Karvių sergančių slaptuoju mastitu diagnostika, gydymas ir profilaktika ; Bovine subclinical mastitis diagnostics, treatment and prophylaxis
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LITHUANIAN VETERINARY ACADEMY J ŪRAT Ė KLIMAIT Ė BOVINE SUBCLINICAL MASTITIS DIAGNOSTICS, TREATMENT AND PROPHYLAXIS Summary of doctoral dissertation Biomedical sciences, veterinary medicine (12B) KAUNAS, 2005 The research work has been carried out in the Lithuanian Veterinary Academy, in 2001-2005, Kaunas, Lithuania. The dissertation is written in Lithuanian. The research supervisor: Assoc. Prof. Dr. Eugenijus Aniulis (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine – 12B). Scientific adviser: Assoc. Prof. Dr. Albina Aniulien ė (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine – 12B). Chairman of the veterinary medicine council: Prof. at Incumbent Dr. Antanas Sederevi čius (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine – 12B). Members: Prof. Habil. Dr. Henrikas Žilinskas (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B); Prof. at Incumbent Dr. Bronius Bakutis (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine – 12B); Prof. Habil. Dr. Aniolas Sruoga (VU Ecology Institute, biomedical sciences, biology – 01B); Prof. Habil. Dr. Justinas Antanas Dobilas (LVA Veterinary Institute, biomedical sciences, veterinary medicine – 12B). Opponents: Prof. Habil. Dr. Algimantas Matusevi čius (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine – 12B); Dr.
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| Publié par | lithuanian_veterinary_academy |
| Publié le | 01 janvier 2005 |
| Nombre de lectures | 110 |
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LITHUANIAN VETERINARY ACADEMY J Ū RAT Ė KLIMAIT Ė BOVINE SUBCLINICAL MASTITIS DIAGNOSTICS, TREATMENT AND PROPHYLAXIS Summary of doctoral dissertation Biomedical sciences, veterinary medicine (12B) KAUNAS, 2005
The research work has been carried out in the Lithuanian Veterinary Academy, in 2001-2005, Kaunas, Lithuania. The dissertation is written in Lithuanian. The research supervisor: Assoc. Prof. Dr. Eugenijus Aniulis (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B). Scientific adviser: Assoc. Prof. Dr. Albina Aniulien ė (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B). Chairman of the veterinary medicine council: Prof. at Incumbent Dr. Antanas Sederevi č ius (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B). Members: Prof. Habil. Dr. Henrikas ilinskas (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B); Prof. at Incumbent Dr. Bronius Bakutis (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B); Prof. Habil. Dr. Aniolas Sruoga (VU Ecology Institute, biomedical sciences, biology 01B); Prof. Habil. Dr. Justinas Antanas Dobilas (LVA Veterinary Institute, biomedical sciences, veterinary medicine 12B). Opponents: Prof. Habil. Dr. Algimantas Matusevi č ius (Lithuanian Veterinary Academy, biomedical sciences, veterinary medicine 12B); Prof. Habil. Dr. Henrikas Stankevi č ius (Lithuanian Food Institute, biomedical sciences, zootechny 13B). Public defence of doctoral dissertation in Veterinary medicine science council will take place at the Lithuanian Veterinary Academy I auditorium 2 pm LT on 20 th September of 2005. Address: Til ė s 18, LT 47181 Kaunas, Lithuania. The abstract of doctoral dissertation has been send on 20 th August 2005 according to confirmed address list. This dissertation is available at the libraries of the Lithuanian veterinary Academy and LVA Veterinary Institute.
