Copper residues in meat from wild artiodactyls hunted with two types of rifle bullets manufactured from copper

Copper content in muscle and the presence of bullet fragments were assessed in samples from red deer (Cervus elaphus), roe deer (Capreolus capreolus), fallow deer (Dama dama) and wild boar (Sus scrofa) (total of 46 animals) which had been hunted with two types of solid copper bullets. Also, the release of copper from bullets or fragments remaining in muscle was tested. For bullet type “B”, a fragment was detected in only 1 out of 34 carcasses whereas for type “A”, fragments were detected in all 12 carcasses, with up to 24 fragments (maximum size, 5?×?7?×?2 mm). Median copper concentrations around the shot wound (0–30 mm distance) were 1.25 and 1.77 mg/kg fresh weight for bullet types B and A, respectively, and thus in the expected range for venison. Around bullet fragments that had remained in muscles, the copper content increased significantly. In roe deer longissimus muscle that had been exposed for 7 days at +7 °C to bullets of type B, up to 1,000 mg/kg copper (fresh weight) was found in a distance of 0–2 mm. However, in a distance of 10–20 mm, maximum copper contents were <10 mg/kg fresh weight. Bullet fragments can constitute physical hazards and will release copper under acidic conditions as those prevailing in meat. Removal of bullet fragments prior to culinary preparation should ensure that a recommended dietary copper intake of 1.25 mg per adult consumer per day is not exceeded. From a food hygiene viewpoint, non-fragmenting bullets seem to be preferable.     

Recreational Shooting of Prairie Dogs: A Portal for Lead Entering Wildlife Food Chains

Abstract: Although lead shot has been banned for waterfowl hunting in North America, some predators continue to exhibit elevated lead burdens, which has been attributed to ingesting metallic lead from other projectiles. Few studies have investigated residual lead fragments in hunted upland animals. Therefore, specific portals for lead entering wildlife food chains remain largely unknown. Prairie dogs (Cynomys spp.) are shot for recreation with minimal regulation in western North America. Because recreational shooters mostly use expanding bullets and rarely remove or bury carcasses, shot prairie dogs could make lead accessible to predators and scavengers. To determine whether and to what degree shot prairie dogs carry lead fragments, we analyzed carcasses shot by recreational shooters with 2 bullet types. Bullet type influenced the probability of bullet fragments being retained in carcasses; 87% of prairie dogs shot with expanding bullets contained bullet fragments, whereas 7% of carcasses shot with non-expanding bullets did. The amount of bullet fragments per carcass also differed between bullet types; carcasses shot with expanding bullets contained a mean of 228.4 mg of the lead-containing bullet core and 74.4 mg of the copper-alloy jacket, whereas carcasses shot with non-expanding bullets averaged only 19.8 mg of the core and 23.2 mg of the jacket. Lead fragments in carcasses shot with expanding bullets were small in size; 73% of all lead mass in each carcass was from fragments that weighed <25 mg each, small enough to be easily ingested and absorbed by secondary consumers. The amount of lead in a single prairie dog carcass shot with an expanding bullet is potentially sufficient to acutely poison scavengers or predators. Therefore, shot prairie dogs may provide an important portal for lead entering wildlife food chains and may pose risks to raptors and carnivores. Managers should consider measures, such as using non-expanding or lead-free ammunition, to reduce the likelihood of lead consumption and poisoning in upland wildlife.     

Ammunition Type and Shot Placement Determine Lead Fragmentation in Deer

Lead remnants from spent ammunition introduce health risks to humans and wildlife that consume game harvested with firearms. Most current research has focused on effects of high-velocity rifle bullets, whereas low-velocity lead ammunition has received little attention. We examined whether fragmentation characteristics differed between 3 common low-velocity ammunition types when shot into the thoracic cavity or shoulder of white-tailed deer (Odocoileus virginianus) culled in Indiana, USA, from May–September of 2009. We shot and subsequently radiographed 43 deer to determine the number of fragments, fragment size, and distance traveled by individual fragments. We also radiographed deer post-evisceration to determine the proportion of fragments available to humans and wildlife scavengers for consumption in muscle and visceral tissue, respectively. All radiographed deer had evidence of fragmentation, with a geometric mean of 13.1 (95% CI = 10.3, 16.8) fragments/deer. Most fragments (89%) were <5 mm from wound channels, and no fragment traveled beyond 205 mm from a wound channel. Fragments were often retained within the muscle tissue of deer with a geometric mean rate of 0.55 (95% CI = 0.48, 0.65). Muzzleloader bullet fragments were larger than those generated by rifled and sabot slugs, and sabot slug fragments had the shortest dispersal from wound channels. Shoulder-shot placement and bone contact for all ammunition resulted in a greater number of fragments (P < 0.01). Shoulder-shots also generated more small fragments and higher fragment retention in muscle tissue. The overall mean number of lead fragments detected across our ammunition treatments was less than previous studies. Our results indicate ammunition type and shot placement may be considerations for hunters wishing to limit their potential exposure to lead from harvested big game. Additionally, when compared to high-velocity rifle bullets, significantly fewer lead fragments are made available to humans and wildlife that consume game harvested with low-velocity ammunition types, such as those tested here. Complete elimination of lead ingestion by humans and wildlife, however, is only possible by using non-toxic ammunition alternatives. © 2020 The Wildlife Society.     

