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As climate change intensifies, heat stress mitigation for pigs becomes more important. Trials involving induced heat waves are useful to test several measures (e.g. reduced stocking density) at a faster rate, but only when accurately evaluated and validated. In the present study, we investigated the suitability of an artificial heating protocol at different pig weights (experiment 1). The impact of different stocking densities on fattening pigs during an artificial heat wave (experiment 2) was also investigated. Experiment 1: Forty 20-week-old pigs weighing 96.5 ± 7.3 kg (W100) and forty 17-week-old pigs weighing 72.7 ± 9.9 kg (W70) were housed in two compartments. An artificial heat wave (heat load) was induced for 3 days. During 3-day periods before, during and after the heat load, physiological parameters (respiration rate (RR), rectal temperature (Trectal), skin temperature (Tskin) and behavior) were measured and average daily feed intake was observed. Ambient temperature, relative humidity and temperature-humidity index (THI) were monitored. Experiment 2: A total of 150 fattening pigs were randomly divided into three treatment groups: SD1.3 (1.3 m2/pig), SD1.0 (1.0 m2/pig) and SD0.8 (0.8 m2/pig). All pens had a total pen surface of 4.88 m2, corresponding with 4, 5 and 6 fattening pigs in the SD1.3, SD1.0 and SD0.8 groups, respectively. The heat load was induced for 7 days on week 21. Respiration rate and Trectal were observed as in experiment 1. Average daily gain and average daily feed intake were also noted. During the heat load, THI reached ≥ 75 (78.4 (experiment 1) and 78.6 (experiment 2)), even when relative humidity decreased to ± 45%. Every physiological parameter showed significant increases during the heat load. The prolonged heating protocol in experiment 2 also provoked significant decreases in average daily feed intake (15%) and average daily gain (19%) for all groups. Weight within the studied range of 70–100 kg did not have a significant impact on any of the parameters. However, Tskin was affected by both weight and heat load (P

The impact of pork production on global livestock’s greenhouse gas emissions is substantial. Understanding the factors influencing these emissions is crucial in achieving a more sustainable pig husbandry. In two independent experiments, the impact of slaughter weight on the carbon footprint (CFP) of pig feed intake (CFPFI) was evaluated for growing-finishing pigs of different sexes (entire males (EM), barrows (BA), immunocastrates (IC) and gilts (GI)). In experiment 1 118 animals were raised individually in experimental conditions. In experiment 2 384 animals were housed in group (four pigs per pen), in controlled commercial farm circumstances. All animals were fed ad libitum in a three-phase feeding regime and slaughtered at different BW, ranging from 110 to 148 kg (experiment 1) and from 99 to 138 kg (experiment 2). When only the fattening period was considered, the CFPFI was expressed per kg carcass growth. When the production of piglets was also taken into account, the CFPFI was calculated per kg carcass weight. For all sexes, the heavier the pig, the higher the CFPFI per kg carcass growth (P

For many years, pig production has focused on maximizing performance by selecting for maximal muscle growth and feeding diets that allow the animals to express their genetic potential. However, it is unclear whether this selection for muscle deposition has affected the capacity of pigs to cope with by-product-based diets, which rely on fat as the primary energy source instead of starches and sugars. Therefore, an experiment was set up to investigate if different types of boars affect how their progeny cope with alternative ingredients in the diet, with a possible need for adapted breeding schemes. Two types of boars within the Piétrain sire line were used based on either a high or low estimated breeding value for daily feed intake (HFI: high feed intake, low feed intake). When their progeny reached 14 weeks of age, two dietary strategies were compared: a control (CON) vs a by-product-based diet high in fat and fiber (HFF). The CON diet was mainly based on cereals (corn, wheat, barley) and soybean meal. The HFF diet was formulated to contain the same net energy, CP and digestible amino acid levels without any cereals or soybean meal. In total 192 animals were included in the experiment (48 animals/type of boar/diet) and performance, digestibility, carcass and meat quality were compared. None of the parameters showed a significant interaction (P

Incremental levels of the phytase, OptiPhos® Plus, were fed to 150 weaned piglets (five treatments of 30 pigs) to determine the effect on piglet performance, nutrient digestibility and bone mineralisation. Piglets were fed a basal diet from four to six weeks and a pre-starter diet from six to ten weeks of age. The basal diets contained reduced digestible phosphorus (P) and calcium (Ca) (1.5 g/kg digestible P, and Ca 5.5 g/kg and 5.6 g/kg for weaner and pre-starter, respectively) to provide a negative control. Phytase was added at 0, 125, 250, 500 and 1000 phytase units (FTU)/kg feed. Faecal samples were collected at nine weeks of age and analysed for P, Ca and crude protein (CP) content to calculated digestibility. Metacarpal bone IV mineralisation was assessed by ash content and the weight and length of the bone. Incremental increases of phytase improved final body weight (BW) (P

The practice of surgical castration of piglets and its alternatives is still under debate. Production of boars may impair meat quality due to boar taint and reduced tenderness compared to meat from surgically castrated male pigs, while immunocastration reduces boar taint and may improve meat quality but seems to be less accepted by the pig chain. In this study, we aimed to evaluate the consumer’s sensory appreciation of meat from barrows (BAs), immunocastrates (ICs) and boars (BOs) in six European countries, taking into account the selection of tainted carcass and consumers’ appreciation of boar taint. Loin chops of 30 BAs, 30 ICs and 30 BOs were evaluated by 752 consumers in six countries: Belgium, Czech Republic, Poland, Portugal, Romania and Spain. Consumers rated odour, flavour, tenderness, juiciness, overall liking and willingness to buy and sensitivity to and liking of androstenone (AND) and liking of skatole (SKA) was also tested. In each of the six countries, consumers liked the odour of the BO samples less than that of BA, and IC intermediate. For flavour, tenderness, juiciness, overall liking and willingness to buy, liking scores given by the Czech, Polish and Portuguese consumers significantly differed between the BA, BO and IC. Willingness to buy was highest for BA by Czech and Polish consumers and for BA and IC by Portuguese consumers. The frequency of the negative check all terms that apply terms also differed, with a higher frequency of disgusting for BO compared to BA and IC and of off-flavour, irritating, manure, sweat, disappointing compared to BA, and intermediate for IC. 31% of the consumers disliked the odour of AND (NEGAND), and 36% of them were not sensitive; in contrast, 77% of the consumers disliked SKA (NEGSKA). The decrease in flavour liking score for BO compared to BA and IC was more outspoken by the NEGAND consumer, while NEGSKA consumers gave an overall lower liking score independent of the type of male pig. The results of this study indicate that IC can be a valid alternative for surgical castration.

