The influence of plating technique and incubation temperature on bacterial count from fish and fishery products

For bacterial sampling of raw unprocessed fish and frozen fishery products, spread plate method is preferable to pour plate method; incubation of plates at 30°C gives a higher count than incubation at 37°C. Analysis of variance of the data shows that sample variation between different types of fishes is highly significant whereas the variations between triplicate plates are not significant at 5 % level.

Incubation temperature for total bacterial count of frozen sea foods

Effect of incubation temperatures of 37°C ambient and 5-10°C on total plate count of commercial frozen prawns, squids, cuttle fish and froglegs were studied. Results indicate that incubation at ambient temperature gives the best results.

Environmental monitoring of bacterial contamination and antibiotic resistance patterns of the fecal coliforms isolated from Cauvery River, a major drinking water source in Karnataka, India

The present study focuses prudent elucidation of microbial pollution and antibiotic sensitivity profiling of the fecal coliforms isolated from River Cauvery, a major drinking water source in Karnataka, India. Water samples were collected from ten hotspots during the year 2011-2012. The physiochemical characteristics and microbial count of water samples collected from most of the hotspots exhibited greater biological oxygen demand and bacterial count especially coliforms in comparison with control samples (p <= 0.01). The antibiotic sensitivity testing was performed using 48 antibiotics against the bacterial isolates by disk-diffusion assay. The current study showed that out of 848 bacterial isolates, 93.51 % (n=793) of the isolates were found to be multidrug-resistant to most of the current generation antibiotics. Among the major isolates, 96.46 % (n=273) of the isolates were found to be multidrug-resistant to 30 antibiotics and they were identified to be Escherichia coli by 16S rDNA gene sequencing. Similarly, 93.85 % (n=107), 94.49 % (n=103), and 90.22 % (n=157) of the isolates exhibited multiple drug resistance to 32, 40, and 37 antibiotics, and they were identified to be Enterobacter cloacae, Pseudomonas trivialis, and Shigella sonnei, respectively. The molecular studies suggested the prevalence of blaTEM genes in all the four isolates and dhfr gene in Escherichia coli and Sh. sonnei. Analogously, most of the other Gram-negative bacteria were found to be multidrug-resistant and the Gram-positive bacteria, Staphylococcus spp. isolated from the water samples were found to be methicillin and vancomycin-resistant Staphylococcus aureus. This is probably the first study elucidating the bacterial pollution and antibiotic sensitivity profiling of fecal coliforms isolated from River Cauvery, Karnataka, India.

Bacterial quality and depuration of the green mussel Perna viridis from natural beds

Green mussel (Perna viridis) and sea water from their natural beds on the coastal areas of Porto Novo were studied between April and August 1996 for their bacterial quality. Water samples from the beds were also analysed for their physico-chemical parameters. The total bacterial count of mussels from natural beds as well as bed waters ranged 10³ organisms per gram of mussel meat suspension and per milliliter of sea water. The faecal coliforms were found to be within the permissible limits. Pathogenic bacteria such as Salmonella spp., Streptococcus spp. and Staphylococcus spp. were absent. The variations in pH, temperature, salinity and dissolved oxygen of the seawater samples were insignificant. The mussels were subjected to depuration by different methods among which chlorination was found to be most effective.

Seasonal variations of bacterial flora of fresh oil sardines (Sardinella longiceps)

An elaborate study was made on the qualitative and quantitative seasonal variations in the bacterial flora of fresh oil sardines and their biochemical reactions. It was observed that the total bacterial loads and their phosphorescent and biochemical characters were influenced by changes in seasons. During monsoon season total bacterial count was high. Mesophiles predominated during summer, but phosphorescent bacteria were less. Winter favoured the selection of biochemically less active groups of bacteria.

SEM analysis of the in situ early bacterial colonization on two novel feldspathic ceramics submitted to different types of glazing

Aim: The aim of this study was to evaluate in situ, the early bacterial colonization on feldspar-ceramics submitted to different glazing. Methods and Materials: Fourteen standardized disc specimens (diameter: 5 mm, thickness: 1.5 mm) of each of two micro-particulate feldspathic ceramics (VM7 and VM13, Vita) were produced according to manufacturers' specifications for a total of 28 specimens (24 for the analysis of biofilm and 4 for topographic analysis analyzing the ceramic surfaces). Specimens from each type of ceramic were submitted to two different glazing methods composing four groups: VM7 glazed using glazing liquid Vita Akzent® 25 (G1) and glaze firing (G2), VM13 glazed using glazing liquid (G3) and glaze firing (G4). Six individuals (n=6) wore oral appliances with four ceramic specimens, fixed on the buccal face of the appliances. After 8 hours, each sample was evaluated for the presence (1) or absence (0) of bacterial colonization under a scanning electron microscope (SEM) on five randomly selected fields. The value for each sample was cumulative of the results observed in the fields. One sample from each group was evaluated under a SEM to verify the topographic pattern. Results: There was no difference with regard to bacterial colonization between the feldspar-ceramics and between the glazing types (Kruskal-Wallis non-parametric test). Conclusion: Feldspar-ceramics submitted to firing or glaze firing with Vita Akzent® 25 present a similar condition for in situ bacterial colonization. The similar topographic pattern of the ceramic surfaces seems to have influenced the bacterial colonization.