Nickel and copper residues in meat from wild artiodactyls hunted with nickel-plated non-lead rifle bullets

A nickel (Ni)-plated copper-solid bullet type released up to 93 μg Ni /10 g bullet mass when immersed into meat juice for 7 days (to simulate fragments remaining in venison). A non-nickel-plated counterpart of identical construction released no Ni, but up to 250 μg copper. During thermal processing of pork cubes with embedded bullets, an average of 2.8 and up to 4.3 (maximum) μg Ni were released from the Ni-plated bullet to the surrounding meat. Average nickel and copper content in meat samples (taken in 2–3 cm distance from the shot wounds) from 30 roe deer (Capreolus capreolus) and 3 sika deer (Cervus nippon) killed with nickel-plated copper bullets did not differ significantly from those in controls (roe deer haunch). Contamination scenarios would Cu and Ni contents per portion increase moderately by 20 and 3.3 μg, respectively. In order to limit alimentary Ni uptake, the technological need for Ni-plating of bullets should be carefully evaluated.     

Widespread Lead Exposure in Golden Eagles Captured in Montana

Lead poisoning threatens many species of raptors, including golden eagles (Aquila chrysaetos). Much of this lead likely comes from bullet fragments that remain in the carcasses of animals killed by hunters. The likelihood of lead exposure may peak during fall hunting seasons and early winter until carcasses from hunting become scarce. From 2011 to 2018 in western Montana, USA, we captured 91 golden eagles in winter, tested their blood lead levels (BLL), and outfitted a subset of birds (n = 29) with global positioning system [GPS] transmitters. Nearly all golden eagles (94.5%) had elevated BLL (≥10 μg/dL), and 8 of them had BLL above clinical exposure (>60 μg/dL), where they may lose coordination and experience a host of other neurological and physiological disorders. Golden eagles caught late in winter tended to have lower BLL than those caught earlier. At least 69% of the golden eagles equipped with GPS transmitters migrated northward, spending the summer throughout Alaska, USA, and northwestern Canada. Blood lead levels did not differ between migratory and nonmigratory golden eagles. Overall, elevated BLL are widespread among golden eagles throughout winter in western Montana. Promoting nonlead hunting ammunition in areas with high densities of golden eagles will reduce the birds' lead exposure. © 2020 The Wildlife Society.     

Potential Hazard to Human Health from Exposure to Fragments of Lead Bullets and Shot in the Tissues of Game Animals

Background

Lead is highly toxic to animals. Humans eating game killed using lead ammunition generally avoid swallowing shot or bullets and dietary lead exposure from this source has been considered low. Recent evidence illustrates that lead bullets fragment on impact, leaving small lead particles widely distributed in game tissues. Our paper asks whether lead gunshot pellets also fragment upon impact, and whether lead derived from spent gunshot and bullets in the tissues of game animals could pose a threat to human health.

Methodology/Principal Findings

Wild-shot gamebirds (6 species) obtained in the UK were X-rayed to determine the number of shot and shot fragments present, and cooked using typical methods. Shot were then removed to simulate realistic practice before consumption, and lead concentrations determined. Data from the Veterinary Medicines Directorate Statutory Surveillance Programme documenting lead levels in raw tissues of wild gamebirds and deer, without shot being removed, are also presented. Gamebirds containing ≥5 shot had high tissue lead concentrations, but some with fewer or no shot also had high lead concentrations, confirming X-ray results indicating that small lead fragments remain in the flesh of birds even when the shot exits the body. A high proportion of samples from both surveys had lead concentrations exceeding the European Union Maximum Level of 100 ppb w.w. (0.1 mg kg⁻¹ w.w.) for meat from bovine animals, sheep, pigs and poultry (no level is set for game meat), some by several orders of magnitude. High, but feasible, levels of consumption of some species could result in the current FAO/WHO Provisional Weekly Tolerable Intake of lead being exceeded.

