Entomological Methods Research Articles

Evaluation of the Human IgG Antibody Response to Aedes albopictus Saliva as a New Specific Biomarker of Exposure to Vector Bites

BackgroundThe spread of Aedes albopictus, a vector for re-emergent arbovirus diseases like chikungunya and dengue, points up the need for better control strategies and new tools to evaluate transmission risk. Human antibody (Ab) responses to mosquito salivary proteins could represent a reliable biomarker for evaluating human-vector contact and the efficacy of control programs.Methodology/Principal FindingsWe used ELISA tests to evaluate specific immunoglobulin G (IgG) responses to salivary gland extracts (SGE) in adults exposed to Aedes albopictus in Reunion Island. The percentage of immune responders (88%) and levels of anti-SGE IgG Abs were high in exposed individuals. At an individual level, our results indicate heterogeneity of the exposure to Aedes albopictus bites. In addition, low-level immune cross-reactivity between Aedes albopictus and Aedes aegypti SGEs was observed, mainly in the highest responders.Conclusion/SignificanceAb responses to saliva could be used as an immuno-epidemiological tool for evaluating exposure to Aedes albopictus bites. Combined with entomological and epidemiological methods, a “salivary” biomarker of exposure to Aedes albopictus could enhance surveillance of its spread and the risk of arbovirus transmission, and could be used as a direct tool for the evaluation of Aedes albopictus control strategies.

Assessment of Decomposition Studies Indicates Need for Standardized and Repeatable Research Methods in Forensic Entomology

The National Research Council (NRC) released a report in 2009 discussing the “science” used in forensics. Specifically, the report outlined concerns regarding the state of the forensic sciences and what was needed to raise the level of rigor and reliability of these fields in a court of law. In response to this NRC document, the present paper examined several parameters used in decomposition studies that have implications for forensic entomology. Specifically, this analysis was conducted to determine the degree of repeatability in methods across studies as related to general conclusions drawn in court, entomology and forensic investigations. Forensic entomologists primarily analyze insect evidence recovered from decomposing remains to estimate a portion of the Period of Insect Activity (PIA), which encompasses the time of colonization, to infer a minimum Postmortem Interval (PMI). One method utilized by entomologists relies on succession data from published studies to generate estimates of the minimum PMI of a decedent. For this article, approximately 75 publications on arthropod succession on vertebrate carrion were reviewed for 13 criteria that are known to play instrumental roles in governing this process: 1) animal model, 2) time of actual death, 3) euthanasia method, 4) storage method, 5) storage time, 6) time of removal from storage to placement in the field, 7) time of day remains placed in the field, 8) catalog of arthropods associated with the remains over time 9) time of initial insect contact, 10) time of initial colonization (i.e., arthropod offspring located on the remains), 11) study site, 12) number of replicate carcasses and 13) months and season of study. These criteria were selected as they can directly impact arthropod colonization and succession patterns on vertebrate carrion and are easily recorded. Data indicated that such information is highly fragmented, and that key criteria necessary to repeat studies (a core principle of the scientific method) are often lacking in the published literature. As an example, among the studies included in this analysis, we could not find significant associations between arthropod taxa richness and carcass model or carcass size. In the case of carcass size, island biogeography theory predicts that larger areas/resources will hold higher diversity. We suspect that the high degree of reporting variability in taxonomic resolution and taxa-specific study focus precluded such relationships that have been reported for other disciplines of biology. Consequently, we suggest that, to date, because of these issues there has not been a comprehensive analysis (e.g., meta-analysis) to provide general inference of arthropod succession patterns on carrion to predict a minimum PMI. In order to begin standardizing such studies, we suggest that future research endeavors examining arthropod succession on carrion record detailed data for all of the suggested criteria. Doing so can result in data amassed over time for use in comprehensive and strong meta-analyses. Such results could allow for greater appreciation of variation associated with arthropod succession on carrion.

