Elemental composition of game meat from Austria

Concentrations of 26 elements (B, Na, Mg, P, S, K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Rb, Sr, Mo, Cd, Sb, Ba, Hg, Pb, U) in wild game meat from Austria were analysed using an inductively coupled plasma mass spectrometer. All investigated animals were culled during the hunting season 2012/2013, including 10 chamois (Rupicapra rupicapra), 9 hare (Lepus europaeus), 10 pheasant (Phasianus colchicus), 10 red deer (Cervus elaphus), 12 roe deer (Capreolus capreolus) and 10 wild boar (Sus scrofa). In 19 out of 61 meat samples lead concentrations were higher than 0.1 mg/kg, the maximum limit in meat as set by the European Commission (Regulation EC No 1881/2006), which is most likely caused by ammunition residues. Especially, pellet shot animals and chamois show a high risk for lead contamination. Despite ammunition residues all investigated muscle samples show no further health risk with respect to metal contamination.     

Fatty acid profile of meat (Longissimus lumborum) from female roe deer (Capreolus capreolus L.) and red deer (Cervus elaphus L.)

The aim of this study was to compare the fatty acid profile of intramuscular fat (IMF) in female roe deer (Capreolus capreolus L.) and red deer (Cervus elaphus L.) living in the wild. The experimental materials comprised samples of the Longissimus lumborum (LL) muscle collected from 20 carcasses of does aged 3–5 years and 15 carcasses of hinds aged 4–6 years. All animals were hunter-harvested in the forests of North-Eastern Poland. The IMF of does had considerably higher concentrations of monounsaturated and polyunsaturated fatty acids. Therefore, it was characterized by more desirable values of quality parameters, and provided more health benefits. The observed differences in the fatty acid composition of IMF between does and hinds are important in view of both the nutritional value of meat and its susceptibility to lipid oxidation and rancidification.     

Real-time PCR for detection and quantification of red deer (Cervus elaphus), fallow deer (Dama dama), and roe deer (Capreolus capreolus) in meat mixtures

A rapid real-time polymerase chain reaction (PCR) technique using SYBR Green detection system, has been developed for the quantification of red deer, fallow deer, and roe deer DNAs in meat mixtures. The method combines the use of cervid-specific primers that amplify a 134, 169, and 120 bp of the 12S rRNA gene fragment of red deer, fallow deer and roe deer, respectively, and universal primers that amplify a 140 bp fragment on the nuclear 18S rRNA gene from eukaryotic DNA. The C_t (threshold cycle) values obtained with the 18S rRNA primers are used to normalize those obtained from each of the cervid-specific systems, serving as endogenous control for the total content of PCR-amplifiable DNA in the sample. Analysis of experimental raw and heat treated binary mixtures of red deer, fallow deer or roe deer meat in a swine meat matrix demonstrated the suitability of the assay for the detection and quantification of the target cervid DNAs in the range 0.1–0.8%, depending on the species and treatment of the meat samples analyzed.     

Authentication of a traditional game meat sausage (Alheira) by species-specific PCR assays to detect hare,rabbit, red deer,pork and cow meats

Alheira is a traditional meat product that is typical from the Northeast region of Portugal and much appreciated. It is a sort of sausage produced industrially or by small artisanal producers, having wheat bread and meats as main ingredients. Game meat Alheira (Alheira de caça) is considered one of the most attractive products since it should include different game meats. The aim of the present work was to identify the species of origin of meats added to game meat Alheira samples to verify their compliance with labelling. Species-specific PCR assays targeting mitochondrial genes of rabbit, hare, red deer, cow and pork were optimised and applied to industrial and artisanal samples. The assays revealed adequate specificity for each of the targeted species, with sensitivities of 0.01–0.1%. Results of the evaluation of 18 commercial samples identified several inconsistencies with labelling, namely the absence of declared game species (red deer, hare and rabbit) in ten samples and the presence of undeclared cow species in nine of the analysed samples. These findings indicate the occurrence of misleading labelling, suggesting the adulteration by substitution of game meats by cow meat to reduce production costs and the need to protect and valorise this kind of traditional food product.     

