Growth of gram-positive mastogenic bacteria in normal, simulated bulk tank, and mastitic milk held at simulated fluctuating temperatures of farm bulk tank

Growth of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus bovis, and Streptococcus uberis was studied in normal milk, simulated bulk tank milk, and aseptic mastitic milk held at simulated fluctuating temperatures of farm bulk tank for 48 h. With the exception of S. bovis, growth rates of the other five bacteria were similar in both normal and simulated bulk tank milk. Mastitic milk inhibited growth of all bacteria studied. A 24-h adjustment period occurred before most of the bacteria started growing. The mastitis level in a dairy herd may be monitored by cultures of bulk tank milk samples and by calculations as discussed in this study.     

Microbiological quality of bulk tank raw milk in Prince Edward Island dairy herds

The objectives of this study were to evaluate microbiological quality of bulk tank milk in Prince Edward Island, to evaluate correlation among milk quality criteria, and to determine seasonal effects on milk quality parameters. Bulk tank raw milk quality was evaluated on all Prince Edward Island dairy herds (n = 235) over a 2-yr period (March 2005 to March 2007). Biweekly total aerobic (TAC), preliminary incubation (PIC), laboratory pasteurization, and coliform (CC) counts were determined using a Petrifilm culture system. Additionally, bulk tank somatic cell count was determined weekly. The mean and median values were 12.8 × 10³ and 4.9 × 10³ cfu/mL for TAC, 29.6 × 10³ and 13 × 10³ cfu/mL for PIC, 87 and 12 cfu/mL for laboratory pasteurization count, 21 and 5 cfu/mL for CC, and 218 × 10³ and 187 × 10³ cells/mL for somatic cell count. There was moderate correlation (0.57) between TAC and PIC. All other correlation coefficients were low (<0.26). Correlation results suggest that a single quality parameter could not predict others used in this study. Seasonal data indicate that 1) in general, all counts tended to be low in winter, 2) the CC and somatic cell count were always high in summer, and 3) TAC tended to be high during summer.     

Prevalence,antimicrobial resistance,and molecular characterization of Campylobacter spp. in bulk tank milk and milk filters from US dairies

Campylobacter spp. are frequently isolated from dairy cows as commensal organisms. Sporadic Campylobacter infections in humans in the United States are generally attributed to poultry, but outbreaks are also commonly associated with dairy products, particularly unpasteurized or raw milk. Bulk tank milk samples and milk filters from US dairy operations were collected during the National Animal Health Monitoring System Dairy 2014 study and analyzed using real-time PCR and traditional culture techniques for the presence of thermophilic Campylobacter species. The weighted prevalence of operations from which we detected Campylobacter spp. in either bulk tank milk or milk filters was 24.9%. We detected Campylobacter spp. in a higher percentage of operations with 100–499 cows (42.8%) and 500 or more cows (47.5%) than in operations with 30–99 cows (6.5%). Campylobacter spp. were also more frequently detected in operations in the west than the east (45.9 and 22.6%, respectively). We isolated Campylobacter spp. from approximately half of PCR-positive samples, representing 12.5% (weighted prevalence) of operations. The majority (91.8%) of isolates were C. jejuni, but C. lari and C. coli were also isolated. We detected resistance to tetracycline in 68.4% of C. jejuni isolates, and resistance to ciprofloxacin and nalidixic acid in 13.2% of C. jejuni isolates. Based on pulsed-field gel electrophoresis, we found that dairy-associated C. jejuni were genotypically diverse, although clonal strains were isolated from different geographic regions. These results suggest that bulk tank milk can be contaminated with pathogenic Campylobacter spp., and that the consumption of unpasteurized or raw milk presents a potential human health risk.     