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LIETUVOS VETERINARIJOS AKADEMIJA
J Ū RAT Ė KLIMAIT Ė KARVI Ų SERGAN Č I Ų SLAPTUOJU MASTITU DIAGNOSTIKA, GYDYMAS IR PROFILAKTIKA
Daktaro disertacijos santrauka Biomedicinos mokslai, veterinarin ė medicina (12B) Kaunas, 2005
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Darbas atliktas 2001-2005 metais Lietuvos veterinarijos akademijoje. Disertacija parayta lietuvi ų kalba. Mokslinis vadovas: Doc. dr. Eugenijus Aniulis (Lietuvos veterinarijos akademija, biomedicinos mokslai, veterinarin ė medicina 12B) Mokslinio darbo konsultantas: Doc. dr. Albina Aniulien ė (Lietuvos veterinarijos akademija, biomedicinos mokslai, veterinarin ė medicina 12B) Disertacija ginama Lietuvos veterinarijos akademijos Veterinarin ė s medicinos mokslo krypties taryboje: Pirmininkas: E. prof. p. dr. Antanas Sederevi č ius (Lietuvos veterinarijos akademija, biomedicinos mokslai, veterinarin ė medicina 12B). Nariai: Prof. habil. dr. Henrikas ilinskas (Lietuvos veterinarijos akademija, biomedicinos mokslai, veterinarin ė medicina 12B); E. prof. p. dr. Bronius Bakutis (Lietuvos veterinarijos akademija, biomedicinos mokslai, veterinarin ė medicina 12B); Prof. habil. dr. Aniolas Sruoga (VU Ekologijos institutas, biomedicinos mokslai, biologija 01B); Prof. habil. dr. Justinas Antanas Dobilas (Lietuvos veterinarijos akademijos Veterinarijos institutas, biomedicinos mokslai, veterinarin ė medicina 12B). Oponentai: Prof. habil. dr. Algimantas Matusevi č ius (Lietuvos veterinarijos akademija, biomedicinos mokslai, veterinarin ė medicina 12B); Prof. habil. dr. Henrikas Stankevi č ius (Lietuvos maisto institutas, biomedicinos mokslai, zootechnika 13B). Disertacija bus ginama vieame Veterinarin ė s medicinos mokslo krypties tarybos pos ė dyje 2005 m. rugs ė jo 20 d. 14 val. Lietuvos veterinarijos akademijos I auditorijoje. Adresas: Til ė s g. 18, 47181 Kaunas, Lietuva Disertacijos santrauka isiuntin ė ta 2005 m. rugpj ūč io 20 d. Disertacij ą galima peri ū r ė ti Lietuvos veterinarijos akademijos ir LVA Veterinarijos instituto bibliotekose.
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INTRODUCTION Bovine mastitis in the meat processing industry has been and will be one of the most important interferences in producing high quality milk products. If the clinical form is registered amongst 2-5% of lactating and dry cows, then the incidence of sub-clinical mastitis (SCM) can be as high as in 50% of cows. In Lithuania, this form affects 44-47% of cows (Japertas, 2000). Such milk has an increased Somatic Cell Count (SCC), its technological characteristics change: it weakens fermentation, lactic acid and rennet enzyme effects, milk is less thermostabile. The quality of cows milk depends not only on the healthiness of the udder, but also the animals nutrition, husbandry and care conditions, age, lactation and season (Sender, 1995; Koldeweij et al., 1999). Sub-clinical mastitis can be prolonged and progress to the clinical form. Sub-clinical mastitis is diagnosed 15-40 times more often than clinical mastitis, though the sub-clinical form induces 60-70% of losses. ( Ивашура , 1991; Jukna, et al., 1994). From mastitis-affected milk, no single causative agent is identified, but about 150 various micro-organisms. Usually mastitis causative agents are S. aureus, S. agalactiae, S. dysgalactiae and S. uberis. Less common causative agents are E. coli , Corynobacteria , Pseudomonas , yeasts and mycoplasma (Wats, 1988; Siugzdait ė , 1997). Due to common and erroneous use of antibiotics and sulfamides, resistant strains of micro-organisms arise, upsetting the udders non-pathogenic micro-organism proportions. An important role in SCM etiology is attributed to yeasts. During 1980-1985, from mastitis affected milk, 1% of the samples grew pure cultures of Candida genus yeasts (Zelba, 1985; Sestakauskas, 1987). In some countries an increase of yeast-caused mastitis