Assessment of exposure to Plasmodium falciparum transmission in a low endemicity area by using multiplex fluorescent microsphere-based serological assays

BackgroundThe evaluation of malaria transmission intensity is a crucial indicator for estimating the burden of malarial disease. In this respect, entomological and parasitological methods present limitations, especially in low transmission areas. The present study used a sensitive multiplex assay to assess the exposure to Plasmodium falciparum infection in children living in an area of low endemicity. In three Senegalese villages, specific antibody (IgG) responses to 13 pre-erythrocytic P. falciparum peptides derived from Lsa1, Lsa3, Glurp, Salsa, Trap, Starp, Csp and Pf11.1 proteins were simultaneously evaluated before (June), at the peak (September) and after (December) the period of malaria transmission, in children aged from 1 to 8 years.ResultsCompared to other antigens, a high percentage of seropositivity and specific antibody levels were detected with Glurp, Salsa1, Lsa3NR2, and Lsa1J antigens. The seropositivity increased with age for all tested antigens. Specific IgG levels to Glurp, Salsa1, Lsa3NR2, and Lsa1J were significantly higher in P. falciparum infected children compared to non-infected and this increase is significantly correlated with parasite density.ConclusionThe multiplex assay represents a useful technology for a serological assessment of rapid variations in malaria transmission intensity, especially in a context of low parasite rates. The use of such combined serological markers (i.e. Glurp, Lsa1, Lsa3, and Salsa) could offer the opportunity to examine these variations over time, and to evaluate the efficacy of integrated malaria control strategies.

Presença de mosquitos (Diptera: Culicidae) em piscinões na zona leste de São Paulo

Alterations in the environment contribute to changes in weather patterns, which cause an increase in rainfall, which causes flooding. Flooding has been addressed by building reservoirs, called piscinões, to contain the excess rain. In this report, we evaluated the Culicidae fauna of epidemiological importance in the reservoirs of Caguaçu and Inhumas. Monthly collections were performed in both reservoirs, which are situated in the Eastern section of the City of São Paulo. Monthly collections of Culicidae were undertaken using the entomological scoop method and a battery aspirator during the period between March 2006 and February 2007. Descriptive statistical and simple linear regression analyses of the data were carried out. A total of 8,917 Culicidae mosquitoes were collected. The majority of the insects captured were of Culex (Culex) quinquefasciatus, representing 98.9% of the specimens identified in the Inhumas and 95.2% of those identified in Caguaçu. In the Caguaçu reservoir, the greatest frequency of immature mosquitoes was observed in the spillway (61%). In the Inhumas reservoir, the greatest occurrence of immature Culicidae was detected in the channel (42.6%). The linear regression analysis showed that 87% of the larvae in the third and fourth stages and 60% of the pupae were collected during periods of rainfall. In the Inhumas reservoir, 36% of the larvae and 18% of the pupae were collected during periods of rainfall. Culex quinquefasciatus, a vector for agents of filariasis and arbovirus diseases and a nuisance to the human population, was frequent in both environments. Methods of control of the species in the studied dams are necessary to reduce their epidemiological potential.

Bionomics of the malaria vector Anopheles farauti in Temotu Province, Solomon Islands: issues for malaria elimination.

BackgroundIn the Solomon Islands, the Malaria Eradication Programmes of the 1970s virtually eliminated the malaria vectors: Anopheles punctulatus and Anopheles koliensis, both late night biting, endophagic species. However, the vector, Anopheles farauti, changed its behaviour to bite early in the evening outdoors. Thus, An. farauti mosquitoes were able to avoid insecticide exposure and still maintain transmission. Thirty years on and the Solomon Islands are planning for intensified malaria control and localized elimination; but little is currently known about the behaviour of the vectors and how they will respond to intensified control.MethodsIn the elimination area, Temotu Province, standard entomological collection methods were conducted in typical coastal villages to determine the vector, its ecology, biting density, behaviour, longevity, and vector efficacy. These vector surveys were conducted pre-intervention and post-intervention following indoor residual spraying and distribution of long-lasting insecticidal nets.ResultsAnopheles farauti was the only anopheline in Temotu Province. In 2008 (pre-intervention), this species occurred in moderate to high densities (19.5-78.5 bites/person/night) and expressed a tendency to bite outdoors, early in the night (peak biting time 6-8 pm). Surveys post intervention showed that there was little, if any, reduction in biting densities and no reduction in the longevity of the vector population. After adjusting for human behaviour, indoor biting was reduced from 57% pre-intervention to 40% post-intervention.ConclusionIn an effort to learn from historical mistakes and develop successful elimination programmes, there is a need for implementing complimentary vector control tools that can target exophagic and early biting vectors. Intensified indoor residual spraying and long-lasting insecticide net use has further promoted the early, outdoor feeding behaviour of An. farauti in the Solomon Islands. Consequently, the effectiveness of IRS and the personal protection provided by bed nets is compromised. To achieve elimination, any residual transmission should be targeted using integrated vector control incorporating complementary tools such as larviciding and/or zooprophylaxis.