Immunocastration, a technique consisting of two vaccinations against gonadotropin-releasing hormone (GnRH), can be used as alternative to surgical castration of piglets. It reduces boar taint and allows higher economic and ecological efficiency compared to barrows. The feeding strategy of immunocastrates, however, can still be improved. After second vaccination, when immunisation becomes fully effective, feed intake of immunocastrates increases sharply. This study aimed to investigate whether energy intake of immunocastrates after second vaccination could be reduced by lowering the dietary energy level of the finishing phase, without negatively affecting animal performance and quality of pork production. We hypothesised that immunocastrates already reach their limits in voluntary feed intake after second vaccination, and therefore would not be able to compensate the lower dietary energy level, in contrast to barrows. Therefore, this study aimed to assess the effect of high-energy diet (HE, net energy (NE) = 10.2 MJ/kg) compared to low-energy diet (LE, NE = 8.8 MJ/kg) in barrows and immunocastrates and as a reference, gilts and entire male pigs on a standard high-energy diet were included. CP and standardised ileal digestible amino acid levels were similar in both diets. For each treatment, eight pen replicates of six pigs per pen were evaluated on performance, carcass quality, meat and fat quality, digestibility, economic and ecological sustainability, behaviour and effectiveness of immune response. No difference in feed intake of immunocastrates between LE and HE could be demonstrated. As a result, daily energy intake of immunocastrates was higher on HE compared to LE, which resulted in a higher daily gain on HE. Feed conversion ratio (FCR) of immunocastrates on HE did not differ significantly with FCR of entire males. Barrows did not show higher average daily gain on HE compared to LE. Nitrogen efficiency was better in HE compared to LE, without negative effects on digestibility, carcass quality, economic parameters, behaviour or immune response. Small positive effects on the palatability of the meat of immunocastrates on HE were observed, although consumers did not prefer one of both feeds. Immunocastration was successful in reducing sexual and aggressive behaviour as well as in lowering the prevalence of boar taint from 15% in EM to 0% in immunocastrates. However, in two out of 96 immunocastrates (one on HE and one on LE), the immunocastration was not fully effective. In conclusion, this study did not show advantages of feeding immunocastrates or barrows a low-energy diet.

Economic margins on pig farms are small, and changing slaughter weights may increase farm profitability. However, one can question if the optimal slaughter weight is the same for each sex. On three farms, crossbred pigs (n = 1128) were used to determine the effect of sex and slaughter weight on performance, carcass quality and gross margin per pig place per year. On each farm, an equal number of entire males (EMs), barrows (BAs), immunocastrates (IC) and gilts (GIs) were housed separately in group pens. Pens were randomly divided into three categories of different slaughter weights: 105, 117 and 130 kg BW. In BA, the high average daily feed intake (ADFI) and the lower capacity to gain muscle led to a higher feed conversion ratio (FCR) and lower lean meat percentage in comparison to EM and IC. In all sexes, ADFI and FCR increased with an increasing slaughter weight but the effect of slaughter weight on carcass quality varied between sexes. In BA and GI, slaughter weight had no effect on carcass quality, but in EM and IC, carcass quality improved at higher slaughter weights. Gross margin per pig place per year was calculated as gross margin per pig × barn turnover per year, taking into account fixed costs per round, feed costs and output price per pig. The slaughter weight that gained the highest gross margin per year differed between sexes. Slaughtering BA and GI at 130 kg BW, compared to 105 or 117 kg BW, decreased the gross margin per pig place per year due to the lower margin per pig and barn turnover at higher weights. In IC and EM, no difference in gross margin per pig place per year could be demonstrated between slaughtering at 105, 117 or 130 kg BW. In IC, the increasing gross margin per pig with increasing slaughter weights counteracted with the lower barn turnover. In EM, gross margin per pig did not differ between slaughter weights, but the effect of barn turnover was too small to demonstrate significant differences between slaughter weights on gross margin per pig place per year. In conclusion, slaughter weight has an impact on profitability in BA and GI: they should not be slaughtered at 130 kg BW but at lower weights, but no effect could be demonstrated in EM and IC.

Boar taint is an unpleasant taste and odor that can occur in entire male pigs and is caused by androstenone, skatole, and to a lesser extent indole accumulating in fat tissue. In the present observational study, we evaluated an extensive list of such potential risk factors which influence boar taint: social hierarchy and puberty attainment, housing, health, preslaughter conditions, season, feed, carcass composition, slaughter weight or age, and breed. Details on these factors were collected by interviews with the participating farmers, observations on each farm by trained observers and farmers, as well as slaughterhouse data. Twenty-two farms (in West- and East-Flanders, ranging from 160 to 600 sows, selected on suitability) raising entire male pigs were included in the study to evaluate the link between boar taint and potential risk factors related to the farm and slaughter batch (114 slaughter batches and 16 791 entire male pigs in total). Average olfactory boar taint prevalence was 1.8 ± 0.8%. Boar taint prevalence varied also within farms up to a maximum range between slaughter batches of 9.1% which suggests an effect of factors varying between slaughter batches such as season or other variables varying between slaughter batches. Less aggressive behavior at the end of fattening as well as lower skin lesion scores at fattening as well as at slaughter could be associated with less boar taint. The same might be said for sexual behavior, though less convincingly from this study. Measures that reduce aggression and stress have therefore have the potential to lower boar taint prevalence. The same might be said for sexual behavior, though less convincingly from this study. Furthermore, boar taint prevalence was generally higher in winter than in summer, which is relevant from a planning perspective for the slaughterhouses to seek alternative markets. Finally, increased CP gave significantly lower boar taint prevalences. This may to some extent be explained by the negative association between boar taint and lean meat percentage, as increased dietary CP levels promote the carcass lean meat percentages which can then be associated with lower boar taint levels.

