The Inhibitory Effect of Some Natural Essential Oils upon Prototheca Algae in vitro Growth

The aim of this paper was to present the in vitro evaluation of the inhibitory effect of some natural essential oils against unicellular algae from Prototheca genus. Ten P. zopfii isolates from cows mastitic milk samples and one P. wickerhamii referent strain (RE-4608014ATCC16529), from American Type Collection, were submitted to antifungal susceptibility testing by classical diffusimetric method. The natural products tested were represented by fir (Abies alba), savory (Satureja hortensis), peppermint (Mantha piperita), tea tree (Maleleuca alternifolia), grape seed and oregano (Oreganum compactum) essential oils. Comparative the efficiency of antimicrobial drugs (Itraconasole) was tested. For Prototheca zopfii tested isolates the highest efficiency were registered for: savory, mint and tea tree oils while P. wickerhamii proved to be sesitive for all natural products. Difficulties in treating protothecosis in humans and animals with conventional drugs and the potent in vitro activity of essentials oils demonstrated here raise the interest in further investigations on the therapeutic use of these non-conventional natural products

RESEARCH REGARDING THE DEVELOPMENT OF UNICELLULAR ALGAE FROM PROTOTHECA GENUS ISOLATED FROM MASTITIC COWS

Eighteen strains of Prototheca zopfii, an unicellular alga of Chlorococales order, Chlorellaceae family, Prototheca genus were isolated from subclinical mastitis cows. In our study, the Prototheca was able to develop on liquid and solid glucosed media, blood agar, potato agar, glucosed media with different pH values (from 2 to 9), media with antibiotics (lincomycin and spectinomycin). The Prototheca shows a good development in aerobic conditions at 37oC and the culture appears to be visible after 36 to 48 hours. The liquid media appears to be clear, but at the bottom of the culture tubes a granular sediment easy homogenisable is formed. At the surface a thin coating could appear. On solid media the colonies have small size, irregular ice crystal shape, but after 5 to 7 days at laboratory temperature the colonies grow to 3-4 mm, have matte whitish color, increased height and their surface has a blackberry or cauliflower appearance, aspects which are characteristic for Prototheca. On blood agar the colonies present the same aspect, without hemolysis. It is recommended to examine culture plates under a stereo microscope (x1.6 or x4). Acid and antibiotic based media have selective effect, and will eliminate bacteria and yeast growth

RESEARCHES REGARDING IMMUNOGENITY OF UNICELLULAR ALGAE FROM PROTOTHECA GENUS

In order to obtain hyperimmun serum two rabbit groups were used; each group was constituted of two animals, 6 months old, with an approximate body mass of 2-2.5 kg. From both Prototheca species (zopfii and wickerhamii), an antigen was prepared by ultrasonication, using a DUS P 150 ultrasounds disintegrator. Rabbits were inoculated on the abdomen sides; the blood collection was made from marginal auricular vein 7 days after each inoculation. For quantification, the antibody concentrations against both P. zopfii and P. wickerhamii from hyperimmun serum, slow agglutination reaction in tubes was performed. This reaction was made after each blood collection. P. zopfii is more immunogenic then P. wickerhamii

Researches Concerning the Sensibility to Neomycin of Some Bacterial Strains Isolated from Chicken

In this study we aimed to determine neomycin effect on Gram negative bacterial strains of the species Escherichia coli, Salmonella spp, Proteus spp and Klebsiella pneumoniae isolated from chickens with different diseases. Sensitivity to neomycin was also appreciated for standard reference strains: Bacillus anthracis 1190 R, Staphylococcus aureus 6538P, Bacillus cereus ATCC 14579, Pseudomonas aeruginosa ATCC 27853, E. coli ATCC 10536 and Salmonella enteritidis ATCC 13076. The experiment took place at the department of Microbiology of the Faculty of Veterinary Medicine Cluj-Napoca within October 2010 - March 2011, realizing the sensitivity tests to neomycin using the diffusimetric method. After testing a total of 33 bacterial strains, the resistance to neomycin was observed for 3 species (Escherichia, Proteus and Klebsiella) while for genus Salmonella no resistant strains were registered. Neomycin resistance was of 20% for strains of Escherichia, 25% for Proteus and 33.3% in those from genus Klebsiella. Sensitivity percentage of the tested strains ranged between 25% and 66.6%. Percentage of moderate sensitive strains ranged from 0 to 50%. Of the 6 standard reference strains tested, one was classified as sensitive (Staphylococcus aureus 6538P), three resistant (Pseudomonas aeruginosa ATCC 27853, E. coli ATCC 10536 and Salmonella enteritidis ATCC 13076) and two moderate sensitive (Bacillus anthracis 1190 R and Bacillus cereus ATCC 14579). The results obtained reveals that resistance to neomycin for the strains isolated from chickens varied from one bacterial species to another. Infections involving Escherichia coli, Proteus spp, Pseudomonas aeruginosa and Klebsiella pneumoniae should be considered more carefully, neomycin being recommended for the treatment only when the outbreak strains were tested and classified as sensitive to this antibiotic

