Development of a microsatellite-based method for the differentiation of European wild boar (Sus scrofa scrofa) from domestic pig breeds (Sus scrofa domestica) in food

Twenty microsatellites (simple sequence repeats, SSR) were used to discriminate wild boar from domestic pig and to identify mixtures of the two. Reference groups of wild boar and pig samples were collected from the UK and Europe for genetic assignment tests. Bayesian Analysis of Populations software (BAPs) gave 100% correct assignment for blind wild boar and pig samples and correctly identified mixed samples. DNA was extracted from 12 commercial food samples (11 labeled as containing wild boar) including patés, salamis, and sausage, and good SSR profiles were obtained. Eleven samples were correctly assigned as pig, and two as mixed meats. One sample sold as wild boar meat was clearly assigned as pig. A further 10 blind samples of meat cuts were analyzed, eight wild boar and two pig, and all were correctly assigned.     

Identification of a fungi-derived terrestrial halogenated natural product in wild boar (Sus scrofa)

In this study, we identified and quantitated a tetrachlorinated compound found at high concentrations in some samples of the meat of free-ranging wild boar (Sus scrofa) from Southern Germany. Mass spectrometric analysis indicated that the compound was a tetrachloromethoxyphenol isomer, and the subsequently synthesized tetrachloro-p-methoxyphenol was identical with the unknown compound in wild boar. Tetrachloro-p-methoxyphenol is a known secondary metabolite of basidiomycetous fungi, which in turn are regular feed items of the wild boar. It is extremely likely that the wild boar have accumulated tetrachloro-p-methoxyphenol by exploiting basidiomycetes. The highest concentration in the samples (n = 22) was ~1 mg/kg lipids tetrachloro-p-methoxyphenol. This concentration was higher than that of polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT) in any of the samples. Some samples did not contain tetrachloro-p-methoxyphenol, which indicates varied preferences in fungi by wild boars. Our data suggest that during their entire evolution, humans have been in contact with the natural product tetrachloro-p-methoxyphenol by consuming wild boars.     

Microsatellite DNA Analysis of Wild Hops, Humulus lupulus L.

To study the relationships and genetic diversity among wild hops, Humulus lupulus, we analyzed 133 samples of wild hops collected from Europe, Asia and North America using polymorphism on 11 microsatellite loci. Although only three primers showed bands in Japanese hops, all other samples showed polymorphic bands at most loci. There were no duplicate genotypes among samples of European, Chinese and North American hops, and each individual hop could be distinguished completely. The phylogenetic tree constructed from DA distance with the UPGMA method showed a large cluster comprised of European hops, although Russian hops from the Caucasus and Altai regions were separate from the European cluster. Chinese and North American samples gave distinct clusters suggesting genetic differentiation. This study has indicated that hop microsatellite DNA is differentiated, and is dependent upon the origin in regions of Europe, Asia and North America.     

Direct and highly species-specific detection of pork meat and fat in meat products by PCR amplification of mitochondrial DNA

Highly species-specific primers for pork D-loop mtDNA have been designed. Use of these and restrictive PCR amplification conditions has improved a reliable and rapid method for detecting a PCR-amplified 531 bp band from pork. It has been proved useful for detecting both pork meat and fat in meat mixtures, including those dry-cured and heated by cooking. Absence of response in PCR-amplified samples or mixtures from bovine, ovine, chicken, and human was also demonstrated. Furthermore, wild boar and pork samples can be also easily distinguished by a simple AvaII restriction analysis.     

Livestock grazing activities and wild boar rooting affect alpine earthworm communities in the Central Pyrenees (Spain)

