Pesticide intoxications in the Centre of Portugal: three years analysis

Pesticides are used in most countries around the world to protect agricultural and horticultural crops against damage. Poisoning by these toxicant agents occurs as a result of misuse or accidental exposure, and also by oral ingestion (voluntary or not). In Portugal, pesticide intoxications are still a cause of death, found in a considerable number of cases. The authors retrospectively examined the cases of pesticide poisoning in the Centre of Portugal, from autopsies performed in the Forensic Pathology Service of Coimbra's Delegation of the National Institute of Legal Medicine (NILM) and from other autopsies carried out in the Centre of Portugal, as well as some samples taken in hospitals in cases of suspected intoxication. In this study, the positive cases have been especially studied, in order to identify the pesticide used, as well as the etiology. The frequency of intoxications and its distribution by sex and age were also analyzed. Between January 2000 and December 2002, the Forensic Toxicology Laboratory received 639 pesticide analysis requests. In 2000, in a total of 149 analysis requests, 30 cases were positive, 63.3% from male individuals and 36.7% from female. In 2001, the analysis requests increased to 240 as well as the positive cases (43), 74.4% from male individuals and 25.6% from female and in 2002, the total cases analyzed also increased to 250, with 38 positive (73.6% from male individuals and 26.4% from female). Among the pesticides, organophosphorus insecticides still constitute the most important class detected in forensic intoxications, representing 63% of the total positive cases, followed by herbicides, with 33% of the positive results. Quinalphos is the most important organophosphorus insecticide, present in 32 of the 111 positive cases, followed by the herbicide paraquat, detected in 31 cases. The study emphasizes the increasing number of pesticide analyses, particularly relevant for the organophosphorus compounds and herbicides. Intoxication suspicion, accidental or voluntary, seems to be the most common cause of the incidents, for which analyses are requested, but it is also evident that the putative cause is unknown in a large number of cases. Therefore, more stringent legislation and enforcement regarding the sale and distribution of these toxic substances are needed.     

The Special Case of Pesticides: Science and Regulation

Pesticide regulation is a data-intensive activity that is strongly controlled at the federal level. The pesticide law FIFRA controls pesticide sale, use, and disposal. Normal use of pesticides is excluded under RCRA and CERCLA. There are approximately 1,000 pesticide active ingredients with highly diverse functions ranging from insecticides to disinfectants and pheromones (insect sex attractants). There is a relatively large database on many pesticides, which helps reduce uncertainty in risk assessments. Witnesses subject to Daubert should be familiar with the data as well as the computer models designed for pesticides. “Inert” ingredients that are formulated with the pesticide active ingredients can become issues in torts cases. Federal courts have ruled both in favor and against FIFRA preemption, i.e., the ability of a plaintiff to seek relief in a torts case may be blocked by FIFRA. Golf course permitting provides a special land use whereby fundamental pesticide risk and regulatory issues are debated at the level of local planning and zoning commissions.     

Trends in domestic animal medico-legal pathology cases submitted to a veterinary diagnostic laboratory 1998-2010*

Abstract: Pathologists at veterinary diagnostic laboratories receive medico‐legal cases from a variety of animal species for postmortem examination. A search of computerized records of the Animal Health Laboratory, University of Guelph, Guelph, Ontario, Canada from 1998 to 2010 identified 1706 medicolegal cases. These were categorized according to the history as criminal investigations, anesthetic‐related deaths, insurance, litigation, malpractice cases, and regulatory cases. Statistically significant linear trends in the proportion of medicolegal cases for all animals and criminal cases for companion animals were identified over the 12 year period. Companion animals had significantly greater odds of being a medicolegal case in all categories except for insurance and regulatory cases, compared to noncompanion animals. Based on pathology reports for the 271 criminal cases, 43.1% were consistent with neglect, 29.2% were compatible with non‐accidental injury, 4.80% were poisonings, 10.7% were deemed to be due to natural disease, and 11.43% were inconclusive.     