was observed (H. Krukowski, 2001; E. Malinowski et al., 2002). Polish dairy farm statistics showed that 14.4% of milk samples grew Candida genus yeasts. In prophylaxis and treatment of bovine SCM, prolonged action antibiotics act on gram-positive, gram-negative microbes and mycoplasma, however they are ineffective against yeasts. In weakening microbe activity, the inflammation process is upheld by yeasts. For treatment of yeast and micro-organism caused mastitis, a single imported preparation is currently offered Neomastipra Fung (amoxicillin trihydrate 1 mg, nistatine 5 mln. IU, hydrocortizone 20 mg; Laboratory Hipra S.A., Spain). Our develpoed preparation (in consultancy with other specialists), conditionally named Gentafung is manufactured by Norfachema (Lithuanian & Norwegian Joint Stock Company). This preparation has been used to treat cows with SCM, having causative agents of gram-positive and gram-negative micro-organisms and yeasts. Treatment and prophylaxis efficacy depended on correct identification of causative agents as their sensitivity to various antibacterial medications differ, especially as the current supply is quite broad. This is why it is advisable to determine the market-offered antimicrobial efficacy in treating cows with SCM. Aims of the study: To determine Candida genus yeasts and micro-organism caused distribution and diagnostics of bovine SCM. To compare the preparation Gentafung and Neomastipra Fung treatment efficacy in treating cows with SCM. To confirm the offered commercial antibacterial preparation injection frequency and duration of use. Goals of the study: 1. Determine the distribution of Candida genus yeasts and micro-organisms causing SCM; 2. Determine identification of yeasts, their biochemical and physiological characteristics; 3. Perform a comparative evaluation of Candida genus yeast commercial identification systems with classical diagnostic methods; 4. Determine preparations Gentafung and Neomastipra Fung treatment efficacy; 5. Determine efficacy of homeopathic preparations in treating SCM caused by Candida genus yeasts and micro-organisms; 6. Assess antimicrobial preparations in treating SCM caused by S. aureus and micro-organisms; 7. Confirm prophylactic medications efficacy on drying cows; 8. Determine Candida genus caused SCM morphological changes in the udders parenchyme. Novelty of the study: The distribution of yeast fungi has been ascertained as their influence on SCM etiology. Our developed preparation for treatment for yeast and microbe caused mastitis Gentafung is composition of wide spectrum antibiotics and antifungal medications. An evaluation was performed of commercially available homeopathic preparations in treating SCM. A new recommended preparation OrbeSeal was evaluated for drying cows. A comparison of Candida genus yeast commercial identification systems diagnostic accuracy was performed via classical methods.
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Practical assignment significance: Gentafung preparation was evaluated in cow herds for its efficacy against gram positive, gram negative microbes and yeast fungi. The distribution of yeast and micro organism caused SCM was clarified in cow herds. An evaluation of commercially newly available preparations was performed for drying cows. Volume and structure of the study: This dissertation is written in the Lithuanian language, contains 138 pages and includes: an introduction, literature review, materials and methods, results, discussion, conclusions, suggestions, a summary in the English language, a list of used literature including 209 references, acknowledgements, 11 tables, 21 pictures, 18 appendix. RESEARCH METHODS Time, location, and conditions of the research During 2001-2005, scientific research was performed at the Lithuanian Veterinary Academys Non-Contagious Disease Dept. and Livestock Reproduction Laboratory, 28 agricultural companies and 30 dairy farms. Experiments were performed using 2-6 yr. old lactating and pregnant dry Lithuanian Black & White and Lithuanian Red breed cows. Cows were fed according to the farms determined ration, water was offered ad libitum. For bacteriological testing, milk samples were placed in sterile test tubes according to aseptic procedures. 