Development of resistance to antibiotics in bacteria Staphylococcus spp. Isolated from milk samples in the sheep breedings on east of Slovakia

During the last three years (2015 to 2017), the frequency of occurrence of bacteria Staphylococcus spp. were examined in total 3466 individual and 12 pool milk samples. Experiment was carried out in two herds of breed of sheep Improved Valaska in region of East Slovakia. Were isolated and taxonomically identified 15 species of the genus Staphylococcus spp. (n = 444). From the coagulase positive staphylococci (CPS), S. aureus was isolated during the reporting period, however, most often in the first year (45). The incidence of S. intermedius and S. hyicus has been irregular. From the coagulase negative staphylococci (CNS) (n = 288), were isolated S. epidermidis present in 37.5% (108), S. scheiferi 25.69% (74) and S. chromogenes 23.61% (68), while other species occurred only rarely. The bacteria S. aureus (n = 117) showed the highest resistance to novobiocine 14.5%, to erythromycin 12.8%, lincomycin 7.69% and also 7.69% to penicillin. In the framework of the CNS sensitivity we tested 108 strains of S. epidermidis, from which it was 11.1% resistant to novobiocine and 8.3% to erythromycin. Statistical comparison of the incidence of resistance to penicillin and novobiocine in S. aureus and S. epidermidis was performed using Chi square test. There was statistically significant dependence of tracked characters was confirmed on significance level α = 0.05. Also, the incidence of intermediate sensitivity of bacteria in the Staphylococcus spp., indicates the unfavourable development of resistance to the most commonly used antibiotics to treat the inflammation of the udder in sheep. It is therefore recommended to regularly check the resistance to antibiotics and often isolated bacteria CNS

Occurrence of some pathogenity factors in coagulase negative Staphylococci isolated from mastitis milk in dairy cows

Experiment we carried out on the herd of 115 Slovak Pied dairy cows. Were realized three collections of individual milk samples for isolation and identification of coagulase-negative staphylococci (CNS). We collected 341 milk samples. Coagulase - negative staphylococci were isolated in 18.8 % (64). Highest number of CNS we isolated from acute, subclinical, subacute and latent forms of mastitis, respectively. S. warneri (2.88 %), S. chromogenes (17.19 %), S. cohnii (15.63 %) and S. xylosus (14.06 %) was isolated in highest percentage. Was detected presence of 7 virulence factors, include production of staphylococcal enterotoxins, and resistance for antibiotics. S. chromogenes, S. warneri and S. xylosus were detected as most virulent species carrying most virulence factors, and were resistant for most species of tested antibiotics (S. xylosus resistant for N, NV, L, P, E; S. chromogenes, S. sciuri for 4 species of antibiotics). There pathogens were isolated from acute forms of mastitis.

Influence of selenium and vitamin e supplementation during pregnancy on udder health and milk quality in dairy cows at parturition

Selenium and vitamin E ranks among very important antioxidant agents protecting the organism from the effect of reactive oxygen forms. The deficiency of both nutrients during pregnancy in cows often result in metabolic disorders and increased of cases of related diseases (mastitis, retained placenta and other reproductive disorders). The aim of the present work was to study the influence of different dose of parenteral administration selenium and vitamin E in dairy cows prior to parturition on selected metabolic parameters, udder health and milk quality. A total in herd of 270 Holstein cattle in east of Slovakia in a two-four lactation-gestation cycle the control group (C) and 2 experimental groups (D, D1) were selected. All groups were similarly housed, managed and fed with the diet containing from 36 to 42 mg vitamin E and 0.2 mg.kg-1 Se of DM through the study period. In group D a products containing vitamin E and selenium were administered IM four weeks prior to the expected date of parturition in total dose of 1000 mg of dl-α-tocopherol acetate and of 44 mg sodium selenite per cow, respectively. In group D1 the same products were administered twice, four and two weeks prior to parturition. Blood samples were 4 weeks prior to predicted calving date (the time of treatment), on parturition day and at 14th day after calving for assessment of plasma vitamin E and selenium concentrations. Blood samples of the calves were drawn from jugular vein at birth and first colostrum was also collected. The occurrence of the mastitis and retained placenta during the first 14th day after calving were evaluated in all groups. Higher plasmatic and colostral concentrations of selenium and vitamin E were found only in group with repeat application of Se and vitamin E (D1) collected on the day of parturition. At the 14th day of postpartal period a trend of lower occurrence of mastitis was observed in group D1 compared to D group, administered IM once and control group. Parenteral supplementation of selenium and vitamin E during pregnancy had no impact on their transmission into the milk and on the presence of bacterial agents in raw milk obtained from dairy cows diagnosed with mastitis.&nbsp

