Lipolysis during ripening of Emmental cheese considering organization of fat and preferential localization of bacteria

This study followed the progression of lipolysis in Emmental cheese by quantifying the concentrations of individual free fatty acids (FFA) released during ripening in each of the different rooms: 12 days at 12 degrees C, 28 days at 21 degrees C, and 8 days at 4 degrees C. Lipolysis, which corresponded to 1.56% of fat, mainly occurred in the 21 and 4 degrees C rooms, with 68 and 16.5% of total FFA, respectively. The nonselectivity of lipolytic enzymes was evidenced: all fatty acids were released with level of > or =1%. Differential scanning calorimetry experiments showed that the thermal properties of cheese were affected by (i) lipolysis of fat, that is, the monoacylglycerols, diacylglycerols, and FFA that may be localized at the fat/whey interface, and/or by (ii) hydrolysis of high-melting-point triacylglycerols constituted mainly by long-chain saturated fatty acids (e.g., palmitic acid). Analysis of the cheese microstructure was performed using confocal laser scanning microscopy. Fat globules were mainly disrupted after pressing of curd grains, leading to the release of the milk fat globule membrane (MFGM); fat inclusions were surrounded by pockets of whey, delimited by casein strands. Moreover, colonies of bacteria were preferentially localized in situ at the fat/protein interface. This study showed that both the localization of bacteria and the supramolecular organization of fat which was not protected by the MFGM can help the accessibility of milk fat to lipolytic enzymes and then contribute to the quality of cheese.     

Development of a real time PCR for the detection of Taylorella equigenitalis directly from genital swabs and discrimination from Taylorella asinigenitalis

A discriminatory real time PCR for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM), and the related species T. asinigenitalis was developed for the direct examination of genital swabs. The 112bp amplicons produced from the two species were discriminated from each other using TaqMan probes labelled with different fluorophores. The TaqMan PCR was shown to be specific for the 16S ribosomal DNA of the two species of taylorella and did not cross-hybridise with the 16S ribosomal DNA of other bacteria tested. Direct amplification from genital swabs was shown to be equally sensitive to that of culture methods. Prevalence in a sample set from The Netherlands was shown to be equivalent to that demonstrated by culture. A companion real time PCR that amplified a fragment of the 16S rDNA gene of equine commensal bacteria was developed to ensure bacterial DNA was extracted from swab material supplied for testing. The use of a rapid and reliable real time PCR for the organism causing CEM should aid the control of this disease.     

Sensory properties and aromatic composition of baked snacks containing brewer's spent grain

The incorporation of brewer's spent grain (BSG) into baked snacks (crispy-slices) was investigated in terms of nutritional, sensory and aromatic properties. Wheat flour was replaced with BSG at the levels of 10, 15 and 25%. Snacks containing 10% BSG exhibited high Crispiness index (Ci), low Crispiness work (Wc) and a high number of peaks during texture analysis, indicating that the crispiness of the samples was not negatively affected. However higher levels of BSG affected the texture and the crumb structure of snacks and the results were significantly different to the 100% wheat control. Addition of BSG altered the odour profile of the snacks as shown by the volatile profiles, however sensory results indicated that BSG-containing snacks at a level of 10% were highly acceptable and highlighted the possibility of using BSG as a baking ingredient in the formulation of enhanced fibre baked snacks.     

Genetic characterization and antimicrobial susceptibility of Mannheimia haemolytica isolated from the nasopharynx of feedlot cattle

A surveillance study was undertaken to examine the population dynamics and antimicrobial resistance of Mannheimia haemolytica isolated from feedlot cattle. A total of 416 isolates were collected from the nasopharynx either upon entry or exit from two feedlots in southern Alberta, Canada. Isolates were serotyped, characterized by pulsed-field gel electrophoresis and tested for susceptibility to ten antimicrobial agents via disk diffusion. Resistant isolates were screened by PCR for select antimicrobial-resistance gene determinants. Isolates were highly diverse, with 335 unique pulsed-field profiles identified among 147 strongly related clusters (similarity ≥ 85%). Clonal spread of isolates throughout the feedlots was limited and no clear association was found between genetic relatedness of M. haemolytica and sampling event (entry or exit). Pulsed-field profiles sharing a common serotype and resistance phenotype tended to cluster together. The majority of isolates were identified as serotype 2 (74.5%) although both serotype 1 (11.9%) and 6 (12.7%) were detected. Only 9.54% of isolates exhibited antimicrobial resistance. Resistance to oxytetracycline was most prevalent (n=16), followed by ampicillin (n=10), and amoxicillin/clavulanic acid (n=7). Multi-drug resistance was observed in five isolates. The tetH gene was detected in all but two oxytetracycline resistant isolates. Other detectable resistance determinates included ermX and bla(ROB-1). In the two feedlots examined, M. haemolytica exhibited considerable genetic diversity and limited resistance to common veterinary antibiotics. Garnering further information on the linkage between genotype and phenotype should contribute toward a better understanding of the pathogenesis and dissemination of M. haemolytica in feedlots.