Genetic evaluation of Piétrain sires in Flanders occurs under standardized conditions, on test stations with fixed dam breeds, standardized diets and uniform management practices. As environmental conditions vary on commercial farms and differ from the test stations, this study aimed at understanding to what extent the sire, the dam breed and the interaction between both affects the translation of breeding values to practice. Dams of two commercial breeds were inseminated with semen from one of five different sires selected for contrasting breeding values (daily gain, feed conversion ratio and carcass quality). For each sire by dam breed combination, six pen replicates (with three gilts and three barrows per pen) were evaluated for growth performance from 9 weeks of age (20 kg) to slaughter (110 kg), and for carcass and meat quality. In our experimental setup, both sire and dam breed affected growth, carcass and meat quality traits. No significant sire×dam breed interactions on performance could be detected. Though a tendency for interaction on average daily feed intake between 20 and 110 kg (P=0.087), and on pork colour (lightness) (P=0.093) was present. In general, offspring of all tested sires behaved similarly in both dam breeds, indicating that estimated breeding values for Piétrain sires determined in one dam breed are representative in other dam breeds as well.

Background: The Global Anticoagulant Registry in the FIELD–Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. Methods and results: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012–2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P

The perspective of a possible ban on surgical castration of male pigs in the EU is a real challenge for pork production systems aiming at (very) high-quality products. Information was collected from a total of 272 situations in 16 European countries, including 170 situations related to EU protected designations (Database of Origin & Registration (DOOR) database) and 102 other situations related to high-quality products or differentiated production systems, in order to evaluate their potential sensitivity to the use of entire male pigs along four dimensions: BT_Inc, likelihood of increased levels of boar taint compounds compared with conventional production of entire males; BT_Per, extent to which (some of) the associated pork product(s) are susceptible to perception of boar taint by consumers; FatQQ, likelihood that the quality of (some of) the related products is decreased due to the lower fat quantity and quality in entire males; Manag, increased likelihood of animal management and welfare problems compared with conventional production of entire males. Situations corresponding to EU protected designations (DOOR situations) were on average more sensitive to entire male production but 11% of the non-DOOR situations were highly potentially sensitive, whereas one-third of the DOOR situations had low potential sensitivity. In total, 37% of the situations where castration is not formally specified as mandatory exhibited high potential sensitivity to entire male production. Three main patterns of situations were identified via ascending hierarchical clustering. A first pattern including 31% of the DOOR situations and 74% of the other ones, had potentially no increased risk compared with conventional production of entire males. A second pattern including 28% of the DOOR situations and 16% of the other ones had a high, moderate and low potential sensitivity for FatQQ, BT_Inc and Manag, respectively. The third pattern including 41% of the DOOR situations and 11% of the other situations had high potential sensitivity for BT_Inc and FatQQ, associated with moderate to high sensitivity for Manag. The approach used to evaluate the sensitivity to entire male pig production from the limited information collected for this study has many limitations. More precise approaches using more specific information are needed to evaluate the actual sensitivity of individual situations to the use of entire male pigs. Still, the present study provides a first global insight on the capacity of European production systems aiming at high-quality products to use entire male pigs as an alternative to surgical castration.

During growth, pigs convert plant protein into animal protein. The major part of the ingested protein is excreted via manure, with potential nitrogen (N) losses to the environment. To limit N losses and increase sustainability of pork production, the efficiency of protein conversion should be maximized. The aim of this paper is to critically evaluate diet and management strategies linked with N efficiency. Besides nutrition, we discuss three management strategies observed in science and in practice to be linked with improved N efficiency: genetic selection, castration and slaughter weight. Because diet has a marked effect on eventual N losses, it must also be taken into account when evaluating management strategies. A reductionist approach, such as feeding the same diet across all management treatments, may overestimate the effect of a management strategy and eventually lead to incorrect conclusions. The amount of excreted N depends on the amount of ingested N, the amount of absorbed N, the amino acid (AA) balance in the diet and the animal’s N and AA requirements. Daily multiphase feeding adapted to the individual animal’s AA needs is likely to be the most N efficient. For animals housed in groups, phase feeding is necessary. When combined with periods of temporary AA restriction, N efficiency can be further improved. Specific AA consumption must be balanced by applying the ideal protein concept. With better knowledge of the requirements of individual animals and the commercial availability of certain AAs, the total dietary CP level can be lowered within limits. Further research is needed on the minimal CP level that allows maximal performance. For this end a useful parameter may be the ratio of standardized ileal digestible (SID) lysine : apparent total tract digestible CP level. By combining optimal nutrition and management, a whole body N efficiency approaching 60% may be achievable in the near future.

Sequence variation of the hypervariable segments (HVS) I/II of mitochondrial DNA (mtDNA) and the haplogroup affiliation were determined in a sample of 271 Italian subjects. This analysis showed that 42% of the individuals could be ascribed to H, the most frequent haplogroup in European Caucasian populations. This fraction was then screened for specific single nucleotide polymorphisms located in the coding region to identify H subclades H1–H15. We set up two multiplex polymerase chain reactions and specific SNaPshot assays to investigate the frequency distribution of these subgroups in our population sample and to examine their usefulness in discriminating among commonly shared HVS I/II sequences. This allowed the assignment of a large portion of the mtDNAs (∼70%) to specific subhaplogroups, with H1 and H5 being the most represented. About two-thirds of the individuals sharing common HVS I/II sequences were subdivided and ascribed to specific H subhaplogroups with a significant reduction of the frequencies of the most common mtDNA haplotypes. Haplogroup H subtyping could thus be extremely useful in forensic identification when many samples have to be analysed and compared, avoiding excessive time-consuming and labor-intensive sequencing analysis. [ABSTRACT FROM AUTHOR]

There is a commitment by the European pig sector to ban surgical castration of male piglets in the European Union in 2018. One alternative to castration is to raise entire male pigs, with an increased risk of boar taint. A field study was performed to: (1) evaluate inter- and intra-farm variation in boar taint prevalence, (2) investigate factors measured at slaughter influencing boar taint and (3) evaluate the relationship between sensorial scoring by a trained panel and the concentration of boar taint components. From 34 farms, neck fat samples were collected from all entire male pigs in at least two slaughter batches per farm (78 batches; 9167 animals). In addition to olfactory boar taint analysis, data were also collected on fresh skin lesions (score 0 to 3) at the slaughter line, slaughter weight, lean meat percentage, duration of transport, time spent in lairage, total delivery duration, day length, shortening of days and outdoor mean temperature. Using the hot iron method, neck fat samples were scored (eight-point scale) for boar taint. Average boar taint prevalence (score ≥3) was 5.6±2.5% and the mean difference between the maximum and minimum prevalence per farm was 4.3±3.2%. Androstenone (AND), skatole (SKA) and indole concentrations were measured for a subset (n=254) of the samples. According to binomial univariate mixed models, entire male pigs with a higher skin lesion score had higher odds of having boar taint (P=0.031), as did fatter entire male pigs (P