Forensic entomology: applications and limitations

Forensic entomology is the science of collecting and analysing insect evidence to aid in forensic investigations. Its main application is in the determination of the minimum time since death in cases of suspicious death, either by estimating the age of the oldest necrophagous insects that developed on the corpse, or by analysing the insect species composition on the corpse. In addition, toxicological and molecular examinations of these insects may help reveal the cause of death or even the identity of a victim, by associating a larva with its last meal, for example, in cases where insect evidence is left at a scene after human remains have been deliberately removed. Some fly species can develop not only on corpses but on living bodies too, causing myiasis. Analysis of larvae in such cases can demonstrate the period of neglect of humans or animals. Without the appropriate professional collection of insect evidence, an accurate and convincing presentation of such evidence in court will be hampered or even impossible. The present paper describes the principles and methods of forensic entomology and the optimal techniques for collecting insect evidence.

Evaluation of oviposition traps as an entomological surveillance method for Aedes aegypti (Diptera, Culicidae)

This study aimed to describe the behavior of oviposition traps for Aedes aegypti over time, to compare it with the larval survey and to investigate the association with climatic variables. It was conducted in Sao Jose do Rio Preto city, Sao Paulo. Daily climatic data and fortnightly measurements for oviposition traps and larval infestation were collected from October 2003 to September 2004. Three different periods were identified in the behavior of oviposition traps' positivity and mean number of eggs: increase, plateau and decrease in values. These measurements followed the variation of climatic data from the first and third periods. High correlation was obtained between the positivity and the mean number of eggs. The oviposition traps showed higher capacity to detect the vector than did larval survey. It was observed that the first (October to December) and third (May to September) periods were considered to be the most suitable to use oviposition traps than larval surveys.

Holocene dynamics of vegetation and ecological conditions in the southern Yamal Peninsula according to the results of comprehensive analysis of a relict peat bog deposit

A comprehensive analysis of samples from a frozen peat deposit in the upper reaches of the Khadytayakha River by palynological, carpological, entomological, and radiocarbon methods has been performed to reconstruct changes in the regional and local vegetation and climatic conditions during the Holocene. The results show that this peat deposit was formed from 8000 to 5000 years BP. During that period, the climate in the southern Yamal Peninsula was considerably warmer, and the present-day subarctic shrub tundra zone was occupied by plant communities of spruce, birch, and larch forests.

Human IgG response to a salivary peptide, gSG6-P1, as a new immuno-epidemiological tool for evaluating low-level exposure to Anopheles bites

BackgroundHuman populations exposed to low malaria transmission present particular severe risks of malaria morbidity and mortality. In addition, in a context of low-level exposure to Anopheles vector, conventional entomological methods used for sampling Anopheles populations are insufficiently sensitive and probably under-estimate the real risk of malaria transmission. The evaluation of antibody (Ab) responses to arthropod salivary proteins constitutes a novel tool for estimating exposure level to insect bites. In the case of malaria, a recent study has shown that human IgG responses to the gSG6-P1 peptide represented a specific biomarker of exposure to Anopheles gambiae bites. The objective of this study was to investigate if this biomarker can be used to estimate low-level exposure of individuals to Anopheles vector.MethodsThe IgG Ab level to gSG6-P1 was evaluated at the peak and at the end of the An. gambiae exposure season in children living in Senegalese villages, where the Anopheles density was estimated to be very low by classical entomological trapping but where malaria transmission occurred during the studied season.ResultsSpecific IgG responses to gSG6-P1 were observed in children exposed to very low-level of Anopheles bites. In addition, a significant increase in the specific IgG Ab level was observed during the Anopheles exposure season whereas classical entomological data have reported very few or no Anopheles during the studied period. Furthermore, this biomarker may also be applicable to evaluate the heterogeneity of individual exposure.ConclusionThe results strengthen the hypothesis that the evaluation of IgG responses to gSG6-P1 during the season of exposure could reflect the real human contact with anthropophilic Anopheles and suggest that this biomarker of low exposure could be used at the individual level. This promising immuno-epidemiological marker could represent a useful tool to assess the risk to very low exposure to malaria vectors as observed in seasonal, urban, altitude or travellers contexts. In addition, this biomarker could be used for the surveillance survey after applying anti-vector strategy.

