Efficacy of levamisole, ivermectin and moxidectin against Capillaria spp. in European hedgehogs (Erinaceus europaeus).

This randomised study aimed to assess and compare the efficacy of treatment protocols containing levamisole, ivermectin, or moxidectin against Capillaria spp. in naturally infected European hedgehogs (Erinaceus europaeus) presented to a British wildlife rehabilitation centre. Faecal analysis, consisting of wet mount and flotation, was performed for 229 hedgehogs weighing ≥200g. Animals testing positive for Capillaria spp. (81%), excluding pregnant females, were randomly allocated a treatment protocol. Initially, hedgehogs (n = 50) received one of six 'pilot' protocols, whereas the remaining animals (n = 97) received one of three 'main' protocols. Faecal analysis was repeated on day 8 and day 12 after treatment initiation. Efficacy of each treatment was assessed based on Capillaria reduction rate (CRR), weight gain, presence of respiratory clinical signs, and outcome. Pilot protocols containing only moxidectin had a significantly lower CRR (≥28.1%) compared to those with levamisole or ivermectin (≥86.6%), whereas the main protocols containing levamisole had a significantly higher CRR (≥93.0%) compared to those containing only ivermectin (≥69.3%). Clinical parameters did not differ significantly between treatments, but animals with respiratory clinical signs at the end of the trial were significantly more likely to have lower CRR and test positive for Crenosoma striatum. C. striatum often appeared refractory to treatment, and managing these infections requires additional anthelmintic therapy. Based on the formulations and dosages trialled, moxidectin is not recommended for treating capillariosis in European hedgehogs, whereas levamisole given orally for two consecutive days at 25-35 mg/kg is suggested as the treatment of choice.

From Materials to Cell: State-of-the-Art and Prospective Technologies for Lithium-Ion Battery Electrode Processing.

Electrode processing plays an important role in advancing lithium-ion battery technologies and has a significant impact on cell energy density, manufacturing cost, and throughput. Compared to the extensive research on materials development, however, there has been much less effort in this area. In this Review, we outline each step in the electrode processing of lithium-ion batteries from materials to cell assembly, summarize the recent progress in individual steps, deconvolute the interplays between those steps, discuss the underlying constraints, and share some prospective technologies. This Review aims to provide an overview of the whole process in lithium-ion battery fabrication from powder to cell formation and bridge the gap between academic development and industrial manufacturing.

Aqueous Ni-rich-cathode dispersions processed with phosphoric acid for lithium-ion batteries with ultra-thick electrodes.

Lithium-ion battery (LIB) production can benefit both economically and environmentally from aqueous processing. Although these electrodes have the potential to surpass electrodes conventionally processed with N-methyl-2-pyrrolidone (NMP) in terms of performance, significant issues still exist with respect to ultra-thick cathodes (≫4 mAh/cm2 areal capacities). A major concern for these types of electrodes with high-nickel active material stems from lithium leaching from active material, which drives the pH of the dispersion in excess of 12 and subsequently corrodes the current collector interface. As this corrosion reaction proceeds, hydrogen generation at the interface creates bubbles which cause severe cracking in the dried electrode surface. When areal loadings are increased, this effect becomes more pronounced and is detrimental to both mechanical and electrochemical properties of these electrodes. Herein, a technique for mitigating corrosion at the current collector by adjusting the pH of the dispersion with the addition of phosphoric acid is investigated. Phosphoric acid was added in 0.5 wt% increments between 0.0 and 1.5 wt%, and effects on rheology, adhesion, corrosion, and electrochemical performance were investigated. A technique is reported for producing aqueous processed cathodes with areal loadings of 6-8 mAh/cm2 with reduced surface cracking and superior high-rate discharge capacity (i.e. high-power performance) for this class of cathode loadings.

Administration of an oxytocin receptor antagonist attenuates sexual motivation in male rats.