The forensic deaths caused by pesticide poisoning between the years 2006 and 2009 in Izmir, Turkey

Pesticide poisoning is still a significant health problem in Turkey. We conducted a retrospective study of autopsy cases at Izmir Branch of the Council of Forensic Medicine to describe the characteristics of deaths caused by pesticide poisoning between 2006 and 2009. The distributions of the cases according to gender and age were as follows: men 74.1% (n = 40, mean [±SD] age, 44.7 ± 14.1), women 25.9% (n = 14, mean [±SD] age, 39.2 ± 18.9). The majority of pesticide-poisoning deaths were suicides (n = 43, 80%) followed by accidents (n = 4, 8%) and homicide (n = 1, 2%). The manner of death could not be determined in six cases (11%). Suicides mostly occurred at home (n = 26, 63%) (p < 0.05). Methomyl was the most frequent pesticide (n = 9, 17%) among the all cases. This study reported that most of the pesticides found in poisoning cases were highly hazardous types. Combined efforts of medical professionals and law makers are needed for enacting strict laws against highly hazardous pesticides.     

Cytochrome b gene for species identification of the conservation animals

A partial DNA sequence of cytochrome b gene was used to identify the remains of endangered animals and species endemic to Taiwan. The conservation of animals species included in this study were: the formosan gem-faced civets, leopard cats, tigers, clouded leopards, lion, formosan muntjacs, formosan sika deers, formosan sambars, formosan serows, water buffalo, formosan pangolins and formosan macaques. The control species used included domestic cats, domestic dogs, domestic sheeps, domestic cattles, domestic pigs and humans. Heteroplasmy was detected in the formosan macaque, domestic pig and domestic cats. The frequencies of heteroplasmy in these animals were about 0.25% (1 in 402bp). Sequences were aligned by Pileup program of GCG computer package, and the phylogenetic tree was constructed by the neighbor-joining method. The results of sequence comparison showed that the percentage range of sequence diversity in the same species was from 0.25 to 2.74%, and that between the different species was from 5.97 to 34.83%. The results of phylogenetic analysis showed that the genetic distance between the different species was from 6.33 to 40.59. Animals of the same species, both the endangered animal species and domestic animals, were clustered together in the neighbor-joining tree. Three unknown samples of animal remains were identified by this system. The partial sequence of cytochrome b gene adopted in this study proved to be usable for animal identification.     

利用Q-TOFLC/MS建立二级谱图数据库快速检测90种农药的初步研究

目的建立90种农药的数据库（含二级谱图）,以达到快速筛选检验该范围内的未知样品目的。方法优化一种Q-TOF LC/MS分析条件,将90种农药分别依次进行Q-TOF LC/MS-MS二级谱图分析,利用PCDL软件建立数据库。并同时实验了血样中10种不同浓度农药（100ng/mL、500ng/mL）的添加实验,对建库方法进行了验证。结果阳性识别率在80%以上。结论本方法简便灵敏、快速实用。     

有机磷农药中毒的气相色谱检测方法

本文对17种有机磷农药进行了气相色谱定性分析,对其中11种有机磷农药作了定量分析方法的研究,筛选出七根色谱柱及有关色谱最佳条件,全面系统地建立了微量有机磷农药气相色谱定性、定量分析方法。为了解决生物样品中各种有机磷农药的提取方法,本文设计了从胃、肝、血、尿中提取、净化11种有机磷农药的方法并测定其回收率。用本方法提取、净化生物样品中农药,其回收率大部分超过80%。     

Poisoning deaths in Central China (Hubei): A 10-year retrospective study of forensic autopsy cases

A retrospective study of autopsy cases was conducted at the Department of Forensic Medicine, Tongji Medical College (DFM-TMC), in Hubei, China to describe the characteristics of poisoning deaths from 1999 to 2008. A total of 212 poisoning deaths were investigated by DFM-TMC during the 10-year period. The poisoning deaths ranged from 17 cases in 1999 to 27 cases in 2008. Of the 212 cases, 82 deaths (38.7%) were from pesticides, 36 deaths (17.0%) from carbon monoxide, 34 deaths (16.0%) from drugs, 22 deaths (10.4%) from alcohol, 17 deaths (8.0%) from other chemicals, 15 deaths (7.1%) from poisonous plants and animals, and six deaths (2.8%) from heavy metals. Of the 82 pesticide poisoning deaths, 43 (52.4%) cases were caused by rodenticides, mainly tetramine (N = 39). The majority of poisoning deaths were accidents (63.7%), followed by suicides (25.9%) and homicides (3.8%). The manner of death could not be determined in 14 cases (6.6%).     