Game meat consumption by hunters and their relatives: a probabilistic approach

This study aimed to estimate the consumption of meat and products derived from hunting by the consumer population and, specifically, by hunters and their relatives. For this purpose, a survey was conducted on the frequency of consuming meat from the four most representative game species in Spain, two of big game, wild boar (Sus scrofa) and red deer (Cervus elaphus), and two of small game, rabbit (Oryctolagus cuniculus) and red partridge (Alectoris rufa), as well as of processed meat products (salami-type sausage) made from those big game species. The survey was carried out on 337 habitual consumers of these types of products (hunters and their relatives). The total mean game meat consumption, per capita in this population group, is 6.87 kg/person/year of meat and 8.57 kg/person/year if the processed meat products are also considered. Consumption of rabbit, red partridge, red deer and wild boar, individually, was 1.85, 0.82, 2.28 and 1.92 kg/person/year, respectively. It was observed that hunters generally registered a larger intake of game meat, this being statistically significant in the case of rabbit meat consumption. Using probabilistic methods, the meat consumption frequency distributions for each hunting species studied were estimated, as well as the products made from big game species and the total consumption both of meat by itself and that including the products made from it. The consumption frequency distributions were adjusted to exponential ones, verified by the test suitable for it according to Akaike Information Criterion (AIC), Bayesian Information Criterion (BIC), the Chi-squared and Kolmogorov–Smirnov statistics. In addition, the consumption percentiles of the different distributions were obtained. The latter could be a good tool when making nutrition or contaminant studies since they permit the assessment of exposure to the compound in question.     

Identification of meats from red deer (Cervus elaphus), fallow deer (Dama dama), and roe deer (Capreolus capreolus) using polymerase chain reaction targeting specific sequences from the mitochondrial 12S rRNA gene

Polymerase chain reaction (PCR) based on oligonucleotide primers targeting the mitochondrial 12S rRNA gene was applied to the specific identification of meats from red deer (Cervus elaphus), fallow deer (Dama dama), and roe deer (Capreolus capreolus). The use of a common reverse primer, together with forward specific primers for red deer, fallow deer, and roe deer, allowed the selective amplification of the desired cervid sequences. The specificity of each primer pair was verified by PCR analysis of DNA from various game and domestic meats. The assay can be useful for the accurate identification of meats from cervid species, avoiding mislabeling or fraudulent species substitution in meat products.     

Assessment of levels of bacterial contamination of large wild game meat in Europe

The variations in prevalence and levels of pathogens and fecal contamination indicators in large wild game meat were studied to assess their potential impact on consumers. This analysis was based on hazard analysis, data generation and statistical analysis. A total of 2919 meat samples from three species (red deer, roe deer, wild boar) were collected at French game meat traders’ facilities using two sampling protocols. Information was gathered on the types of meat cuts (forequarter or haunch; first sampling protocol) or type of retail-ready meat (stewing meat or roasting meat; second protocol), and also on the meat storage conditions (frozen or chilled), country of origin (eight countries) and shooting season (autumn, winter, spring). The samples were analyzed in both protocols for detection and enumeration of Escherichia coli, coagulase + staphylococci and Clostridium perfringens. In addition, detection and enumeration of thermotolerant coliforms and Listeria monocytogenes were performed for samples collected in the first and second protocols, respectively. The levels of bacterial contamination of the raw meat were determined by performing statistical analysis involving probabilistic techniques and Bayesian inference. C. perfringens was found in the highest numbers for the three indicators of microbial quality, hygiene and good handling, and L. monocytogenes in the lowest. Differences in contamination levels between game species and between meats distributed as chilled or frozen products were not significant. These results might be included in quantitative exposure assessments.     