Improving farm management by modeling the contamination of farm tank milk with butyric acid bacteria

Control of contamination of farm tank milk (FTM) with the spore-forming butyric acid bacteria (BAB) is important to prevent the late-blowing defect in semi-hard cheeses. The risk of late blowing can be decreased via control of the contamination level of FTM with BAB. A modeling approach was applied to identify an effective control strategy at the farm level. The simulation model developed was based on a translation of the contamination pathway into a chain of unit operations. Using various simulations, the effects of factors related to feed quality, feed management, cattlehouse hygiene, and milking practices on the contamination level of FTM were evaluated. Contamination level of silage was found to be the most important factor. When silage contains on average less than 3 log₁₀ BAB/g, a basic pretreatment of udder teats before milking (∼75% removal of attached spores) is sufficient to assure an FTM contamination level below 1 BAB/mL. When silage contains more than 5 log₁₀ BAB/g, it should not be fed, because it then becomes almost impossible to assure an FTM contamination level below 1 BAB/mL. Measures aimed at improving cattlehouse hygiene, the contamination via soil, and the contamination level of other feeds contribute only marginally to the control of the contamination of FTM with BAB. Application of the modeling methodology could be beneficial for the control of the contamination of FTM with other microorganisms such as Bacillus cereus.     

Minimizing the level of butyric acid bacteria spores in farm tank milk

A year-long survey of 24 dairy farms was conducted to determine the effects of farm management on the concentrations of butyric acid bacteria (BAB) spores in farm tank milk (FTM). The results were used to validate a control strategy derived from model simulations. The BAB spore concentrations were measured in samples of FTM, feces, bedding material, mixed corn and grass silage fed to cows in the barn, and soil. In addition, a questionnaire was used to gather farm management information such as bedding material used and teat cleaning method applied. The average BAB spore concentration in FTM was 2.7 log₁₀ spores/L, and 33% of the FTM samples exceeded a concentration of 3 log₁₀ spores/L. Control of the average spore concentration in mixed silage fed was the only aspect of farm management that was significantly related to the concentration of BAB spores in FTM. Farms that fed mixed silage with the lowest average BAB spore concentrations (3.4 log₁₀ spores/g) produced FTM with the lowest average concentration (2.1 log₁₀ spores/L). The efficiency of farm management in controlling the BAB spore concentration in FTM depended to a large extent on the ability of farmers to prevent incidents with elevated BAB spore concentrations in mixed silage (>5 log₁₀ spores/g) and not on the average BAB spore concentration in mixed silage across the year. The survey showed that farmers should aim for a concentration in mixed silage of less than 3 log₁₀ spores/g and should prevent the concentration from exceeding 5 log₁₀ spores/g to ensure a concentration in FTM of less than 3 log₁₀ spores/L. These results correspond with the previously reported model simulations.     

Prevalence, antimicrobial resistance, and molecular characterization of methicillin-resistant Staphylococcus aureus from bulk tank milk of dairy herds

It was the objective of the study to estimate the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in bulk tank milk from German dairy herds and to characterize isolates from bulk tank milk with respect to their Staph. aureus protein A (spa) and staphylococcal cassette chromosome mec (SCCmec) type, their phenotypic antimicrobial resistance and resistance- resp. virulence-associated genes using broth microdilution and a microarray for Staph. aureus. Bulk tank milk samples (25 mL) were tested for MRSA using a 2-step selective enrichment protocol. Presumptive MRSA were confirmed by PCR. Thirty-six isolates collected from bulk tank milk of dairy herds in 2009 and 2010 were included in the characterization. All isolates displayed spa-types assigned to the clonal complex CC398. Based on the epidemiological cut-off values for the interpretation of minimum inhibitory concentrations isolates were resistant to tetracycline (100%), clindamycin (58%), erythromycin (52%), quinupristin/dalfopristin (36%), and kanamycin (27%). Isolates did not carry genes associated with typical virulence factors for Staph. aureus such as the Panton-Valentine leukocidin. However, they did carry hemolysin genes. Livestock-associated MRSA of CC398 does occur in German dairy herds and the strains have similar properties as described for strains from pigs.     