2374 milk samples were tested. Diagnostic tests for SCM were performed on 1742 cows, 6968 udder quarters. Cows were selected having >500 10 3 /cm 3 general Somatic Cell Counts (SCC). Determining treatment efficacy, the medications were injected according to instructions for use. Assessment of general milk bacterial contamination and SCC For these investigations, samples were placed in special tubes from aggregate milk and presented to the State Company Pieno Tyrimai. SCC was determined using the following instruments: Somascope MK2 (Delta Instruments, Holland) and Fossomatic (Foss Electric, Denmark). Fat, protein and lactose amounts in milk were determined via Lactoscope 550 (Delta Instruments Holland), and canned milk general bacterial contamination with the equipment Cobra 2024-Asterias (Biocom, France). Bacteriological milk sample testing From the affected cows udder quarter, milk samples were aseptically placed in sterile tubes at the end of milking. Mastitic milk samples were sown on Mac-Conkey (enterobacteria) (Oxoid, England), Edvardso (streptococci) (Oxoid, England), sheeps blood (staphylococci) (Oxoid, England), BairdParker (Liofilchem, Italy), Biggy (Oxoid, England), OGYE (Liofilchem, Italy), Sabouraud dextrose (Remel, USA) agar. To resow cultures, we used meat peptone agar and boullion (Oxoid, England). Sheeps blood, meat peptone and boullion, Mac-Conkey, OGYE, Sabouraud dextrose agars were autoclaved for 15 minutes at 121 ° C, and Edvards media for 20 minutes at 115 ° C. Milker outwash samples were obtained post-milking using transportable media (Transport Swab, Italy). Yeast identification Research was performed at the Botanical Institutes Biodestructor Laboratory. Sowing the milk samples on Biggy, OGYE, Sabouraud dextrose agar, we placed the dishes in a (24±1)º C thermostat under aerobic conditions for 5-7 days. Pure yeast cultures from mature colonies were examined and selected according to typical characteristics. They were microscopically examined and confirmed as yeasts. Later they were renown on bevelled yeast extract, dextrose and oxytetracycline agar and grown for 3 days at 24±1º C temperature. Yeast morphological, physiological and biochemical characteristic examinations were performed using Kreger van Rij (1984) methods. Yeast species were identified via Kreger-van Rij (1984) established characteristics. Comparative evaluation of yeast identification via classic methods and commercial express systems Yeast identification systems (Integral system yeast, Liofilchem, Italy) kit is comprised of 12 biochemical tests and 6 antimycotic preparations. Fungichrom I (International microbio, France) kit is comprised of 16 biochemical tests.
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Veterinary medication efficacy in treating cows with SCM Therapeutic effect of Gentafung and Neomastipra fung Preparation Gentafung (gentamycin 1.5 g, fluconazole 0.5 g; Norfachema, Lithuania) was used in treating 20 cows with 52 infected udder quarters. 