Variation in growth kinetics and phenotype of Aeromonas spp. from clinical, meat processing and fleshfood sources.

Sixty-four strains of motile aeromonads (A. hydrophila, A. sobria and A. caviae), isolated from clinical meat processing and ready-to-eat fleshfood sources, and the A. hydrophila type strain were tested with respect to their growth kinetics at 4 degrees C and 37 degrees C, and the reported indicators of pathogenicity: autoagglutination and haemolysis (tested using a CAMP reaction). Between the species, A. caviae grew the fastest at 37 degrees C and had the highest percentage of strains not able to grow at 4 degrees C (after 200 h incubation). Within the species, food-derived strains of A. hydrophila were better adapted to growth at lower temperatures than those from clinical or meat processing sources. Clinical strains of A. hydrophila autoagglutinated more frequently than those from other sources, but not differences in CAMP reactions were noted. Aeromonas caviae and A. sobria isolates appeared to be homogeneous with respect to growth kinetics at the temperatures tested. A comparison of the growth kinetics of the A. hydrophila type strain and a food-derived A. hydrophila strain clearly reflected the latter's enhanced ability to grow at low temperatures.     

CHARACTERIZATION OF AEROMONAS SPECIES ISOLATED FROM FOOD

Characterization of ten Aeromonas spp. isolates from local foods and A. hydrophila ATCC 7965, revealed that all strains of A. hydrophila, A. sobria and one strain of A. caviae showed strong hemolytic activity. All of them were protease producers except one A. caviae strain. Some A. hydrophila and A. caviae strains fermented lactose, coagulated milk and reduced litmus. The majority of strains were resistant to at least 5 of 14 antibiotics tested. All the strains grew over the range of 5–42C except those of A. caviae which failed to grow at 42C. Frozen storage at -16C greatly decreased the viability of the tested strains. Radiation D₁₀values of strains ranged from 0.109–0.253 kGy in nutrient broth and increased when some strains were irradiated in ground meat at room temperature. Both strains of A. hydrophila ( ATCC andFI ), A. sobria FI and A. caviae FI grew normally in the presence of 1 and 3% salt. Only A. hydrophila FI tolerated 6.5% salt. These four strains were able to grow in the presence of 0.05–0.3% potassium sorbate at pH 7.0, while at pH 5.0 the presence of sorbate prevented their growth .     

Antibiotic resistance pattern of motile aeromonads from farm raised fresh water fish

Motile aeromonads isolated from the intestines of farm-raised freshwater fish such as Catla catla, Labeo rohita and Ctenopharyngodon idella have been characterized to species level. Morphological and physiological grouping revealed 61% Aeromonas hydrophila, 30% Aeromonas caviae, 7% Aeromonas sobria and 2% which remained unidentified. Hemolytic activity was detected mostly in A. hydrophila, while only half of the A. sobria and A. caviae showed this activity. Antibiotic resistance patterns of the strains revealed that they had acquired a relatively higher resistance to oxytetracycline, amoxycillin, ampicillin, novobiocin and polymixin-B, implicating possible use of these antibiotics in the aquaculture systems.     

Some virulence properties and characterization of motile Aeromonas species from milk and white cheese

Three hundred and thirty-eight samples of milk and milk products in Ankara were screened for the presence of motile Aeromonas species. The overall frequency of aeromonads in these samples was 27%. As expected, raw milk samples were more contaminated with aeromonads than were other products. Sixty-five (49.2%) of the 132 bulk raw milk samples were contaminated with aeromonads. In 35 of these samples, populations of aeromonads from 1.5 × 10² to 3.0 × 10³ cfu/mL were counted in ampicillin dextrin agar (ADA). Ten (40%) of the 25 raw milk samples sold in the street were contaminated with aeromonads. In eight of these samples, 1.2 × 10²−3.0 × 10³ cfu/mL were counted. Five (16%) of the 31 pasteurized milk and 12 (8%) of the 150 white cheese samples were contaminated with Aeromonas spp., but no countable aeromonads population was noted in ADA. The incidence of A. hydrophila, A. sobria and A. caviae in all samples was found to be 90.2%, 4.3% and 5.4%, respectively. The majority of the strains identified as A. hydrophila and A. sobria were able to produce haemolysin, protease and DNase, while strains identified as A. caviae were only positive for DNase. All isolated Aeromonas species ( A. hydrophila, A. sobria and A. caviae ) were positive for uptake of Congo red dye. Nevertheless, only strains identified as A. hydrophila and A. sobria showed a high rate of positive results when tested for the production of the Voges–Proskauer reaction and lysine decarboxylase. The results of this work show that motile Aeromonas spp., especially A. hydrophila , are frequently found in these samples. As these products are usually commonly consumed in Ankara, they can pose a risk especially for children, the elderly and immunocompromised individuals.     