A study on the prevalence of gram-negative bacteria in bulk tank milk

Bulk tank milk from 131 dairy herds in eastern South Dakota and western Minnesota were examined for coliforms and noncoliform bacteria. Coliforms were detected in 62.3% of bulk tank milk samples. Counts ranged from 0 to 4.7 log10 cfu/ml. The mean count was 3.4 log10 cfu/ml. Gram-negative noncoliform bacteria were observed in 76.3% of bulk tank milk. Counts ranged from 0 to 6.2 log10 cfu/ml. The mean count was 4.8 log10 cfu/ml. A total of 234 isolates from bulk tank milk were examined to species level; 205 isolates belonged to 28 species. Coliforms and gram-negative noncoliform bacteria accounted for 32.9 and 67.1% of the total isolates, respectively. Organisms such as Agrobacterium radiobacter, Bordetella spp., Comamonas testosteroni, Listonella damsela, Ochrobactrum anthropi, and Oligella urethralis were isolated from bulk tank milk in this study. These organisms have not been reported previously in bulk tank milk. A total of 116 isolates of Pseudomonas spp. were isolated from raw milk; 98 isolates belonged to nine Pseudomonas spp., and the remaining 18 isolates could not be identified to their species level. Pseudomonas was the most predominant genus. Pseudomonas fluorescens was the most predominant species isolated from bulk tank milk and accounted for 29.9% of all isolates examined. The results of the study suggest that counts of coliforms and noncoliform bacteria in bulk tank milk vary considerably. The isolates represent a wide variety of Gram-negative bacterial species. Examination of bulk tank milk for coliforms and noncoliform bacteria could provide an indication of current and potential problems associated with bacterial counts and milk quality.     

A survey of foodborne pathogens in bulk tank milk and raw milk consumption among farm families in pennsylvania

A 2-part study was conducted to determine the risk of exposure to human pathogens from raw milk. The first part of the study focused on determining raw milk consumption habits of dairy producers. A total of 248 dairy producers from 16 counties in Pennsylvania were surveyed. Overall, 105 (42.3%) of the 248 dairy producers consumed raw milk and 170 (68.5%) of the 248 dairy producers were aware of foodborne pathogens in raw milk. Dairy producers who were not aware of foodborne pathogens in raw milk were 2-fold more likely to consume raw milk compared with dairy producers who were aware of foodborne pathogens. The majority of dairy producers who consumed raw milk indicated that taste (72%) and convenience (60%) were the primary factors for consuming raw milk. Dairy producers who resided on the dairy farm were nearly 3-fold more likely to consume raw milk compared with those who lived elsewhere. In the second part of the study, bulk tank milk from the 248 participating dairy herds was examined for foodborne pathogens. Campylobacter jejuni (2%), Shiga toxin-producing Escherichia coli (2.4%), Listeria monocytogenes (2.8%), Salmonella (6%), and Yersinia enterocolitica (1.2%) were detected in the milk samples. Salmonella isolates were identified as S. enterica serotype Typhimurium (n = 10) and S. enterica serotype Newport (n = 5). Of the 248 bulk tank milk samples, 32 (13%) contained ≥1 species of bacterial pathogens. The findings of the study could assist in developing farm community-based educational programs on the risks of consuming raw milk.     

Short communication: Genetic characterization of antimicrobial resistance in Acinetobacter isolates recovered from bulk tank milk

A total of 176 Acinetobacter isolates, including 57 Acinetobacter baumannii originally obtained from 2,287 bulk tank milk (BTM) samples in Korea was investigated for the genetic basis of antimicrobial resistance using molecular methods. In addition, the occurrence and cassette content of integrons were examined and the genetic diversity of A. baumannii strains identified was evaluated. Aminoglycoside-modifying enzyme genes were detected in 15 (88.2%) of the 17 aminoglycoside-resistant Acinetobacter isolates tested. The most common aminoglycoside-modifying enzyme gene identified was adenylyltransferase gene aadB (n = 9), followed by phosphotransferase genes aphA6 (n = 7) and aphA1 (n = 5). Of the 31 isolates resistant to tetracycline, tet(39) was detected in 20 of them. The genetic basis of resistance to sulfonamide was identified in 15 (53.6%) of 28 trimethoprim-sulfamethoxazole-resistant isolates and 9 (32.1%) of them carried both sul1 and sul2 genes. A bla_ADC-7-like gene was detected in 1 β-lactam-resistant A. baumannii. Furthermore, class 1 integron was identified in 11 Acinetobacter isolates. Two gene cassettes dfrA15, conferring resistance to trimethoprim, and aadA2, conferring resistance to aminoglycosides, were identified in 8 Acinetobacter isolates. None of the isolates was positive for class 2 or class 3 integrons. Pulsed-field gel electrophoresis revealed that most of the A. baumannii strains from BTM samples were genetically diverse, indicating that the occurrence of A. baumannii strains in BTM was not the result of dissemination of a single clone. Elucidation of resistance mechanisms associated with the resistance phenotype and a better understanding of resistance genes may help in the development of strategies to control infections, such as mastitis, and to prevent further dissemination of antibiotic resistance genes. To the best of our knowledge, this is the first report of molecular characterization of antimicrobial-resistant Acinetobacter spp. from milk.     