20ml was injected into each affected udder quarter 3 times q12h. 20ml of preparation Neomastipra fung (amoxicillin trihydrate 1 mg, nistatin 5 mln. IU, hydrocortisone 20 mg; Laboratory Hipra S.A, Spain) was applied to each infected udder quarter 3 times q12h. 59 cows were treated with 152 infected udder quarters. The efficacy of preparations was judged by the milk samples bacteriological qualitative compositions pre and post treatment. To determine SCCs, general bacterial contamination, protein, fat and lactose amounts, milk samples were taken from the aggregate prior to, and Day 14 and 21 post-treatment. Therapeutic effect of Noroclav, Synulox LC and Synulox RTU Preparation Noroclav (clavulanic acid 35 mg, amoxicillin 140 mg; Pfizer animal health, Belgium) was used in treating 10 cows with 30 infected udder quarters, was injected 5 days into muscle (1xd). Preparation Synulox LC (clavulanic acid 50 mg, amoxicillin 200 mg, prednisolone 10 mg; Pfizer animal health, Belgium) was used in treating 9 cows with 12 infected udder quarters, was injected in to all teat canal 5 time after milking. Preparation Synulox LC (clavulanic acid 50 mg, amoxicillin 200 mg, prednisolone 10 mg; Pfizer animal health, Belgium), was injected in to all teat canal 5 time after milking and preparation Synulox RTU (clavulanic acid 35 mg, amoxicillin 140 mg; Pfizer animal health, Belgium) was injected 5 days into muscle (1xd), was used in treating 10 cows with 40 infected udder quarters The efficacy of preparations was judged by the milk samples bacteriological qualitative compositions pre and post treatment. To determine SCCs, protein, fat and lactose amounts, milk samples were taken from the aggregate prior to, and Day 21 and 60 post-treatment. Effect of Biomast For the purpose of treatment, 45 lactating cows were selected which had somatic cell counts higher than 400 10 3 /cm 3 . 3 ml. of preparation was injected SQ, laterally to the udders lymphnodes on both sides. Milk samples for bacteriological testing, general bacterial contamination, somatic cell counts, proteins, fats and lactose amounts were obtained prior to preparation injections and Day 14, 21, 60 post-treatment. Leukograms were used to evaluate immune response from the blood of 5 cows diagnosed with subclinical mastitis. Bovine SCM caused by yeast fungi and microbe treatment using homeopathical preparations Traumeel ® ad us.vet, PhosphorHomaccord and Traumeel ® S Gel Using preparation Traumeel ® ad us.vet (Aconitum napellus D4 0.3 ml; Arnica montana D4 0.5 ml; Atropa belladonna D4 0.5 ml; Bellis perennis D4 0.25 ml; Calendula officinalis D4 0.5 ml; Chamomilla recutita D5 0.5 ml; Echinacea angustifolia D4 0.125 ml; Echinacea purpurea D4 0.125 ml; Hamamelis virginiana D3 0.05 ml; Achillea millefolium D5 0.5 ml; Hypericum perforatum D4 0.15 ml; Symphytum officinale D8 0.5 ml; Hepar sulfuris D6 0.5 ml; Hahnemanni soluble mercury D8 0.25 ml; sodium chloride q. S. 5 ml; Biologische Heilminttel Heel GmbH, Germany) and PhosphorHomaccord (Phosphorus D10 0.02 ml; Phosphorus D30 0.02 ml; Phosphorus D200 0.02 ml; Argentum nitricum D10 0.015 ml; Argentum nitricum D30 0.015 ml; Argentum nitricum D200 0.015 ml; Paris quadrifolia D6 0.015 ml; Paris quadrifolia D10 0.015 ml; Paris quadrifolia D30 0.015 ml; Paris quadrifolia D200 0.015 ml; Biologische Heilmittel Heel GmbH, Germany) we treated 6 cows having 15 SCM-affected udder quarters. 5 ml was injected IM. Repeat injections were administered after 24 h and after 4 days. Milk samples for bacteriological testing, determining general contamination, SCC, protein, fat and lactose amounts were obtained prior to application of the preparations on Day 14 and 21 post-treatment. Preparation Traumeel ® S Gel (Arnica montana D3 1.5g; Symphylum officinale D4 0.1 g; Hypericum perforatum D6 0.09 g; Mercurius solubilis D6 0.04 g; Hepar sulphur D6 0.025 g; Aconitum napellus D1 0.0025 g; Belladonna D1 0.05 g; Calendula officinalis ǿ ; Hamamelis virginica ǿ aà 0.45 g; Echinacae angustifolia ǿ ; Echinacae purpurae ǿ ; Chamomilla ǿ aà 0.15 g; Bellis perenuis ǿ 0.1 g; Millefolium ǿ 0.09 g; exc q.s. ad 100g; Homeoden Heel, Belgium) was injected to six cows having 16 affected udder quarters, 10 ml each, 3 times, 12h after milking. To determine aggregate milk bacterial contamination, SCC, protein and lactose amounts, samples were obtained prior to injecting preparations, 14 and 21 days post-treatment. Milk sample bacteriological tests were performed prior to, and 14, 21 days post-treatment.
