Cultivation and molecular identification of Ehrlichia canis and Ehrlichia chaffeensis from a naturally co-infected dog in Venezuela

Background: Diagnosis of canine ehrlichiosis in Venezuela is normally performed by examination of buffy coat smears (BCS). Characteristic inclusion bodies are frequently observed in leukocytes and platelets from dogs with clinical signs of the disease. Objective: The purpose of this study was to investigate the co-infection of a dog with Ehrlichia canis and E hrlichia chaffeensis using microbiological and molecular techniques. Methods: Primary cultures of monocytes from a dog showing signs of ehrlichiosis were performed. Ehrlichial inclusions in blood cells were demonstrated by BCS and in cultured cell smears with direct immunofluorescence and Dip Quick staining. Nested PCR analysis was performed with DNA from blood samples and cultures, using primers specific for E. canis and E. chaffeensis. The amplified DNA fragments were sequenced to confirm the specificity of the amplifications. Results: The BCS of the naturally infected dog contained intracellular morulae. Ehrlichial inclusions were observed 9 days after inoculation of the primary cultures. After 3 passages with monocytes from a healthy dog, 65% of infected cells, and cells with >60 morulae were observed. A healthy female German Shepherd dog, seronegative for E. canis and E. chaffeensis antigens and without contact to ticks, was inoculated with an infected culture. The animal developed signs of canine monocytic ehrlichiosis and became seropositive. Nested PCR results and sequencing of amplified DNA fragments demonstrated the simultaneous presence of E. canis and E. chaffeensis in both dogs. Conclusions: This is the first report of E. chaffeensis in dogs in South America. This organism was previously identified in dogs by PCR only in the United States.     

Quantification of regulating ecosystem services provided by weeds in annual cropping systems using a systematic map approach

Ecosystem services have received increasing attention in life sciences, but only a limited amount of quantitative data are available concerning the ability of weeds to provide these services. Following an expert focus group on this topic, a systematic search for articles displaying evidence of weeds providing regulating ecosystem services was performed, resulting in 129 articles. The most common service found was pest control and the prevailing mechanism was that weeds provide a suitable habitat for natural enemies. Other articles showed that weeds improved soil nutrient content, soil physical properties and crop pollinator abundance. Weeds were found to provide some important ecosystem services for agriculture, but only a small number of studies presented data on crop yield. Experimental approaches are proposed that can: (i) disentangle the benefits obtained from ecosystem services provisioning from the costs due to weed competition and (ii) quantify the contribution of diverse weed communities in reducing crop competition and in providing ecosystem services. Existing vegetation databases can be used to select weed species with functional traits facilitating ecosystem service provisioning while having a lower competitive capacity. However, for services such as pest control, there are hardly any specific plant traits that have been identified and more fundamental research is needed.     

A Case of Hydrochemical Characterization of an Alluvial Aquifer Influenced by Human Activities

21 physico-chemical parameters have been determined in 47 groundwater samples collected in October 1994 from the alluvial aquifer of the Pisuerga river, located at the north-east of Valladolid (north-western Spain). Conventional hydrochemical methods and multivariate statistical analysis have been used to discriminate natural phenomena and pollution processes in the area. It has been demonstrated the existence of a heterogeneity in groundwater mineral contents related to anthropic actions such as irrigation, source of irrigation water and location of polluting sources. This study demonstrates the existence of two sub-units in the alluvial aquifer (left and right river banks) with different hydrochemical characteristics, and identifies three different singular areas in the left bank. Groundwater from this bank showed higher levels of dissolved ions caused by concentration processes derived from the predominant use of groundwater for crop irrigation, whilst in the right bank of the aquifer less saline water from canals is used. Degradation of groundwater quality has been observed in some areas of the left aquifer sub-unit (north, Santovenia and south-central sites) where anomalous high concentrations of sulphate, sodium, chloride and nitrate were measured, and can be related to the presence of potentially polluting sources such as leachates from industrial waste disposals, sewage effluents and use of organic and inorganic fertilizers.     

Recent advances in lipid nutrition in fish larvae

Due to the importance of dietary lipid utilization for larval rearing success, increasing attention has been paid during the last years to different aspects of larval lipid nutrition such as digestion, absorption, transport and metabolism, which are frequently studied by different research groups. The present study reviews the published information on these aspects, including some recent results obtained in our laboratory, that contribute to a better understanding of larval lipid nutrition.Neutral lipase activity was found in the digesta of larval gilthead seabream as early as first feeding, followed by a significant increase which reached up 8 times the initial levels at day 15 and was clearly influenced by the fatty acid composition of dietary lipids. Accordingly, the capacity for lipid absorption by the intestinal epithelium has been also observed at the onset of exogenous feeding, although the specific location in the different digestive tract segments differ with species. Whereas the capacity to absorb lipid increases with development in live prey-fed larvae, this improvemment is delayed in larvae fed formulated diet. Increasing dietary phosphatidyl cholines levels enhanced lipid absorption regardless of whether it is of soybean or marine origin, but the latter improved hepatic lipid utilization. Enzymatic, histological and biochemical evidences suggest that marine fish larvae are able to effectively digest and absorb n-3 HUFA-rich triacylglycerols, but feeding with phosphoacylglycerols, particularly if they are rich in n-3 HUFA, would enhance phosphoacylglycerols digestion and specially lipid transport alowing a better n-3 HUFA incorporation into larval membrane lipids and promoting fish growth. Although the essentiality of n-3 HUFA for larval marine fish has been studied extensively, only recently has the importance of dietary arachidonic acid in the larvae of few species been recognised. Evidences for competitive interactions among these essential fatty acids suggest that besides a minimum dietary requirement for each essential fatty acid, their relative ratios must also be considered.     