Predominant microflora of downgraded Danish bulk tank milk

The microflora of downgraded Danish bulk tank milk was examined to identify the main causes of increased microbial counts. Seventy-five representative samples with a microbial count exceeding 3.0 x 10(4) cfu/mL were selected for a more detailed microbial examination. A total of 1237 isolates from these samples were identified. Gram-negative, oxidase-positive bacteria were found in 72% of the samples. Coliforms were found in 20% of the samples, and non-coliforms were found in 49% of the samples. Coryneforms, other gram-positive rods, Lactococcus spp., Micrococcus spp., and coagulase-negative Staphylococcus spp. were found in 28 to 53% of the samples. Bacillus spp., Enterococcus spp., Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus uberis, and yeasts were found in <25% of the samples. Additionally, the isolates were divided into 3 groups, based on the main cause of an elevated microbial count. Microorganisms primarily associated with poor hygiene dominated the microflora in 64% of the samples; bacteria also related to poor hygiene, but in addition associated with growth at low temperatures (psychrotrophic bacteria) dominated the microflora in 28% of the samples; and bacteria mainly associated with mastitis dominated the microflora in 8% of the samples. A bulk tank milk storage period of 48 h instead of 24 h did not affect the proportion of downgraded milk samples and could not be associated with a specific group of microorganisms. Further, no relationship was found between somatic cell counts and the presence of mastitis bacteria.     

Somatic cell counts,chemical composition and coagulation properties of goat and sheep bulk tank milk

This study focused on the need for bulk milk tank somatic cell count (BMTSCC) thresholds and cut-off levels indicating a decrease in milk quality that consequently influences product quantity and quality. First, 226 ewes and 231 goat bulk tank milk samples were collected from different Israeli herds and coagulation properties were determined. Second, soft cheese was produced. No correlation of coagulation properties was found with BMTSCC for sheep milk up to 3264 × 10³ and goat milk up to 6452 × 10³ cells mL⁻¹. Coagulation properties of goat milk with cell count higher than the latter resulted in a significant decrease in curd firmness. For breeds and management system in Israel, 2500 × 10³ cells mL⁻¹ is suggested as the cut-off level for sheep and 3500 × 10³ cell mL⁻¹ for goats. The cell count cut-off level and milk price according to BMTSCC should be tested and then determined for every breed and management and final dairy product.     

Prevalence of foodborne pathogens in bulk tank milk.

Bulk tank milk from 131 dairy herds in eastern South Dakota and western Minnesota was examined for the presence of for foodborne pathogens. Campylobacter jejuni, shiga-toxin producing Escherichia coli, Listeria monocytogenes, Salmonella spp., and Yersinia enterocolitica were detected in 9.2, 3.8, 4.6, 6.1, and 6.1% of bulk tank milk samples, respectively. Thirty-five of 131 (26.7%) bulk tank milk samples contained one or more species of pathogenic bacteria. Isolates of Salmonella belonged to group D (n = 4), B (n = 2), C (n = 1), and E (n = 1) "O" serogroups. All six isolates of Listeria monocytogenes were identified as O antigen type 1. Four of five isolates of E. coli encoded for the shiga-toxin 2 gene, while one strain encoded for the shiga-toxin 1 gene. Escherichia coli O157:H7 was not isolated from bulk tank milk samples. Based on autoagglutination testing, it was inferred that all eight isolates of Yersinia enterocolitica were likely to be virulent. Non A-grade (manufacturing grade) raw milk producers were at a higher risk (odd's ratio, 4.98; confidence interval, 1.96 to 12.22) of having one or more pathogens in their bulk tank milk than were Grade A producers. It was observed that 21 of 79 (26.6%) dairy producers who consumed raw milk had one or more pathogenic bacteria in their bulk tank milk. The findings of the study warrant the need for educational programs for dairy producers about the risks associated with consumption of raw milk.     