Effect of peroral supplementation with selenium and vitamin e during late pregnancy on udder health and milk quality in dairy cows

The aim of the experiment was to study selenium and vitamin E sources in the diet of dairy cows in late phase of pregnancy and their effects on udder health and milk quality during the first two weeks after calving. The experiment included 48 cows of Holstein breed divided into four equal groups (n = 12). The first experimental group (D1) was fed with addition of vitamin E in total dose of 1020 dl-a-tocopherol acetate Se/cow per day. The second group (D2) was added the selenium at a dose of 0.3 mg.kg-1 of DM in form of sodium selenite. The third group (D3) was supplemented with addition of vitamin E in combination with sodium selenite in total dose of 1020 dl-a-tocopherol acetate Se/cow per day and of 5.0 mg Se/cow per day, respectively. The control group (C) was without the addition of selenium and vitamin E. In group (D2) with addition of selenium at a dose of 0.3 mg.kg-1 of DM and vitamin E a dose of 50 mg dl-a-tocopherol acetate/kg of DM in diet, increased the plasmatic concentration of selenium and vitamin E and reduced the incidence of mastitis by 13.3% and number of somatic cells during peripartal period in comparison with other groups

Antibiotic resistance in bacteria Staphylococcus spp. isolated from samples of raw sheep's milk

From samples of raw sheep's milk were determined results of bacteriological examination from two herds in region of Eastern Slovakia in three years lasting study. The occurrence of Staphylococcus spp. 41.6% (124) was determined from 298 samples. The seven species of staphylococci were on a regular basis isolated: S. epidermidis (34), S. chromogenes (26), S. aureus (16). Alternately have been recorded S. warneri (16), S. schleiferi (15), S. haemolyticus (9) and S. xylosus (8). All isolated pathogens were tested by in vitro test on Mueller-Hinton agar by disc methods on resistance to 10 types of antibiotics.  Highest value of resistance was determined to Penicilin 21.0%, Neomycin 10.5% and Novobiocin 9.7%. Lower resistance was in to Oxacilin 7.2% and Amoxicilin 6.5%. Minimal resistance was founded to Cefoxitin 0.8%, Linkomycin 2.4%, Erytromycin, and Streptomycin 3.2%. Was founded total resistance (21.0%) to all antibiotics in S. epidermidis (34) during the three years, S. chromogenes (26) showed resistance to 8 types of antibiotics (12.9%), S. aureus (16) to 6 antibiotics (10.5%) and S. warneri (16) to 4 antibiotics (5.6%). It was confirmed that sheep's milk remains a major source of staphylococci. Bacteria in comparison with isolates from cows' raw milk, showed lower values of resistance, but were resistant to more than two antibiotics. Recorded occurrence of resistance in staphylococci may be connected with a minimum use of antibiotics in the treatment of mastitis and other diseases in sheep herds. Reported resistance to the tested antibiotics became the basis for the recommendation to use preparations to treat mastitis in sheep principally by the detection of resistance to antibiotics contained

Mastitis pathogens and their resistance against antimicrobial agents in herds of dairy cows situated in marginal parts of Slovakia

Marginal regions are relatively large part of the area Slovakia which in terms of the economy breeding ruminants can efficiently produce animal commodities only occasionally. Geographic, social and economic stability of these regions is strongly influenced by breeding of ruminants with market milk production. Mastitis is a disease complex that assumes highest clinical and economic significance in milk animals particularly medium to high yielding dairy cattle, usually in and around periparturient period. The objective of this study was to evaluate the effectiveness of different antibiotics against mastitis causing microorganisms during first month of lactation in two herds of 230 and 310 dairy cows situated in marginal parts of Slovakia. Milk samples from quarters were cultured and identified bacteria were subjected to antimicrobial susceptibility test by disc diffusion method to a large number of antibiotics. The prevalence of mastitis in the monitored herds of dairy cows was 26.1% to 17.6%, respectively. A total of 1663 milk samples from udder quarters were investigated, 446 (21.3%) samples were positive. No pathogens were isolated from 1663 (78.4%) milk samples. From all tested bacteria Staphylococcus spp. and Streptococcus spp. which were isolated from subclinical and clinical mastitis, were found amoxicillin + clavulanat and tetradelta to be most effective drug followed by ceftiofur and rifaximin. The significant difference was confirmed between the Staph. aureus and coagulase-negative staphylococci (CoNS) isolates with respect to their susceptibility to the various antibiotics. Antibiotic susceptibility tests should be done to determine the effectiveness of drug that can be used for successful treatment of diseases. Proper isolation and identification of the causative organism play significant role in prevention and control of the diseases

Assessment of lipid peroxidation in dairy cows with subclinical and clinical mastitis