Lead,cadmium and organochlorine pesticide residues in hunted red deer and wild boar from northern Italy

The objectives of the present study were to assess heavy metal cadmium (Cd), lead (Pb) and organochlorine pesticide concentrations in tissues of red deer (Cervus elaphus) and wild boar (Sus scrofa) from nine hunting areas and to evaluate related risk factors for the host animal. Over a period of 2 years, a total of 1055 and 210 masseters, 424 and 201 livers, 642 and 152 kidneys were collected from wild boar and red deer, respectively, and concentrations of Cd, Pb and organochlorine pesticides were determined. Comparing the two species, Cd concentration in the kidney (3.72 mg/kg), liver (0.67 mg/kg) and muscle (0.02 mg/kg) of wild boar was found to be significantly higher than in the organs of red deer (1.02 mg/kg in the kidneys, 0.07 mg/kg in the liver and 0.006 mg/kg in muscle). Mean Pb concentrations were found to be similar in both animals, with 0.39, 0.52 and 2.60 mg/kg detected in the wild boar kidney, liver and muscle, respectively, and 0.24, 0.21 and 2.04 mg/kg in the respective organs of the red deer. No difference in concentrations were found based on age class, location of tissue sample or contaminant in the case of wild boar. By contrast, a significantly lower Cd concentration was found in the kidney of the young red deer. The search for organochlorine pesticides in both red deer and wild boar produced negative results with values below the limits of detection. Due to the high levels of renal Cd and muscle Pb detected in wild boar and red deer, further research needs to be carried out in an effort to identify the source of contamination and preserve the health of animals and humans.     

Biogenic amines and polyamines in liver,kidney and spleen of roe deer and European brown hare

Cadaverine (CAD), histamine (HIS), tyramine (TYR) and the polyamines putrescine (PUT), spermidine (SPD) and spermine (SPM) were determined in liver, kidney and spleen of hunted roe deer (n = 39) and European brown hare (n = 20) 3 and 6 h post mortem, respectively. Median concentrations (mg/kg fresh weight) of CAD and TYR were <5 mg/kg for organs of both species, whereas median HIS concentrations were higher in spleen (14.6 and 11.6 mg/kg for hare and roe deer, respectively) and lowest in liver. Maximum PUT concentration was 72.9 mg/kg, but median values were ≤7.8 mg/kg. Mean SPD concentrations were 8.5, 10.7 and 42.9 mg/kg for liver, kidney and spleen of roe deer, and 37.2, 24.4 and 52.0 mg/kg for hare. Mean SPM concentrations were higher (94.6, 79.9, 102.2 and 111.2, 82.8, 91.1 mg/kg, respectively). SPM concentrations in the organs of the male roe deer were significantly higher than in those of females. SPM:SPD ratios (weight base) were in the order of 2:1 for spleen, and higher for liver and kidney. SPM and SPD were correlated significantly (r = 0.42–0.47). Although variations of polyamine concentrations are partially associated with species, organ, age and gender, the relative contribution of individual factors deserves further study.     

The influence of environment, mode of nutrition and animal species on level of nitrosamine contamination in venison

The work aimed to research occurrence of nitrosamines in raw venison from different game species and mode of nutrition effect on their meat concentrations. Samples of venison from male and female elks, male and female red deer, male and female roe deer, male and female fallow deer, male and female wild boars, male and female brown hares and males and females of wild rabbits shot in Poland during the permissible hunting seasons were used for the experiments after their jointing and cutting. In the experiments conducted on eight species and 16 genera of wild male and female game animals a total of 336 analytical assays were conducted. Twenty-one meat samples of 1 kg each (each from a different animal) were collected respectively from males and females of eight animal species and taken to a laboratory. Meat contents of nitrosamines [dimethylonitrosamine (DMNA) and diethylonitrosamine (DENA)] were assessed by Pancholy's method adapted to nitrosamine determination in meat and meat products by Scanlan and Ryes. The levels of DMNA and DENA were determined using a Varian 3400 gas chromatograph coupled to a mass spectrometer (Finnigan MAT ITD. 800). Quantitative and qualitative analysis was conducted by comparison with N-nitrosamine standard solution chromatograms.     