The effect of storage conditions on the composition and functional properties of blended bulk tank milk

The objective of this study was to investigate the effects of storage temperature and duration on the composition and functional properties of bulk tank milk when fresh milk was added to the bulk tank twice daily. The bulk tank milk temperature was set at each of 3 temperatures (2, 4, and 6°C) in each of 3 tanks on 2 occasions during two 6-wk periods. Period 1 was undertaken in August and September when all cows were in mid lactation, and period 2 was undertaken in October and November when all cows were in late lactation. Bulk tank milk stored at the 3 temperatures was sampled at 24-h intervals during storage periods of 0 to 96 h. Compositional parameters were measured for all bulk tank milk samples, including gross composition and quantification of nitrogen compounds, casein fractions, free amino acids, and Ca and P contents. The somatic cell count, heat stability, titratable acidity, and rennetability of bulk tank milk samples were also assessed. Almost all parameters differed between mid and late lactation; however, the interaction between lactation, storage temperature, and storage duration was significant for only 3 parameters: protein content and concentrations of free cysteic acid and free glutamic acid. The interaction between storage temperature and storage time was not significant for any parameter measured, and temperature had no effect on any parameter except lysine: lysine content was higher at 6°C than at 2°C. During 96 h of storage, the concentrations of some free amino acids (glutamic acid, lysine, and arginine) increased, which may indicate proteolytic activity during storage. Between 0 and 96 h, minimal deterioration was observed in functional properties (rennet coagulation time, curd firmness, and heat stability), which was most likely due to the dissociation of β-casein from the casein micelle, which can be reversed upon pasteurization. Thus, this study suggests that blended milk can be stored for up to 96 h at temperatures between 2°C and 6°C with little effect on its composition or functional properties.     

羊乳与牛乳理化特性比较

以莎能奶山羊羊乳为样品,利用乳样自动分析仪,氨基酸自动分析仪、等离子发射光谱、色谱分析对其基本成分、氨基酸组成及含量进行了分析和检测,并与牛乳理化特性进行比较,为科学合理地利用羊乳,生产具有独特功能的羊乳制品提供了理论依据。     

Guidelines for monitoring bulk tank milk somatic cell and bacterial counts

This study was conducted to establish guidelines for monitoring bulk tank milk somatic cell count and bacterial counts, and to understand the relationship between different bacterial groups that occur in bulk tank milk. One hundred twenty-six dairy farms in 14 counties of Pennsylvania participated, each providing one bulk tank milk sample every 15 d for 2 mo. The 4 bulk tank milk samples from each farm were examined for bulk tank somatic cell count and bacterial counts including standard plate count, preliminary incubation count, laboratory pasteurization count, coagulase-negative staphylococcal count, environmental streptococcal count, coliform count, and gram-negative noncoliform count. The milk samples were also examined for presence of Staphylococcus aureus, Streptococcus agalactiae, and Mycoplasma. The bacterial counts of 4 bulk tank milk samples examined over an 8-wk period were averaged and expressed as mean bacterial count per milliliter. The study revealed that an increase in the frequency of isolation of Staphylococcus aureus and Streptococcus agalactiae was significantly associated with an increased bulk tank somatic cell count. Paired correlation analysis showed that there was low correlation between different bacterial counts. Bulk tank milk with low (<5000 cfu/mL) standard plate count also had a significantly low level of mean bulk tank somatic cell count (<200,000 cells/mL), preliminary incubation count (<10,000 cfu/mL), laboratory pasteurization count (<100 cfu/mL), coagulase-negative staphylococci and environmental streptococcal counts (<500 cfu/mL), and noncoliform count (<200 cfu/mL). Coliform count was less likely to be associated with somatic cell or other bacterial counts. Herd size and farm management practices had considerable influence on somatic cell and bacterial counts in bulk tank milk. Dairy herds that used automatic milking detachers, sand as bedding material, dip cups for teat dipping instead of spraying, and practiced pre-and postdipping had significantly lower bulk tank somatic cell and/or bacterial counts. In conclusion, categorized bulk tank somatic cell and bacterial counts could serve as indicators and facilitate monitoring of herd udder health and milk quality.     