THE OCCURRENCE, HEMOLYTIC ACTIVITY AND ANTIBIOTIC SUSCEPTIBILITY OF MOTILE AEROMONAS SPP. ISOLATED FROM MEAT AND MILK SAMPLES IN TURKEY

Research was carried out to detect the incidence of motile Aeromonas species (A. hydrophila, A. caviae, A. sobria) in a variety of raw meat products (minced meat and chicken) and milk samples (raw and pasteurized) commonly consumed in Ankara. Motile Aeromonas species were isolated from 51.8% of samples detected; these were 40 (67.7%) of the 59 samples of minced meat, 20 (86.9%) of the 23 samples of chicken, 75 (47.7%) of the 157 samples of raw milk and 5 (16.0%) of the 31 samples of pasteurized milk. The dominant species were A.hydrophila and A.sobria in the meat samples, while A. hydrophila was the only isolated species in the milk samples. The strains identified as A. sobria or A. hydrophila were the stronger producers of hemolysin, whereas the A. caviae strains were nonhemolytic. The resistance of isolates to eight antibiotics was determined using the Kirby Bauer disc diffusion assay (Ciprofloxacin, Ceftazidime, Ampicillin, Cefixime, Cefoperazone, Erythromycin, Trimethoprim/sulfamethoxazole, Ceftriaxone). All aeromonads isolates (100%) were susceptible to ciprofloxacin but resistant to (100%) ampicillin and erythromycin. We found out that motile Aeromonas species are commonly present in milk and meat samples investigated in Turkey, which may pose a health problem to consumer. In addition, ciprofloxacin, cefoperozone, and ceftriaxone are suitable drugs that can be used in the treatment of Aeromonas-associated infections, particularly in the immunocompromised, elderly people and young children.     

Growth of and toxin production by Aeromonas hydrophila and Aeromonas sobria at low temperatures

The effects of different temperatures on the growth and toxin production of Aeromonas hydrophila and Aeromonas sobria were studied. The results showed that these Aeromonas species are not only able to grow at low temperatures (e.g. at 4 and 10 degrees C) but may also produce cytotoxin, hemolysin and enterotoxin under suitable growth conditions.     

Aeromonas spp. in foods: a significant cause of food poisoning?

From a total of 563 samples of various foodstuffs purchased from retail outlets in the Reading area 287 were found to contain mesophilic Aeromonas spp. The types of sample which were most frequently contaminated were poultry (79.3%) and offal (84.3%). Of three media compared for their efficiency in recovering Aeromonas spp. after enrichment in alkaline peptone water, Difco Aeromonas was the most efficient. Cytotoxin was produced by approximately 50% of the A. hydrophila and A. sobria strains but by none of A. caviae strains. It is concluded that both raw and cooked foods are potential sources for infecting human beings with Aeromonas spp.     

Effect of grass or concentrate feeding systems and rate of growth on triglyceride and phospholipid and their fatty acids in the M. longissimus thoracis of lambs

