her calls only serve to sing her loneliness?

OK . . . now for the technical . . . I wrote this with a documentary in mind. I have used a sampled Finnish/Estonian Kantele, a touch of an acoustic guitar, female vocals, percussion, and a couple of synths.

Baltic winter 42 seconds ethereal sweeping and haunting with female vocals

Baltic Winter Shorts 03 - a Slavic mermaid rests upon a rock on the edge of the Baltic Sea. It is night time during the depths of winter. She risks not being able to return to the sea as it could freeze in the time that she drapes the snow crested pine trees with her ethereal calls. Her voice travels through the Medieval forest in search of her nymph . . . will he hear her? Or will her calls only serve to sing her loneliness?

OK . . . now for the technical . . . I wrote this with a documentary in mind. I have used a sampled Finnish/Estonian Kantele, a touch of an acoustic guitar, female vocals, percussion, and a couple of synths.

Baltic winter ethereal cinematic and haunting

Baltic Winter (No Vocals - a Slavic mermaid rests upon a rock on the edge of the Baltic Sea. It is night time during the depths of winter. She risks not being able to return to the sea as it could freeze in the time that she drapes the snow crested pine trees with her ethereal calls. Her voice travels through the Medieval forest in search of her nymph . . . will he hear her? Or will her calls only serve to sing her loneliness?

OK . . . now for the technical . . . I wrote this with a documentary in mind. I have used a sampled Finnish/Estonian Kantele, a touch of an acoustic guitar, percussion, and a couple of synths.

Comparative biology and production costs of Podisus nigrispinus (Hemiptera: Pentatomidae) when fed different types of prey

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)