In male rats, oxytocin impacts both sexual arousal and certain types of consummatory sexual behaviors. However, the role of oxytocin in the motivational aspects of sexual behavior has received limited attention. Given the role that oxytocin signaling plays in consummatory sexual behaviors, it was hypothesized that pharmacological attenuation of oxytocin signaling would reduce sexual motivation in male rats. Sexually experienced Long-Evans male rats were administered either an oxytocin receptor antagonist (L368,899 hydrochloride; 1mg/kg) or vehicle control into the intraperitoneal cavity 40min prior to placement into the center chamber of a three-chambered arena designed to assess sexual motivation. During the 20-minute test, a sexually experienced stimulus male rat and a sexually receptive stimulus female rat were separately confined to smaller chambers that were attached to the larger end chambers of the arena. However, physical contact between test and stimulus rats was prevented by perforated dividers. Immediately following the sexual motivation test, test male rats were placed with a sexually receptive female to examine consummatory sexual behaviors. Although both drug and vehicle treated rats exhibited a preference for the female, treatment with an oxytocin receptor antagonist decreased the amount of time spent with the female. There were no differences between drug and vehicle treated rats in either general activity, exploratory behaviors, the amount of time spent near the stimulus male rat, or consummatory sexual behaviors. Extending previous findings, these results indicate that oxytocin receptors are involved in sexual motivation in male rats.

The role of estrogen G-protein coupled receptor 30 (GPR30) and sexual experience in sexual incentive motivation in male rats.

Male rats exhibit reductions in sexual motivation following systemic administration of drugs that inhibit the conversion of testosterone to estrogen, which indicates that estrogen signaling plays a role in male rat sexual motivation. Given that estrogen G-protein coupled receptor 30 (GPR30) is expressed in brain areas that are important for male sexual behaviors and endocrine function, the primary aim of the current study was to examine the role that GPR30 plays in sexual motivation in both sexually naïve and sexually experienced male rats. Following the final treatment with either a GPR30 antagonist (G-15) or vehicle control, male rats were placed into the center chamber of a larger three-chambered testing arena that was designed to assess sexual incentive motivation. A sexually receptive stimulus female rat and a stimulus male rat were individually confined to one of the two smaller chambers that were each separated by a perforated partition from the larger end chambers, which test rats had access to. Relative to vehicle treated rats, male rats treated with G-15 exhibited a reduction in the percentage of time spent in the vicinity of a sexually receptive female rat. Although G-15 reduced sexual incentive motivation independent of sexual experience, only sexually-naïve rats treated with G-15 did not exhibit a preference for the sexually receptive stimulus female rat. Collectively, these results indicate that interference with estrogen signaling at GPR30 reduces sexual motivation and that the lack of preference for a sexually receptive female rat over a male rat following G-15 treatment is abrogated by previous sexual experience.

Impact of sustained use of insecticide-treated bednets on malaria vector species distribution and culicine mosquitoes.

Insecticide-treated bednets (ITNs) significantly reduce malaria vector populations. Susceptibility to ITNs differs by vector species, and culicine mosquitoes have not been shown to be significantly affected by the use of ITNs. We examined the impact of 2-4 yr of ITN use on malaria vector species distribution and culicine mosquitoes. Routine entomological surveillance was conducted in adjacent areas with and without ITNs from November 1999 to January 2002. Use of ITNs reduced the proportion of Anopheles gambiae Giles relative to Anopheles arabiensis Giles. The number of culicines per house was significantly lower in the ITN area than in the neighboring area. Changes in the An. gambiae sibling species distribution may help to explain apparent mosquito behavioral changes attributed to ITNs. Reductions in culicines by ITNs may have implications for community perceptions of ITN effectiveness and for control of other diseases such as lymphatic filariasis.

Progression of stunting and its predictors among school-aged children in western Kenya.

