The survival of bacteria exposed to desiccation on surfaces associated with farm buildings

The survival of 11 species of Gram-negative and Gram-positive bacteria was examined on different surfaces exposed to desiccation. There were large variations between species; Pseudomonas spp. and Rhizobium leguminosarum biovars survived for less than 2 d, whilst Enterococcus spp. survived for more than 11 weeks. The type of surface on to which the bacteria were deposited affected survival, but with different effects between species. In addition the survival of spontaneous nalidixic acid-resistant (Nal-r) mutants of a natural Escherichia coli isolate were compared. Overall the differences were slight, but of seven resistant mutants, five survived better than the parent whilst one survived less well. Nine transposon insertion derivatives of one of the Nal-r mutants (ECO80) which survived better than the parent were compared; all survived similarly to the parent except ECO883 which survived less well. The growth characteristics of ECO883 and ECO80 were compared; at high osmotic pressures (>0.4 mol l^-1 NaCl) ECO883 grew more slowly and showed a longer lag time than the parent. Of the osmoregulatory functions studied, ECO883 appeared to be altered with respect to K⁺ transport or accumulation, although the transposon insertion had occurred in a gene distant from known K⁺ transport genes.     

Occurrence of ceftriaxone-resistant commensal bacteria on a dairy farm and a poultry farm

Approximately 40 samples of animal feces, drinking water, feed, bedding, pine wood shavings, compost, and manure slurry were collected from two animal research farms (one dairy and one poultry) and analyzed for ceftriaxone-resistant bacteria. Our study revealed that the total percentage of aerobic bacteria with reduced susceptibility to ceftriaxone (minimal inhibitory concentration (MIC) > or = 16 micro g/mL) ranged from 0.9% to 10.8% in dairy feces and from 0.05% to 3.93% in chicken feces. The percentages of ceftriaxone-resistant bacteria (MIC > or = 64 micro g/mL) were in the range of 0.01% - 2.3% in dairy feces and 0.01% - 0.79% in chicken feces. Environmental samples contained a wide range of ceftriaxone-resistant bacterial populations. Among those environmental samples, fresh pine wood shavings used as chicken bedding contained the highest percentages (41.5%) of ceftriaxone-resistant bacteria, as determined by a plating method. A total of 105 ceftriaxone-resistant (MIC > or = 128 micro g/mL) bacterial isolates were isolated from the above samples and tested for resistance to nine antibiotics: ampicillin, ceftriaxone, streptomycin, kanamycin, gentamicin, chloramphenicol, tetracycline, ciprofloxacin, and nalidixic acid. The most prevalent resistance pattern (34.3%) among isolates included resistance to all nine antibiotics. Results from this study suggest that ceftriaxone-resistant bacteria exist in farm environments, and the ceftriaxone resistance was frequently associated with resistance to multiple antibiotics. Environmental sources such as pine wood shavings used as bedding can be a potential reservoir for transmitting the multidrug-resistant bacteria.     

Influence of substrate wettability on the attachment of marine bacteria to various surfaces

The effect of the initial substrate surface condition, as indicated by the critical surface tension for wetting, on the rate of attachment of marine bacteria to a variety of solid surfaces has been measured. The techniques used to determine the number of bacteria attached per unit surface area were a lipopolysaccharide test utilizing Limulus lysate and direct examination of the surface by scanning electron microscopy. The results obtained by the two techniques are compared and their significance to the control of microbiological slime film formation (microfouling) is discussed.     

Influence of substratum wettability on attachment of freshwater bacteria to solid surfaces

[This corrects the article on p. 811 in vol. 45.].     