Total bacterial counting and the response to treatment of bacterial vaginosis with metronidazole

Objective: To evaluate if the total bacterial count of vaginal samples with bacterial vaginosis assessed by flow cytometry influences the response to treatment with metronidazol. Methods: In this cross-sectional study, 273 low-risk reproductive aged women were enrolled. Vaginal samples were taken to evaluate the pattern of vaginal flora according to Nugent’s criteria, as well as the presence of trichomoniasis and candidosis. Cases identified of bacterial vaginosis were treated with metronidazole and controlled after 45 days. Cervical infection by Chlamydia trachomatis and Neisseria gonorrhoeae were also assessed. Flow cytometry for total bacterial counting was performed in propidium iodide stained cervicovaginal samples, using fluorescent beads at a known concentration. Non-parametric Mann-Whitney test was used to compare total bacterial count between groups of interest, at p<0.05. Results: From the total of 273 women enrolled, 50 were excluded as they presented at least one of the infections investigated. Bacterial vaginosis was detected in 79 women (35.4%), of which 33 (41.8%) returned for re-evaluation after treatment, being 21 cases successfully treated and 12 with persistent abnormal vaginal flora. Flow cytometric data showed that total bacterial counting does not differ between normal flora and bacterial vaginosis samples (p=0.14). Also, no difference was found between the cases of treated and persistent bacterial vaginosis (p=0.48). Conclusion: Total bacterial counting does not influence the response to metronidazole treatment of bacterial vaginosis

Quantitative assessment of total and Gram-positive aerobic bacteria in fresh and ambient-temperature-stored sub-tropical marine fish

The current study was undertaken to enumerate Gram-positive bacteria in fresh sub-tropical marine fish and determine the effect of ambient storage (25°C) on the Gram-positive bacterial count. Total and Gram-positive bacteria were enumerated in the muscles, gills and gut of fresh and stored Pseudocaranx dentex, Pagrus auratus and Mugil cephalus on tryptone soya agar (TSA) and TSA with 0.25% phenylethyl alcohol (PEA), respectively. Initial studies indicated that PEA significantly reduced total aerobic bacterial count (TABC) whereas control Gram-positive bacteria were not affected by 0.25% PEA. TABC significantly increased in all fish body parts, whereas Gram-positive aerobic bacterial count (GABC) significantly increased only in the muscles and gills during ambient storage for 15 h. The TABC of the fish species increased from 4.00, 6.13 and 4.58 log cfu g-1, respectively in the muscles, gills, and gut to 6.31, 7.31 and 7.23 log cfu g-1 by the end of storage. GABC increased from 2.00, 3.52 and 2.20 log cfu g-1 to 4.70, 5.85 and 3.36 log cfu g-1. Within each species, TABC were significantly higher in the gills compared to that of muscles and gut; however, no significant differences were found in GABC between muscles and gills. This study demonstrated the potential importance of Gram-positive bacteria in sub-tropical marine fish and their spoilage.

Äidinmaito - terveysjuomaa ja normaalibakteereita

The chemical composition of breast milk has been studied in detail in the past decades. Hundreds of new antibacterial and antiviral components have been found. Several molecules have been found to promote the proper function of neonatal intestine. However, microbiological studies of breast milk have been, until recently, focused mainly on detecting harmful and pathogenic bacteria and viruses. Natural microbial diversity of human milk has not been widely studied before the work reported in this thesis. This is mainly because breast milk has traditionally been thought to be sterile - even if a certain amount of commensal bacteria have usually been detected in milk samples. The first part of this licentiate thesis contains a short literature review about the anatomy and physiology of breast feeding, human milk chemical and microbiological composition, mastitis, intestinal flora and bacteriocins. The second part reports on the experiments of the licentiate work, concentrating on the microbial diversity in the milk of healthy breast-feeding mothers, and the ability of these bacteria to produce antibacterial substances against pathogenic bacteria. The results indicate that human milk is a source of commensal bacteria for infant intestine. 509 random isolates from 40 breast milk samples were isolated and identified by 16S rRNA sequencing. Median bacterial count was about 600 colony forming units per milliliter. Over half of the isolates were staphylococci, and almost one third streptococci. The most common species were skin bacteria Staphylococcus epidermidis and oral bacteria Streptococcus salivarius and Streptococcus mitis. Lactic acid bacteria, identified as members of Lactobacillus-, Lactococcus- and Leuconostoc -genera, were found in five milk samples. Enterococci were found in three samples. A novel finding in this study is the capability of these commensal bacteria to inhibit the growth of pathogens. In 90 precent of the milk samples commensal bacteria inhibiting the growth of Staphylococcus aureus were found. In 40 precent of samples the colonies could block the growth completely. One fifth of the isolated Staph. epidermidis strains, half of Str. salivarius strains, and all lactic acid bacteria and enterococci could inhibit or block the growth of Staph. aureus. In further study also Listeria innocua- and Micrococcus luteus active isolates were found in 33 and 11 precent of milk samples (out of 140). Furthermore, two Lactococcus lactis isolates from the breast milk were shown to produce bacteriocin nisin, which is an antimicrobial molecule used as a food preservative. The importance of these human milk commensal bacteria in the development of newborn intestinal flora and immune system, as well as in preventing maternal breast infections, should be further explored.