Rapid Assessment of Malaria Transmission Using Age-Specific Sero-Conversion Rates

BackgroundMalaria transmission intensity is a crucial determinant of malarial disease burden and its measurement can help to define health priorities. Rapid, local estimates of transmission are required to focus resources better but current entomological and parasitological methods for estimating transmission intensity are limited in this respect. An alternative is determination of antimalarial antibody age-specific sero-prevalence to estimate sero-conversion rates (SCR), which have been shown to correlate with transmission intensity. This study evaluated SCR generated from samples collected from health facility attendees as a tool for a rapid assessment of malaria transmission intensity.Methodology and Principal FindingsThe study was conducted in north east Tanzania. Antibodies to Plasmodium falciparum merozoite antigens MSP-119 and AMA-1 were measured by indirect ELISA. Age-specific antibody prevalence was analysed using a catalytic conversion model based on maximum likelihood to generate SCR. A pilot study, conducted near Moshi, found SCRs for AMA-1 were highly comparable between samples collected from individuals in a conventional cross-sectional survey and those collected from attendees at a local health facility. For the main study, 3885 individuals attending village health facilities in Korogwe and Same districts were recruited. Both malaria parasite prevalence and sero-positivity were higher in Korogwe than in Same. MSP-119 and AMA-1 SCR rates for Korogwe villages ranged from 0.03 to 0.06 and 0.07 to 0.21 respectively. In Same district there was evidence of a recent reduction in transmission, with SCR among those born since 1998 [MSP-119 0.002 to 0.008 and AMA-1 0.005 to 0.014 ] being 5 to 10 fold lower than among individuals born prior to 1998 [MSP-119 0.02 to 0.04 and AMA-1 0.04 to 0.13]. Current health facility specific estimates of SCR showed good correlations with malaria incidence rates in infants in a contemporaneous clinical trial (MSP-119 r2 = 0.78, p<0.01 & AMA-1 r2 = 0.91, p<0.001).ConclusionsSCRs generated from age-specific anti-malarial antibody prevalence data collected via health facility surveys were robust and credible. Analysis of SCR allowed detection of a recent drop in malaria transmission in line with recent data from other areas in the region. This health facility-based approach represents a potential tool for rapid assessment of recent trends in malaria transmission intensity, generating valuable data for local and national malaria control programs to target, monitor and evaluate their control strategies.

Evaluación de estrategias comunitarias para el control de Aedes aegypti en Cali, Colombia

Dengue viruses transmitted principally by the urban mosquito Aedes aegypti, cause one of the major public health problems confronting tropical cities. Insecticide spraying has been the mainstay of mosquito control; however, its continuous use has selected for resistance. Other important methods of control involve community participation. This study evaluated two control methods for Ae. aegypti that can be used by the community: Lethal ovitraps (LOs) and Bacillus thuringiensis var israeliensis (Bti) briquettes. The project study was carried out in four similar neighborhoods within a representative district in the city of Cali, Colombia. Three interventions (LO, Bti, LO+Bti plus education and one control (education only) area were evaluated for efficacy in post-intervention entomological surveys. Additionally, entomological indices were also compared to results from a pre-intervention survey carried out on a sample of city blocks in the same neighborhoods. Relative vector abundance in relation to weather conditions using the same entomological sampling methods was compared. The interventions did not achieve significant differences in vector abundance among the treatments. However, the interventions achieved a significant reduction in entomological indices compared with those observed during the pre-intervention survey: House index 15.1% vs. 8.5%, mean pupae per house 1.15 vs. 0.073, and Adult index 56.3% vs. 34.8% (p < 0.05). The lack of significant differences among the interventions, and between treated and control blocks suggested that educational activities together with periodic visits to the houses produced similar reductions of immature and adult Aedes aegypti.

The multiplicity of malaria transmission: a review of entomological inoculation rate measurements and methods across sub-Saharan Africa.