OBJECTIVE: The objectives of this study were (1) to assess whether a cohort of school-aged children experiences progression of stunting over a 2-y-period of observation and (2) to identify baseline nutritional and body composition risk factors for the progression of stunting. METHODS: As part of a large-scale, randomized controlled trial assessing the impact of insecticide-treated bednets (ITNs) on nutritional status, we longitudinally followed a cohort of school-aged children over a 2-y-period in western Kenya. Anthropometric measurements were made at four time points from which Z-scores for height-for-age (HAZ), weight-for-age (WAZ), and body mass index (BMIZ) were calculated. Two measures of body composition, upper arm fat area and upper arm muscle area, were derived from mid-upper arm circumference (MUAC) and triceps skinfold thickness. RESULTS: Subjects experienced a mean change in HAZ from baseline to 9 months of -0.16 [-0.19, -0.13], from baseline to 16 months of -0.18 [-0.22, -0.15], and from baseline to 24 months of -0.36 [-0.41, -0.31]. Thus, the average individual's change in HAZ at the three follow-up time points is significantly less than zero, meaning that, on average, the cohort is deviating further from NCHS reference medians over time. The baseline nutritional measure that explained the greatest amount of variance in the progression of stunting was the upper arm muscle area Z-score (F=8.1; P=0.005). CONCLUSIONS: This longitudinal study provides further evidence from a distinct ecological setting regarding the progression of undernutrition during middle childhood in the developing world. It suggests that school-aged children in the developing world do not experience catch-up growth or remain stable. Rather, they continue to deviate from NCHS standards, accruing greater height deficits with age. In addition, absolute lean body mass explained the most variability in the progression of stunting, which supports cross-sectional findings from other studies.

Population Structure of Anopheles gambiae in Africa.

The population structure of Anopheles gambiae in Africa was studied using 11 microsatellite loci in 16 samples from 10 countries. All loci are located outside polymorphic inversions. Heterogeneity among loci was detected and two putative outlier loci were removed from analyses aimed at capturing genome-wide patterns. Two main divisions of the gene pool were separated by high differentiation (F(ST) > 0.1). The northwestern (NW) division included populations from Senegal, Ghana, Nigeria, Cameroon, Gabon, Democratic Republic of Congo (DRC), and western Kenya. The southeastern (SE) division included populations from eastern Kenya, Tanzania, Malawi, and Zambia. Inhospitable environments for A. gambiae along the Rift Valley partly separate these divisions. Reduced genetic diversity in the SE division and results of an analysis based on private alleles support the hypothesis that a recent bottleneck, followed by colonization from the NW populations shaped this structure. In the NW division, populations possessing the M rDNA genotype appeared to form a monophyletic clade. Although genetic distance increased with geographic distance, discontinuities were suggested between certain sets of populations. The absence of heterozygotes between sympatric M and S populations in the DRC and the high differentiation in locus 678 (F(ST)>0.28) contrasted with low differentiation in all other loci (-0.02

Extent of digestion affects the success of amplifying human DNA from blood meals of Anopheles gambiae (Diptera: Culicidae).

The success of distinguishing blood meal sources of Anopheles gambiae Giles through deoxyribonucleic acid (DNA) profiling was investigated by polymerase chain reaction (PCR) amplification at the TC-11 and VWA human short tandem repeats (STR) loci. Blood meal size and locus had no significant effect on the success of amplifying human DNA from blood meals digested for 0, 8, 16, 24 and 32 h (P = 0.85 and 0.26 respectively). However, logistic regression found a significant negative relationship between time since ingestion and the success probability of obtaining positive PCR products among meals digested for between 8 and 32 h (P = 0.001). Approximately 80% of fresh blood meals were successfully profiled. After 8 h, the proportion of blood meals that could be successfully profiled decreased slowly with time after ingestion, dropping to below 50% after approximately 15 h. There was no significant difference in the success of amplifying human DNA from blood meals of mosquitoes killed at time 0 and 8 h after ingestion (P = 0.272).

Plasmodium falciparum genotypes, low complexity of infection, and resistance to subsequent malaria in participants in the Asembo Bay Cohort Project.

To assess the relationship between the within-host diversity of malaria infections and the susceptibility of the host to subsequent infection, we genotyped 60 children's successive infections from birth through 3 years of life. MSP-1 Block2 genotypes were used to estimate the complexity of infection (COI). Malaria transmission and age were positively associated with the number of K1 and Mad20 alleles detected (COI(KM)) (P < 0.003). Controlling for previous parasitemia, transmission, drug treatment, parasite density, sickle cell, and age, COI(KM) was negatively correlated with resistance to parasitemia of > 500/microl (P < 0.0001). Parasitemias with the RO-genotype were more resistant than those without this genotype (P < 0.0000). The resistance in low COI(KM) infections was not genotype specific. We discuss the impact of genotype-transcending immunity to conserved antigenic determinants. We also propose a diversity-driven immunomodulation hypothesis that may explain the delayed development of natural immunity in the first few years of life and suggest that interventions that decrease the COI(KM) could facilitate the development of protective immunity.