In 102 conditionally healthy volunteers aged from 19 to 30 years (average age is 19,53 ± 11 years) the volatility of heart rate variability (HRV) spectral parameters depending on body mass index (BMI) were evaluated. According to WHO recommendations on the calculation and interpretation of BMI were such groups of volunteers: underweight, normal body weight, overweight, obesity I degree, obesity II degree, obesity III degree. Among HRV parameters were evaluated total power (TP, ms2), power of very low frequency (VLF, ms2), low frequency (LF, ms2) and high frequency (HF, ms2) domains of HRV spectrum in the 5-minute intervals of ECG in I standard lead. The data were processed by methods of nonparametric statistics. It was established that spectral characteristics of HRV in volunteers with normal BMI have a high TP with harmonious relations between VLF, LF and HF domains; decreased or increased BMI provokes TP reduction by decreasing power of all domains of HRV (VLF, LF, HF) with a predominance of VLF proportion and this effect increases with the degree of deviation of the parameter.

Immunocastration or vaccination against boar taint can be used as alternative for surgical castration of male piglets. The vaccine is administrated twice. After the second vaccination (V2), the pigs behave like barrows instead of boars and their feed intake increases which may result in a lower lean meat percentage. The timing of V2 is therefore crucial to find the right balance between the advantages of entire males and barrows. In this study, we evaluated the effect of time post second injection within the advised time frame (4 v. 6 weeks before slaughter) on behaviour, performance, carcass and meat quality of immunocastrated male pigs. In total, 180 animals (hybrid sow×Piétrain): 60 gilts, 60 male pigs vaccinated 6 weeks before slaughter (IM-6) and 60 male pigs vaccinated 4 weeks before slaughter (IM-4), all slaughtered at comparable slaughter weights. After 20 weeks of age, IM-6 showed more inactive behaviour at the expense of playing and aggressive behaviour. Daily feed intake (DFI), daily gain (DG) and feed conversion ratio (FCR) did not differ significantly between IM-6 and IM-4. Gilts had a lower DFI and DG in the late finishing phase and a higher FCR overall compared with both IM groups. Gilts showed a higher lean meat content compared with both IM groups. Earlier vaccination increased dressing percentage, which could partly be explained by the lower weight of the gastrointestinal tract, but not by testes weight. Meat quality traits and palatability did not differ significantly between IM-6 and IM-4. Vaccination of immunocastrates at 6 compared with 4 weeks before slaughter improved the calmness in the stable and the dressing percentage, while maintaining performance and carcass characteristics.

Heavy metal pollution presents a serious problem for human health and ecosystems. During this survey we were collected a total number of 16 surface water samples. The analyses of four heavy metals, Cadmium, Chromium, Copper and Lead, were performed at the Institute of Applied Nuclear Physics, University of Tirana, using Graphite Furnace Atomic Absorption Spectrometry. The current study reports the presence of heavy metals Cd, Cr, Cu and Pb in selected samples. Their concentrations are resulted to be quite low. Average concentrations of metals in the surface water samples were: Pb-2.14μg/L; Cd-0.04 μg/L; Cr-9.44 μg/L; Cu-3.28 μg/L. The concentration of heavy metals in surface water samples is compared with maximum contaminant levels recommended by World Health Organization and Environmental Protection Agency. From the results obtained, none of the analyzed samples contained heavy metals concentrations above the MCL determined from EPA. One of all surface water samples contained the chromium element 6.8% higher than the MCL determined from WHO.

Societal pressure to ban surgical castration of male piglets is rising due to animal welfare concerns, thus other methods to prevent boar taint need to be explored. Genetic selection against boar taint appears to be a long-term sustainable alternative. However, as boar taint is linked to reproductive hormones, it is important to consider possible negative side effects such as delayed sexual maturity or changes in behaviour. We reported earlier that the melanocortin-4 receptor (MC4R) marker can be used to reduce boar taint levels in fat of boars. The objective of this study was to evaluate whether MC4R marker-assisted selection for lower boar taint prevalence affects plasma levels of boar taint compounds and testosterone; sexual maturity; behaviour; skin lesions; and lameness in boars and gilts. Using an intervention study with a 2×2 design, 264 boars and gilts differing on position 893 of the MC4R gene (AA v. GG) were compared. The MC4R polymorphism did not affect the plasma concentration of either androstenone or testosterone at different time points, whereas the concentration of skatole was significantly lower (P=0.003) and the concentration of indole tended to be lower (P=0.074) in GG compared with AA boars. A higher percentage of gilts of the GG genotype were in puberty at slaughter age compared with AA gilts (P

Male piglets are castrated to reduce boar taint and also to reduce aggressive and sexual behaviour. However, the procedure as traditionally performed is painful and negatively affects performance. Large-scale results about the consequences of implementing alternatives on farms are lacking. We, therefore, investigated the practical applicability of the following five alternatives that can be implemented in the short term: surgical castration (1) without pain relief (CONT, control group), (2) with analgesia (MET, Metacam, 0.2 ml, 10 to 15 min before castration), (3) with general anaesthesia (CO2, inhalation, 100% CO2, 25 s, 3 l/min), (4) vaccination against boar taint (IM, two injections with Improvac) and (5) production of entire males (EM). The study consisted of the following two trials: (1) an experimental farm trial with 18 animals/treatment and (2) a large field trial on 20 farms with ~120 male pigs/farm per treatment and all treatments performed on each farm. Performance results as well as data on carcass traits, boar taint (hot-iron method) and testes development and weight were collected in both trials. Neither castration nor administration of analgesia or anaesthesia had an effect on daily gain of the piglets in the farrowing crates (P>0.05). Farmer records indicated that mortality in the farrowing crates (1.1%), nursery pens (1.8%) and fattening stable (2.2%) was not influenced by MET or CO2 compared with EM, IM or CONT (P>0.05). No significant differences were found for daily gain (P>0.05) nor slaughter age (P>0.05). Immunocastrates and EM had a better gain-to-feed ratio (P