Risk assessment of the lead intake by consumption of red deer and wild boar meat in Southern Spain

The presence of heavy metals in big game meat may pose a risk to human health. The main objective of this paper is to carry out a risk assessment study (using a probabilistic and point-estimate approach) of lead intake by consumption of red deer and wild boar meat in Southern Spain based on Spanish data collected in the period 2003–2006. In general, the concentration levels found for wild boar meat (mean?=?1291?µg?kg^?1) were much higher than those observed in red deer meat (mean?=?326?µg?kg^?1). The results from a point-estimate risk assessment showed that the estimated average intake of lead among different exposure scenarios varied from 0.1 to 6.5 and from 0.3 to 38?µg?kg^?1?week^?1 for red deer and wild boar meat, respectively; and from 0.3 to 35?µg?kg^?1?week^?1 for individuals consuming both red deer and wild boar meat, and that the estimated intake of lead by consumption of big game meat differed significantly between hunters and non-hunters, it being higher for hunters. Besides this, results from the probabilistic risk assessment study corroborated the fact that risk is greater in hunter populations, reaching a maximum in individuals consuming only wild boar and both types of meat, with 0.4% and 0.2% of the population above the provisional tolerable weekly intake (PTWI), respectively. Likewise, the hunter populations consuming wild boar and both types of big game meat (red deer and wild boar meat) were exposed to the maximum lead level (56?µg?kg^?1?week^?1), which corresponded approximately to 224% of the PTWI. Further data and studies will be needed to give a complete risk estimation in which it will be crucial to consider the contribution to the lead intake level of other foods in the diet of both population groups.     

Wild game consumption habits among Italian shooters: relevance for intakes of cadmium,perfluorooctanesulphonic acid,and 137cesium as priority contaminants

ABSTRACT

The consumption habits of 766 Italian shooters (96% males, 4% females), on average 52 years old, have been investigated, in Italy, through the distribution of questionnaires delivered during shooters’ attendance to training and teaching courses, in compliance with 853/2004/EC Regulation provisions on food hygiene. The most consumed wild species recorded were pheasant > woodcock > choke among feathered animals, and wild boar > hare > roe deer among mammals, respectively. An average of 100–200 g game per serving (four servings per month) was consumed, with highest intakes of 3000 g per month; meat, liver, and heart were the preferred food items. Mammalian and feathered game was regularly consumed with friends and relatives in 83% and in 60% of cases, respectively. Accounting for an inventoried population of 751,876 shooters in Italy, it is estimated that there is regular consumption of wild game in around the 3% of the Italian population. More than 80% of responders were aware of health risks related to game handling and to food safety issues. Due to the occurrence in wild boar meat and liver of the heavy metal cadmium (Cd), the persistent organic pollutant perfluorooctan sulphonic acid (PFOS), and the radionuclide ¹³⁷cesium (¹³⁷Cs), it was possible to demonstrate the usefulness of such a food consumption database for intake assessment in this sensitive group of consumers. In high consumers of wild boar, threshold concentrations for intakes have been estimated in the ranges of 48–93 ng g^–1 for Cd, 35–67 ng g^–1 for PFOS and 0.20–0.34 Bq kg^–1 for ¹³⁷Cs.     

Identification of hare meat by a species-specific marker of mitochondrial origin

Meat species identification in food has gained increasing interest in recent years due to public health, economic and legal concerns. Following the consumer trend towards high quality products, game meat has earned much attention. The aim of the present work was to develop a DNA-based technique able to identify hare meat. Mitochondrial cytochrome b gene was used to design species-specific primers for hare detection. The new primers proved to be highly specific to Lepus species, allowing the detection of 0.01% of hare meat in pork meat by polymerase chain reaction (PCR). A real-time PCR assay with the new intercalating EvaGreen dye was further proposed as a specific and fast tool for hare identification with increased sensitivity (1 pg) compared to end-point PCR (10 pg). It can be concluded that the proposed new primers can be used by both species-specific end-point PCR or real-time PCR to accurately authenticate hare meat.     

Altersbestimmung beim Reh (Capreolus capreolus [L.] und beim Rothirsch (Cervus elaphus L.) mit Hilfe der Trockengewichtsbestimmung der Augenlinse

Zusammenfassung Bei den Ergebnissen haben sich bedeutende Unterschiede im Trockengewicht der Augenlinse zwischen verschieden alten Tieren gezeigt. Die Augenlinse kann beim Reh (Capreolus capreolus L.) sehr gut als Altersindikator verwendet werden. Beim Reh steigt das Trockengewicht der Augenlinse zwischen einem 1 Monat alten Tier und einem 2- bis 3jährigen Tier um das Dreifache und zwischen einem 2- bis 3jährigen und einem 10 bis 11 Jahre alten Tier nur um 1/3 des Ausgangsgewichts an. Beim Rothirsch (Cervus elapbus L.) zeigen sich ähnliche Verhältnisse wie beim Reh (Capreolus capreolus L.). Leider stand mir mit dem Linsenmaterial von 19 Rothirschen zu wenig Material zur Verfügung, um die Unterschiede besonders bei alten Tieren klarer herausheben zu können. Auch beim Rothirsch steigt das Gewicht der Augenlinse bis zum 3. Lebensjahr stark an.