Conclusions/Significance

The potential health hazard from lead ingested in the meat of game animals may be larger than previous risk assessments indicated, especially for vulnerable groups, such as children, and those consuming large amounts of game.     

Embedded lead shot in European starlings Sturnus vulgaris: an underestimated hazard for humans and birds of prey

The poisoning of wild animals by lead (Pb) ammunition fired by hunters has been known for many decades, especially in the case of waterbirds. More recently, it has been demonstrated that raptors are also exposed to the risk of plumbism when feeding on unretrieved quarry that was wounded or killed by hunters. Further studies reveal that even humans can be subject to a significant Pb dose while consuming game animals killed by traditional ammunition. Given the relevance of this issue, several pieces of research have been carried out to assess frequency, dimension, and the number of Pb fragments embedded in the carcasses of ungulates, partridges, ducks, and other birds to evaluate the risk related to the consumption of game meat. In spite of their great importance as quarry species across southern Europe, until now, no data have been available on small passerines. To assess the quantity and type of Pb embedded in songbirds, we x-rayed 196 starlings shot in Italy and found Pb pellets and/or visible fragments in 118 carcasses (60.2 %). We counted 128 shotgun pellets in 85 carcasses. In 28 birds, we detected both whole pellets and lead fragments; in 33, we found only small fragments. By excising and weighing a sample of 20 shotgun pellets (diameter 1.35–1.99 mm), we calculated a Pb load of 3.75 g in the whole sample of 196 starlings, corresponding to an average of 27.32 mg/100 g of body weight. This is a conservative estimation, because fragments were not considered. Compared to game birds of a larger size, the starlings in our study had a lower amount of embedded Pb, but the shot pellets and fragments embedded in their tissues were abundant and tiny. Given the results of previous studies, the quantity and level of fragmentation suggest that the risk of Pb poisoning cannot be ruled out for humans and birds of prey consuming the meat from songbirds killed with traditional ammunition.     

The role of ethics in the management of New Zealand's wild mammals

Abstract

Wild deer are used to illustrate the conflicts that arise when an animal is both a pest and a valued resource. At present there are c. 250 000 wild deer in New Zealand. These are pursued by c. 37 000 hunters who spend more than $20 million annually to harvest c. 70 000 deer. Nearly two-thirds of the harvest is taken by recreational hunters; the remainder forms the basis of a small industry for venison export. Despite their recreational and commercial value, wild deer continue to be a significant conservation pest. Conservationists generally would like wild deer to be eradicated, or, failing that, their densities held at the lowest levels possible with the control resources available. Hunters, however, would like deer to remain, and at higher rather than lower densities, to provide better prospects for hunting success. We discuss ways in which these apparently incompatible aspirations can be partially reconciled.     

Lead Toxicity Risks in Gunshot Victims

Background

Gunshot wounds require surgeons to decide whether to remove or leave bullet fragments in the body. Surgeons also decide how to follow up with patients who have lead fragments retained in their body. Current literature recommends to remove only intra-articular fragments without the need for a follow-up for patients with the metal retained. Therefore, this study investigates chronic lead toxicity for gunshot wounds.

Methods

The study was performed in the metropolitan area of Rio de Janeiro/Brazil, between 2013 and 2015. It was a case-control study that included 45 victims of gunshot lesions with metallic fragments retained for more than 6 months. The 45 controls were matched for gender, age, and race. We compared the lead blood levels and frequency of symptoms.

Results

The control group had average blood lead levels of 2.17 μg/dL (95% Confidence Interval [CI]; 1.71–2.63) and median 2.1 μg/dL. The case group had average values of 9.01 μg/dL (CI; 6.07–11.96) and median values of 6.5 μg/dL with p-values < = 0.001. The case group reported the following more frequently: irritancy, bad mood, headache, memory losses, daylight drowsiness, myalgia, weakness, abdominal pain, joint pain, trembling, tingling limbs. There was statistical significance for the differences of symptoms frequencies and for odds ratio between groups.