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Preparation prophylactic efficacy on drying cows Effect of Biomaston drying cows For prophylactic purposes against SCM we used the biostimulator Biomast (Corynebacterium uberis 22, SDT Abiotest, Poland). 5 end-lactation cows received 3 mls of the preparation, injected SQ lateral to the supramammarian lymphnodes on both sides 14 days prior to end-lactation. Another group of 5 end-lactation cows received 3 mls of biostimulator SQ lateral to lymphnodes on both sides and 10 mls of Evetsel IM (tocopherol acetate (Vit.E), 750 mg., sodium selenite 10.95 mg, Pliwa Krakow S.A., Poland). Milk samples for bacteriological testing, general bacterial contamination, proteins, fat and lactose amount determination were taken prior to preparation injections and postpartum once colostrums production has stopped. Leukograms were used to determine immune responses in both groups of cows. Drying cow prophylaxis using preparations OrbeSeal and Albadry Plus In accordance to research statistics (according to SCC), cows were divided into three groups: 1)- SCC up to 200.000/ cm 3 , 2)- from 201.000 to 500.000/ cm 3 , 3) > 501.000/ cm 3 . The first group of six cows were treated in all udder quarters after final milking with OrbeSeal (65% bismuth subnitrate, Pfizer Animal Health, Great Britain); Group 2- 23 cows, and Group 3 12 cows were treated in all udder quarters after final milking Albadry Plus (novobiocin 400 mg, benzylpenicillin procaine salt 200.000 IU; Pharmacia N.V, Belgium) and OrbeSeal (65% bismuth subnitrate; Pfizer Animal Health, Great Britain). Milk composition (State Company Pieno Tyrimai) and bacterial tests were performed from post-partum cows, after the colostrum period. Histological investigation of udder parenchyme From two slaughtered cows, 8 udder quarters, in which it was confirmed that mastitis was caused by yeasts, tissue parenchyme samples were obtained for histological investigation. Samples were fixed in 10% neutral formalin solution. Slides were prepared using O. Kublickien ė s method (1978). Statistical result analysis Test results and statistical data were calculated via computer program SPSS. Statistical data arithmetic averages were calculated (Xa), average error margins (± Se), standard deviation (S), variable coefficient (Cv), cow number in group (N), average difference reliability coefficient (td). According to StjudentoGaseto Table, we determined the average difference reliability (p). The difference was accepted as statistically reliable if p<0,05. RESEARCH RESULTS Udder quarter tests using reagent CMT For express diagnostics we used reagent CMT. Our test data showed that cow udders confirmed having SCM in one quarter 519 cases (7.44 % ), two quarters 1 396 cases (20.03 % ), three quarters 1 275 cases (18.30 % ), all quarters 2 156 cases (30.94%). In evaluating which side of the udder shows developing inflammation, we determined that it is near equal: right side - 49.55 % , left 50.45 % . However we noticed that the right rear quarter is more commonly affected with developing inflammations than the other quarters. At least 41.15 % of inflammations were confirmed in the right frontal quarter Bacteriological milk sample test results Having performed bacteriological tests from 2 374 affected cows, the following microflora were identified: pure cultures 26.37%, mixed cultures 64.20%. No pathogenic microflora identified 9.43% of samples. Pure microflora were comprised of: yeast fungi 7.98%, S. aureus 19.97%, CNS 58.15%, Streptococcus spp . 5.43%, Enterobacteriaceae family 8.47%. Most common mixed infections were: Enterobacteriaceae family bacteria 96.65%, Staphylococcus genus 96.59%, amongst these S. aureus 14.83%, Streptococcus genus 41.27% , Candida genus yeasts 60.76%. From milk samples from cows with SCM, 2 microorganisms were identified in 43.44% cases, 3 microorganisms 41.47%, 4 microorganisms 15.09% cases (Fig. 1).
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Gryna kult ū ra / Pure cultures Miri kult ū ra / Mixed cultures Fig. 1. Milk sample bacteriologic test results 1 pav. Pieno m ė gini ų bakteriologinio tyrimo rezultatai Milker outwash bacteriological testing Examining 72 samples of outwash from milkers, 16 samples (22.22%) grew pure cultures, mixed 50 samples (69.44%) and only 6 samples (8.34%) showed no micro-organism growth. Pure culture microflora was comprised of yeast fungi 25.00%, CNS 50.00%, enterobacteria 25.00%. Mixed microfloras was comprised of yeast fungi 30.00%, CNS 90.00%, streptococci 10.00%, enterobacteria 16.00% (3 samples). Our research shows that milker disinfection pre and post milking is an important factor in reducing micro-organism transfer during milking. Identified yeast species description Performing classical method identification of 49 isolated yeast species, their biochemical and physiological characteristics were determined. Test results identified: C. krusei 14 28.57%, C. famata 3 6.12%, C. glabrata 1 2.04%, Rhodotorula rubra 9 18.37%, C. tropicalis 10 20.41%, Cr. laurentii 1 2.04%, C . inconspicua 3 6.12%, C. parapsilosis 5 10.20%, Geotrichum candidum 3 6.12% (Fig. 2).