THE ANTIMYCOTIC EFFECT OF SOME BEE PRODUCTS AND VEGETAL EXTRACTS ON GENUS MALASSEZIA FUNGI

For both antimicrobial and antimycotic therapy, the researches of the latest years are mentioning an increased resistance of the bacterial and fungal strains to these substances. A natural alternative to classical therapy was aimed, so 14 Malassezia pachydermatis strains isolated from dogs suffering with dermatitis and otitis in Cluj-Napoca were tested regarding the capacity of some bee products to inhibit their “in vitro” development. From the products tested, the best results were registered for propolis tincture, polioel 3 and royal jelly. The media of the diameter inhibit area in mm, was of 22,21 mm for propolis tincture, 14,92 mm for polioel 3 and respectively of 8,78 mm for royal jell

Anatomical and Histopathological Researches in Experimental Infections with Prototheca Algae in Mice

In order to realize this study, three BALB mice groups were used: one inoculated with P. zopfii, one with P. wickerhamii suspension and one control group. Each group was constituted of twelve animals, 2 months old, with an approximate body mass of 20 g. Both Prototheca species: one referent strain Prototheca wickerhamii (RE-4608014 ATCC 16529) and one Prototheca zopfii isolate (from cow mastitic milk sample), were cultured in glucose broth medium, 48 hours, at 37ºC. Except control group, each animal was i.p. inoculated with 0,1 ml algal suspension. Although any clinical disease sign was not observed in 10, 15 and 20 days from the beginning of the experiment for both inoculated groups, four mice were slaughted. Microscopic lesions caused by Prototheca algae were represented by granuloma and piogranuloma found in different internals as confirmed by identification of organism by staining with periodic acid-Schiff (PAS) and culture. So appreciated illness severity we utilized a score system in order to obtained a lesional systemic score for each animal. We appreciated: limph nodes reaction, the presence or absence of granuloma in skin, liver, pancreas, diaphragm, kidney, spleen and intestine. The data were statistically process using “t” test. In group inoculated with P. zopfii the lesions were more severs than those evidenciated in P. wickerhamii infected group

CULTURE CHARACTERIZATION OF DERMATOPHYTES FROM GENUS TRICHOPFHYTON IN CATTLE

In domestic ruminants (calves, cattle) dermatophytoses (ringworm) are caused mainly by T. verrucosum and sometimes by T. mentagrophytes (1). These fungi develop in the superficial layer of the skin, nails, and hair.This skin disease is spread worldwide in cattle and is responsable for high economic losses in cattel farm

STUDY REGARDING RESPONSE TO ANTIMICROBIAL AGENTS OF SALMONELLA ENTERICA STRAINS, ISOLATED FROM VARIOUS ECOLOGICAL AREAS

Between 2006 - 2008, we collected 106 Salmonella from birds (79 isolates) and pigs (27 isolates). These isolates were tested for susceptibility by antimicrobial disk diffusion method (Kirby –Bauer), according to NCCLS guidelines (1). The most sensitive drug for S. enterica tested were enrofloxacine (95.2%), ceftriaxone (93.3%), trimetroprim-sulfametoxazole (86.7%), ceftazidime (85.8%), ampiciline - sulbactam (83.9%), gentamycin (81.1%) and minimal values of sensibility were recorded for nitrofurantoine (3.8%), furazolidon (4.7%), nalidixic acid (5.6%), neomycin (6.6 %), flumequin (6.6%). tetracycline (7.5%)

SALMONELLA ENTERITIDIS: A MAJOR HAZARD FOR THE BIRD BREADING FARMS

During July 2005 - August 2006, 31 Salmonella enteritidis strains were isolated from 2 farms, mostly from chikens, in the first days after hatching, fact that indicates the vertical transmission of the infection. Isolation and identification was made following the AHDI – Bucharest standards and IEO reccomendations. The bone samples were positive for 24 strains (77,41%) and those of liver for 7 samples (22,58%). The results were confirmed by the refference laboratory for Salmonella, from Cantacuzino Institute, Bucharest

ANTIBACTERIAL EFFECT OF ALBIENS ALBA AND LAVANDULA AUGUSTIFOLIA OILY EXTRACTS ON VARIOUS BACTERIAL SPECIES

The aim of the study is to evaluate the antibacterial action of Albiens alba and Lavandula augustifolia oily extracts on bacterial species such as: Staphylococcus intermedius, Streptococcus pyogenes, Escherichia coli and Pseudomonas aeruginosa. Essential extracts of the two plants in 3 dilutions (20%, 10%, 1%) were used for each bacterial specie. Bacterial strains were isolated from dogs with dermatitis. For germs sensibility tests natibiograma by difusimethrical method was used, utilising a culture of 24 h with 2 optical density on MacFarland scale