Plasmodium falciparum malaria is a serious tropical disease that causes more than one million deaths each year, most of them in Africa. It is transmitted by a range of Anopheles mosquitoes and the risk of disease varies greatly across the continent. The "entomological inoculation rate" is the commonly-used measure of the intensity of malaria transmission, yet the methods used are currently not standardized, nor do they take the ecological, demographic, and socioeconomic differences across populations into account. To better understand the multiplicity of malaria transmission, this study examines the distribution of transmission intensity across sub-Saharan Africa, reviews the range of methods used, and explores ecological parameters in selected locations. It builds on an extensive geo-referenced database and uses geographical information systems to highlight transmission patterns, knowledge gaps, trends and changes in methodologies over time, and key differences between land use, population density, climate, and the main mosquito species. The aim is to improve the methods of measuring malaria transmission, to help develop the way forward so that we can better assess the impact of the large-scale intervention programmes, and rapid demographic and environmental change taking place across Africa.

Results and experiences from the first EU proficiency test for the detection of Phytophthora ramorum1

The Crop Protection unit of the Institute for Agricultural and Fisheries Research offers plant disease diagnostic services through its ‘Diagnostic Centre for Plants’ (ILVO‐DCP). ILVO‐DCP has requested accreditation (ISO 17025) for a number of diagnostic detection methods in bacteriology, mycology, entomology and nematology. Accreditation forms an essential part of quality control programs such as ISO17025, which can in part be realized by proving the laboratory's competence in inter‐laboratory proficiency or ring tests. In 2006, ILVO‐DCP organized such a proficiency test for the detection of Phytophthora ramorum. The protocol was developed using the standard ILAC‐G13:2000, and defined rules for participation, sample preparation and transport, communication, fraud prevention and reporting. Eight European diagnostic centres participated in the proficiency test including ILVO‐DCP. Each participant received one set of 10 coded samples, each sample consisting of either leaves or stems that were artificially inoculated with either the target or an alternative organism. Participants could use one or more methods listed in the EPPO diagnostic protocol PM 7/66. They had to report within a specific timeframe and received a detailed report of their performance. The success rate of the proficiency test was 100%. This paper lists some of the experiences gained from organising this type of proficiency test.

Occurrence ofCulicoidesspp. (Diptera: Ceratopogonidae) in Tunisia, with emphasis on the bluetongue vectorCulicoides imicola

Following the bluetongue (BT) outbreaks in Tunisia from 1999 to 2002, BTV (bluetongue virus) serotype 2 was isolated; however, no entomological investigation was performed. In the study presented here, we assessed the Culicoides species populations (particularly C. imicola) in proximity to the BT outbreaks locations, both as a retrospective analysis and to update the list of Culicoides species present in Tunisia. The insects were caught using light traps and the species identification was performed according to the standard entomological methods. This study reveaaled the presence of significant numbers of C. imicola in all the tested locations. In addition, we reported a new Culicoides species for the Tunisian fauna C. punctatus.

Zur Anwendung von Computern für die Determination in der Entomologie

On the use of computer determinating methods in entomology. In this article the present situation of a computer-system for diagnostics of the systematical position of biological objects is examined. There different stages are shown, in which the research in this field of diagnostics have developed. In concrete examples of the list of scheme of destination for two groups of beatles the rules of preparation are shown how to inform the computer-systems and peculiarities of its working method are examined. It is reported about a Soviet system of such way, that is realized in a small computer. The importance of a computer-diagnosis for entomology is shown by it.

Application of DNA-Based Methods in Forensic Entomology

A forensic entomological investigation can benefit from a variety of widely practiced molecular genotyping methods. The most commonly used is DNA-based specimen identification. Other applications include the identification of insect gut contents and the characterization of the population genetic structure of a forensically important insect species. The proper application of these procedures demands that the analyst be technically expert. However, one must also be aware of the extensive list of standards and expectations that many legal systems have developed for forensic DNA analysis. We summarize the DNA techniques that are currently used in, or have been proposed for, forensic entomology and review established genetic analyses from other scientific fields that address questions similar to those in forensic entomology. We describe how accepted standards for forensic DNA practice and method validation are likely to apply to insect evidence used in a death or other forensic entomological investigation.