In alpine areas, shifts in traditional grazing activities are globally affecting ecosystem properties and rural livelihoods. The ongoing decrease in extensive husbandry, with a decline in sheep numbers and a relative increase in cattle stocking rates, has resulted in the abandonment of large alpine grazing areas. This pastoral change has been recently associated with increased disturbances of wild boar (Sus scrofa), mainly within cattle-stocked ranges. In turn, cattle areas favor earthworm communities, a preferred trophic resource for wild boars in mountain environments. However, it is unknown whether wild boar disturbances, together with grazing activities, can affect earthworm communities. Our aim is to analyze the abundance, richness and ecological categories of earthworms and soil parameters (soil C and N concentrations, moisture, and C:N ratio) in relation to the occurrence of wild boar disturbances and grazing activities at different stocking pressures. We sampled two different grazing scenarios differing in the distribution of cattle along a grazing gradient, which was represented by three levels of stocking pressure (high, intermediate and low). Our results showed a complex effect of grazing activities and disturbances on the abundance and richness of earthworms, along with variations in C:N ratio and soil moisture, especially with increasing cattle presence. At high-stocking pressures differences in earthworm abundance and richness between disturbed and undisturbed areas were limited, whereas at intermediate-stocking pressures earthworms were favored by wild boar disturbances. Ecological categories of earthworms responded differently; endogeic species were the most affected by grazing pressures and wild boar rooting, with highest occurrence at high-stocking pressures and within boar disturbed areas. In sum, pastoral use and soil disturbances affected earthworm community structure and composition in complex ways. These results indicate an interaction of processes that is relevant to understand current changes in alpine ecosystems.     

Detecting hybridization in wild (Coturnix c. coturnix) and domesticated (Coturnix c. japonica) quail populations

Two subspecies of Coturnix coturnix have allopatric distributions in Europe, Maghreb and western Asia (the Common quail C. c. coturnix), and in eastern Asia (the Japanese quail C. c. japonica), except for sympatric breeding areas in the Baikal (Russia) and Kentei (Mongolia) regions, where they could hybridize. Japanese quails have been reproduced in captivity for centuries and domesticated. Massive releasing of Japanese domesticated quails, or hybrids, in west European countries might threaten with hybridization the gene pool of wild quail populations. Here, we used mitochondrial (mtDNA) and nuclear DNA markers (microsatellites), and Bayesian admixture analyses aiming to assess subspecies distinction and identify hybrids in wild and captive-reared quails. A phylogenetic tree identified two major mtDNA clades, corresponding to the two subspecies, and allowed detecting 12 hybrids in wild Common quails sampled in Spain. Additional hybrids were identified in wild quails sampled in Spain, Italy and Senegal using microsatellite markers. Wild quails sampled in Mongolia showed japonica mtDNAs, but their microsatellite genotypes were admixed, suggesting that Common and Japanese quails can hybridize in nature. Some captive-reared stocks of Japanese quails were also admixed. Introgression of domesticated Japanese quail genes in wild Common quail populations might affect the phenotypic expression of functional traits, as body size, feather colours, sexual calls and migratory behaviours. Hence, restocking with captive-reared non-native quails should be banned.     

Razorbacks: Feral Pigs as Agricultural Pests,Disruptors of Ecosystems,Reservoirs of Contagion,and Favored Game for Sport and Subsistence Hunters: A Review of the Literature, 2005–2011

Whether they are called “razorbacks,” as is common in the southern United States (and where they are notably the official sports mascots of the University of Arkansas), or “wild boar,” in much of the rest of the world, feral pigs are interbreeding subtypes of Sus scrofa, the same pig domesticated over 5,000 years ago. Once they have become established in the wild, these formerly manageable animals rapidly undergo a dramatic reversion to a wild appearance, may display aggression toward humans, engage in agriculturally and ecologically damaging behaviors, and spread diseases to both livestock and humans. They are the targets of many ongoing, increasingly sophisticated, science-based, eradication efforts.     

Identification and genetic structure of wild Italian <Emphasis Type="Italic">Humulus lupulus</Emphasis> L. and comparison with European and American hop cultivars using nuclear microsatellite markers

Nine genic SSR loci were used to evaluate the genetic diversity and identify accessions in wild Italian Humulus lupulus L., in comparison with widely cultivated European and U.S. commercial cultivars. A collection of 80 wild hop samples from Italy and 43 hop cultivars from Europe and U.S., were characterized. Allelic frequency analysis revealed 65 distinct Italian genotypes and differentiated all the commercial cultivars; moreover, specific alleles were observed for wild and cultivated hops. The number of alleles identified in the wild population were 104 and 123 within all the accessions. The maximum polymorphic information content was evidenced for locus HlGA23 in the Italian wild population and in the whole set of accessions (0.905 and 0.902 respectively). The dendrogram constructed from Euclidean distance with the UPGMA method showed two main clusters, one including commercial American and European accessions and one mostly composed by wild Italian accessions. Model-based clustering (Bayesian method) placed the accessions into five germplasm groups, one of which was characterized by Italian genotypes only. The study showed for the first time the great biodiversity present in Italy, and the remarkable differences with European and American hops. It was also found that within the population of north-central Italy a large genetic variability is present, suited to be studied and exploited; this genetic wealth could be used in future breeding programs in order to develop new hop varieties carrying characteristics useful for brewers.     