The aim of this study was to evaluate the performance of various boar taint detection methods, measure the relationship between them and identify possible points of improvement for boar taint detection. The methods used to evaluate boar taint in the carcasses of 448 entire male pigs and 17 barrows were the hot iron method (n = 442), a standardised (n = 323) and home (n = 58) consumer meat-evaluation panel, an expert panel assessment of meat and fat (n = 464) and laboratory analysis of skatole, androstenone and indole in fat (n = 464). The axillary odour of a number of slaughtered entire male pigs was also investigated (n = 231). As correlation coefficients were generally weak, a positive result for one of these detection methods did not per se result in a positive result for all other methods. Results of one detection method could not be generalised. The choice to use one or more detection methods deserves consideration depending on the aim of the study. In this paper, we suggest some possible improvements for evaluating boar taint with a consumer panel based on our results and experience. The home consumer evaluation was correlated with the concentration of indole (r = 0.27) but not with skatole or androstenone. We therefore recommend that lab analyses include indole testing. The hot iron method seems to be an easy and fast detection method, which yields comparable or better correlation coefficients with the other detection methods than an expert panel evaluating fat samples. However, the reliability of the hot iron method depends on the training and reliability of one or two assessors. Efforts should be made to further optimise this method by evaluating the effect of testing conditions. The axillary odour score was moderately correlated with the other detection methods (up to 0.32). More research is needed to evaluate the possibilities of axillary odour as a boar taint detection method.

Piétrain (P), Large White (LW) and Belgian Landrace stress negative (BN) boars were slaughtered at 50, 70, 90 or 110 kg live weight to investigate breed differences and the effect of slaughter weight on boar taint prevalence. Boar taint was quantified by four different methodologies: sensory evaluation of neckfat heated with a hot iron in the slaughterhouse, sensory evaluation of meat by consumer panels, sensory evaluation of fat and meat by expert panels and laboratory analysis of indole, skatole and androstenone in backfat. Skatole levels in backfat were significantly higher for LW and BN than for P boars. The androstenone levels and the hot iron method revealed a significant interaction between breed and slaughter weight. On the other hand, experts detected an effect of weight on the androstenone odour perception, which was significantly higher in fat from boars slaughtered at 90 kg compared with 50 kg, and significantly higher in meat from boars slaughtered at 110 kg compared with 50 kg. Consumers did not detect differences in the sensory characteristics among breeds or slaughter weight. These results indicate opportunities to minimise the risk of boar taint in entire male pigs by carefully selecting a combination of breed and slaughter weight. Along with the optimal slaughter weight, the effectiveness of reducing boar taint by lowering slaughter weight appeared to be breed dependent.

Objective: Eribulin mesylate (Halaven®) is a non-taxane inhibitor of microtubule indicated as monotherapy in patients with metastatic breast cancer (MBC), which progresses after anthracycline and taxanes therapy. In this retrospective observational study, we want to evaluate the efficacy of Eribulin in elderly women with MBC pretreated with anthracyclines and taxanes., Patients and Methods: 40 elderly patients > 70 years of age were enrolled, and the median age was 76 years (range 70-82). Overall survival (OS), Progression Free Survival (PFS), Objective Response Rate (ORR) were primary endpoints, tolerability, carcinoembryonic antigen levels 15.3 (Ca 15.3), before and after treatment, and Quality of Life (QoL) were secondary endpoints., Results: Eribulin treatment was well tolerated, produced a good level of disease control, a manageable toxicity profile and a significant impact on QoL. Median OS was 12.8 months and median PFS was 3.2 months. A significant correlation was observed between reduction of Ca 15.3 and PFS with a value of 0.59 (p = 0.002)., Conclusions: Despite a limited number of patients and a modest manageable toxicity, Eribulin is a chemotherapy treatment that has showed to be an effective and well-tolerated therapeutic option in elderly patients with MBC. Further analysis should focus on the elderly patients in our setting of study.

Violence against women is a violation of human rights, crossing all cultures, classes, levels of education, earnings, ethnic and age groups. We conducted a retrospective study to review forensic records of sexual assault examinations carried out in different Italian health facilities and to correlate these findings with the results of the forensic DNA analyses. The goal was to determine which factors could have affected the obtained results, to identify the fundamental aspects to search for while examining a sexual assault victim in order to gather useful evidence to identify the offender and reconstruct the dynamics of the fact. We analysed 102 cases that occurred between 2006 and 2017, coming from ten participating laboratories. Despite a relatively limited number of cases, this study shows that the ability to ascertain the presence of male biological material in the samples collected is not a problem for forensic laboratories and seems to be influenced by other factors, such as how much time elapsed between the event and the sampling, the availability of the aggressor's biological material on the victim and the identification of biological fluids/stains. Therefore, the need for health structures to adopt specific protocols has been highlighted. It is necessary for health structures to define specific pathways and adopt homogeneous procedures or operational protocols, and it is essential to provide adequate training for health personnel. The results of the study could be useful in drafting and revising protocols/guidelines implemented in Italian hospital. Issues related to the limited number of analyses requested by Italian Authorities are also discussed., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Storage conditions influence the integrity of the recoverable DNA from forensic evidence in terms of yield and quality. FTA cards are widely used in the forensic practice as their chemically treated matrix provides protection from the moment of collection to the point of analysis with current STR typing technology. In this study, we assess the recoverability and the integrity of DNA from 11-year-old saliva on FTA cards using a forensic quantitative real-time polymerase chain reaction (qPCR) commercial assay. The quality after long-term storage was investigated in order to evaluate if the FTA device could assure enough stability over time, applying some internally validated quality criteria of the STR profile. Furthermore, we used a 3D interpolation model to combine the quantitative and qualitative data from qPCR to calculate the minimum optimal DNA input (MODI) to add to the downstream PCR reaction based on the quantitative and qualitative data of a sample. According to our results, when saliva sample is properly transferred onto FTA cards and then correctly stored according to the manufacturer's instructions, it is possible to recover sufficient amounts of DNA for human identification even after more than a decade of storage at ambient temperature. Degradation affected the quality of results especially when the Degradation Index exceeds the value of 2.12, requiring modifications of the standard internal workflow to improve the genotyping quality. Above this value, the application of a "corrective factor" to the PCR normalization process was necessary in order to adjust the recommended manufacturer's PCR DNA input taking into account the degradation level. Our results demonstrated the importance to consider in predictive terms the parameters obtained with the real-time quantification assay, both in terms of quantity (DNA concentration) and of quality (DI, inhibition). Informatics predictive tools including qPCR data together with the variables of storage duration and conditions should be developed in order to optimize the DNA analysis process.