Sampling of frozen prawns for bacteriological estimation

Experiments were conducted to study the significance of difference between samples taken from the surface and interior of a frozen shrimps block, as well as to determine the size of sample necessary to represent the whole block, with respect to bacterial count determination. The results showed that the surface samples and interior samples did not differ significantly at 5% level of significance and that the minimum quantity representative of the block was 21-26 gms in the case of a block weighing about 1300 gms. The procedure adopted for taking the bacterial count was the normal standard plate count method.

Evaluation of chemical tests for the quality of prawns

The object of this study was to determine the value of physical, bacteriological and chemical tests used to find out and compare the indices of quality of prawns stored at 0°C and at 18°C. pH value, nature of drip, the total bacterial count, presence or absence of tryptophan, trimethylamine content, glycogen, lactic acid, vitamins such as thiamin, riboflavin and niacin were estimated periodically to evaluate the quality of prawns stored at 0°C, whereas in addition to organoleptic changes, pH, bacterial count, nature of growth in peptone water, soluble protein in salt solution and loss of moisture, glycogen, lactic acid, and changes in vitamin B contents were noted periodically for prawns stored at -l8°C. Riboflavin and niacin were not affected appreciably but the retention of thiamin in prawns was very low.

Effect of different media on the determination of the number of micro organisms in frozen prawns (shrimp)

The Indian standards Institution have recently laid down specifications for the grading of frozen shrimp of which the bacterial examination of the product forms a significant part. Work carried out in this regard in our laboratories have shown that various improvements could be made in the composition of the medium to obtain a more true picture of the total bacterial count. Changes in the medium for total bacterial count have, therefore, been suggested and the reasons for the same discussed.

Ice storage characteristics of cultured silver carp

Cultured silver carp (Hypopthalmichthys molitrix 800-1000 g) was stored in ice (fish to ice ratio 1:1) in a plywood box insulated with one inch thick expanded polystyrene and subjected to detailed examination of quality by chemical, microbiological and organoleptic evaluation at regular intervals to assess the storage life in good acceptable form. Alpha-amino nitrogen, non-protein nitrogen and pH values showed no positive correlation as spoilage index. Total volatile base nitrogen was not high at the end of the storage period although the fish became unacceptable during the period. There was steep decrease in total bacterial count during initial stages of storage and then increased steadily on further storage. Organoleptic evaluation of raw and cooked meat revealed that fish was in good acceptable form up to 14 days in ice.

Effects of gamma radiation on nutritional and microbial quality of Pampus chinensis (Euphrasen 1788)

To evaluate the efficiency of gamma radiation in combination with low temperature Chinese pomfret, Pampus chinensis were preserved by the treatment of different doses of gamma radiation (3, 5 and 8 KGy) at freezing temperature (-20°C) during 90 days of storage period. Quality assessments for fish were carried out at an interval of 15 days during the storage period. Quality assessments were done by organoleptic, chemical (Total Volatile Nitrogen, TVN and Trimethylamine, TMA) and microbiological (Total Bacterial Count, TBC and Total Mould Count, TMC) evaluation. From the analysis of all parameters, maximum shelf-life was observed for irradiated (8 KGy) sample. It remained acceptable up to 75 days and that was the highest duration among 4 types of samples.

Estimation of micro-flora associated with different stages of Macrobrachium rosenbergii (de Man)

The bacterial flora occurring in muscle, haemolymph, hepatopancreas and gill of brood, juveniles, water, eggs, larvae and rearing water were estimated by selective plate count technique for Entrobacteriaceae, Streptococaceae and Vibrionaceae members. The total viable bacterial count was estimated by total plate count technique on nutrient agar. The total viable counts of bacteria were lowest in water from 6.10x10² CFU/mL) and highest in egg (6.06x10super(8) CFU/g). In brood the total counts were varying from 1.62x10² CFU/g in muscle to 2.20x10super(5) CFU/g in gills. In juveniles, the total plate counts were varying from 2.8x10super(4) CFU/g in muscles to 3.67x10 super(8) CFU/g in hepatopancreas. Selective plate counts show that Enterobacteriaceae members dominate in egg and gills of brood and hepatopancreas of juveniles. Vibrios were found to be dominant in water and larvae of rearing tank. Haemolymph of brood was sterile and did not contain any bacteria while muscle of juvenile was having very low count of total viable bacteria.