Longitudinal assessment of dairy farm management practices associated with the presence of psychrotolerant Bacillales spores in bulk tank milk on 10 New York State dairy farms.

The ability of certain spore-forming bacteria in the order Bacillales (e.g., Bacillus spp., Paenibacillus spp.) to survive pasteurization in spore form and grow at refrigeration temperatures results in product spoilage and limits the shelf life of high temperature, short time (HTST)-pasteurized fluid milk. To facilitate development of strategies to minimize contamination of raw milk with psychrotolerant Bacillales spores, we conducted a longitudinal study of 10 New York State dairy farms, which included yearlong monthly assessments of the frequency and levels of bulk tank raw milk psychrotolerant spore contamination, along with administration of questionnaires to identify farm management practices associated with psychrotolerant spore presence over time. Milk samples were first spore pasteurized (80°C for 12 min) and then analyzed for sporeformer counts on the initial day of spore pasteurization (SP), and after refrigerated storage (6°C) for 7, 14, and 21 d after SP. Overall, 41% of samples showed sporeformer counts of >20,000 cfu/mL at d 21, with Bacillus and Paenibacillus spp. being predominant causes of high sporeformer counts. Statistical analyses identified 3 management factors (more frequent cleaning of the bulk tank area, the use of a skid steer to scrape the housing area, and segregating problem cows during milking) that were all associated with lower probabilities of d-21 Bacillales spore detection in SP-treated bulk tank raw milk. Our data emphasize that appropriate on-farm measures to improve overall cleanliness and cow hygiene will reduce the probability of psychrotolerant Bacillales spore contamination of bulk tank raw milk, allowing for consistent production of raw milk with reduced psychrotolerant spore counts, which will facilitate production of HTST-pasteurized milk with extended refrigerated shelf life.

Identification and characterization of psychrotolerant coliform bacteria isolated from pasteurized fluid milk.

The presence of coliform bacteria in pasteurized fluid milk typically indicates that product contamination occurred downstream of the pasteurizer, but it may also indicate pasteurization failure. Although coliform detection is frequently used as a hygiene indicator for dairy products, our understanding of the taxonomic and phenotypic coliform diversity associated with dairy products is surprisingly limited. Therefore, using Petrifilm Coliform Count plates (3M, St. Paul, MN), we isolated coliforms from high-temperature, short-time (HTST)-pasteurized fluid milk samples from 21 fluid milk processing plants in the northeast United States. Based on source information and initial characterization using partial 16S rDNA sequencing, 240 nonredundant isolates were obtained. The majority of these isolates were identified as belonging to the genera Enterobacter (42% of isolates), Hafnia (13%), Citrobacter (12%), Serratia (10%), and Raoultella (9%); additional isolates were classified into the genera Buttiauxella, Cedecea, Kluyvera, Leclercia, Pantoea, and Rahnella. A subset of 104 representative isolates was subsequently characterized phenotypically. Cold growth analysis in skim milk broth showed that all isolates displayed at least a 2-log increase over 10 d at 6°C; the majority of isolates (n=74) displayed more than a 5-log increase. In total, 43% of the representative isolates displayed lipolysis when incubated on spirit blue agar at 6°C for 14 d, whereas 71% of isolates displayed proteolysis when incubated on skim milk agar at 6°C for 14 d. Our data indicate that a considerable diversity of coliforms is found in HTST-pasteurized fluid milk and that a considerable proportion of these coliforms have phenotypic characteristics that will allow them to cause fluid milk spoilage.

Identification of dairy farm management practices associated with the presence of psychrotolerant sporeformers in bulk tank milk.

Some strains of sporeforming bacteria (e.g., Bacillus spp. and Paenibacillus spp.) can survive pasteurization and subsequently grow at refrigeration temperatures, causing pasteurized fluid milk spoilage. To identify farm management practices associated with different levels of sporeformers in raw milk, a bulk tank sample was obtained from and a management and herd health questionnaire was administered to 99 New York State dairy farms. Milk samples were spore pasteurized [80°C (176°F) for 12 min] and subsequently analyzed for most-probable number and for sporeformer counts on the initial day of spore pasteurization (SP), and after refrigerated storage (6°C) at 7, 14, and 21 d after SP. Management practices were analyzed for association with sporeformer counts and bulk tank somatic cell counts. Sixty-two farms had high sporeformer growth (≥3 log cfu/mL at any day after SP), with an average sporeformer count of 5.20 ± 1.41 mean log10 cfu/mL at 21 d after SP. Thirty-seven farms had low sporeformer numbers (<3 log cfu/mL for all days after SP), with an average sporeformer count of 0.75 ± 0.94 mean log10 cfu/mL at 21 d after SP. Farms with >25% of cows with dirty udders in the milking parlor were 3.15 times more likely to be in the high category than farms with ≤10% of milking cows with dirty udders. Farms with <200 cows were 3.61 times more likely to be in the high category than farms with ≥200 cows. Management practices significantly associated with increased bulk tank somatic cell count were a lack of use of the California mastitis test at freshening and >25% of cows with dirty udders observed in the milking parlor. Changes in management practices associated with cow cleanliness may directly ensure longer shelf life and higher quality of pasteurized fluid milk.

The ability of common mastitis-causing pathogens to survive an ensiling period.