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Fig. 2. Yeast species 2 pav. Mieli ų r ū ys
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30%
6% 2%
C. krusei C. famata C. glabrata Rh. rubra C. tropicalis Cr. laurenti C. inconspicua C. parapsilosis G. candidum
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Comparative evaluation of yeast via classical and express methods Classical yeast identification methods were compared to express method systems and Fungichrom I . Comparing the classical identification method with the express yeast identification method we determined that of 49 yeasts isolated 81.63% coincided, and comparing with Fungichrom I 91.84% results coincided. In comparing express diagnostic methods with Fungichrom I, 88.89% of results coincided. Veterinary preparation efficacy in treating cows with SCM Therapeutic Gentafung efficacy Having performed milk tests from 20 cows having 52 infected udder quarters, it was determined that the general SCC was quite high (1031.90 ± 143.23 10 3 /cm 3 ). The aggregate bacterial contamination exceeded standard margins and which comprised 370.5 ± 43.84 10 3 /cm 3 (example 66). Prior to applying medication in the udder quarters, the milk composition test results showed: proteins 3.60 ± 0.11%, fat 4.01 ± 0.11%, lactose 4.33 ± 0.09%. Bacteriological tests from all the collected milk samples from affected quarters resulted in mixed microflora which comprised: enterobacteria and yeast fungi 3 samples (15%), CNS, enterobacteria and yeast fungi 7 samples (35%), CNS, streptococci and yeast fungi 5 samples (25%), CNS, enterobacteria, streptococci and yeast fungi 1 sample (5%), CNS and yeast fungi 4 samples (20%). 14 days post treatment of affected quarters, tests of milk samples constituted positive medication affects on the milk gland. A positive result also was shown by a 4.92% (p>0.5) reduction in SCC. The general milk bacterial contamination decreased by 25.51% (0,1<p<0.05). A change was noted in milk composition also. A slight decrease in proteins was observed 4.73% (p>0.4), though fat increased 11.32% (p<0.01) as did lactose 10.73% (p<0.005). A positive affect of the medication was noted on the milk gland and affected quarters by the identified microorganisms. Gentafung ingredients (gentamycin) acted positively against grampositive and gramnegative microbes and yeast fungi (flukonazole). In testing the milk samples from previously affected quarters, even 50% did not grow microbe or yeast fungi cultures. The remaining samples grew mixed microflora (50%), which comprised CNS and enterobacteria (30%) 3 samples, CNS and yeast fungi 4 samples (40%), CNS, streptococci and yeast fungi 3 samples (30%). 21 days post treatment the SCC decreased by 69.05% (p<0.001) in comparison to the amount prior to treatment. Total bacteria count decreased by a remarkable 83.97% (p<0.001). That the udder had healed and returned to a physiological norm is shown by milk quality indicators. Protein amounts stabilised at 3.60% (0,1<p<0.05), fat and lactose increased by 12.22% (p<0.001) and 10.81% (p<0.001) respectively. In testing milk bacteriologically, 5 cows in 7 quarters (13.46%) grew mixed microflora, comprised of CNS and enterobacteria. The results of milk bacteriological and milk composition tests show that gentamycin was effective against gram positive and gram negative bacteria and against yeast flukonazole, thereby the treatment results were rather satisfactory. Post-treatment SCC was 319.40 ± 40.89 10 3 /cm 3 (p<0.001), and the general bacterial contamination was 59.40 ± 9.11 10 3 /cm 3 (p<0.001). Of the treated 20 cows and 52 infected quarters 85% healed completely. Therapeutic Neomastipra Fung efficacy 59 cows were selected for treatment of 152 infected udder quarters having an aggregate SCC of 1616.03 ± 155.80 10 3 /cm 3 , and the total bacterial count was 399.94 ± 27.04 10 3 /cm 3 . Fat, protein and lactose amounts were within physiological parameters: 4.53 ± 0.10%, 3.42 ± 0.07%, 4.59 ± 0.04%. Prior to treatment we bacteriologically tested milk samples from the infected udder quarters identifying S. aureus , enterobacetria and yeast fungi in 5 samples (8.47%), CNS, enterobacteria and yeast fungi in 44 samples (74.58%), CNS and yeast fungi in 9 samples (15.25%), enterobacteria and yeast fungi 1 sample (1.70%). -The medications component amoxicillin is effective against gram positive and gram negative bacteria, and nistatine against yeast fungi. 14 days post treatment, after three applications of the medication, the general milk composition from the affected udder quarters had changed: SCC decreased by 49.65% (p<0.001), BBU 47.02% (p<0.001). A positive effect on the milk gland was a slight increase in fat and lactose amounts repectively: 10.11% (p>0.5) and 10.07% (p>0.5). Protein amounts remained unchanged. From the affected quarters, bacteriological testing of samples identified mixed microflora S. aureus and yeast fungi 8.47%, CNS, enterobacteria 49.16%. No microbes grew in 25 of the samples (42.37%). After 21 days, aggregate milk from affected quarters showed a decrease in SCC 77.71% and was 371.58 ± 32.95 10 3 /cm 3 (p<0.001). The milk from 59 treated cows and 152 infected quarters was within the standard raw milk requandement values < 400,000/cm 3 (p<0.001). Protein, fat and lactose: 3.49 ± 0.06% (p>0.2), 4.70 ± 0.10% (p>0.2) and 4.63 ± 0.04% (p>0.2) respectively. In bacteriologically examining milk samples we noticed that in 43 of the samples (72.88%) microbes and yeast fungi were not observed. Only 5 samples (8.47%) showed S. aureus and 11 samples CNS and enterobacteria (18.65%).