Thirty-two male Ile-de-France lambs were used in a factorial 2×2 design to analyse the effects of feeding systems (grass outdoor, G, or concentrate and hay indoor: stall, S) and of growth rate (low, L, or high, H) on total lipids, triglyceride (TG) and phospholipid (PL) contents and their fatty acid composition in the longissimus thoracis muscle (L.T.). Contents were lower for TG (10.4 vs. 15.8 mg/100 g fresh tissue, P<0.05) and higher for PL (6.4 vs. 5.8 mg/100 g fresh tissue, P <0.05) in grass-fed lambs compared to stall-fed ones. TG of grass fed lambs displayed lower proportions of palmitic acid (C16:0), monounsaturated fatty acids (MUFAs), linoleic acid (C18:2n-6) and other (n-6) polyunsaturated fatty acids (PUFA) and higher proportions of stearic acid (C18:0), linolenic acid (C18:3n-3), cis 9, 11 trans C18:2 and trans monounsaturated fatty acids. In PL of the same lambs only lower MUFA, C18:2n-6 and (n-6) PUFA and higher C18:3n-3, (n-3) PUFA and cis 9, 11 trans C18:2 were observed. Growth rate had no effect on lipid, TG or PL contents of longissimus thoracis. However C18:0 proportions were higher in TG and lower in PL for low growth rate lambs. Low growth rate lambs had also lower cis 9, 11 trans C18:2 in TG. Thus, irrespective of growth rate, the muscle lipids characteristic of grass fed lambs fulfilled the recommended features of human food components much better than that of stall fed lambs, namely for CLA and C18:3n-3. The lower ratios of (n-6) to (n-3) PUFA displayed in grass fed lambs both in TG and in PL were also useful to discriminate all the grass fed lambs from all the stall fed animals.     

Cow colostrum and early milk enriched with eicosapentaenoic and docosahexaenoic fatty acid

The objective of the study was to explore whether it is possible to alter cow colostrum and early milk fatty acid composition with a low level of fat supplement, high in docosahexaenoic (DHA) and eicosapentaenoic (EPA) fatty acid. Diets included a control diet and a diet supplemented with DHA- and EPA-enriched fat supplement. Addition of fat supplement significantly decreased saturated fatty acids, C14:0 and C16:0 and increased the values of monounsaturated fatty acids, polyunsaturated fatty acids (PUFA), n3 fatty acids, EPA, DHA, C18:1n9cis and C18:1n11trans. The percentage of short-chain fatty acids significantly increased with the progress of lactation, while the percentage of PUFA, n3 and n6 significantly decreased. These results showed that fat supplement, high in DHA and EPA, modified the fatty acid profile of colostrum and milk fat and increased the proportion of beneficial fatty acids for human health.     

毛叶山桐子果实化学成分及脂肪酸动态分析与评价

对不同生长期的重庆1号、重庆2号品种的毛叶山桐子果实的整果、果肉和种子中的基本化学成分含量进行比较,并运用气相色谱法对油脂中脂肪酸组成及相对含量进行分析.结果表明,2个品种果实在不同生长期和不同部位中粗脂肪、蛋白质、总糖、灰分、总酚、总黄酮等成分含量差异显著(P＜0.05);2个品种毛叶山桐子作为油脂原料的最佳采收期分...     

Hemolytic and proteolytic activities of Aeromonas hydrophila and Aeromonas veronii biovar sobria in broth and salmon extract at different temperatures

Expression of hemolytic and proteolytic activities throughout the growth cycle was investigated with two enterotoxic aeromonad strains assigned to the species Aeromonas hydrophila and Aeromonas veronii biovar sobria. Although growth kinetic data were dependent on strain, temperature, and substrate, maximum populations attained were higher than 9 log CFU/ml in aerated tryptone soya broth plus yeast extract (TSBYE) and salmon extract within the range 4 to 28 degrees C. For both strains in TSBYE, variable amounts of hemolytic activity were first detected at any temperature when aeromonad counts were over 9 log CFU/ml. Afterwards, this activity increased up to similar levels (109 to 112 hemolytic units per ml) without a significant increase in populations. Salmon extract supported hemolysin synthesis at 28 but not 4 degrees C. Proteolytic activity of the A. hydrophila strain was only expressed in salmon extract at 28 degrees C, whereas A. veronii biovar sobria did at 28 degrees C in both substrates and at 10 degrees C in TSBYE.     