Lactic acid bacteria: hydrophobicity and strength of attachment to meat surfaces

The hydrophobicity and strength of a ttachment of several lactic acid bacteria with antimicrobial activity were studied. Hydrophobicity was determined by bacterial adherence to hydrocarbons (BATH; octane or xylene), adhesion to nitrocellulose filters (NCF), salt aggregation test (SAT) and adherence to phenyl–Sepharose beads (PSB). The relative hydrophobicity of lactic acid bacteria depended markedly on the method used. No correlation between either SAT or BATH (octane) and strength of attachment (Sr value) existed. However, a significant relationship between strength of attachment and BATH (xylene), NCF and PSB, respectively, was observed, showing the highest correlation coefficient ( r = 0·778) for BATH (xylene).     

Effect of milk proteins on adhesion of bacteria to stainless steel surfaces.

Stainless steel coupons were treated with skim milk and subsequently challenged with individual bacterial suspensions of Staphylococcus aureus, Pseudomonas fragi, Escherichia coli, Listeria monocytogenes, and Serratia marcescens. The numbers of attached bacteria were determined by direct epifluorescence microscopy and compared with the attachment levels on clean stainless steel with two different surface finishes. Skim milk was found to reduce adhesion of S. aureus, L. monocytogenes, and S. marcescens. P. fragi and E. coli attached in very small numbers to the clear surfaces, making the effect of any adsorbed protein layer difficult to assess. Individual milk proteins alpha-casein, beta-casein, kappa-casein, and alpha-lactalbumin were also found to reduce the adhesion of S. aureus and L. monocytogenes. The adhesion of bacteria to samples treated with milk dilutions up to 0.001% was investigated. X-ray photoelectron spectroscopy was used to determine the proportion of nitrogen in the adsorbed films. Attached bacterial numbers were inversely related to the relative atomic percentage of nitrogen on the surface. A comparison of two types of stainless steel surface, a 2B and a no. 8 mirror finish, indicated that the difference in these levels of surface roughness did not greatly affect bacterial attachment, and reduction in adhesion to a milk-treated surface was still observed. Cross-linking of adsorbed proteins partially reversed the inhibition of bacterial attachment, indicating that protein chain mobility and steric exclusion may be important in this phenomenon.     

Dietary influence on protein level in milk and milk yield in dairy cows

Diet can influence the yield of milk protein more than it can influence milk protein content. Providing sufficient dietary crude protein in forms that will maximize amounts of amino acids required for milk protein synthesis to the gastrointestinal tract, in forms that can be digested and absorbed, will provide for optimal milk protein yield and content. Maximizing ruminal microbial protein synthesis is an important part of this strategy. The rest of the strategy involves providing sufficient amounts of the remaining required amino acids as ruminally protected proteins, or amino acids in forms that can be digested in the gastrointestinal tract. A diet deficient in protein will reduce milk protein content 1 to 2 g kg⁻¹ and may substantially reduce yields of milk and milk protein. A diet containing high amounts of readily fermentable carbohydrates may increase milk protein content 1 to 2 g kg⁻¹, and may increase yields of milk and protein, but may also result in digestive and metabolic upsets. Diets containing supplemental fat will increase yield of milk protein, but not as extensively as the increase in yield of milk, because milk protein content is usually reduced 1 to 2 g kg⁻¹. The increased efficiency of milk fat and lactose synthesis is likely to be the reason for this depression in milk protein content. A means of overcoming this problem is a continuing research challenge.     

The use of milk progesterone profiles to characterise components of subfertility in milked dairy cows