Analysis of volatiles in meat from Iberian pigs and lean pigs after refrigeration and cooking by using SPME-GC-MS

The volatile compounds generated in meat from Iberian and lean pigs after four different treatments (raw, refrigerated, cooked, and refrigerated cooked meat) were analyzed. The different treatments showed different volatile profiles. Methyl alcohols and ketones (such as 2-ethyl-hexan-1-ol, 2-methyl-butan-1-ol, 3-methyl-butan-1-ol, and 3-hydroxy-butan-2-one) were the most representative in refrigerated meat because of the degradation of carbohydrates and proteins together with the Strecker degradation pathway. Lipid-derived volatiles were the most abundant in cooked meat and refrigerated cooked meat. Meat from different pig breeds presented different volatile profiles, probably due to different enzymatic and oxidative deterioration susceptibility. Otherwise, the fat content and its compositional characteristics also played an important role in the generation of volatiles. As compared to samples from lean pigs, muscles from Iberian pigs showed a higher content of heme iron that may have promoted the generation of higher content of total lipid-derived volatiles during the refrigeration of cooked meat. Despite that, the formation of volatiles with low thresholds and related to intense rancidity perception likely to be derived from polyunsaturated fatty acids was higher in lean pork than in meat from Iberian pigs. This might be expected to lead to a more intense development of a warmed over flavor during refrigeration of cooked samples from lean pigs.     

Improved recovery of fecal streptococci from tropical (Caribbean) marine recreational waters by anaerobic-MF method using m-Enterococcus agar

The possible advantage of anaerobic membrane filtration (anaerobic-MF) for fecal streptococci (FS) enumeration in tropical (Caribbean) marine recreational waters was evaluated with samples from the beaches of Barbados. Selected samples were also analyzed for fecal coliforms in a comparison. Up to 80% of presumptive FS colonies on m-Enterococcus agar in aerobic-MF were false-positive. Anaerobic-MF reduced presumptive FS and background bacterial counts by 2 to 5 orders of magnitude, increased proportions of FS verification in 54 of the 80 samples, but conferred no obvious advantage in the FC counts. 94 to 100% of blue colonies on MFC agar in 24 hr aerobic- and anaerobic-MF cultures were confirmed as FC. Presumptive and background bacteria in FS counts, proportions of verified FS, and presumptive FC counts correlated with one another. The MF and most probable number (MPN) methods were compared. In a modified MPN confirmation, cultivation of FS colonies in Brain heart infusion agar prior to esculin hydrolysis test gave substantially higher counts than anaerobic-MF at three stations. The overall FS and FC data apparently indicate only low contamination levels occur. Beaches exceeding the region's quality (USEPA) standards are associated with municipal wastewate outfalls.     

公猪舍夏季温度和流场数值CFD模拟及验证

为研究夏季全漏缝地板公猪舍湿帘风机蒸发降温效果及舍内环境分布规律,该文利用计算流体力学CFD(computational fluid dynamics)对北京养猪育种中心SPF(Specific Pathogen Free Swine)公猪舍进行模拟研究并通过实测数据进行验证。研究中将漏缝地板作为多孔介质简化,基于标准k-?湍流模型对空载及装猪猪舍内的风速场和温度场进行模拟,通过模拟值与实测值的对比验证模型的合理性。结果表明采用该模型模拟空载时猪舍,风速场模拟值与实测值误差较小,相对误差范围在0.25%~30.8%。模拟温度与实测温度最大绝对误差为0.48 K,平均绝对误差为0.11 K,平均相对误差为0.5%。模拟装猪时的猪舍,温度分布结构与装猪前相似,但整体温度略有上升。该研究可对当前常用的含漏缝地板猪舍建模研究提供参考,并为畜禽舍内改造和建筑实践提供理论依据。     

Thermal susceptibility of Salmonella in the Moroccan food snail, Helix aspersa.