Experimental evidence for toxin production by Aeromonas hydrophila and Aeromonas sobria in a meat extract at low temperatures

The ability of enterotoxigenic strains of Aeromonas hydrophila and Aeromonas sobria to produce exotoxins (enterotoxin and haemolysin) in a meat extract at low temperatures (5 and 12 degrees C) was investigated. All three strains incubated at 12 degrees C were enterotoxigenic and haemolytic in the meat extract after 5 days. At 5 degrees C, five of the six strains tested were able to produce these exotoxins after 8 days incubation while one strain was neither enterotoxigenic nor haemolytic after 5, 8 and 11 days. The possible involvement of performed toxin(s) in Aeromonas gastroenteritis is also discussed.     

Enumeration and confirmation of Aeromonas hydrophila, Aeromonas caviae, and Aeromonas sobria isolated from raw milk and other milk products in Northern Greece.

A total of 138 raw cow's and 57 raw ewe's milk samples; 80 pasteurized cow's milk samples; 39 Anthotyros cheese, 36 Manouri cheese, and 23 Feta cheese samples; and 15 rice pudding samples were examined for the presence and any countable population of Aeromonas species. Twenty-two (15.9%) of the 138 cow's milk samples analyzed were contaminated with A. hydrophila. In 13 of these samples, populations of 3.0x10(2) to 5.0x10(3) CFU/ml were counted in starch ampicillin agar (SAA). Eighteen cow's milk samples (13.0%) were contaminated with A. caviae, and in eight of these samples, populations of 2.0x10(2) to 3.0x10(3) CFU/ml were counted in SAA. Five cow's milk samples (3.6%) were contaminated with A. sobria, and in two of these samples, populations of 2.5x10(3) and 5.0x10(3) CFU/ml were counted in SAA. Eleven cow's milk samples (7.9%) were contaminated with other Aeromonas spp. not classified. Eight (14.0%) of the 57 ewe's milk samples analyzed were contaminated with A. hydrophila. In these samples, populations of 5.0x10(2) to 5.0x10(3) CFU/ml were counted in SAA. Six ewe's milk samples (10.5%) were contaminated with A. caviae, and populations of 1.5x10(2) to 1.0x10(3) CFU/ ml were counted in SAA. Two ewe's milk samples (3.5%) were contaminated with A. sobria, and populations counted in SAA were 5.0x10(2) and 1.0x10(3) CFU/ml. Four samples (7.0%) were contaminated with other Aeromonas spp. not classified. A. hydrophila was recovered in 4 (10.2%) and 3 (8.3%) of the Anthotyros and Manouri cheese samples analyzed, respectively, but no countable populations were noted in SAA. None of the pasteurized milk, Feta cheese, and rice pudding samples yielded Aeromonas spp. The results of this work indicate that motile Aeromonas are common in raw milk in Greece. Also, the presence of A. hydrophila in the whey cheeses Anthotyros and Manouri indicates that postprocessing contaminations of these products with motile Aeromonas may occur during production.     

Influence of food system conditions on N-acyl-L-homoserine lactones production by Aeromonas spp

Eleven of 13 Aeromonas strains were shown to produce AHLs. Results of TLC showed that N-butanoyl-L-homoserine lactone (C4-HSL) was the main AHL produced in LB medium at 30 degrees C. The influence of different carbon sources, temperature, pH values and salt concentrations on AHL production was determined in eight A. hydrophila and one A. caviae strain. Additionally a quantitative study of C4-HSL production by A. hydrophila strain 519 under different conditions was performed. Positive results were found in the AHL induction assay for some Aeromonas strains in cultures in LB agar incubated at 12 degrees C after 72-96 h. The induction of the sensor strains by Aeromonas spp. occurred in LB medium supplemented with all carbon sources in a concentration of 0.5%. The production of C4-HSL by A. hydrophila 519 was found until 3.5% (w/v) of NaCl. For pHs close to the neutrality the C4-HSL production by A. hydrophila was evident after 24-48 h of incubation. A. hydrophila 519 produced C4-HSL under anaerobic conditions. Also, the AHL production by Aeromonas strains was studied in simulate agar of shrimp, fish and some vegetables. The production of AHLs was evident by almost all the test strains in shrimp simulated agar. In fish agar only for one of three fish species tested, positive results were found. Induction assay in vegetables simulated agar showed principally negative results, probably because of the presence of inhibitory compounds in these vegetables.     

