Animal species identification is one of the primary duties of official food control. Since ostrich meat is difficult to be differentiated macroscopically from beef, therefore new analytical methods are needed. To enforce labeling regulations for the authentication of ostrich meat, it might be of importance to develop and evaluate a rapid and reliable assay. In the present study, a loop-mediated isothermal amplification (LAMP) assay based on the cytochrome b gene of the mitochondrial DNA of the species Struthio camelus was developed. The LAMP assay was used in combination with a real-time fluorometer. The developed system allowed the detection of 0.01% ostrich meat products. In parallel, a direct swab method without nucleic acid extraction using the HYPLEX LPTV buffer was also evaluated. This rapid processing method allowed detection of ostrich meat without major incubation steps. In summary, the LAMP assay had excellent sensitivity and specificity for detecting ostrich meat and could provide a sampling-to-result identification-time of 15 to 20 minutes.

Entomophagy is an ancient and actually African tradition that has been receiving renewed attention since edible insects have been identified as one of the solutions to improve global nutrition. As any other foodstuff, insects should be regulated by the government to ensure product quality and consumer safety. The goal of the present paper was to assess the current legal status of edible insects in Africa. For that, corresponding authorities were contacted along with an extensive online search, relying mostly on the FAOLEX database. Except for Botswana, insects are not mentioned in national regulations, although the definitions for “foodstuff” allow their inclusion, i.e., general food law can also apply to insects. Contacted authorities tolerated entomophagy, even though no legal base existed. However, insects typically appear in laws pertaining the use of natural resources, making a permit necessary (in most cases). Pest management regulation can also refer to edible species, e.g., locusts or weevils. Farming is an option that should be assessed carefully. All this creates a complex, nation-specific situation regarding which insect may be used legally to what purpose. Recommendations for elements in future insect-related regulations from the food hygiene point of view are provided.

In the present study, a robust TaqMan real-time PCR amplifying the F57 and the ISMav2sequences ofMycobacterium aviumsubsp.paratuberculosisfrom bovine fecal samples was developed and validated. The validation was based on the recommendations of International Organization for Standardization protocols for PCR and real-time PCR methods. For specificity testing, 205 bacterial strains were selected, including 105M. aviumsubsp.paratuberculosisstrains of bovine, ovine, and human origin and 100 non-M. aviumsubsp.paratuberculosisstrains. Diagnostic quality assurance was obtained by use of an internal amplification control. By investigating six TaqMan reagents from different suppliers, the 100% detection probability was assessed to be 0.1 picogramM. aviumsubsp.paratuberculosisDNA per PCR. The amplification efficiency was 98.2% for the single-copy gene F57 and 97.8% for the three-copy insertion sequence ISMav2. The analytical method was not limited due to instrument specificity. The triplex real-time PCR allowed the reliable detection ofM. aviumsubsp.paratuberculosisDNA using the ABI Prism 7000 sequence detection system, and the LightCycler 1.0. TaqMan_mgband locked nucleic acid fluorogenic probes were suitable for fluorescent signal detection. To improve the detection ofM. aviumsubsp.paratuberculosisfrom bovine fecal samples, a more efficient DNA extraction method was developed, which offers the potential for automated sample processing. The 70% limit of detection was assessed to be 10²CFU per gram of spiked bovine feces. Comparative analysis of 108 naturally contaminated samples of unknownM. aviumsubsp.paratuberculosisstatus resulted in a relative accuracy of 98.9% and a sensitivity of 94.4% for fecal samples containing <10 CFU/g feces compared to the traditional culture method.

Clostridium (C.) perfringens is the causative agent of several diseases in animals and humans, including histotoxic and enteric infections. To gain more insight into the occurrence of its different toxin-genotypes in dairy herds, including those toxin genes previously associated with diseases in cattle or humans, 662 isolates cultivated from feces, rumen content and feed collected from 139 dairy farms were characterized by PCR (detecting cpa, cpb, iap, etx, cpe, and both allelic variants of cpb2). Isolates from feces were assigned to type A (cpa positive, n = 442) and D (cpa and etx positive, n = 2). Those from rumen content (n = 207) and feed (n = 13) were all assigned to type A. The consensus and atypical variants of the cpb2 gene were detected in 64 (14.5 %) and 138 (31.22 %) of all isolates from feces, and 30 (14.5 %) and 54 (26.1 %) of all isolates from rumen content, respectively. Both allelic variants of cpb2 occurred frequently in animals without signs of acute enteric disease, whereby the atypical variant dominated. Five (0.8 %) of all type A isolates were positive for the cpe gene. Therefore, the present study indicates that dairy cows are no primary source for potentially human pathogenic enterotoxin gene positive strains.