Avian foraging studies: an overlooked source of distribution data for macroecological and conservation studies

ABSTRACTMacroecological and biogeographical studies and their applicability for biological conservation vitally depend on distribution data. These are usually taken from distribution atlases and databases or maps found in identification guides. A previous study pointed to another little explored source of data — local faunistic studies. Here, I would like to draw attention to another potential source. Papers that analysed the food composition of some taxa (e.g. insectivorous birds) are an overlooked source of rich information on taxa distributions. These studies frequently include an ‘Appendix’ with a list of food items determined to the species level. These studies also contain data on abundances, number of samples, sampling time, and geographical location as a rule. Foraging birds naturally provide data on invertebrate distributions with good spatio‐temporal coverage and reasonably large samples. Importantly, birds frequently collect rare and by entomological methods hardly detectable species (i.e. those living high, in tree canopies, in very dense vegetation, or with secretive lifestyle). Data from bird dietary studies may help to ameliorate one of the most serious problems in distribution studies — zero inflation. I briefly discuss pros and cons of this so far neglected source of biogeographical data.

PCR and dissection as tools to monitor filarial infection of Aedes polynesiensismosquitoes in French Polynesia

BackgroundEntomological methods may provide important tools for monitoring the transmission of filariasis in French Polynesia. In order to standardize our PCR method and refine our protocol to assess filarial infection levels in mosquitoes, we compared dissection of the vector, Aedes polynesiensis, with the poolscreening polymerase chain reaction (PS-PCR) assay.Methods(1) Mosquitoes were collected in human landing catches in five areas in Moorea island, French Polynesia. (2) A fraction of the captured mosquitoes was dissected for Wuchereria bancrofti larvae. (3) Laboratory-reared mosquitoes (uninfected as well as experimentally infected ones) were repeatedly tested to optimize a PS-PCR protocol (DNA extracts from 1–50 pooled mosquitoes were tested with an internal standardized system and primers specific for the Ssp1 repeat sequence. PCR products were analysed by gel electrophoresis). (4) Another fraction of the captured mosquitoes was assayed by PS-PCR according the optimized protocol.ResultsThe prevalence of field-mosquito infection with W. bancrofti ranged from 1 to 8 % by dissection (L1–L3) and point estimates of infection prevalence, as assayed by PS-PCR, ranged from 0.4 to 3.7 %. There was a moderately strong correlation between larval infection rates as determined by dissection and PCR.DiscussionOur results suggest that the PS-PCR assay is specific and highly sensitive for detecting parasite DNA. We obtained similar although not identical results with dissections of mosquitoes. PS-PCR appears to be adequate for testing large numbers of mosquitoes in the context of filariasis elimination programs. The role and advantages of using entomologic methods to monitor filariasis programs are discussed.

Evaluation of the antibody response to Anopheles salivary antigens as a potential marker of risk of malaria

The evaluation of human immune responses to arthropod bites may be a useful marker of exposure to vector-borne diseases, with applications to malaria, the most serious parasitic infection in humans. The specific antibody (Ab) IgG response to saliva obtained from Anopheles gambiae mosquitoes was evaluated in young children from an area of seasonal malaria transmission in Senegal. Specific IgG was higher in children who developed clinical Plasmodium falciparum malaria within the 3 months that followed than in those who did not (P<0.05), and it increased significantly (P<0.0001) with the level of Anopheles exposure, as evaluated by conventional entomological methods. These results suggest that evaluation of antisalivary Ab responses could be a useful approach for identifying a marker for the risk of malaria transmission.

Forensic entomology.

Necrophagous insects are important in the decomposition of cadavers. The close association between insects and corpses and the use of insects in medicocriminal investigations is the subject of forensic entomology. The present paper reviews the historical background of this discipline, important postmortem processes, and discusses the scientific basis underlying attempts to determine the time interval since death. Using medical techniques, such as the measurement of body temperature or analysing livor and rigor mortis, time since death can only be accurately measured for the first two or three days after death. In contrast, by calculating the age of immature insect stages feeding on a corpse and analysing the necrophagous species present, postmortem intervals from the first day to several weeks can be estimated. These entomological methods may be hampered by difficulties associated with species identification, but modern DNA techniques are contributing to the rapid and authoritative identification of necrophagous insects. Other uses of entomological data include the toxicological examination of necrophagous larvae from a corpse to identify and estimate drugs and toxicants ingested by the person when alive and the proof of possible postmortem manipulations. Forensic entomology may even help in investigations dealing with people who are alive but in need of care, by revealing information about cases of neglect.