The occurrence of HIV antibodies in drug addicts autopsied at the University Institute of Forensic Medicine in Copenhagen in 1987

In 1987 a total of 89 intravenous drug addicts were admitted to medico-legal autopsy at the University Institute of Forensic Medicine in Copenhagen. Totally 10 cases were reactive by the Elisa test and five of these, or 7% of the total material tested, could be confirmed by the immuno-blotting test. The other five cases must be considered as false positive, possibly due to antibodies against cellular antigens. Among the five real positive cases, four were known before-hand while in one case the HIV-positivity was not known. These findings - together with the possibility of false negative reactions - especially regarding persons with acute HIV-infection, requires special precautionary measures when autopsies of drug addicts are performed.     

Pesticide levels in head hair samples of Cretan population as an indicator of present and past exposure

In the present work we assessed chronic exposure of different working population groups of Messara and Sitia districts, Crete, Greece, to common currently used pesticides (diazinon, fenthion, methyl parathion and malathion) and two banned pesticides hexachlorocyclohexane (HCH) and DDT. The study population (211 persons, 110 females and 101 males) was divided to three groups; people working in greenhouses, animal breeders and people working in open cultivations. Methanolic extraction of pulverized hair was used for organophosphate pesticides extraction, followed by liquid-liquid extraction with water-ethyl acetate as a clean up step. The extraction of organochlorine pollutants was performed by acidic hydrolysis of the hair matrix followed by liquid-liquid and solid phase extraction. The levels of the aforementioned pesticides were measured by GC-ECD and gas chromatography-mass spectrometry (GC-MS). The median concentrations of a-HCH, HCB, lindane, opDDE, ppDDE, opDDD, ppDDD + opDDT and ppDDT were determined at 7.2, 2.2, 70.2, 2.7, 5.7, 3.1, 2.6 and 23.2 pg/mg. The median concentration of total HCHs and DDTs detected in the three working groups were: 95.0 and 8.9 pg/mg for the greenhouse workers, 38.2 and 3.3 pg/mg for the animal breeders and 24.1 and 5.2 pg/mg for the open cultivation group. Ten head hair samples were positive for diazinon at a mean concentration of 2.8 pg/mg. Fenthion, methyl parathion and malathion were not detected. Our results demonstrated the ability to assess chronic human past pesticides exposure, offering valuable information to epidemiological clinical studies.     

Reconciling Autonomy and Beneficence in Treatment Decision-Making for Companion Animal Patients

This article explores how the concept of consent to medical treatment applies in the veterinary context, and aims to evaluate normative justifications for owner consent to treatment of animal patients. We trace the evolution of the test for valid consent in human health decision-making, against a backdrop of increased recognition of the importance of patient rights and a gradual judicial espousal of a doctrine of informed consent grounded in a particular understanding of autonomy. We argue that, notwithstanding the adoption of a similar discourse of informed consent in professional veterinary codes, notions of autonomy and informed consent are not easily transferrable to the veterinary medicine context, given inter alia the tripartite relationship between veterinary professional, owner and animal patient. We suggest that a more appropriate, albeit inexact, analogy may be drawn with paediatric practice which is premised on a similarly tripartite relationship and where decisions must be reached in the best interests of the child. However, acknowledging the legal status of animals as property and how consent to veterinary treatment is predicated on the animal owner’s willingness and ability to pay, we propose that the appropriate response is for veterinary professionals generally to accept the client’s choice, provided this is informed. Yet such client autonomy must be limited where animal welfare concerns exist, so that beneficence continues to play an important role in the veterinary context. We suggest that this ‘middle road’ should be reflected in professional veterinary guidance.     

The Sooner the Better: The Arguments for the Use of Extended Welfare Assessment Grids in Animal Welfare Cases