Thirty samples of 10-15 Helix aspersa food snails were examined for Salmonella by a surface rinsing method and by analysis of whole snails rinsed with 70% ethanol. Thirteen samples were positive by the rinsing method and 6 were positive by whole snail analysis, with this difference being significant (P less than 0.01). Although Salmonella contamination in H. aspersa appeared to be predominantly surface, the pathogen was also found within the snail meat. The ability of surface and subsurface Salmonella organisms to survive in cooked snails was determined in 90 samples. Thermocouple readings indicated that an internal temperature of at least 200 degrees F (93 degrees C) was reached within the snail meat during cooking by a typical recipe. This temperature was sufficient to kill both surface and subsurface Salmonella in 29 samples positive for the pathogen prior to heating. A variety of serotypes representing several somatic groups was isolated.     

三品种小型猪与云南野猪白细胞抗原复合物(SLA)微卫星的遗传多样性

为研究明光小耳猪、滇南小耳猪、巴马香猪遗传多样性以及命名,本研究利用猪白细胞抗原复合物(SLA)区域内18个微卫星,探讨了3品种小型猪和云南野猪(Sus scrofa silvianus)的观测杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)等遗传参数;且从单倍型分型、聚类分析以及主成分分析进一步研究了其亲缘关系。结果表明,云南野猪多态性最高(PIC=0.694),巴马香猪最低(PIC=0.585),3品种小型猪均具有丰富的遗传多样性。在检测群体中SLAⅠ、Ⅱ区域分别有12种和14种单倍型,其中明光小耳猪、滇南小耳猪和云南野猪共享4个单倍型H1、H3、H4和H03,巴马香猪仅与明光小耳猪共享单倍型H2。通过系统聚类和主成分分析发现,明光小耳猪、滇南小耳猪与云南野猪亲缘关系较近,巴马香猪与之关系较远。研究结果提示,明光小耳猪和滇南小耳猪具有相似的遗传背景,明光小耳猪可能是滇南小耳猪的一个品系,这为解决同种异名问题提供了分子证明。     

Influence of combined grazing by wild boar and small ruminant on soil and plant nutrient contents in a coppice oak forest

Limited information is available about the impact of grazing on soil and plant nutrient contents in Mediterranean deciduous oak forests grazed by both livestock and wild life. Thus, the main objective of this study was to investigate the effect of wild boar and small ruminants grazing in a coppice oak forest in N Greece. Eight even‐aged stands of similar canopy cover and site quality were selected: four stands with long‐time grazing mainly by wild boar and four stands with long‐time grazing by sheep and goats. A plot of 150 m² in each of the stands was fenced in order to be protected from grazing. A similarly sized plot open to grazing was assigned next to the fenced plot. Species composition was recorded in fenced and grazed plots. Soil samples and individual plants of the dominant palatable grass species Dactylis glomerata and Brachypodium sylvaticum were collected in every fenced and grazed plot. These samples were analysed for contents of nutrients. The results indicated that grazing increased organic matter, C, and total N, P and Ca, while it had no effect on the plant nutrients Mg, Na, K and all micronutrients, and pH as well. Wild boar grazing had a significantly negative impact on the concentrations of Na, Fe, Zn, and Mn in the soil. Beside a decrease in K and Mn, grazing had minimal effects on the contents of plant nutrients. Conversely, wild boar grazing increased the contents of several nutrients such as N, Na, Cu and Fe in plant tissue. It seems that species composition was more affected by the topography and management than by the soil features as revealed by RDA analysis. The results of this study indicate that it is important to maintain moderate small ruminant grazing intensity for the sustainable multi‐disciplinary management of coppice oak forests. It is also important to control wild boar population in order to facilitate ecosystem process, soil preservation and forage quality.     

Wild boar and red deer affect soil nutrients and soil biota in steep oak stands of the Eifel

In sloping oak forests of the German low mountain range high wild boar (Sus scrofa) and red deer (Cervus elaphus) population densities may affect soil ecological processes by grubbing, grazing, trampling and dunging. We simulated wild boar grubbing in a fenced exclosure and an unfenced replicate and established two adjacent control plots, one fenced, the other unfenced. We evaluated if repeated soil bioturbation and game exclusion by fencing influence soil texture, soil chemical and soil biotic properties in the upper soil over a time period of 2 years. Soil bioturbation was conducted in November 2000 and 2001 creating a grubbing pattern similar to that found in naturally grubbed areas. Soil and fauna sampling was performed in spring and fall of the years 2001 and 2002.Soil bioturbation did not affect soil texture, pH and the contents of organic carbon and nitrogen. In contrast, the contents of potassium and magnesium, the microbial activity and the abundance of saprophageous and predatory soil arthropods were generally lower in grubbed plots compared to ungrubbed control plots (p≤0.05).The exclusion of game did not improve soil quality. On the contrary, microbial activity and the contents of organic carbon and total nitrogen were elevated outside the fenced exclosure (p≤0.05) which may be related to the deposition of urine and dung.Our study found that large mammals affect soil nutrient cycling in sloping oak forests either directly by the deposition of urine and dung or indirectly by accelerating nutrient leaching and disturbing the decomposer system in the soils.     