Two microalgae Crypthecodinium cohnii and Phaeodactylum tricornutum as alternative source of essential fatty acids in starter feeds for seabream (Sparus aurata)

Despite oils extracted from algae and other microorganisms that may constitute excellent sources of HUFAs, few studies have determined the nutritional value of different microalgal species for young marine fish. Six thousand gilthead seabream (Sparus aurata) postlarvae (73 mg body weight) were fed for 57 days diets containing either fish oil as a single lipid source or 2 and 4% of Cryptecodinum cohnii or 5% Phaeodactylum tricornutum. Fish oil substitution by C. cohnii resulted in improved fish survival and a very good growth performance, in agreement with a higher proportion of DHA in diets and in total lipids of fish. Incorporation of DHA and other fatty acids was proportional to their contents in diet suggesting the good nutritional utilization of homogenized C. cohnii. Lower survival rates were found in fish fed P. tricornutum and could be related to an epithelial degeneration observed in the anterior intestine. This degeneration could be related to a higher lipid content in these fish or to the strong hornlike cornutate processes found in the valves of the diatom P. tricornutum.     

Incidence of systemic granulomatosis is modulated by the feeding sequence and type of enrichment in meagre (Argyrosomus regius) larvae

Systemic granulomatosis is the most frequent disease in juvenile and adult meagre, but studies regarding the first appearance of granulomas in larvae do not exist. In order to evaluate this, meagre larvae were fed four different feeding regimes as follows: RS and RO (rotifer enriched with Easy DHA Selco or Ori‐Green from 3 to 30 dph respectively), RAS and RAO (rotifer enriched with Easy DHA Selco or Ori‐Green from 3 to 21 dph and Artemia enriched with Easy DHA Selco or Ori‐Green from 12 to 30 dph respectively). All treatments were also fed with commercial microdiet from 20 to 30 dph. At 30 dph weight, length, specific growth rate and survival were significantly higher in Artemia‐fed larvae, regardless of the enrichment. Microscopic first appearance of granulomas was observed in 20 dph larvae fed RS and RO. At 30 dph granulomas and thiobarbituric acid reactive substances (TBARS), values were significantly higher in RS and RO‐fed larvae than in RAS and RAO‐fed larvae. The results showed that granulomas first appeared in meagre larvae at 20 dph when fed rotifers only. Conversely, a reduced appearance of granulomas and lipid peroxidation occurs when Artemia is included in the feeding sequence reinforcing the hypothesis of a nutritional origin of the systemic granulomatosis.     

The determination of lipase and phospholipase activities in gut contents of turbot (Scophthalmus maximus) by fluorescence-based assays

To study the potential of fluorescence based assays in the study of lipid digestion in fish, acyl esters of 4-methylumbelliferone and 1-acyl-2-[6 (7 nitro-1,3 benzoxadiazol-4-yl)amino]caproyl labeled phosphatidylcholine compounds (NBD-PC) were used as substrates for the assay of neutral lipase and phospholipase, respectively, in the gut contents of turbot. 4-Methylumbelliferyl hepatanoate (4-MUH) was hydrolysed at a higher rate than the butyrate or oleate esters whilst the hexanoic (C6) ester of NBD-PC was a more convenient substrate for the phospholipase assay than the dodecanoic (C12) ester. Neutral lipase activity was almost 10% higher when 50 mm potassium phosphate buffer pH 7.8 was used instead of 0.01 m citrate/sodium phosphate buffer pH 7.2. Both assays were very sensitive: neutral lipase and phospholipase activities were detectable at a minimum protein concentration in the digesta of 0.04 and 1.25 mg/ml, respectively. When the variations in lipolytic activities with gut segment and with size of fish were examined neutral lipase activity was always found to be higher in the hindgut and rectum segments than in the foregut. Although phospholipase activity was also found to be highest in the hindgut of the largest fish examined (av. wt. 182.3g), in fish of average weight 8g fish the activity was similar in all three segments. In the digesta from the whole gut of smaller fish (av. wt. 0.2, 0.6 and 1.43g) neutral lipase and phospholipase activities increased with increasing body mass when expressed as per ml of digesta. It is concluded that fluorescence-based assays are applicable to the study of lipid digestion in fish of different size.     