Conclusions

Although the mean lead levels found were lower than the current laboratory references, low levels have been associated with both rising morbidity and mortality. The WHO stated: “There is no known level of lead exposure that is considered safe”. In conclusion, this work showed that bullets retained in the body are not innocuous. There are impacts in the blood lead levels and symptoms related to it, even with few fragments, extra-articular located or existing with low blood lead levels.     

Trophic Facilitation or Limitation? Comparative Effects of Pumas and Black Bears on the Scavenger Community

Scavenging is a widespread behaviour and an important process influencing food webs and ecological communities. Large carnivores facilitate the movement of energy across trophic levels through the scavenging and decomposition of their killed prey, but competition with large carnivores is also likely to constrain acquisition of carrion by scavengers. We used an experimental approach based on motion-triggered video cameras at black-tailed deer (Odocoileus hemionus columbianus) carcasses to measure the comparative influences of two large carnivores in the facilitation and limitation of carrion acquisition by scavengers. We found that pumas (Puma concolor) and black bears (Ursus americanus) had different effects on their ecological communities. Pumas, as a top-level predator, facilitated the consumption of carrion by scavengers, despite significantly reducing their observed sum feeding times (165.7 min±21.2 SE at puma kills 264.3 min±30.1 SE at control carcasses). In contrast, black bears, as the dominant scavenger in the system, limited consumption of carrion by scavengers as evidenced by the observed reduction of scavenger species richness recorded at carcasses where they were present (mean = 2.33±0.28 SE), compared to where they were absent (mean = 3.28±0.23 SE). Black bears also had large negative effects on scavenger sum feeding times (88.5 min±19.8 SE at carcasses where bears were present, 372.3 min±50.0 SE at carcasses where bears were absent). In addition, we found that pumas and black bears both increased the nestedness (a higher level of order among species present) of the scavenger community. Our results suggest that scavengers have species-specific adaptions to exploit carrion despite large carnivores, and that large carnivores influence the structure and composition of scavenger communities. The interactions between large carnivores and scavengers should be considered in future studies of food webs and ecological communities.     

The extent of lead fragmentation observed in deer culled by sharpshooting

Sharpshooting is a proven management technique to lower white-tailed deer (Odocoileus virginianus) densities in areas where hunting is precluded. A donation program that allows for the consumptive use of these culled deer is often necessary to gain public approval for such a program. We culled 40 deer in Indiana using sharpshooting methods (head and neck shot placement) and radiographed the carcasses to determine if lead fragmentation spread throughout the skeletal muscle system. In 30 deer where shot placement was between the cranium and fourth cervical vertebrae, we observed no lead fragments in any thoracic or crural muscle tissue. Of 10 deer where shot placement was between the fifth and seventh cervical vertebrae, 8 deer experienced lead fragments in the extensor spinae muscle. Deer culled with highly frangible bullets via sharpshooting in the cranium or upper cervical spine have minimal risk of experiencing lead fragmentation in the thoracic or crural muscle systems. Deer shot in the lower neck may experience lead fragmentation in the anterior extensor spinae muscle, and up to 40 cm of that muscle should be removed before consumption. © 2011 The Wildlife Society.     

Adaptive management of sika deer populations in Hokkaido, Japan: theory and practice

We investigated the utility of adaptive management (AM) in wildlife management, reviewing our experiences in applying AM to overabundant sika deer (Cervus nippon) populations in Hokkaido, Japan. The management goals of our program were: (1) to maintain the population at moderate density levels preventing population irruption, (2) to reduce damage to crops and forests, and (3) to sustain a moderate yield of hunting without endangering the population. Because of significant uncertainty in biological and environmental parameters, we designed a “feedback” management program based on controlling hunting pressure. Three threshold levels of relative population size and four levels of hunting pressure were configured, with a choice of four corresponding management actions. Under this program, the Hokkaido Government has been promoting aggressive female culling to reduce the sika deer population since 1998. We devised a harvest-based estimation for population size using relative population size and the number of deer harvested, and found that the 1993 population size (originally estimated by extrapolation of aerial surveys) had been underestimated. To reduce observation errors, a harvest-based Bayesian estimation was developed and the 1993 population estimate was again revised. Analyses of population trends and harvest data demonstrate that hunting is an important large-scale experiment to obtain reliable estimation of population size. A serious side effect of hunting on sika deer was inadvertent lead poisoning of large birds of prey. The prohibition of the use of lead bullets by the Hokkaido Government was successful in reducing the lead poisoning, but the problem still remains. Two case studies on sika population irruption show that the densities set by maximum sustainable yield may be too high to prevent damage to agriculture, forestry, and/or ecosystems. Threshold management based on feedback control is better for ecosystem management. Since volunteer hunters favor higher hunting efficiency in resource management (e.g., venison), it is necessary to support the development of professional hunters for culling operations for ecosystem management, where lower densities of deer should be set for target areas. Hunting as resource management and culling for ecosystem management should be synergistically combined under AM.     