Minimizing the level of Bacillus cereus spores in farm tank milk

In a year-long survey on 24 Dutch farms, Bacillus cereus spore concentrations were measured in farm tank milk (FTM), feces, bedding material, mixed grass and corn silage, and soil from the pasture. The aim of this study was to determine, in practice, factors affecting the concentration of B. cereus spores in FTM throughout the year. In addition, the results of the survey were used in combination with a previously published modeling study to determine requirements for a strategy to control B. cereus spore concentrations in FTM below the MSL of 3 log₁₀ spores/L. The B. cereus spore concentration in FTM was 1.2 ± 0.05 log₁₀ spores/L and in none of samples was the concentration above the MSL. The spore concentration in soil (4.9 ± 0.04 log₁₀ spores/g) was more than 100-fold higher than the concentration in feces (2.2 ± 0.05 log₁₀ spores/g), bedding material (2.8 ± 0.07 log₁₀ spores/g), and mixed silage (2.4 ± 0.07 log₁₀ spores/g). The spore concentration in FTM increased between July and September compared with the rest of the year (0.5 ± 0.02 log₁₀ spores/L difference). In this period, comparable increases of the concentrations in feces (0.4 ± 0.03 log₁₀ spores/g), bedding material (0.5 ± 0.05 log₁₀ spores/g), and mixed silage (0.4 ± 0.05 log₁₀ spores/g) were found. The increased B. cereus spore concentration in FTM was not related to the grazing of cows. Significant correlations were found between the spore concentrations in FTM and feces (r = 0.51) and in feces and mixed silage (r = 0.43) when the cows grazed. The increased concentrations during summer could be explained by an increased growth of B. cereus due to the higher temperatures. We concluded that year-round B. cereus spores were predominantly transmitted from feeds, via feces, to FTM. Farmers should take measures that minimize the transmission of spores via this route by ensuring low initial contamination levels in the feeds (<3 log₁₀ spores/g) and by preventing growth of B. cereus in the farm environment. In addition, because of the extremely high B. cereus spore concentrations in soil, the contamination of teats with soil needs to be prevented.     

Microbiological and physicochemical stability of raw,pasteurised or pulsed electric field-treated milk

Pulsed electric field (PEF) processing was investigated as an alternative dairy preservation technology that would not compromise quality yet maintain safety. PEF processing of raw whole milk (4% fat) was conducted at two processing conditions (30 kV/cm, 22 μs, at either 53 or 63 °C outlet temperature) and compared with two thermal treatments (15 s, at either 63 or 72 °C) and a raw milk control and replicated twice. Milk bottles (2 L) from each treatment were incubated for two weeks, at 4 and 8 °C, and assessed for total plate count, pH, colour, rennetability, plasmin activity and lipid oxidation. The microbial quality of the thermal (72 °C/15 s) and PEF (63 °C) were similar. A drop in pH occurred after a change in counts was observed. Rennetability was not different between the treatments. Short chain acids dominated the volatile profile of the milk samples. Concentration of volatiles derived from microbial activity, namely 2,3-butanedione, acetic acid and other milk lipid derived short chain free fatty acids (e.g. butanoic and hexanoic acids), followed the trend of microbial activity in milk samples.Industrial relevanceResearch on the application of PEF to control spoilage and pathogenic microorganisms and enzyme systems in milk spans a wide array of processing equipment and reaction conditions. While industrial scale PEF processing of liquid milk for preservation and improved quality seems generally possible, substantiation of lower thermal damage under safe and scalable PEF conditions is required to enable economic feasibility.     