Modification of the bile salts-Irgasan-brilliant green agar for enumeration of Aeromonas species from food

The present study evaluated the productivity of BIBG medium for the isolation of Aeromonas spp. from food and describes a modification of the BIBG medium (mBIBG) (increased pH (8.7), replacement of xylose by soluble starch as a carbon source, decreased concentration of bile salts) to increase its selectivity and electivity. Using the mBIBG medium, growth of the majority of the Enterobacteriaceae (9/10) was suppressed except for Citrobacter freundii. The mBIBG medium supported growth of Pseudomonas species but a clear distinction between Aeromonas and Pseudomonas colonies could be made. Interpretation of the mBIBG medium should be performed after 24 h of incubation. It was noted that three of the 27 Aeromonas strains tested did not develop on the mBIBG medium. The ability or inability to grow on a selective medium is strain-dependent. Enumeration of Aeromonas species (A. hydrophila LMG 3771, A. caviae LMG 3775, A. veronii biovar veronii LMG 9075, A. veronii biovar sobria LMG 13071) from artificially contaminated foods (shrimp, minced meat (beef/pork), precut leek, and shredded carrots) confirmed that the mBIBG medium is suitable for quantitative recovery of aeromonads (ca. 10(2)-10(7) cfu/g) in the presence of a high background flora (10(5)-10(6) cfu/g). Screening of naturally contaminated foods (vegetables, seafood, meat) for the presence of Aeromonas resulted in three out of 14 food samples showing presumptive Aeromonas colonies on mBIBG.     

PREVALENCE AND RESISTANCE TO ANTIBIOTICS FOR AEROMONAS SPECIES ISOLATED FROM RETAIL FISH IN TURKEY

Mesophilic aeromonads are among the most common bacteria in water habitats throughout the world, and these bacteria frequently cause disease in fishes. They are also causative agents of acute diarrheal disease in man following ingestion water. In this study, a total of 132 market fish (64 freshwater and 68 seawater) samples representing were collected in Ankara (Turkey) and investigated for the presence of Aeromonas spp. They were isolated from 106 (80.3%) out of the 132 fish samples tested. The distribution of Aeromonas spp. varied depending on the samples (gill, intestine, liver, kidney) examined. In freshwater samples, the predominant species was A. caviae (66.0%), followed by A. hydrophila (22.6%) and A. veroni bv. sobria (11.6%). In seawater samples, the predominant species was found A. veroni bv. sobria (41.5%), followed by A. hydrophila (30.1%) and A. caviae (28.3%). The 132 isolated Aeromonas spp. strains were further examined for hemolytic, lipolytic and proteolytic activity. Of the present isolates, more than 80% (A. veroni bv. sobria, A. hydrophila) were positive for hemolysin activities. Lipolytic and proteolytic activity of identified strains were found in more lower incidence. All aeromonads (A. hydrophila, A. veroni bv. sobria, A. caviae) strains showed resistance to ampicillin, cephalothin and trimethoprim. The least resistance was found for chloramphenicol (9.0%). In contrast, all the strains were susceptible to ciprofloxacin and ceftriaxone. In Turkey, there have been few studies on Aeromonas and its virulence factors. This study therefore highlights an important incidence of Aeromonas spp., with virulence and antibiotic resistance, isolated from fish intended for human consumption in Turkey.     