Lead Exposure in Bald Eagles from Big Game Hunting,the Continental Implications and Successful Mitigation Efforts

Studies suggest hunter discarded viscera of big game animals (i.e., offal) is a source of lead available to scavengers. We investigated the incidence of lead exposure in bald eagles in Wyoming during the big game hunting season, the influx of eagles into our study area during the hunt, the geographic origins of eagles exposed to lead, and the efficacy of using non-lead rifle ammunition to reduce lead in eagles. We tested 81 blood samples from bald eagles before, during and after the big game hunting seasons in 2005–2010, excluding 2008, and found eagles had significantly higher lead levels during the hunt. We found 24% of eagles tested had levels indicating at least clinical exposure (>60 ug/dL) during the hunt while no birds did during the non-hunting seasons. We performed driving surveys from 2009–2010 to measure eagle abundance and found evidence to suggest that eagles are attracted to the study area during the hunt. We fitted 10 eagles with satellite transmitters captured during the hunt and all migrated south after the cessation of the hunt. One returned to our study area while the remaining nine traveled north to summer/breed in Canada. The following fall, 80% returned to our study area for the hunting season, indicating that offal provides a seasonal attractant for eagles. We fitted three local breeding eagles with satellite transmitters and none left their breeding territories to feed on offal during the hunt, indicating that lead ingestion may be affecting migrants to a greater degree. During the 2009 and 2010 hunting seasons we provided non-lead rifle ammunition to local hunters and recorded that 24% and 31% of successful hunters used non-lead ammunition, respectively. We found the use of non-lead ammunition significantly reduced lead exposure in eagles, suggesting this is a viable solution to reduce lead exposure in eagles.     

Exposure of California Condors to Lead From Spent Ammunition

ABSTRACT The scientific evidence that California condors (Gymnogyps californianus) are frequently sickened and killed by lead poisoning from spent ammunition supports the conclusion that current levels of lead exposure are too high to allow reintroduced condors to develop self-sustaining populations in the wild in Arizona and, by inference, in California. The evidence for lead poisoning and its source comes from the following sorts of data: 1) 18 clinical necropsies revealing high levels of lead in body tissues and (or) presence of lead shotgun pellets and bullet fragments in digestive tracts; 2) moribund condors showing crop paralysis and impending starvation with toxic levels of lead in their blood; 3) widespread lead exposure among free-flying condors, many with clinically exposed or acute levels; 4) temporal and spatial correlations between big game hunting seasons and elevated lead levels in condors; and 5) lead isotope ratios from exposed condors showing close similarity to isotope ratios of ammunition lead but isotope ratios in less exposed condors being similar to environmental background sources, which are different from ammunition lead. Simple population models reveal harmful demographic impacts of unnatural mortality from lead on population trajectories of reintroduced condors. Recent innovations in the manufacture of nonlead shotgun pellets and bullets with superior ballistics now provide for a simple solution to the problem of lead ingestion by condors, many other species of wildlife, and human beings: substitute nontoxic forms of ammunition for traditional lead-based ammunition. The substitution of nontoxic ammunition would be highly efficacious for hunting, economically feasible, and the right thing to do.     

Risk Factors for Death among Children Less than 5 Years Old Hospitalized with Diarrhea in Rural Western Kenya, 2005–2007: A Cohort Study

Background

Diarrhea is a leading cause of childhood morbidity and mortality in sub-Saharan Africa. Data on risk factors for mortality are limited. We conducted hospital-based surveillance to characterize the etiology of diarrhea and identify risk factors for death among children hospitalized with diarrhea in rural western Kenya.