Investigations of pipe-borne pollution in Benin City,Nigeria

Two important environmental parameters could be identified as being the main causes of pipe borne water contamination in the City of Benin. These are the combined effects of rainfall and excessive construction work and, secondly, the lack or inadequacy of plumbing codes. To study the combined effects of rainfall and excessive construction on the contamination of water, samples were tested for total coliform using the multiple-tube fermentation test. Sites were chosen to allow for the comparison of the coliform counts in areas of comparatively insignificant construction to areas of excessive public construction. Results show that in the areas of heavy public construction, the curves for both the rainfall and the coliform counts in the consumer lines were skewed in the same manner. The peak rainfall was in September (437.I mm) which corresponds to peak coliform number in the consumer lines for both Zone A (62 coliforms/100 ml) and Zone B (60 coliforms/100 ml) for the same month. When compared to the area of insignificant public construction as shown in Zone C, rainfall had little or no effect on the coliform counts either at the pumping station or in the consumer line. The maximum number of coliforms recorded for the consumer line was 5/100 ml during the month of June. Results suggest that the high counts of coliforms in the Benin City water supply could be attributed primarily to the effects of construction which resulted in pipe breakage. During heavy rainfall and flooding, water under flood pressure enters the pipes through cracks, improperly tightened joints and damaged pipes. The improper enforcement of plumbing codes or lack of such codes was also identified as another cause of the water contamination as shown in Zone D. A study of the effects of water pipes passing through septic tank drain fields also shows that such effluents served as another source of major contamination. The highest recorded number for coliforms for the pipe-borne water just before entering the septic tank drain field was 12/100 ml and after it passed through the field the number had gone up to 60/100 ml. Artificial contamination of the septic tank effluents to try to detect leaks in the water pipes passing through the absorption field was investigated. The bacterial contaminant could be recovered in the pipe-borne water and the quantity was skewed in the same manner as total rainfall between periods of sampling.     

Volatile profiles of dry-cured meat products from three different Iberian x Duroc genotypes

Volatile profiles of two Iberian dry-cured products, dry-cured loin and ham, from three different Iberian x Duroc genotypes, was assessed. Three groups of 10 pigs, each (5 males and 5 females) from different genotypes, were studied: GEN1 = male Iberian x female Duroc1; GEN2 = male Duroc1 x female Iberian; and GEN3 = male Duroc2 x female Iberian. The genotype Duroc1 (DU1) corresponded to pigs selected for the production of dry-cured meat products (hams, loins, and shoulders), with a high level of fattening, while the genotype Duroc2 (DU2) corresponded to animals selected for meat production. Genotype slightly affected the volatile profiles of both dry-cured meat products, although dry-cured loin from GEN3 showed higher hexanal content. Dry-cured loin showed a volatile profile very different to that found in dry-cured ham. Volatile compounds of dry-cured meat products were mainly originated by lipid and protein degradation. Most of the volatile detected in both meat products came from lipid oxidation such as acids, aldehydes, ketones, alcohols, and hydrocarbons. In addition, a high proportion of volatile compounds from the Maillard reaction was found. Branched aldehydes and some sulfur and nitrogen compounds have their origin in the amino acids degradation by the Strecker reaction, while branched alcohols and acids come from the lipid oxidation of branched aldehydes. Dry-cured ham showed a higher number and a higher level of compounds with origin in protein and lipid degradation than dry-cured loin, which agrees with the longer ripening of the hams (24 months) with respect to the loins (4 months). In dry-cured loins, apart from these compounds, seasoning mixture provides high amount of volatiles, such as terpenes (from paprika and oregano) and sulfur compounds (from garlic), which have great importance in the overall aroma of this product.     