Milk progesterone (P₄) concentrations of 1682 postpartum (PP) dairy cows during 2503 lactations were used to define and quantify the incidence of atypical ovarian patterns and to assess their impact on reproductive performance. A total of 257 animals (10.94%) with their first significant luteal activity after day 44 PP were considered a result of delayed ovulation type I (DOVI). Prolonged luteal activity (P₄>3 ng/ml for at least 19 days) observed in 170 (7.3%) and 161 (6.35%) animals during first and subsequent cycles was considered a result of the presence of a persistent corpus luteum (CL), respectively denoted as PCLI and PCLII. Following the demise of an oestrous cycle CL, a total of 322 (12.85%) animals showed a delayed ovulation Type II (DOVII) with P₄<3 ng/ml for >12 days. In 238 inseminated animals (9.92%) prolonged luteal activity was followed by the CL demise which may indirectly indicate the incidence of a late embryo to early foetal mortality (LEM). In this study animals during 794 (31.7%) lactations had at least one atypical ovarian pattern before insemination that, in comparison to those with typical P₄ patterns, contributed to a delayed conception (88.2 vs. 106.2 days), higher number of services per conception (1.49 vs. 1.8), lower first service conception rate (60.9 vs. 43.7%) and a reduced total conception rate (92.6 vs. 82.1%), all of which were significantly different at P<0.001. The incidence of PCLI and PCLII before insemination resulted in a higher level of LEM. Milk progesterone monitoring offers an accurate and objective measurement of factors associated with PP ovarian activity which will assist in investigating the genetic and environmental factors affecting fertility.     

Attachment of bacteria to model solid surfaces: oligo(ethylene glycol) surfaces inhibit bacterial attachment

Abstract An NADP(H)-specific glutamate dehydrogenase of Haloferax mediterranei has been purified to apparent homogeneity and characterised. The purified enzyme was stabilized by glycerol in absence of salt. Glutamate dehydrogenase from Hf. mediterranei is a hexameric enzyme with a native molecular mass of 320 kDa composed of monomers each with a molecular mass of 55 kDa. At pH 8.5 the enzyme has K _ms of 0.018, 0.34 and 4.2 mM for NADP+, 2-oxoglutarate and ammonium, respectively. Amino acid composition and sequence of the first 16 residues of the N-terminus have been determined.     

ATP level variations in heterotrophic bacteria during attachment on hydrophilic and hydrophobic surfaces.

A survey of the extracellular ATP levels of 86 heterotrophic bacteria showed that gram-negative bacteria of the genera Sulfitobacter, Staleya, and Marinobacter secreted elevated amounts of extracellular ATP, ranging from 6.0 to 9.8 pM ATP/colony forming unit (cfu), and that gram-positive bacteria of the genera Kocuria and Planococcus secreted up to 4.1 pM ATP/cfu. Variations in the levels of extracellular and intracellular ATP-dependent luminescence were monitored in living cells of Sulfitobacter mediterraneus ATCC 700856T and Planococcus maritimus F 90 during 48 h of attachment on hydrophobic (poly[tert-butyl methacrylate], PtBMA) and hydrophilic (mica) surfaces. The bacteria responded to different polymeric surfaces by producing either intracellular or extracellular ATP. The level of intracellular ATP in S. mediterraneus ATCC 700856T attached to either surface was as high as 50-55 pM ATP/cfu, while in P. maritimus F 90 it was 120 and 250 pM ATP/cfu on PtBMA and mica, respectively. S. mediterraneus ATCC 700856T generated about 20 and 50 pM of extracellular ATP/cfu on PtBMA and mica, respectively, while the amount generated by P. maritimus F 90 was about the same for both surfaces, 6 pM ATP/cfu. The levels of extracellular ATP generated by S. mediterraneus during attachment on PtBMA and mica were two to five times higher than those detected during the initial screening. High-resolution atomic force microscopy imaging revealed a potentially interesting correlation between the porous cell-surface of certain alpha- and gamma-proteobacteria and their ability to secrete high amounts of ATP.     

Effects of niacin on milk yield,milk composition,and blood components of dairy cows in hot weather

Twenty multiparous Friesian cows, 90 ± 30 days postpartum, were allocated to two groups of ten cows according to calving date, lactation number and daily milk yield, and assigned randomly to one of two diets in a cross-over experiment. The experimental diet consisted of concentrate and maize silage in the proportions 1:1 (dry matter basis) containing either 0 or 10 g niacin per cow per day, which was handmixed into the concentrate. The diets were offered individually as total mixed rations in two equal proportions at 09:00 and 20:00 h in amounts sufficient to ensure 10% refusals. Dry matter, metabolisable energy and crude protein intakes, as well as milk yield, milk protein proportion and yield, milk lactose, total solids and solids-not-fat proportions were not affected by niacin supplementation. In contrast, niacin supplementation increased milk fat proportion and yield. No differences were observed in blood serum concentrations of glucose, total protein, triglycerides, cholesterol, Na, K, Ca, P or Mg. However, the serum concentration of urea was lower when the cows were supplemented with niacin.     