Incidence and seasonal variation of Listeria species in bulk tank goat's milk

Four hundred and fifty raw goat's milk samples obtained from the bulk tanks of 39 goat farms were analyzed for Listeria spp. over a 1-year period. Modified versions of the U.S. Department of Agriculture Food Safety and Inspection Service (USDA-FSIS) and Food and Drug Administration (FDA) protocols were used for recovery of Listeria. Overall, 35 (7.8%) samples yielded Listeria spp. with Listeria monocytogenes identified in 17 of the 35 (3.8%) Listeria-positive samples. Listeria innocua was detected in 26 (5.8%) samples. Eight milk samples contained both L. monocytogenes and L. innocua. Milk samples from 18 of the 39 (46.2%) farms were positive for Listeria at least once during this 1-year study. The modified USDA-FSIS method, which used Listeria repair broth rather than University of Vermont (UVM) broth for primary enrichment followed by a 4-h nonselective incubation period, yielded more Listeria-positive samples (77.1%) than the FDA method (51.4%). All L. monocytogenes isolates belonged to serotypes 1 (62.6%) or 4 (37.4%). Moreover, five different Listeria ribotypes were identified from 34 selected L. monocytogenes isolates, 2 of which were deemed to be of clinical importance. Listeria isolation rates were markedly higher during winter (14.3%) and spring (10.4%) as compared to autumn (5.3%) and summer (0.9%) with these trends similar to those previously reported for cow's milk.     

Identification and characterization of elevated microbial counts in bulk tank raw milk

The bacterial composition of bulk tank milk from 13 farms was examined over a 2-wk period to characterize sudden elevations in the total bacterial count referred to as "spikes." Bulk tank milk samples collected at each pick-up were analyzed for standard plate count, Petrifilm aerobic count, somatic cell count, gram-negative organisms, and streptococci. Twenty standard plate count spikes were observed: 12 associated with streptococci, 4 associated with gram-negative organisms, 2 associated with streptococci and gram-negative organisms, and 2 that were not definitively characterized. Spikes ranged from 14,000 to 600,000 cfu/ml. Streptococcus uberis was isolated as the predominant organism from 11 spikes, and Escherichia coli was isolated from 4 spikes. Statistical analysis of total bacterial counts indicated a high correlation (r = 0.94) between standard plate counts and Petrifilm aerobic count. Regression analysis of standard plate counts and Petrifilm aerobic counts yielded the equation log10 (standard plate count) = 0.73 + 0.85log10 (Petrifilm aerobic count), indicating that the correlation, although strong, is not one to one. In a related pilot study, triplicate bulk tank milk samples were collected and analyzed for total bacterial count and presumptive streptococcus, gram-negative, and staphylococcus counts. Two-way ANOVA of these triplicate data indicated a lack of significant variation among the triplicate samples, suggesting that one sample can reliably gauge the microbial status of the entire bulk tank.     

Association of minimum inhibitory concentration cluster patterns with dairy management practices for environmental bacteria isolated from bulk tank milk

Environmental bacteria have emerged over the past few years to become significant causes of mastitis. Bacteria in this group are often reported by practicing veterinarians to be increasingly resistant to intramammary therapy and responsible for elevated bulk tank somatic cell counts. The purpose of this study was to determine the extent of association of the minimum inhibitory concentrations for selected antimicrobial agents with environmental bacteria isolated from bulk tank milk on California dairies and their housing facilities, husbandry practices, and antimicrobic-use strategies. Bulk tank milk samples were collected from 2 dairy cooperatives that had their milk cultured at the Milk Quality Laboratory, University of California Davis, Veterinary Medicine Teaching and Research Center in Tulare, CA. Samples were collected from July 2001 through March 2002 on 88 d; and 404 environmental bacteria isolated from 93 dairies were found. Minimum inhibitory concentrations were determined on 337 of the isolates for 10 antimicrobial agents. Cluster analysis was performed on the minimum inhibitory concentration values for each organism, and 4 antimicrobial clusters with varying degrees of resistance were found.A 69-question survey questionnaire was completed on-farm for 49 of the 73 dairies that had at least 3 environmental bacterial isolates. The questionnaire sought information on housing facilities, milking management, mastitis prevention, antimicrobial usage strategies, and owner/veterinary involvement in disease control and prevention. Multinomial logistic regression analysis found significant associations between the antimicrobial agent-resistance cluster groups and some of the housing and bedding practices, failure to dry udders before milking, and antimicrobial treatment of nonmastitis conditions. No association was noted for antimicrobial agent treatment of mastitis and the resistance cluster patterns.