The objective of this study was to assess the presence of a Listeria monocytogenes-containing biofilm in milking equipment as a potential source of bulk tank milk contamination on a dairy farm where milk contamination had been previously documented. Samples were collected from milking equipment and milking parlor premises on 4 occasions and analyzed for the presence of L. monocytogenes. Pulsed-field gel electrophoresis (PFGE) typing was conducted on L. monocytogenes isolates from the milking equipment, parlor and storage room floors, bulk tank milk, and in-line milk filters. Pieces from milk meters and rubber liners were obtained to visually assess the presence of a biofilm using scanning electron microscopy. A total of 6 (15%), 4 (25%), and 1 (6%) samples were culture-positive for L. monocytogenes in the first, second, and third sample collection, respectively. Two samples were L. monocytogenes hly PCR-positive but were culture-negative in the fourth sample collection. Combined AscI and ApaI restriction analysis yielded 6 PFGE types for 15 L. monocytogenes isolates obtained from milking equipment, parlor, bulk tank milk, and milk filters. A predominant and persistent PFGE type (PFGE type T) was observed among these L. monocytogenes isolates (9/15 isolates). Scanning electron microscopy of samples from the bottom cover of 2 milk meters showed the presence of individual and clusters of bacteria, mainly associated with surface scratches. The presence of a bacterial biofilm was observed on the bottom covers of the 2 milk meters. Prevention of the establishment of biofilms in milking equipment is a crucial step in fulfilling the requirement of safe, high-quality milk.

In recent years, bovine colostrum has gained popularity as a human food because it is an excellent source of bioactive proteins, which have been claimed to inhibit viral and bacterial pathogens, improve gastrointestinal health, and enhance body condition. A study was conducted to determine bacteriological quality and occurrence of Salmonella in colostrum collected from dairy herds (n = 55) in Pennsylvania. Colostrum samples were analyzed for standard plate count, preliminary incubation count, laboratory pasteurization count, Staphylococcus aureus, Streptococcus agalactiae, coagulase negative staphylococci, streptococci, coliforms, and non-coliforms. A standardized polymerase chain reaction assay was used for detection of Salmonella in colostrum. Salmonella were detected in 8 of 55 (15%) of colostrum samples. Streptococcus agalactiae (1000 colony-forming units [CFU]/mL) was detected in one colostrum sample. The mean standard plate count (977,539 CFU/mL), preliminary incubation count (12,094,755 CFU/mL), laboratory pasteurization count (615 CFU/mL), Staphylococcus aureus (306 CFU/mL), coagulase negative staphylococci (164,963 CFU/mL), streptococci (256,722 CFU/mL), coliforms (323,372 CFU/mL), and non-coliforms (111,544 CFU/mL) counts in colostrum were considerably higher than raw bulk tank milk counts reported previously from Pennsylvania. Analysis revealed that farm size did not influence the bacteriological quality of colostrum. Collection, handling, and storage of colostrum need to be addressed to improve bacteriological quality of colostrum intended not only for feeding calves but also for human consumption.

A study was conducted in two parts to determine the prevalence of toxigenic Clostridium difficile in veal calves and retail meat. The first part of the study focused on the veal production continuum (farm to abattoir). Fifty calves from 4 veal herds (n=200) were followed for 18-22 weeks from the time of arrival on the veal farm to the time of slaughter. Fecal samples were collected from calves every 4-6 weeks. Half of the calves included in the study (n=100) were followed to the abattoir where carcass swabs were collected post slaughter. Fecal samples and carcass swabs were screened for genes encoding C. difficile toxins TcdA, TcdB, and CDT by using real-time polymerase chain reaction (PCR). Carcass swabs were also screened for toxigenic C. difficile by using traditional culture methods. In the second part of the study, ground veal products (n=50 samples) purchased from local grocery stores were examined for toxigenic C. difficile by using real-time PCR and traditional culture methods. Fecal samples from 56 of 200 (28%) calves tested positive for C. difficile toxin genes at least once over the course of the study. Calf age (p=0.011) influenced prevalence of C. difficile toxin genes in calf feces. Toxin genes of C. difficile were detected in one carcass swab by multiplex real-time PCR only. Toxigenic C. difficile was detected by PCR and culture in four (8%) and three (6%) ground veal samples, respectively. The findings of the study reveal that toxigenic C. difficile was most prevalent in veal calves (12%) just before slaughter, although viable toxigenic C. difficile was not recovered from veal carcasses. On the contrary, viable toxigenic C. difficle was recovered from 6% retail meat, thus suggesting that contamination occurs either during or after veal fabrication.

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A study was conducted to assess the diversity among fecal Escherichia coli from healthy lactating cattle. E. coli (n = 100) isolates from 10 healthy lactating dairy cows of a Pennsylvania dairy herd were examined for phenotypic and genotypic characteristics. Results revealed 26, 58, 10, and 6 E. coli isolates belonged to phylogenetic groups A, B1, B2, and D respectively. Overall, 63 serotypes, nine antibiotic resistance profiles, and 65 pulsed-field gel electrophoresis (PFGE) profiles were observed among the 100 isolates. Based on the combination of phenotypic and genotypic characteristics, the 100 E. coli isolates were classified into 76 clonal types. The numbers of different phenotypic and genotypic characteristics of E. coli were observed for each cow at ranges of 2-10, 1- 4, 2-10, and 4-10 for serotypes, antibiograms, PFGE profiles, and clonal types, respectively. The Chao1 estimator was used to estimate diversity among fecal E. coli. It was estimated that a range of 3-55, 1- 4, 2-55, and 8-55 fecal isolates from one cow would be required to include all possible types of E. coli based on serotype, antibiotic resistance profile, PFGE profile, and clonal type respectively. Based on the findings of the study it can be inferred that 1) dairy cattle should be considered as a significant reservoir of genotypically and phenotypically diverse E. coli, and 2) epidemiological investigations that focus on commensal bacteria should take into consideration the diversity within the species being investigated; if not addressed adequately, inappropriate sample size could lead to inaccurate study findings.