The influence of cytochalasin B, colchicine, and vinblastine on the attachment of Entamoeba histolytica to glass surfaces (author's transl)]

The attachment of E. histolytica trophozoites from monoxenic TPS-medium to glass surfaces was studied at different temperatures and pH values and in the presence of cytochalasin B, colchicine, and vinblastine. At pH 7.0 and at 36 degrees C optimal adhesion rates of the amebae were observed in the range from 80% to 95%. Marked inhibition of adhesion was noted at higher concentrations of cytochalasin B. The adhesion rate was reduced to 50% by 50 micrograms/ml CB. Higher dosages successively prevented nearly all adhesion. Colchicine was without any effect. Vinblastine had a certain low effect but only at very high dosage. The observations are consistent with the assumption that the adhesion process is at least partially mediated by cytochalasin-sensitive contractile microfilaments, whereas microtubuli are obviously not involved in the cell-substrate adhesion.     

Lactic acid bacteria fermentation of human milk oligosaccharide components, human milk oligosaccharides and galactooligosaccharides

Human milk contains about 7% lactose and 1% human milk oligosaccharides (HMOs) consisting of lactose with linked fucose, N-acetylglucosamine and sialic acid. In infant formula, galactooligosaccharides (GOSs) are added to replace HMOs. This study investigated the ability of six strains of lactic acid bacteria (LAB), Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus reuteri, Streptococcus thermophilus and Leuconostoc mesenteroides subsp. cremoris, to digest HMO components, defined HMOs, and GOSs. All strains grew on lactose and glucose. N-acetylglucosamine utilization varied between strains and was maximal in L. plantarum; fucose utilization was low or absent in all strains. Both hetero- and homofermentative LAB utilized N-acetylglucosamine via the Embden-Meyerhof pathway. Lactobacillus acidophilus and L. plantarum were the most versatile in hydrolysing pNP analogues and the only strains releasing mono- and disaccharides from defined HMOs. Whole cells of all six LAB hydrolysed oNP-galactoside and pNP-galactoside indicating β-galactosidase activity. High β-galactosidase activity of L. reuteri, L. fermentum, S. thermophilus and L. mesenteroides subsp. cremoris whole cells correlated to lactose and GOS hydrolysis. Hydrolysis of lactose and GOSs by heterologously expressed β-galactosidases confirmed that LAB β-galactosidases are involved in GOS digestion. In summary, the strains of LAB used were not capable of utilizing complex HMOs but metabolized HMO components and GOSs.     

Scanning electron microscopy of dairy equipment surfaces contaminated by two milk-borne micro-organisms

Ethanol dehydration followed by argon replacement induced drying (ARID) was found to be a suitable method for the preparation of glass, stainless steel and rubber surfaces which had been in contact with inoculated milk and which were to be examined using scanning electron microscopy (SEM). This technique was used to examine samples of all three materials which had been subjected to both single and repeated inoculation with whole milk containing a Pseudomonas sp. or a Micrococcus sp. and incubated for various periods. Some samples were also prepared for SEM using a cryofixation technique. The Pseudomonas sp. was found to proliferate on glass and stainless steel surfaces but not on rubber. Due to the clumping tendency of the Micrococcus sp. proliferation of this organism was more difficult to assess accurately. In general there was no difference in results obtained between single and repeated inoculation. Various factors which may have aided attachment of micro-organisms to surfaces were identified viz. , surface channels present in stainless steel, milk deposits and the production of extracellular material. The value of using both the cryofixation and chemical preparatory techniques for the identification of artifacts is discussed.