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Decreases were noted in the amount microorganisms in the aggregate milk as did total bacterial count (82.37%) (p<0.001). Of the treated 59 cows, 43 (72.88%) completely healed. Our test results show that the medication Neomastipra Fung component nistatine is effective against yeast fungi, and amoxicillin against most grampositive and gramnegative bacteria. In performining an antibiogram, identified microbes were averagely (16mm) sensitive to amoxicillin In comparing preparations Gentafung and Neomastipra Fung post treatment, it was determined that Gentafung was more effective than Neomastipra Fung (Fig. 3).
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Gentafung Neomastipra fung Fig. 3. Therapeutic effect of Gentafung and Neomastipra fung 3 pav. Gydomasis preparat ų Gentafung ir Neomastipra fung efektyvumas The efficacy of preparations Noroclav, Synulox LC and Synulox RTU The first group of cows (preparation) Noroclav prior to treatment grew pure cultures ( S. aureus ) in 60% of cases from milk samples taken after milking. Mixed microflora ( S. aureus, streptococci, enterobacteria, Candida genus yeasts) were identified in 40% of samples. A rather large SCC was found in aggregate milk (980,80 ± 397,80 10 3 /cm 3 ). 21 days post treatment, bacteriological tests distinguished S. aureus in 20% of samples. A slight increase was identified (10%) of mixed microflora. Judging from bacteriologic test results Noroclav is substantially effective against S. aureus, enterobacteria and streptococci, though ineffective against Candida genus yeasts. 21 days post treatment, no micro-organisms were identified in 30% of samples. 2 months post treatment S. aureus was identified in 30% of samples. During treatment, milk composition changed. 21 days post treatment, the fat amount in milk decreased by 6,16% (p>0,4), after 2 months 3,61% (p>0,4) in comparison with amounts prior to treatment. Lactose amounts decreased also. 21 days post treatment, lactose decreased by 2,50% (p>0,4), after two months 3,34% (p>0,5). Protein decreased by 6,03% (p>0,4), and after two months increased by 10,10% (p>0,4). One of the most important indicators of udder recovery is the SCC during treatment. 21 days post treatment, the SCC decreased by 56,66% (0,1<p<0,05), after 2 months 61,15% (0,1<p<0,05) and thereby reached the milk standard requandements (381,00 ± 109,45 10 3 /cm 3 ). 70% of treated cows fully recovered. Prior to treatment with Synulox LC,44,44% of bacteriologically examined milk samples grew pure cultures (22,22% S. aureus and 22,22% KNS). Mixed microflora grew in 55,56% samples ( S. aureus, KNS, streptococci, enterobacteria, Candida genus yeasts). 21 days post treatment, 22,22% of bacteriologically examined samples grew Candida genus yeasts, after 2 months samples grew 55,56% mixed microflora ( S. aureus, Candida gentus yeasts). During treatment, other indicators fluctuated as well. 21 days post treatment, milk fat decreased by 23,27% (p>0,1), proteins 1,1% (p>0,5), however lactose amounts increased by 10,70% (0,1<p>0,05). Lactose increase 11
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