Methods and Findings

We enrolled all children <5 years old, hospitalized with diarrhea (≥3 loose stools in 24 hours) at two district hospitals in Nyanza Province, western Kenya. Clinical and demographic information was collected. Stool specimens were tested for bacterial and viral pathogens. Bivariate and multivariable logistic regression analyses were carried out to identify risk factors for death. From May 23, 2005 to May 22, 2007, 1,146 children <5 years old were enrolled; 107 (9%) children died during hospitalization. Nontyphoidal Salmonella were identified in 10% (118), Campylobacter in 5% (57), and Shigella in 4% (42) of 1,137 stool samples; rotavirus was detected in 19% (196) of 1,021 stool samples. Among stools from children who died, nontyphoidal Salmonella were detected in 22%, Shigella in 11%, rotavirus in 9%, Campylobacter in 5%, and S. Typhi in <1%. In multivariable analysis, infants who died were more likely to have nontyphoidal Salmonella (adjusted odds ratio [aOR] = 6·8; 95% CI 3·1–14·9), and children <5 years to have Shigella (aOR = 5·5; 95% CI 2·2–14·0) identified than children who survived. Children who died were less likely to be infected with rotavirus (OR = 0·4; 95% CI 0·2–0·8). Further risk factors for death included being malnourished (aOR = 4·2; 95% CI 2·1–8·7); having oral thrush on physical exam (aOR = 2·3; 95% CI 1·4–3·8); having previously sought care at a hospital for the illness (aOR = 2·2; 95% CI 1·2–3·8); and being dehydrated as diagnosed at discharge/death (aOR = 2·5; 95% CI 1·5–4·1). A clinical diagnosis of malaria, and malaria parasites seen on blood smear, were not associated with increased risk of death. This study only captured in-hospital childhood deaths, and likely missed a substantial number of additional deaths that occurred at home.

Conclusion

Nontyphoidal Salmonella and Shigella are associated with mortality among rural Kenyan children with diarrhea who access a hospital. Improved prevention and treatment of diarrheal disease is necessary. Enhanced surveillance and simplified laboratory diagnostics in Africa may assist clinicians in appropriately treating potentially fatal diarrheal illness. Please see later in the article for the Editors'' Summary     

Sensitivity of Five Rapid HIV Tests on Oral Fluid or Finger-Stick Whole Blood: A Real-Time Comparison in a Healthcare Setting

Background

Health authorities in several countries recently recommended the expansion of human immunodeficiency virus (HIV) antibody testing, including the use of rapid tests. Several HIV rapid tests are now licensed in Europe but their sensitivity on total blood and/or oral fluid in routine healthcare settings is not known.

Methods and Findings

200 adults with documented HIV-1 (n = 194) or HIV-2 infection (n = 6) were prospectively screened with five HIV rapid tests using either oral fluid (OF) or finger-stick whole blood (FSB). The OraQuick Advance rapid HIV1/2® was first applied to OF and then to FSB, while the other tests were applied to FSB, in the following order: Vikia HIV 1/2®, Determine HIV 1–2®, Determine® HIV-1/2 Ag/Ab Combo® and INSTI HIV-1/HIV-2®. Tests negative on FSB were repeated on paired serum samples. Twenty randomly selected HIV-seronegative subjects served as controls, and the results were read blindly. Most patients had HIV-1 subtype B infection (63.3%) and most were on antiretroviral therapy (68.5%). Sensitivity was 86.5%, 94.5%, 98.5%, 94.9%, 95.8% and 99% respectively, with OraQuick OF, OraQuick FSB, Vikia, Determine, Determine Ag/Ab Combo and INSTI (p<0.0001). OraQuick was less sensitive on OF than on FSB (p = 0.008). Among the six patients with three or more negative tests, two had recent HIV infection and four patients on antiretroviral therapy had undetectable plasma viral load. When patients positive in all the tests were compared with patients who had at least one negative test, only a plasma HIV RNA level <200 cp/ml was significantly associated with a false-negative result (p = 0.009). When the 33 rapid tests negative on FSB were repeated on serum, all but six (5 negative with OraQuick, 1 with INSTI) were positive. The sensitivity of OraQuick, Determine and Determine Ag/Ab Combo was significantly better on serum than on FSB (97.5%, p = 0.04; 100%, p = 0.004; and 100%, p = 0.02, respectively).

Conclusion

When evaluated in a healthcare setting, rapid HIV tests were less sensitive on oral fluid than on finger-stick whole blood and less sensitive on finger-stick whole blood than on serum.     