Animals are protected under national animal welfare legislation, against intentional acts of cruelty and a failure to act, resulting in neglect and causing an animal to suffer. The Royal Society for the Prevention of Cruelty to Animals (RSPCA) bears the responsibility of investigating and prosecuting the majority of animal welfare offences in England and Wales. In recent years, how they operate has been criticised, and it has been debated whether they should be able to bring private prosecutions, and what their role should actually be. This criticism calls for a change in the way in which the RSPCA approach cases of animal welfare, to strengthen their continuing role in ensuring positive animal welfare is achieved and, where not, prosecuted. This paper outlines the need for a new approach and how it can be managed. Honess and Wolfensohn (Altern Lab Anim 38:205, 2010) have developed an Extended Welfare Assessment Grid (EWAG), a visualisation mapping tool of welfare impact, which has been useful for assessing the welfare of animals used in laboratories. This tool has proven so useful, veterinarians are now using it in veterinary hospitals to help assess whether an animal is likely to further deteriorate, due to disease and illness, and to show any short-term welfare impact on the animal (Williams in UFAW conference, Newcastle upon Tyne, 2018). This paper will explore the potential for the EWAG to be adapted to assess the welfare of animals when owners are not meeting the welfare needs of their companion animals. RSPCA can use it to support their assessments of the current welfare of an animal under a person’s ownership and whether the animal’s welfare will deteriorate should they remain under that ownership. The EWAG will be a useful tool for those working in animal welfare, such as the RSPCA, to help organisations to intervene earlier, work in partnership with an owner, and support their claims of a risk to animal welfare.

     

Simultaneous hair testing for opiates, cocaine, and metabolites by GC-MS: a survey of applicants for driving licenses with a history of drug use

A sensitive GC-MS method for the simultaneous determination of opiates, cocaine, and metabolites in hair at a cut-off level of 0.1 ng/mg was adopted to assess past exposure to these drugs in applicants for driving licenses with a history of drug use. The sampling protocol consisted of collection of one hair (sample A, 5-cm length) and one urine sample. When hair and urine (EMIT Syva, cut-off levels: 0.3 mg/l for opiates, 0.15 mg/l for cocaine, GC-MS confirmation of positives) were both positive or negative the protocol was concluded. In the other cases, the assessment of 'current exposure' to drugs was carried out, in order to avoid seriated random urinalysis, by collecting a second hair sample (sample B) 6 weeks later and analysing the proximal 1-cm segment. Out of the 214 'A' hair samples analyzed, 14 (6.5%) tested positive for morphine and/or 6-acetylmorphine (6AM), and 26 (12%) for cocaine and/or benzoylecgonine (BE), whereas none of the samples tested positive for both drugs. Levels between 0.1 and 1 ng/mg of the single analytes were found in eight out of the 14 morphine-6AM positives (57%) and in 18 out of the 26 cocaine-BE positives (69%). The time course of positive cases showed a progressive decrease of morphine-6AM positives and a corresponding increase of cocaine-BE positives within the study period September 1995-February 1999. No cases with positive urine and negative hair were observed. Among the 40 positive cases, seven (four and three for opiates and cocaine, respectively) were found to be 'currently exposed to drug', four by urinalysis (three and one) and three by analysis of the hair sample B (1 and 2).     

Quantitative assessment of the percent fat in domestic animal bone marrow

Measurement of the amount of fat in femoral bone marrow can provide a quantitative assessment of the nutritional status of an individual animal. An analytical method is presented for quantitating the percent fat in bone marrow from three domestic species: bovine, canine, and equine. In this procedure, fat is extracted from bone marrow using pentane, and the percent fat recovered is determined gravimetrically. Based on analyses from adult animals (normal body condition scores), the average percentage of fat in the bone marrow was >80%. In cases in which animals have been diagnosed as emaciated or exhibit serous atrophy of fat (body scores of 1 or 2), the femoral bone marrow fat was less than 20%. In domestic animals, bone marrow fat analysis can be a useful, quantitative measure that, when used in conjunction with all other data available, can support a diagnosis of starvation or malnutrition.     

The use of serial measurement of plasma cholinesterase in the management of acute poisoning with organophosphates and carbamates

OBJECTIVE: To evaluate the benefits of using serial measurements of plasma cholinesterase (butyrylcholinesterase, BuChE) activity in the management of cholinesterase inhibitor insecticidal poisoning. METHOD: After establishing and validating BuChE activity test, and making it available for clinical service in the toxicology laboratory at Jordan University Hospital. Serial measurements of BuChE were performed on samples taken from 10 symptomatic patients presented with the manifestations of poisoning due to acetylcholinesterase inhibitor insecticides during the year 2001. The number of serial repeats of BuChE activity tests ranged from 2 to 4 and from 8 to 11 for patients with carbamates and organophosphates (OPs) poisoning, respectively. The results of serial measurement of BuChE obtained from each patient's samples were used to draw a curve; three different types of curves were obtained from all patients samples. RESULT: The obtained curves were found to follow our three proposed curves, which support our point view regarding the importance of the proposed curves in the differential diagnosis and treatment of cholinesterase inhibitor pesticides poisoning. CONCLUSION: This study pointed out the importance of utilizing serial measurements of BuChE activity in the diagnosis and the management of organophosphates and carbamates poisoning. The BuChE activity results were used to support diagnostic and prognostic criteria that guided patient management and follow up. Applying those curves to large number of patients' samples will enhance its credibility. The study also demonstrated the importance of direct contract between toxicologist and physician in treatment of the pesticides poisoned patients.     