Mastitis is still one of the major causes of economic losses in dairy sector. The routine application of bacteriologic examination of milk samples is often insufficient and for this reason, alternative parameters are used to identify trends in the development of the udder health. Therefore, the objectives of this study were to determine the relationship of oxidative product levels, using malondialdehyde (MDA) as a marker on occurrence of mastitis and its causing pathogens. Dairy herd of 223 Slovak spotted cattle were tested for etiology and occurrence of mastitis based on assessment of clinical signs, abnormal udder secretions, Californian Mastitis Test (CMT) with subsequent collecting of milk samples for bacteriological examination. From 892 quarter milk samples were selected for MDA detection 51 subclinical (SM) and 26 clinical mastitis (CM) quarters with positive CMT score and positive bacteriological examination of Staphylococcus spp. and Streptococcus spp. as well 40 healthy quarters. Results showed that among the current pathogens of the mammary gland belong CNS, S. aureus, S. sanguinis, S. uberis and E. coli, which were the most frequently isolated from SM and CM. The highest MDA level was observed from clinical cases of mastitis however, increased MDA levels were detectable from subclinical cases. Bacterial isolates from subclinical quarter milk samples are different levels of MDA. In this study, we found that quarter milk samples infected with S. uberis were higher compared to other pathogens. In conclusion, differences in both severity of mastitis and mastitic pathogens were associated with differences of oxidative products in infected udders

THE MICROBIOLOGICAL LOAD OF SHEEP MILK FROM PRIMARY PRODUCTION TO ITS PROCESSING

<div style="mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><table style="height: 434px;" border="0" cellspacing="0" cellpadding="0" width="664" align="left"><tbody><tr><td style="padding-top: 3.1pt; padding-right: 0cm; padding-bottom: 3.1pt; padding-left: 0cm;" height="434" align="left" valign="top"><div style="mso-element: para-border-div; border: none; border-bottom: solid windowtext 2.25pt; padding: 0cm 0cm 1.0pt 0cm;"><p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><span style="font-size: 10.0pt; mso-bidi-font-weight: bold;">In the breeding with the average number of 220 sheep (zo&scaron;ľachten&aacute; vala&scaron;ka) with traditional hand milking in the Eastern Slovakia the microbiological load of milk during the process of primary production, transport, before and after pasteurisation as well as during dairy processing to cheese curd was observed. The results in three seasons were compared to those obtained at finishing of milking in the season before.</span></p> <p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><span style="font-size: 10.0pt; mso-bidi-font-weight: bold;">The microbiological load of milk was observed using the bacteriological methods for determination of the presence of Staphylococcus sp. and other bacteria, and determination of the total number of staphylococci: a)<span style="mso-spacerun: yes;">&nbsp; </span>in milliliter of pool milk sample; b)<span style="mso-spacerun: yes;">&nbsp; </span>the transport control &ndash; smears from transport tank and determination of the total number of staphylococci in the tank milk sample; c) bacteriological examination of bulk tank milk in the dairy plant before and after pasteurisation, including examination of cheese curd.</span></p> <p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><span style="font-size: 10.0pt; mso-bidi-font-weight: bold;">After pasteurisation no staphylococci were recorded as in milk as in cheese.</span></p> <p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><span style="font-size: 10.0pt; mso-bidi-font-weight: bold;">Out of 112 strains of Staphylococcus aureus only four strain produced staphylococcal enterotoxins (SE), but in another 7 strains a gene for production of SE, type C was found.</span></p> <p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><span style="font-size: 10.0pt; mso-bidi-font-weight: bold;">The measures introduced during the following season led to the fact that total numbers of coagulase-positive staphylococci in milk within the process of primary production and transport did not exceed the limit permitted by legislation, and after pasteurisation of milk and cheese curd they were not found at all.<span style="mso-spacerun: yes;">&nbsp;</span></span></p><p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;">&nbsp;</p><p class="MsoNormal" style="text-align: justify; border: none; mso-border-bottom-alt: solid windowtext 2.25pt; padding: 0cm; mso-padding-alt: 0cm 0cm 1.0pt 0cm; mso-element: frame; mso-element-frame-width: 498.1pt; mso-element-frame-height: 319.45pt; mso-element-frame-vspace: 3.1pt; mso-element-wrap: around; mso-element-anchor-vertical: page; mso-element-anchor-horizontal: page; mso-element-left: 52.45pt; mso-element-top: 55.2pt; mso-height-rule: exactly;"><span style="font-size: 10.0pt; mso-bidi-font-weight: bold;"><span style="mso-spacerun: yes;"><span style="font-size: 12px;"><strong>doi:10.5219/58</strong></span></span></span></p></div></td></tr></tbody></table></div