Association of Blood Lead (Pb) and Plasma Homocysteine: A Cross Sectional Survey in Karachi,Pakistan

Background

High blood lead (Pb) and hyperhomocysteinemia have been found to be associated with cardiovascular disease (CVD). Mean blood Pb and mean plasma homocysteine levels have been reported to be high in Pakistani population. The objective of the present study was to assess the relationship of blood Pb to the risk of hyperhomocysteinemia in a low income urban population of Karachi, Pakistan.

Methodology/Principal Findings

In a cross sectional survey, 872 healthy adults (355 males, 517 females; age 18–60 years) were recruited from a low income urban population of Karachi. Fasting venous blood was obtained and assessed for blood Pb and plasma/serum homocysteine, folate, pyridoxal phosphate (PLP, a coenzymic form of vitamin B6) and vitamin B12. The study population had median (IQR) blood Pb of 10.82 µg/dL (8.29–13.60). Prevalence of high blood Pb (levels >10 µg/dL) was higher in males compared to females (62.5% males vs 56% females; p value = 0.05). Mean ± SD/median (IQR) value of plasma homocysteine was significantly higher in the highest quartile of blood Pb compared to the lowest quartile 16.13±11.2 µmol/L vs 13.28±9.7µmol/L/13.15 (10.33–17.81) µmol/L vs 11.09 (8.65 14.31) µmol/L (p value<0.001). Daily consumption of fruit juice had a positive influence on both levels of plasma homocysteine and blood Pb. Compared with the lowest quartile of blood Pb, the OR for hyperhomocysteinemia was 1.69 (95% CI, 1.00 to 2.85) for the fourth quartile when the model was adjusted for age, gender, folate and vitamin B12.

Conclusions/Significance

This study showed a relationship between blood Pb and hyperhomocysteinemia in a general population of Karachi, Pakistan. The harmful effect of Pb on cardiovascular system could be due to its association with hyperhomocysteinemia.     

Accuracy and Reliability of Pallor for Detecting Anaemia: A Hospital-Based Diagnostic Accuracy Study

Background

Anaemia is a common disorder. Most health providers in resource poor settings rely on physical signs to diagnose anaemia. We aimed to determine the diagnostic accuracy of pallor for anaemia by using haemoglobin as the reference standard.

Methodology/Principal Findings

In May 2007, we enrolled consecutive patients over 12 years of age, able to consent and willing to participate and who had a haemoglobin measurement taken within a day of assessment of clinical pallor from outpatient and medicine inpatient department of a teaching hospital. We did a blind and independent comparison of physical signs (examination of conjunctivae, tongue, palms and nailbed for pallor) and the reference standard (haemoglobin estimation by an electronic cell counter). Diagnostic accuracy was measured by calculating likelihood ratio values and 95% confidence intervals (CI) at different haemoglobin thresholds and area under the receiver operating characteristic curve. Two observers examined a subset of patients (n = 128) to determine the inter-observer agreement, calculated by kappa statistics. We studied 390 patients (mean age 40.1 [SD 17.08] years); of whom 48% were women. The haemoglobin was <7 g/dL in 8% (95% confidence interval, 5, 10) patients; <9 g/dL in 21% (17, 26) patients and <12 g/dL in 64% (60, 70) patients. Among patients with haemoglobin <7 g/dL, presence of severe tongue pallor yielded a LR of 9.87 (2.81, 34.6) and its absence yielded a LR of 0. The tongue pallor outperformed other pallor sites and was also the best discriminator of anaemia at haemoglobin thresholds of 7 g/dL and 9 g/dL (area under the receiver operating characteristic curves (ROC area  = 0.84 [0.77, 0.90] and 0.71[0.64, 0.76]) respectively. The agreement between the two observers for detection of anaemia was poor (kappa values  = 0.07 for conjunctival pallor and 0.20 for tongue pallor).

Conclusions/Significance

Clinical assessment of pallor can rule out and modestly rule in severe anaemia.     

Recent Rapid Rise of a Permethrin Knock Down Resistance Allele in Aedes aegypti in México

Background

Aedes aegypti, the ‘yellow fever mosquito’, is the primary vector to humans of dengue and yellow fever flaviviruses (DENV, YFV), and is a known vector of the chikungunya alphavirus (CV). Because vaccines are not yet available for DENV or CV or are inadequately distributed in developing countries (YFV), management of Ae. aegypti remains the primary option to prevent and control outbreaks of the diseases caused by these arboviruses. Permethrin is one of the most widely used active ingredients in insecticides for suppression of adult Ae. aegypti. In 2007, we documented a replacement mutation in codon 1,016 of the voltage-gated sodium channel gene (para) of Ae. aegypti that encodes an isoleucine rather than a valine and confers resistance to permethrin. Ile1,016 segregates as a recessive allele conferring knockdown resistance to homozygous mosquitoes at 5–10 µg of permethrin in bottle bioassays.