A forage-based study was conducted to examine the ability of enterococci, streptococci, and gram-negative bacteria to survive a 3-wk ensiling process. Harvested grass and corn forages were each divided into 3 treatment groups consisting of 2 commercially available silage inoculants and 1 negative control group. Within 24h of harvest, a uniform amount of forage was added to each of 18 vacuum-sealable freezer bags. Inoculants 1 and 2 were applied to each of 6 bags and the remaining 6 bags were not inoculated (negative control). All bags were sealed using an industry-grade vacuum sealer to create a pillow pack. At wk 1, 2, and 3 of the ensiling process, 6 bags were opened, 2 from each of the 3 groups. Dry matter (DM), pH, and bacterial enumeration were determined on the forage before ensiling, as well as at wk 1, 2, and 3 of ensiling. The pH of grass and corn samples decreased to 5.2 and 4.1, respectively, by the end of the ensiling period. Inoculant 2 (7.3 log(10) cfu/g of DM) resulted in a greater enterococcal count in grass samples than did inoculant 1 (4.2 log(10) cfu/g of DM) at 3 wk of ensiling and both were greater than the negative control (1.2 log(10) cfu/g of DM). Counts of streptococci in grass samples at wk 3 were greater in inoculated samples (inoculant 1, 7.6 log(10) cfu/g of DM; inoculant 2, 7.0 log(10) cfu/g of DM) compared with the negative control (4.3 log(10) cfu/g of DM). Treatment differences were observed following 1 wk of ensiling in corn samples. Enterococcal counts on corn samples following the 3-wk ensiling were not significantly different between inoculant 1 (4.3 log(10) cfu/g of DM) and inoculant 2 (4.4 log(10) cfu/g of DM). However, counts in both inoculated groups were greater than in the negative control group (4.0 log(10) cfu/g of DM). No significant treatment differences were observed for streptococci counts at wk 3. Counts for gram-negative bacteria were not detectable by wk 3 of ensiling in either grass or corn samples. The addition of a silage inoculant led to greater counts of enterococci and streptococci in grass silage compared with the negative control at the end of a 3-wk ensiling period. The application of a silage inoculant led to increased enterococci counts, but not streptococci counts, in corn samples following the ensiling period. These data indicate that enterococci and streptococci are able to survive the pH decrease observed during ensiling.

Experimental design applied to the formulation of lipsticks with particular features.

In our work a non-classical experimental design was applied to obtain lipsticks endowed with particular characteristics. Our aim was to formulate lipsticks that leave a brilliant and shiny colour application and have a transparent look. The emollient substances and the waxes (consistency factors) were identified as the main variables of the system. A two phase experimental strategy was thought out: the optimal quantities of consistency factors were selected using a Doehlert experimental matrix, whereas the correct mixtures of emollients were determined using a Scheffé simplex-centroid design. These two design were combined and a set of 49 experiments was obtained. The experiments carried out allowed the definition of a zone of two phases in which the objectives were attained: the correct types and appropriate quantities of emollients and waxes were determined. To find a possible correlation between some mixtures and the lipsticks' sensorial behaviour, differential scanning calorimetry was used. These results, in addition to those obtained using the experimental design allowed us to select the best lipstick formula. (c) Rapid Science Ltd. 1998.

Inflammatory effects of Salmonella typhimurium porins.

The inflammatory activity of porins purified from Salmonella typhimurium has been investigated. Porins (0.3 to 30 micrograms) injected into the rat paw induced a dose-related edema that was not due to lipopolysaccharide contamination and did not appear to be dependent on the activation of the complement system. The edema induced by 30 micrograms of porins was comparable to that caused by 1 mg of carrageenin and was inhibited by indomethacin (5 mg/kg) and dexamethasone (0.1 mg/kg). Porins (1 to 10 micrograms/ml) induced a concentration-related release of histamine from rat peritoneal cells. These results are discussed in the light of the possible pathogenic role of porins in infections.

Epizootiology of epidemic typhus (Rickettsia prowazekii) in flying squirrels.

Vector transmission of Rickettsia prowazekii among wild flying squirrels, Glaucomys volans, was suggested by the occurrence of natural infection of squirrel lice and fleas. Lice, mostly Neohaematopinus sciuropteri Osburn, were found infected in the fall in each of 2 consecutive years; 4 of the 8 pools of this insect tested were infected. Fleas, Orchopeas howardii (Baker), were found infected on two occasions in 1 of the 2 consecutive years. However, only 2 of 14 flea pools were infected. No evidence of infection was found in mites, Haemogamasus reidi Ewing and Androlaelaps fahrenholzi (Berlese). These findings implicate the flying squirrel louse and flea as possible vectors in nature. Serologic tests of flying squirrel sera revealed a maximum incidence of seroconversions in the fall and early winter months, coincident with the maximum increase in abundance of the suspected arthropod vectors. The infection was found to persist form year to year in the same enzootic foci. Infection appeared to spread most rapidly in young, non-immune animals born in the preceding spring and summer after congregating in dense aggregations in the fall. No other animals in the same habitat were found to have been infected. Aspects of the ecology of the ectoparasites associated with the flying squirrels are described, especially seasonal activity and abundance in nests. The potential public health importance of this sylvan disease in flying squirrels and in its ectoparasites, particularly the non-host specific, wide ranging squirrel flea, is noted.