Effect of permethrin-impregnated nets on exiting behavior, blood feeding success, and time of feeding of malaria mosquitoes (Diptera: Culicidae) in western Kenya.

The impact of permethrin-treated bednets on the feeding and house entering/exiting behavior of malaria vectors was assessed in two studies in western Kenya. In one study, matched pairs of houses were allocated randomly to receive bednets or no bednets. Exiting mosquitoes were collected in Colombian curtains hung around half of each house; indoor resting mosquitoes were collected by pyrethrum spray catches. The number of Anopheles gambiae Giles and An. arabiensis Patton estimated to have entered the houses was unaffected by the presence of bednets; Anopheles funestus Giles was less likely to enter a house if bednets were present. Anopheles gambiae and An. funestus were less likely to obtain a blood meal and significantly more likely to exit houses when bednets were present. No difference was detected in An. arabiensis rates of blood feeding and exiting. In a second experiment, hourly night biting collections were done on 13 nights during the rainy season to assess whether village-wide use of permethrin-treated bednets caused a shift in the time of biting of malaria vectors. A statistically significant shift was detected in the biting times of An. gambiae s.l., although the observed differences were small. No change was observed in the hourly distribution of An. funestus biting. Our study demonstrated that, at least in the short-term, bednets reduced human-vector contact and blood feeding success but did not lead to changes in the biting times of the malaria vectors in western Kenya.

All-cause mortality among young children in western Kenya. VI: the Asembo Bay Cohort Project.

Although all-cause mortality has been used as an indicator of the health status of childhood populations, such data are sparse for most rural areas of sub-Saharan Africa, particularly community-based estimates of infant mortality rates. The longitudinal follow-up of more than 1,500 children enrolled at birth into the Asembo Bay Cohort Project (ABCP) in western Kenya between 1992 and 1996 has provided a fixed birth cohort for estimating all-cause mortality over the first 5 yr of life. We surveyed mothers and guardians of cohort children in early 1999 to determine survival status. A total of 1,260 households were surveyed to determine the survival status of 1,556 live births (99.2% of original cohort, n = 1,570). Most mothers (66%) still resided but 27.5% had migrated, and 5.5% had died. In early 1999, the overall cumulative incidence of all-cause mortality for the entire 1992-1996 birth cohort was 26.5% (95% confidence interval, 24.1-28.9%). Neonatal and infant mortality were 32 and 176 per 1,000 live births, respectively. These community-based estimates of mortality in the ABCP area are substantially higher than for Kenya overall (nationally, infant mortality is 75 per 1,000 live births). The results provide a baseline description of all-cause mortality among children in an area with intense Plasmodium falciparum transmission and will be useful in future efforts to monitor changes in death rates attributable to control programs for specific diseases (e.g., malaria and HIV/AIDS) in Africa.

Characteristics of larval anopheline (Diptera: Culicidae) habitats in Western Kenya.

A longitudinal survey of mosquito larval habitats was carried out in Asembo Bay, western Kenya, during the rainy season of 1998. All pools of standing water along a 700-m transect were sampled twice per week. For each habitat, eight environmental variables were recorded and a sample of anopheline larvae was collected for identification. In total, 1,751 Anopheles gambiae s.l. and 2,784 Anopheles funestus Giles were identified. Identification of An. gambiae s.l. by polymerase chain reaction (PCR) indicated that 240 (14.7%) were An. gambiae Giles and 858 (52.4%) were An. arabiensis Patton; PCR failed to identify 539 (32.9%) specimens. Repeated measures logistic regression analysis indicated that An. gambiae and An. arabiensis larvae were associated with small, temporary habitats with algae and little or no aquatic vegetation. Anopheles funestus larvae were associated with larger, semipermanent bodies of water containing aquatic vegetation and algae. Direct comparison of habitat characteristics associated with either An. gambiae or An. arabiensis revealed that algae were associated more commonly with habitats containing An. gambiae; no other differences were detected. Chi-square analysis indicated that these species were collected from the same habitat more frequently than would be expected by chance alone. Together, these results indicate that An. gambiae and An. arabiensis have similar requirements for the larval environment and that, at least in western Kenya, they do not segregate into separate habitats.