Mesophilic aeromonads in wild and aquacultured freshwater fish

Numbers and species of motile Aeromonas were determined in freshly caught freshwater fish, in the surrounding environment, and also during iced chilled storage of fish specimens. Although no significant differences were observed in water samples, initial levels for skin, gill, and intestines were significantly lower in farmed rainbow trout (Oncorhynchus mykiss) than in wild brown trout (Salmo trutta) and pike (Esox lucius). During storage of wild specimens, naturally occurring aeromonads grew fairly well on the surfaces of skin and body cavity. Of 171 strains assigned to the genus Aeromonas, 88% were identified to phenospecies and putative genospecies level by using comprehensive biochemical schemes. The isolates were allocated to putative hybridization groups (HGs) 1 and 3 Aeromonas hydrophila (29%); putative HG 8 Aeromonas veronii biovar sobria (19%); putative HG 2 Aeromonas bestiarum (18%); putative HG 9 Aeromonas jandaei (16%); putative HGs 4 and 5a Aeromonas caviae (2%); putative HG 12 Aeromonas schubertii (2%); and putative HG 11 (unnamed, 0.6%). The remaining 20 isolates (12%) resembled A. schubertii but could not be allocated to currently recognized phenospecies or to putative HGs. Although cultured rainbow trout yielded strains of putative HGs 1, 4, and 8, which appear to be of major clinical importance, most isolates assigned to putative HGs 1 and 8 were recovered from pike. Differences among HGs found in wild animals could be related to their origin (unpolluted rivers for brown trout and urban rivers for pike). The recovery of these aeromonads species was not related to sampling site. The initial levels of motile aeromonads, their behavior during storage, and the strong potential spoilage activity of most isolates confirm that these bacteria can contribute to deterioration of iced wild freshwater fish. Although adequate cooking would inactivate motile aeromonads, the high incidence of isolates belonging to gastroenteritis-associated HGs should be regarded as a potential health concern, particularly for susceptible populations when there is a possibility of cross-contamination.     

Prevalence and resistance to antibiotics for Aeromonas species from retail fish in Malaysia

A total of 87 market fish samples representing five types of fish were evaluated for the presence of Aeromonas spp. Of the samples examined, 69%, 55%, 11.5% and 2.3% harbored Aeromonas spp., A. veronii biovar sobria, A. hydrophila and A. caviae, respectively. The 60 isolated Aeromonas spp. strains were further examined for hemolytic activity, resistance to antimicrobial agents and presence of plasmids. Hemolytic activity varied widely among the isolated strains. Though all the isolates demonstrated resistance to three or more of the antibiotics tested, all were susceptible to ceptazidime. Thirty-four (56.7%) of the sixty isolates harbored plasmids, with sizes ranging from 2.3 to 15.7 kb. These results indicate that hemolytic, multiple antibiotic resistant and genetically diverse aeromonads are easily recovered from fish in this region.     

Numbers and species of motile aeromonads during the manufacture of naturally contaminated Spanish fermented sausages (longaniza and chorizo).

The prevalence of aeromonads in the initial mixes of 14 batches of chorizo and longaniza obtained from three small- and middle-sized factories was 78.5%, with counts ranging from >1.00 to 4.47 log10 CFU/g. Only 2 of 10 mixture samples prepared at a large and modern processing plant yielded these organisms, with levels below 1 log10 CFU/g. The hygienic status of factories significantly affected incidence and counts. Of 39 presumptive isolates from glutamate starch penicillin agar, 36 were confirmed as motile aeromonads and allocated to Aeromonas hydrophila (n = 24), A. veronii biovar sobria (n = 10), and A. caviae (n = 2). All of them were beta-hemolytic and capable of growing at 5 degrees C. Regardless of initial contamination, aeromonads were rapidly inactivated during the early stages of manufacture.     

Prediction of the fatty acid composition of beef by near infrared transmittance spectroscopy

The intramuscular fat content and composition influence consumer selection of meat products. A study predicting the fatty acid (FA) profile of ground beef from the Longissimus thoracis of yearling bulls (n=100) using near infrared transmittance spectroscopy (NIT) was conducted. The samples were scanned using an Infratec 1265 Meat Analyzer which operates in transmittance mode from 850 to 1050nm. NIT technology was able to accurately predict (R(CV)(2) over 0.76) some prominent FAs such as C14:0, C16:0, C16:1cis9, C17:0, C18:1cis9 and C18:1cis11, and minor FAs like C13:0, C15:0, C17:1cis9 and C18:1cis13. When studying FA groups, NIT spectra were able to accurately predict saturated (R(CV)(2)=0.837), branched (R(CV)(2)=0.701) and monounsaturated (R(CV)(2)=0.852) FAs. In addition, NIT spectra provided useful information on the contents of conjugated linoleic acids (CLA) in beef. These results show the potential of NIT technique as a rapid and easy tool to predict the major FAs in beef, especially those located in triglycerides.