Methods and Findings

A total of 81 field collections containing 3,951 Ae. aegypti were made throughout México from 1996 to 2009. These mosquitoes were analyzed for the frequency of the Ile1,016 mutation using a melting-curve PCR assay. Dramatic increases in frequencies of Ile1,016 were recorded from the late 1990''s to 2006–2009 in several states including Nuevo León in the north, Veracruz on the central Atlantic coast, and Yucatán, Quintana Roo and Chiapas in the south. From 1996 to 2000, the overall frequency of Ile1,016 was 0.04% (95% confidence interval (CI95) = 0.12%; n = 1,359 mosquitoes examined). The earliest detection of Ile1,016 was in Nuevo Laredo on the U.S. border in 1997. By 2003–2004 the overall frequency of Ile1,016 had increased ∼100-fold to 2.7% (±0.80% CI95; n = 808). When checked again in 2006, the frequency had increased slightly to 3.9% (±1.15% CI95; n = 473). This was followed in 2007–2009 by a sudden jump in Ile1,016 frequency to 33.2% (±1.99% CI95; n = 1,074 mosquitoes). There was spatial heterogeneity in Ile1,016 frequencies among 2007–2008 collections, which ranged from 45.7% (±2.00% CI95) in the state of Veracruz to 51.2% (±4.36% CI95) in the Yucatán peninsula and 14.5% (±2.23% CI95) in and around Tapachula in the state of Chiapas. Spatial heterogeneity was also evident at smaller geographic scales. For example within the city of Chetumal, Quintana Roo, Ile1,016 frequencies varied from 38.3%–88.3%. A linear regression analysis based on seven collections from 2007 revealed that the frequency of Ile1,016 homozygotes accurately predicted knockdown rate for mosquitoes exposed to permethrin in a bioassay (R² = 0.98).

Conclusions

We have recorded a dramatic increase in the frequency of the Ile1,016 mutation in the voltage-gated sodium channel gene of Ae. aegypti in México from 1996 to 2009. This may be related to heavy use of permethrin-based insecticides in mosquito control programs. Spatial heterogeneity in Ile1,016 frequencies in 2007 and 2008 collections may reflect differences in selection pressure or in the initial frequency of Ile1,016. The rapid recent increase in Ile1,016 is predicted by a simple model of positive directional selection on a recessive allele. Unfortunately this model also predicts rapid fixation of Ile1,016 unless there is negative fitness associated with Ile1,016 in the absence of permethrin. If so, then spatial refugia of susceptible Ae. aegypti or rotational schedules of different classes of adulticides could be established to slow or prevent fixation of Ile1,016.     

High Risk of Lead Contamination for Scavengers in an Area with High Moose Hunting Success

Top predators and scavengers are vulnerable to pollutants, particularly those accumulated along the food chain. Lead accumulation can induce severe disorders and alter survival both in mammals (including humans) and in birds. A potential source of lead poisoning in wild animals, and especially in scavengers, results from the consumption of ammunition residues in the tissues of big game killed by hunters. For two consecutive years we quantified the level lead exposure in individuals of a sentinel scavenger species, the common raven (Corvus corax), captured during the moose (Alces alces) hunting season in eastern Quebec, Canada. The source of the lead contamination was also determined using stable isotope analyses. Finally, we identified the different scavenger species that could potentially be exposed to lead by installing automatic cameras targeting moose gut piles. Blood lead concentration in ravens increased over time, indicating lead accumulation over the moose-hunting season. Using a contamination threshold of 100 µg.L⁻¹, more than 50% of individuals were lead-contaminated during the moose hunting period. Lead concentration was twice as high in one year compared to the other, matching the number of rifle-shot moose in the area. Non-contaminated birds exhibited no ammunition isotope signatures. The isotope signature of the lead detected in contaminated ravens tended towards the signature from lead ammunition. We also found that black bears (Ursus americanus), golden eagles and bald eagles (Aquila chrysaetos and Haliaeetus leucocephalus, two species of conservation concern) scavenged heavily on moose viscera left by hunters. Our unequivocal results agree with other studies and further motivate the use of non-toxic ammunition for big game hunting.