Laboratory and field evaluation of polymerase chain reaction-based forensic DNA profiling for use in identification of human blood meal sources of Aedes aegypti (Diptera: Culicidae).

We modified polymerase chain reaction (PCR)-based forensic DNA profiling for field studies on the feeding behavior of Aedes aegypti, the principal mosquito vector of dengue virus. Human DNA was extracted from oral swabs of human subjects and from blood-engorged mosquitoes, DNA was quantified by slot blot, and alleles at variable number tandem repeats and three short tandem repeats loci were amplified by PCR. Alleles were separated electrophoretically and then visualized by silver staining. A custom software program was written to match DNA fingerprints of potential human hosts to allelic profiles detected in engorged mosquitoes, and to calculate error rates for identification of human hosts of single and multiple-host blood meals. At 29 degrees C in the laboratory, human DNA recovered from mosquito blood meals declined an average of 67% 8 h after feeding and 90% after 24 h. We obtained complete allelic profiles from seven of 10 mosquitoes collected after 24 h. In a field trial, complete DNA profiles were obtained successfully for 43 people living in a rural village in south central Thailand and for 20 of 100 Ae. aegypti that contained blood and were collected in those peoples' homes. Blood imbibed from more than one person was detected in 45% (9 of 20) of the meals. Sixty-five percent of the meals contained blood from nonresidents of the house in which the mosquito was collected or from people who were not profiled; data consistent with the hypothesis that human movement is important for the spread of dengue virus within and among communities. When using alleles at four loci, all of the Thais and nine members spanning three generations of a Chinese-American family had unique allelic profiles. Error rates from classifying possible multiple-host meals as single-host meals were low (1-8%), with the highest error associated with closely related people. Results from our laboratory and field studies indicated that DNA profiling can be used to study the details and epidemiological implications of Ae. aegypti blood-feeding behavior.

The Rift Valley complex as a barrier to gene flow for Anopheles gambiae in Kenya: the mtDNA perspective.

Descriptions of A. gambiae population structure based on microsatellite loci and mitochondrial DNA (mtDNA) were incongruent. High differentiation of populations was measured across the Rift Valley by microsatellites, but no differentiation was detected based on mtDNA. To resolve this conflict, we compared the old data to new mtDNA data using the same specimen previously genotyped in microsatellite loci. Analysis of a larger number of mtDNA sequences resulted in high and significant differentiation between populations across the Rift Valley. We developed a method to assess whether differentiation across the Rift Valley was generated by pure drift rather than mutation-drift, based on DNA sequence data. Applying this method to the mtDNA data suggested that pure drift was the primary force generating differentiation between the populations across the Rift, while mutation-drift generated differentiation across the continent. Given adequate sample size, mtDNA provided congruent results with microsatellite loci.

Effect of Plasmodium falciparum parasitemia density on hemoglobin concentrations among full-term, normal birth weight children in western Kenya, IV. The Asembo Bay Cohort Project.