Aim: The aims of the study are to evaluate the clinical outcomes of first-line treatment with platinum-based chemotherapy and cetuximab in patients with relapsing/metastatic head and neck cancer (RM HNC) and to identify predictors of treatment response., Methods: This is a retrospective, observational, longitudinal, real-world study involving 6 oncology centres in Italy. All consecutive patients with RM HNC treated between January 2007 and December 2016 with a first-line therapy consisting of a platinum-based chemotherapy regimen plus cetuximab were included. The primary objective of the study was to assess overall survival (OS) and progression-free survival (PFS). Secondary objectives included the identification of predictors of treatment response., Results: Overall, 297 patients were identified. Median OS was 10.8 months (95% confidence interval [CI] 9.3-12.2), whereas median PFS was 4.8 months (95% CI 4.3-5.5). On multivariable analysis, independent unfavourable prognostic factors for OS were performance status (PS) Eastern Cooperative Oncology Group (ECOG) >0, presence of residual tumour at primary site, platinum resistance and lack of objective response. Unfavourable predictors for PFS included cancer primary site (paranasal sinuses, hypopharynx), PS ECOG >0, presence of residual tumour at primary site, platinum resistance and lack of objective response. Independent unfavourable predictors of objective response were tumour site (oral cavity, larynx-hypopharynx), residual tumour at primary site and prior chemotherapy., Conclusions: The availability of new treatment modalities and epidemiological changes make the periodic reassessment of prognostic factors of great relevance to guide clinical practice and the design of future randomised clinical trials., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Purpose: Pancreatic carcinoma is the neoplasia with the major mortality, and main standard treatments in this cancer increase survival but do not lead to complete recovery of the patient. The aim of this study was to evaluate the efficacy of Abraxane ® (nab-paclitaxel) in Italian patients with metastatic pancreatic cancer (MPC)., Patients and Methods: We conducted a retrospective analysis of 80 patients. Overall survival (OS) was the primary end point for evaluating the efficacy of nab-paclitaxel in combination with gemcitabine treatment, while carbohydrate antigen 19-9 (CA 19-9) reduction, safety, progression-free survival (PFS), overall response rate and reduction in pain were secondary end points., Results: The median OS was 8 months, and the median PFS was 5 months. A considerable difference in CA 19-9 before and after treatment was observed. Descriptive and correlation analyses were done to examine the relationship between CA 19-9 response and OS. Linear regression analysis between OS and CA 19-9 response revealed that CA 19-9 is an important predictor of OS, showing a positive correlation., Conclusion: Nab-paclitaxel is a well-tolerated and effective treatment for patients affected by MPC. The drug showed an improved tolerability profile, significant pain relief and an increase in survival rate., Competing Interests: Disclosure The authors report no conflicts of interest in this work.

Background: Second-line treatment for advanced non-small-cell lung cancer (NSCLC) patients includes monotherapy with a third-generation cytotoxic drug (CT) or a tyrosine kinase inhibitor (TKI). These options are the actual standard for EGFR wild-type (WT) status, as patients with EGFR mutations achieve greater benefit by the use of TKI in first-line treatment. Some clinical trials and meta-analyses investigated the comparison between CT and TKI in second-line, but data are conflicting., Methods: We designed a retrospective trial to gather information about TKI sensitivity in comparison with CT. We selected from clinical records patients treated with at least 1 line of CT and at least 1 line of TKI. We collected data about age, sex, performance status, comorbidity, smoking status, histotype, metastatic sites, EGFR status, treatment schedule, better response and time-to-progression (TTP) for each line of treatment and overall survival (OS)., Results: 93 patients met selection criteria. Mean age 66,7 (range: 46-84). M/F ratio is 3:1. 39 EGFR-WT and 54 EGFR-UK. All patients received erlotinib or gefitinib as second-line treatment or erlotinib as third-line treatment. No TTP differences were observed for both second-line (HR:0,91; p = 0,6333) and third-line (HR:1.1; p = 0,6951) treatment (TKI vs CT). A trend of a benefit in OS in favor of 3rd-line TKI (HR:0,68; p = 0,11)., Conclusions: This study explores the role of TKIs in EGFR non-mutated NSCLC patients. OS analysis highlights a trend to a benefit in patients who received TKI in third-line, even if this result is statistically non-significant. Further analysis are needed to find an explanation for this observation., Competing Interests: The authors declare no potential conflicts of interests.

Social and cultural factors had a critical role in determining the genetic structure of Europe. Therefore, socially stratified populations may help to focus on specific episodes of European demographic history. In this study, we use uniparental markers to analyse the genetic structure of Partecipanza in San Giovanni in Persiceto (Northern Italy), a peculiar institution whose origins date back to the Middle Ages and whose members form the patrilineal descent of a group of founder families. From a maternal point of view (mtDNA), Partecipanza is genetically homogeneous with the rest of the population. However, we observed a significant differentiation for Y-chromosomes. In addition, by comparing 17 Y-STR profiles with deep-rooted paternal pedigrees, we estimated a Y-STR mutation rate equal to 3.90 * 10(-3) mutations per STR per generation and an average generation duration time of 33.38 years. When we used these values for tentative dating, we estimated 1300-600 years ago for the origins of the Partecipanza. These results, together with a peculiar Y-chromosomal composition and historical evidence, suggest that Germanic populations (Lombards in particular) settled in the area during the Migration Period (400-800 AD, approximately) and may have had an important role in the foundation of this community.