Puffins reveal contrasting relationships between forage fish and ocean climate in the North Pacific

Long‐term studies of predator food habits (i.e., ‘predator‐based sampling’) are useful for identifying patterns of spatial and temporal variability of forage nekton in marine ecosystems. We investigated temporal changes in forage fish availability and relationships to ocean climate by analyzing diet composition of three puffin species (horned puffin Fratercula corniculata, tufted puffin Fratercula cirrhata, and rhinoceros auklet Cerorhinca monocerata) from five sites in the North Pacific from 1978–2012. Dominant forage species included squids and hexagrammids in the western Aleutians, gadids and Pacific sand lance (Ammodytes personatus) in the eastern Aleutians and western Gulf of Alaska (GoA), and sand lance and capelin (Mallotus villosus) in the northern and eastern GoA. Interannual fluctuations in forage availability dominated variability in the western Aleutians, whereas lower‐frequency shifts in forage fish availability dominated elsewhere. We produced regional multivariate indicators of sand lance, capelin, and age‐0 gadid availability by combining data across species and sites using Principal Component Analysis, and related these indices to environmental factors including sea level pressure (SPL), winds, and sea surface temperature (SST). There was coherence in the availability of sand lance and capelin across the study area. Sand lance availability increased linearly with environmental conditions leading to warmer ocean temperatures, whereas capelin availability increased in a non‐linear manner when environmental changes led to lower ocean temperatures. Long‐term studies of puffin diet composition appear to be a promising tool for understanding the availability of these difficult‐to‐survey forage nekton in remote regions of the North Pacific.     

Cuphea growth, yield, and oil characteristics as influenced by climate and soil environments across the upper Midwest USA

Cuphea is a potential new oilseed crop rich in medium-chain fatty acids (C8:0 to C14:0) that may serve as a renewable, biodegradable source of oil for lubricants, motor oil, and aircraft fuel. Impacts of climate and soil environment on cuphea growth and development are not well understood. The objective of this study was to evaluate the influence of climate and soil on growth, seed yield, and seed oil characteristics of two semi-domesticated cuphea genotypes [PSR23 and HC-10 (Cuphea viscosissima Jacq. × C. lanceolata W.T. Aiton)] and three wild species [Cuphea wrightii, Cuphea lutea, and C. viscosissima (VS-6-CPR-1)] that show potential for domestication. The study was conducted in 2007 and 2008 at field sites in North Dakota (ND), Minnesota (MN), Iowa (IA), and Illinois (IL). Cuphea PSR23 and HC-10 were direct seeded in the field, while the three wild species were transplanted. The two plantings were treated as separate experiments. Plant growth, seed yield and oil content for the two direct-seeded lines tended to be distinctly greater in MN and ND than IL and IA, which was related more to growth temperature than soil environment. The three wild species generally performed similarly across the four different environments. C. wrightii had the greatest oil content, ranging from 320 to 360 g kg⁻¹, which was comprised of 59-64% lauric acid. For each genotype, the content of its most prominent saturated medium-chain fatty acid (e.g., C10:0 or C12:0) increased with decreasing latitude of field site. Seed yields for C. wrightii and C. lutea were as high as 1116 kg ha⁻¹. Combined with relatively high seed oil contents (280-350 g kg⁻¹) these species may be good candidates for domestication. Results indicate that PSR23 and HC-10 are more regionally adapted than the wild species studied, which tended to exhibit a greater range of adaptability to climate and soil conditions.     

Biological performance of Octopus vulgaris in ‘integrated aquaculture’ models: effect of two potential fresh diets,sex and sexual maturation during the grow‐out phase

Two fresh diets were tested on the biological performance of males and females of Octopus vulgaris under industrial rearing conditions in floating cages. The presence of females could potentially release paralarvae to the natural environment (‘integrated aquaculture’). Octopuses were fed on a single diet of bogue (aquaculture by‐product), and on a mixed diet (60–40% crab‐bogue), in two trials. In Trial 1 (918 ± 125 g), growth was higher in octopuses fed on the mixed diet (1.9–2.0%d^?1) than in those fed on the bogue diet (1.8–1.9%d^?1) irrespective of sex, and mortality was 3% regardless of sex and diet. In Trial 2 (1483 ± 269 g), growth was higher in males fed on the mixed diet (1.8%d^?1) than in males fed on the bogue diet (1.4%d^?1), both higher than females (1.1–1.3%d^?1), and mortality was 22–28% regardless of sex and diet. Best biological performance in trial 1 was related to the lack of functional maturation in females, evidence by the low average gonadosomatic index (2.0–3.6%) in comparison with trial 2 (8.8–11.4%), regardless of diet. No correlations between final weight and sexual maturity indices were found, so the length of the rearing period to maximize biomass increment or to release paralarvae depends on functional maturation in females.