The relative importance of acute high-density versus persistent low-density Plasmodium falciparum parasitemia in contributing to the public health problem of malarial anemia remains unclear. The Asembo Bay Cohort Project in western Kenya collected monthly hemoglobin (Hb) and parasitologic measurements and biweekly assessments of antimalarial drug use among 942 singleton live births between 1992 and 1996. A mixed-model analysis appropriate for repeated measures data was used to study how time-varying parasitemia and antimalarial drug exposures influenced mean Hb profiles. Incidence of World Health Organization-defined severe malarial anemia was 28.1 per 1,000 person-years. Among children aged less than 24 months, concurrent parasitemia was significantly associated with lower mean Hb, especially when compared to children with no concurrent parasitemia. Increased densities of the 90-day history of parasitemia preceding Hb measurement was more strongly associated with mean Hb levels than concurrent parasitemia density. While the highest quartile of 90-day parasitemia history was associated with lowest mean Hb levels, children in the lowest 90-day exposure quartile still experienced significantly lower Hb levels when compared to children who remained parasitemia-free for the same 90-day period. The results highlight the importance of collecting and analyzing longitudinal Hb and parasitologic data when studying the natural history of malarial anemia.

The Rift Valley complex as a barrier to gene flow for Anopheles gambiae in Kenya.

Recent studies of Anopheles gambiae, the principal mosquito vector of malaria in Africa, suggested that the eastern Rift Valley and its surrounding areas act as a barrier to gene flow. To quantify the unique effect of these areas on gene flow, we measured genetic variation within and between populations from each side of the Rift. Low differentiation was measured between populations on each side of the Rift (mean FST < 0.008, mean RST < 0.002). However, high differentiation was measured across the Rift (mean FST = 0.104; mean RST = 0.032). Genetic diversity within populations was lower in eastern populations, suggesting that the effective population sizes (Ne) of these populations were lower than those of western populations. We partitioned the overall differentiation across the Rift into three factors: variation in Ne between populations contributed 7-20%; distance contributed 10-30%, and the remainder, corresponding to the unique effect of the Rift was 50-80%. The Rift's effect was highly significant based on FST. The greater sensitivity of FST in measuring differentiation indicated that drift and not mutation generated the differences between populations. Restricted gene exchange across several hundred kilometers on the face of intense human transportation implies that active mosquito dispersal is the major form of migration, and that migration is a multistep process, where step length is relatively short.

Elevated oxidase and esterase levels associated with permethrin tolerance in Anopheles gambiae from Kenyan villages using permethrin-impregnated nets.

The permethrin tolerance (PT) of a population of the mosquito Anopheles gambiae (Diptera: Culicidae) increased following the introduction of permethrin-impregnated nets for malaria control in certain villages near Kisumu, western Kenya. Using a biochemical test that indirectly measures oxidases associated with permethrin resistance, we found that this population had higher oxidase levels than a comparison population from villages without impregnated nets. Mosquitoes from a colony of An. gambiae selected for PT, the RSP (reduced susceptibility to permethrin) strain, were exposed to permethrin with or without the oxidase inhibitor piperonyl butoxide (PB). Significantly higher mortality rates occurred when permethrin was synergized by PB, presumably by suppression of oxidases responsible for PT. An unselected (UNS) colony of An. gambiae that was more susceptible than RSP in a permethrin-susceptibility bioassay (i.e. LT50 22 min for UNS, vs. 42min for RSP) was compared with the RSP colony for levels of oxidases and esterases. The levels of both enzymes were very significantly higher in the RSP strain (P<0.0001). We speculate that use of impregnated nets selected for higher oxidase and esterase levels in An. gambiae to metabolize permethrin acquired from the nets. Both oxidase and esterase mechanisms could confer cross-resistance to other pyrethroids.

Analysis of genetic variability in Anopheles arabiensis and Anopheles gambiae using microsatellite loci.

We analysed genetic variability in Anopheles arabiensis and Anopheles gambiae populations using microsatellite loci to determine whether the Rift Valley restricts the flow of genes. Deviations from Hardy-Weinberg expectations were significant, and were most likely to be due to the high frequency of null alleles observed. An. arabiensis populations occurring between 40 and 700 km apart across the Eastern arm of the Rift Valley were not differentiated (pair-wise F(ST) range: 0.0033-0.0265, P > 0.05). Neither were An. gambiae populations from Asembo Bay and Ghana (F(ST): 0.0063, P > 0.05) despite a geographical separation of about 5000 km. In contrast, significant differentiation was observed between An. gambiae populations from Asembo Bay and Kilifi (about 700 km apart; F(ST) = 0.1249, P < 0.01), suggesting the presence of a barrier to gene flow.