The role of DNA damage in PCR processivity/fidelity is a relevant topic in molecular investigation of aged/forensic samples. In order to reproduce one of the most common lesions occurring in postmortem tissues, a new protocol based on aqueous hydrolysis of the DNA was developed in vitro. Twenty-five forensic laboratories were then provided with 3.0 μg of a trial sample (TS) exhibiting, in mean, the loss of 1 base of 20, and a molecular weight below 300 bp. Each participating laboratory could freely choose any combination of methods, leading to the quantification and to the definition of the STR profile of the TS, through the documentation of each step of the analytical approaches selected. The results of the TS quantification by qPCR showed significant differences in the amount of DNA recorded by the participating laboratories using different commercial kits. These data show that only DNA quantification "relative" to the used kit (probe) is possible, being the "absolute" amount of DNA inversely related to the length of the target region (r(2) = 0.891). In addition, our results indicate that the absence of a shared stable and certified reference quantitative standard is also likely involved. STR profiling was carried out selecting five different commercial kits and amplifying the TS for a total number of 212 multiplex PCRs, thus representing an interesting overview of the different analytical protocols used by the participating laboratories. Nine laboratories decided to characterize the TS using a single kit, with a number of amplifications varying from 2 to 12, obtaining only partial STR profiles. Most of the participants determined partial or full profiles using a combination of two or more kits, and a number of amplifications varying from 2 to 27. The performance of each laboratory was described in terms of number of correctly characterized loci, dropped-out markers, unreliable genotypes, and incorrect results. The incidence of unreliable and incorrect genotypes was found to be higher for participants carrying out a limited number of amplifications, insufficient to define the correct genotypes from damaged DNA samples such as the TS. Finally, from a dataset containing about 4500 amplicons, the frequency of PCR artifacts (allele dropout, allele drop-in, and allelic imbalance) was calculated for each kit showing that the new chemistry of the kits is not able to overcome the concern of template-related factors. The results of this collaborative exercise emphasize the advantages of using a standardized degraded DNA sample in the definition of which analytical parameters are critical for the outcome of the STR profiles., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Due to their strategic geographic location between three different continents, Sicily and Southern Italy have long represented a major Mediterranean crossroad where different peoples and cultures came together over time. However, its multi-layered history of migration pathways and cultural exchanges, has made the reconstruction of its genetic history and population structure extremely controversial and widely debated. To address this debate, we surveyed the genetic variability of 326 accurately selected individuals from 8 different provinces of Sicily and Southern Italy, through a comprehensive evaluation of both Y-chromosome and mtDNA genomes. The main goal was to investigate the structuring of maternal and paternal genetic pools within Sicily and Southern Italy, and to examine their degrees of interaction with other Mediterranean populations. Our findings show high levels of within-population variability, coupled with the lack of significant genetic sub-structures both within Sicily, as well as between Sicily and Southern Italy. When Sicilian and Southern Italian populations were contextualized within the Euro-Mediterranean genetic space, we observed different historical dynamics for maternal and paternal inheritances. Y-chromosome results highlight a significant genetic differentiation between the North-Western and South-Eastern part of the Mediterranean, the Italian Peninsula occupying an intermediate position therein. In particular, Sicily and Southern Italy reveal a shared paternal genetic background with the Balkan Peninsula and the time estimates of main Y-chromosome lineages signal paternal genetic traces of Neolithic and post-Neolithic migration events. On the contrary, despite showing some correspondence with its paternal counterpart, mtDNA reveals a substantially homogeneous genetic landscape, which may reflect older population events or different demographic dynamics between males and females. Overall, both uniparental genetic structures and TMRCA estimates confirm the role of Sicily and Southern Italy as an ancient Mediterranean melting pot for genes and cultures.

Great European mountain ranges have acted as barriers to gene flow for resident populations since prehistory and have offered a place for the settlement of small, and sometimes culturally diverse, communities. Therefore, the human groups that have settled in these areas are worth exploring as an important potential source of diversity in the genetic structure of European populations. In this study, we present new high resolution data concerning Y chromosomal variation in three distinct Alpine ethno-linguistic groups, Italian, Ladin and German. Combining unpublished and literature data on Y chromosome and mitochondrial variation, we were able to detect different genetic patterns. In fact, within and among population diversity values observed vary across linguistic groups, with German and Italian speakers at the two extremes, and seem to reflect their different demographic histories. Using simulations we inferred that the joint effect of continued genetic isolation and reduced founding group size may explain the apportionment of genetic diversity observed in all groups. Extending the analysis to other continental populations, we observed that the genetic differentiation of Ladins and German speakers from Europeans is comparable or even greater to that observed for well known outliers like Sardinian and Basques. Finally, we found that in south Tyroleans, the social practice of Geschlossener Hof, a hereditary norm which might have favored male dispersal, coincides with a significant intra-group diversity for mtDNA but not for Y chromosome, a genetic pattern which is opposite to those expected among patrilocal populations. Together with previous evidence regarding the possible effects of "local ethnicity" on the genetic structure of German speakers that have settled in the eastern Italian Alps, this finding suggests that taking socio-cultural factors into account together with geographical variables and linguistic diversity may help unveil some yet to be understood aspects of the genetic structure of European populations.

The 2011 collaborative exercise of the ISFG Italian Working Group GeFI was aimed at validating the five ENFSI/EDNAP miniSTR loci D1S1656, D2S441, D10S1248, D12S391 and D22S1045. The protocol required to type at least 50 multilocus profiles from locally resident individuals and two blind bloodstains in duplicate (i.e., using at least two different commercial kits), and to send the electropherograms to the Organizing Committee. Nineteen laboratories distributed across Italy participated, collecting a total of 960 samples. Full concordance was found for the five new miniSTRs as observed from the comparison of 13,150 alleles. The inspection of the electropherograms allowed the identification of a very limited number of mistypings in the miniSTR genotypes thus contributing to the establishment of an high quality Italian database of frequencies., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

The identification of isolation signatures is fundamental to better understand the genetic structure of human populations and to test the relations between cultural factors and genetic variation. However, with current approaches, it is not possible to distinguish between the consequences of long-term isolation and the effects of reduced sample size, selection and differential gene flow. To overcome these limitations, we have integrated the analysis of classical genetic diversity measures with a bayesian method to estimate gene flow and have carried out simulations based on the coalescent. Combining these approaches, we first tested whether the relatively short history of cultural and geographical isolation of four "linguistic islands" of the Eastern Alps (Lessinia, Sauris, Sappada and Timau) had left detectable signatures in their genetic structure. We then compared our findings to previous studies of European population isolates. Finally, we explored the importance of demographic and cultural factors in shaping genetic diversity among the groups under study. A combination of small initial effective size and continued genetic isolation from surrounding populations seems to provide a coherent explanation for the diversity observed among Sauris, Sappada and Timau, which was found to be substantially greater than in other groups of European isolated populations. Simulations of micro-evolutionary scenarios indicate that ethnicity might have been important in increasing genetic diversity among these culturally related and spatially close populations.