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Summary The results of the studies show significant differences in the dry-weight of the eye lens of roe deer of different ages. In roe deer (Capreolus capreolus L.), the eye-lens is a useful indicator. The dry weight of the eye-lens of the roe increases three-fold between the ages of one month and two or three years. Between the ages of two or three years and ten or eleven years, the dry weight increases by only one fifth of the original weight.Red deer (Cervus elaphus L.) show similar relations to those of the roe deer (Capreolus capreolus L.). Unfortunately the author had the lenses of only 19 red deer at his disposal, a number insufficient to demonstrate significant differences, especially for the older animals. The weight of the eye lens of the red deer increases considerably even in the animal's third year

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Résumé Les résultats laissent apparaître une variation notoire de l'âge des animaux en fonction du poids sec des cristallins de l'oeil. Le cristallin peut très bien être utilisé comme indice de l'âge chez le Chevreuil (Capreolus capreolus L.): en effet, entre l'âge d'un mois et de deux à trois ans, le poids du cristallin varie du simple au triple. A partir de deux à trois ans jusqu'à dix à onze ans, le poids n'augmente que d'un cinquième.Des variations analogues apparaissent chez le Cerf (Cervus elapbus L.) Malgré un matériel réduit de 19 cristallins, on a pu relever une forte variation du poids du cristallin jusqu'á l'âge de trois ans.

     

Development and application of highly specific PCR for detection of chicken (Gallus gallus) meat adulteration

In order to prevent fraud in the sale and strengthen quality assurance, authentic identification of chicken meat is essential. In the present investigation, a chicken (Gallus gallus)-specific polymerase chain reaction (PCR) was developed for the unambiguous identification of chicken meat. The PCR assay employs pair of primers designed against chicken nuclear 5-aminolevulinate (ALA) synthase gene. Highly chicken-specific diagnostic amplicon of 288 bp was established upon PCR and was evident in all the nine breeds/strains of chicken species. Sensitivity of PCR in detecting chicken meat adulteration was established to be at 0.1 % in the foreign meat matrix, while limit of detection (LOD) of chicken DNA was 10 pg. Suitability of the developed chicken-specific PCR was validated and confirmed in raw, cooked/heat treated (60, 80, 100, and 121 °C), and micro-oven cooked meat samples. Possibility of cross-amplification of adulterating DNA was excluded by cross-checking the developed PCR assay with several animal and avian species. The PCR assay developed in this study is highly promising for applications involving circumstances that require authentic identification of chicken meat.     

Toxic and essential metal concentrations in four tissues of red deer ( Cervus elaphus ) from Baranja,Croatia

Kidney (n =?297), liver (n =?52), jawbone (n =?80) and muscle (n =?48) samples collected from red deer (Cervus elaphus) from north-eastern Croatia in the 2002–05 hunting season were analysed for cadmium (Cd), copper (Cu), iron (Fe), mercury (Hg), lead (Pb), selenium (Se) and zinc (Zn) using atomic absorption spectrometry. Statistical evaluation of results showed age-related accumulations of renal cortex Cd and Zn, bone Pb, and muscle Zn. Renal cortex Cd and Zn were significantly associated. In addition, concentrations of Cd and Pb in muscle tissue were significantly correlated with Fe content. Found levels of toxic metals were not likely to affect the health status of animals. A total of 49% of the muscle, 60% of the kidney and 6% of the liver samples were unsuitable for human consumption according to Croatian regulations for Cd in food. However, the calculated intake of Cd through deer meat consumption is small and represents no health risk when consumption is moderate.     