High performance liquid chromatographic method for the analysis of organophosphorus and carbamate pesticides

A method has been developed for the rapid quantitative analysis of organophosphorous and carbamate pesticides using HPLC. Good separation was obtained among the four major groups of organophosphorus pesticides [i.e. aryl phosphorothionate (methyl parathion), alkyl phosphorothionate (malathion), enolphosphate (phosphomidon, monocrotophos, dichlorvos), heterocyclic phosphorothionates (quinalphos)] and carbamates [viz. Carbaryl (Sevin) and Baygon (Dalf)] with a detection limit of 100 ng for all the pesticides. Separation was measured in terms of capacity factor (k') resolution (R) and selectivity factor (alpha ii). The method described can be used for the analysis of biological samples for the presence of organophosphorus and carbamate pesticides in the cases of poisoning. Recovery studies were made in the blood, lung and liver and found to be 85-97% with reproducibility at greater than 95%.     

Incidence of xenobiotics among drivers killed in single-vehicle crashes.

The authors have performed a study of single-vehicle crashes (SVCs) in order to verify a correlation between the loss of vehicle control and the presence of drugs in the body. Overall, 129 cases were recorded and occurred in the catchment area of the Institute of Legal Medicine in Milan between 1986 to 1996. Among the 129 cases under study, respectively 121 men and eight women, 101 were car-drivers and 28 motor-cyclists. The median age was equal to 29 years, while the average age to 32.0 years (range 15-65 years). Fifty eight cases (45.0%) were "positive" for the presence of ethanol > or = 0.8 g/l or other drugs. The sample of "positive cases" was studied according to sex, age, day, hour and type of vehicle. Considering the cases with presence of ethanol, although under the legal limit (20 cases), the total amount of cases (78) becomes even more consistent. The amount of ethanol was found to be respectively 0.34 g/l in daily drivers and 0.87 g/l in nightly drivers (p < 0.01). Our considerations confirm the importance of toxicological analyses in the forensic investigation of traffic deaths being the sample under study recorded following criteria which minimised other possible factors effecting road accidents.     

Estimation of the post-mortem interval in beagle dogs

Estimation of the time that has elapsed between death and the post-mortem examination of a body contributes to the investigation of death by defining the period during which death may have occurred. In a veterinary context, investigations involve a range of incidents including out-of-season shooting of game animals, poaching, death during transportation and cases of neglect or non-accidental injury of domestic animals. This study on dogs of the same breed, of similar weights and held under identical conditions, shows that a combination of rectal temperature, environmental temperature, gross pathology and histological changes can provide a scientifically based estimate of the post-mortem interval during the first 10 h following death, of up to one day, one to three days, three to seven days and one to three weeks.     

Postmortem diagnosis of syphilitic aortitis, including serological verification on postmortem blood

304 postmortem blood samples were serologically examined for syphilis at Statens Seruminstitut. Seventy-six percent of the samples were usable and gave clear-cut results (18% positive, 34% negative and 24% borderline). Twenty-four percent were indeterminable. The blood samples originated from 304 medico-legally examined bodies, 301 of whom were autopsied. The indications for the serological examinations were suspicion of syphilitic aortitis (SA) on the basis of the macroscopical appearance of the aorta at the autopsy--or information on earlier syphilis. Ninety-seven cases of SA were verified macro- and microscopically, 53 being lethal. The serological results were positive in half of these 97 cases, negative in 15, borderline in 21 and indeterminable in 12. A little more than half of the 97 cases were known in advance in the nation-wide syphilis index at the State Serum Institute. The variegated findings in the remaining part of the material are discussed. It is concluded that serological examination of postmortem blood may be a valuable adjuvant for the pathologist for the diagnosis of SA and other syphilitic manifestations.