The analysis of nonhuman biological evidence both animal and botanical to find out the correct species of a sample comes as a great help to crime investigators. Particularly, forensic botany may be useful in many criminal and civil cases, e.g., for linking an individual to a crime scene or physical evidence to a geographic location, or tracking marijuana distribution patterns.Despite many molecular techniques for species identification so far applied, botanical evidences are still overlooked by forensic scientists due to the lack of reproducible and efficient protocols standardized across a wide range of different organisms and among different laboratories.Recently, the term "DNA barcoding" has been coined to describe the use of a short gene sequence from a standardized region of the genome as a molecular tool for species identification. DNA barcodes have been successfully applied to a number of animal groups and introduced in forensic science with the application of the mitochondrial gene COI. Building on this success, ongoing investigations have searched for the best barcode to apply to all land plants. Here we describe the basic protocol based on amplification and sequence analysis of barcoding markers for land plants considering the latest developments of Plant DNA barcoding Project. The aim of this chapter is to provide forensic scientists an accurate and reliable tool for assigning unidentified botanical specimens to the correct species as powerful mainstay in investigations, increasing the contributions from nonhuman DNA to forensics.

Gastric cancer is one of the most common cancers worldwilde. The five-year survival for stage IV gastric cancer is around 5-10% in Western countries. Advanced gastric cancer is sensitive to numerous agents, but there is no generally accepted standard regimen. Here we report on a case of advanced gastric cancer occurring in a 72-year-old man who underwent treatment with capecitabine plus oxaliplatin, achieving a complete response.

Y chromosome variation at 12 STR (the Powerplex® Y system core set) and 18 binary markers was investigated in two major (the Ghegs and the Tosks) and two minor (the Gabels and the Jevgs) populations from Albania (Southern Balkans). The large proportion of haplotypes shared within and between groups makes the Powerplex 12-locus set inadequate to ensure a suitable power of discrimination for the forensic practice. At least 85% of Y lineages in the Jevgs, the cultural minority claiming an Egyptian descent, turned out to be of either Roma or Balkan ancestry. They also showed unequivocal signs of a common genetic history with the Gabels, the other Albanian minority practising social and cultural Roma traditions.

Pegylated liposomal doxorubicin (PLD) has the advantage of delivering active anthracycline directly to the tumor site, while exposing the patient to a lesser degree of doxorubicin-associated toxicities. Recently, a regimen in which paclitaxel is infused weekly over 1 h produced substantial antitumor activity with little myelosuppression. We designed a phase II trial to study the efficacy and toxicity of 10 mg/m(2) PLD on Days 1, 8 and 15, plus 70 mg/m(2) paclitaxel weekly in patients with untreated metastatic breast cancer and a high risk of cardiotoxicity. The study included 35 patients, with 31 (88.5%) evaluable for efficacy and 35 (100%) for toxicity. A total of 28 patients (80%) had two or more sites of disease. Overall, 4 complete and 16 partial responses were noted with an overall response rate of 64.5%, with 6 cases of stable and 5 cases of progressive disease. Toxicity was found to be manageable in that the only grade 3-4 side effects recorded were palmar-plantar erythrodysesthesia, 8.5%; mucositis, 2.8%; leucopenia, 12.5%; anemia, 2.8% and AST/ALT, 2.8%. No cardiotoxicity was observed. In conclusion, weekly PLD plus paclitaxel appears to be a well-tolerated and effective approach for metastatic breast cancer patients with a high risk of cardiotoxicity.

Mitochondrial DNA (mtDNA) U/K and J/T are sister haplogroups within the superhaplogroup R. They are both common in Europe, with a combined overall frequency similar to the one reported for H, the most common European haplogroup (40-50%). In this study, we selected 159 Italian subjects, already ascribed to U/K and J/T by RFLP typing, and assigned each mtDNA to specific clades/subclades by investigating at least one diagnostic coding region SNP. For each sister haplogroup, one multiplex PCR and one SNaPshot minisequencing reaction were set up targeting 16 U/K and 7 J/T coding region SNPs. Each mtDNA sample was clearly assigned to a specific subclade, which could be further subdivided into several minor sub-branches according to peculiar HVS I/II motifs. Such a molecular dissection of haplogroups U/K and J/T could be extremely useful to reduce the overall analysis time and labor intensive sequencing procedures in high volume forensic casework, for example when it is important to rapidly exclude samples in order to restrict the number of suspects.

The present day distribution of Y chromosomes bearing the haplogroup J1 M267(*)G variant has been associated with different episodes of human demographic history, the main one being the diffusion of Islam since the Early Middle Ages. To better understand the modes and timing of J1 dispersals, we reconstructed the genealogical relationships among 282 M267(*)G chromosomes from 29 populations typed at 20 YSTRs and 6 SNPs. Phylogenetic analyses depicted a new genetic background consistent with climate-driven demographic dynamics occurring during two key phases of human pre-history: (1) the spatial expansion of hunter gatherers in response to the end of the late Pleistocene cooling phases and (2) the displacement of groups of foragers/herders following the mid-Holocene rainfall retreats across the Sahara and Arabia. Furthermore, J1 STR motifs previously used to trace Arab or Jewish ancestries were shown unsuitable as diagnostic markers for ethnicity.

Forensic botany can provide significant supporting evidence during criminal investigations. However, it is still an underutilized field of investigation with its most common application limited to identifying specific as well as suspected illegal plants. The ubiquitous presence of plant species can be useful in forensics, but the absence of an accurate identification system remains the major obstacle to the present inability to routinely and correctly identify trace botanical evidence. Many plant materials cannot be identified and differentiated to the species level by traditional morphological characteristics when botanical specimens are degraded and lack physical features. By taking advantage of a universal barcode system, DNA sequencing, and other biomolecular techniques used routinely in forensic investigations, two chloroplast DNA regions were evaluated for their use as "barcoding" markers for plant identification in the field of forensics. We therefore investigated the forensic use of two non-coding plastid regions, psbA-trnH and trnL-trnF, to create a multimarker system for species identification that could be useful throughout the plant kingdom. The sequences from 63 plants belonging to our local flora were submitted and registered on the GenBank database. Sequence comparison to set up the level of identification (species, genus, or family) through Blast algorithms allowed us to assess the suitability of this method. The results confirmed the effectiveness of our botanic universal multimarker assay in forensic investigations.