Abundance of wild prey modulates consumption of supplementary food in the Iberian lynx

Food supplementation is increasingly used as a conservation tool. However, little is known about how much supplemental food is used by target populations or the degree to which the abundance of natural food affects the utilization of supplemental food. Long-term supplementation programmes could cause individuals to rely almost exclusively upon supplemental food and, consequently, lose some skills needed to forage efficiently on natural food. This may result in reduced fitness upon discontinuation of supplemental food. The Iberian lynx (Lynxpardinus) preys almost exclusively upon European wild rabbits (Oryctolagus cuniculus), and some populations are thought to be food limited. We quantified the contribution of supplemented domestic rabbits, whose guard hairs could be distinguished from hairs of wild rabbits, to the diet of the Iberian lynx. We also examined whether the consumption of domestic rabbits varied with the availability of wild rabbits, and with the duration of exposure to supplemental food. Domestic rabbits made up over 50% of the diet. Consumption of domestic rabbits decreased non-linearly as the relative abundance of wild rabbits increased; however, this pattern was true only above a threshold density of one wild rabbit km⁻¹. Below this threshold, supplementation was apparently strictly necessary to retain Iberian lynx. The consumption of domestic rabbits did not increase with the length of the supplementation period. Lynx continued consuming wild rabbits proportionally to their abundance, suggesting lynx did not become dependent upon supplemental food. Understanding how the abundance of natural food modulates consumption of supplemental food may help to adjust supplementation schedules to food availability and to the needs of the target populations.     

Interactions between plant species and mycorrhizal colonization on the bacterial community composition in the rhizosphere

This study assessed the effect of mycorrhizal colonization by Glomus intraradices (Gi) and G. versiforme (Gv) on the bacterial community composition in the rhizosphere of canola, clover and two tomato genotypes (wild type (76R) and its mutant with reduced mycorrhizal colonization (rmc)). Additionally, the effect of light intensity on the rhizosphere bacterial community composition of the tomato genotypes was studied. The bacterial community composition was assessed by denaturing gradient gel electrophoresis (DGGE). In canola, which is considered to be a non-mycorrhizal species, inoculation with Gi increased the shoot dw compared to Gv and the non-mycorrhizal control plants and also induced changes in the bacterial community composition in the rhizosphere. These fungal effects were observed although less than 8% of the root length of canola was colonized. On the other hand, about 50% of the root length of clover was colonized and inoculation with Gv resulted in a higher shoot dw compared to Gi or the control plants but the rhizosphere bacterial community composition was not affected by inoculation. Plant growth, mycorrhizal colonization and bacterial community composition of the two tomato genotypes were affected by a complex interaction between tomato genotype, AM fungal species and light intensity. Low light intensity (photosynthetic photon flux 200–250 μmol m⁻² s⁻¹) increased the shoot–root ratio in both genotypes and reduced colonization in the wild type. The differences in bacterial community composition between the two genotypes were more pronounced at low than at high light intensity (550–650 μmol m⁻² s⁻¹).     

Real-time TaqMan polymerase chain reaction detection and quantification of cow DNA in pure water buffalo mozzarella cheese: method validation and its application on commercial samples

Mozzarella cheese obtained from buffalo (Bubalus bubalis) milk is a typical Italian product certificated by means of the European Protected Designation of Origin (PDO). Mozzarella cheese can also be obtained from bovine milk or bovine/buffalo milk mixtures, but in this case, it cannot be sold as PDO product, and its label must report the actual ingredients. However, bovine milk in PDO products was frequently detected in the past, suggesting fraudulent addition or accidental contamination. Several methods based on end-point polymerase chain reaction (PCR) have been profitably applied in a large number of tests to detect the presence of undeclared ingredients, also in dairy products. In the present study we report a real-time PCR method able to quantify bovine milk addition to pure buffalo cheese products. We validated a normalized procedure based on two targets: bovine mitochondrial cytochrome b (cyt b) to detect and quantify the bovine DNA and nuclear growth hormone (GH) gene used as a universal reference marker. With the use of this real-time PCR assay, 64 commercial mozzarella di bufala cheese samples purchased at local supermarkets, dairy shops, or directly from cheese manufacturers were analyzed. The results obtained demonstrate that most of the commercial samples were contaminated with bovine milk. Therefore, this assay could be conveniently employed to carry out routine and accurate controls aimed not only to discourage any fraudulent behavior but also to reduce risks for consumer health.