Species-specific multiplex real-time PCR assay for identification of deer and common domestic animals

A multiplex real-time PCR method for discriminating deer and common domestic species, including cattle, goat, horse, donkey, pig, and chicken was developed. Species-specific primer pairs were designed and used to produce different size DNA fragments with diverse melting temperature (T _m ) values. The specificity and sensitivity of these primer pairs were separately confirmed using simplex real-time PCR analysis. Multiplex real-time PCR analysis was performed using combined primers, yielding distinct melting curve profiles for each species. The sensitivity limit of the multiplex PCR method was evaluated. Trace DNA of other species in deer DNA could be identified. Common deer products, including blood, meat, and antler were tested using this multiplex PCR method and different species of deer and domestic animals were identified. The rapid multiplex real-time PCR assay described herein is a specific, sensitive, and reliable method for high-throughput authentication of deer and domestic animal products.     

Consumer evaluation of venison sensory quality: Effects of sex,body condition score and carcase suspension method

Intact fallow bucks (n = 20) and non-pregnant fallow does (n = 24) (Dama dama), in the body condition score (BCS) of 2, 3 and 4 (lean, prime and fat) and rising 2 year old red deer stags (n = 26) (Cervus elaphus) of the same BCS range (2–4) were used in this study to determine the impact of sex, BCS and method of carcase suspension on consumer perception of venison quality. Consumers were asked to evaluate cooked meat samples (M. gluteus medius) on an unstructured line scale for colour, flavour, tenderness, juiciness and overall liking. Meat from both fallow deer and red deer was preferred by consumers when carcases had hung by the pelvic suspension (PS) method compared with the Achilles tendon (AT) method of hanging (p < 0.001). Consumers also noted a difference in colour between sexes in fallow deer venison, with venison from 36 month-old does being darker (p = 0.015), and preferred venison from does over 18–24 month-old bucks. There was a significant difference in the consumer scores for tenderness in red deer stags of BCSs 2 and 4 (p = 0.05) with panellists determining BCS 4 animals to be more tender; however no tenderness differences were observed for fallow deer does compared with bucks.     

Microbiological quality of freshly shot game in Germany

In the framework of a project on the hygiene status of freshly shot game 289 samples were microbiologically analysed: 127 samples from wild boars, 95 from roe deer and 67 from red deer. The microbiological parameters evaluated were the mesophilic aerobic count (APC), which showed mean log10-counts of 2.6 cfu/cm² for roe deer, 2.9 cfu/cm² for red deer and 3.2 cfu/cm² for wild boars and the numbers of Enterobacteriaceae, which gave mean log10-values of 2.1 cfu/cm² for all three species with differing ranges. The concentrations of coagulase positive staphylococci were >2.0 log10 cfu/cm² between 3.2 and 6.3%, according to species. Listeria was found in 14 samples and three samples gave a positive result for Campylobacter. Salmonella was not found in any of the samples analysed.     

Determination of mitochondrial cytochrome B gene sequence for red deer (Cervus elaphus) and the differentiation of closely related deer meats

The cytochrome b gene sequence for red deer was determined using the Dye Terminator Cycle Sequencing method and used for identification of deer meat in meat and meat products. Red deer showed a similarity of 94.1, 84.0, 81.1, 85.5 and 85.6% to sika deer (Cervus nippon), bovine, pigs, sheep and goats, respectively. To differentiate the deer meat, oligonucleotide primers RD-1(5′-TCATCGCAGCACTCGCTATAGTACACT-3′), RD-2(5′-ATCTCCAAGTAGGTCTGGTGCGAATAA-3′) were designed for the region of the cytochrome b gene of red deer. The PCR amplified 194 bp fragments from red and sika deer, but no fragments from bovine, pig, chicken, sheep, goat, horse and rabbit DNA. Although cooking the meats reduced the PCR products, red deer could still be detected in meat heated at 120 °C. To discriminate between red and sika deer, these PCR products were digested by a restriction enzyme (EcoRI,BamHI,ScaI) and analyzed by 4% agorose gel electrophoresis. As a result, the red deer fragment was digested by EcoRI to 67/127 bp fragments but not by BamHI and ScaI. The sika deer fragment was digested to 48/146 bp and 49/145 bp fragments with the two other enzymes, and thus it is possible to differentiate between the two kinds of deer from the digestion pattern of restriction enzymes.