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Abstract

This study investigates the effectiveness of intra-mammary ozone administration in the dry period and at the time of delivery for preventing against mastitis in herds with contagious mastitis. The cows were divided into five groups with 10 cows in each. Group 1 was administered an ozone-containing foam preparation via the teat canal into four udder quarters for 5 seconds at the beginning of the dry period; Group 2 was administered ozone at the beginning of the dry period and at the time of delivery; Group 3 was administered ozone at the time of delivery; Group 4 was administered a dry period udder preparation at the beginning of the dry period; and Group 5 was administered only teat seal at the beginning of the dry period. No statistically significant difference was found between the cows with regard to the SCC values at the beginning of the dry period and at the time of delivery (in cows without clinical mastitis, n=25). The SCC values were reported to decrease when the values at the beginning of the dry period and at the time of delivery were compared. All cows except two in Group 1 were detected to have clinical mastitis according to the frequency of microbial isolation in milk at the time of delivery. In conclusion, intra-mammary ozone administration did not prevent mastitis in the dry period or at the time of delivery in herds with contagious mastitis; moreover, it was determined to increase the rate of clinical mastitis in the postpartum period.

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Authors and Affiliations

A. Koseman
I. Seker
A. Risvanli
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Abstract

Great economic losses to the dairy industry are associated with bovine mastitis, which results in poor milk quality and high treatment costs. Anti-inflammatory proteins play an important role in the suppression of the immune response against invading pathogenic microorganisms and are therefore being studied for possible use in the early diagnosis of mastitis. In our study, we used milk samples from 15 cows of Holstein Friesian breed with different health status (5 healthy, 5 subclinical, and 5 clinical animals), and tested them using immunohistochemical (IHC) analysis to evaluate the presence of IL-2, IL-10, TGF-β1, βDEF-2, DEF-3, and Cathelicidin LL37 proteins. The calculation of positively and negatively stained cells for each biomarker was performed using the semiquantitative counting method. We found the presence of all factors with the exception of Cathelicidin LL37, which was almost absent in milk samples of all animal groups. The significant decrease of IL-10, β-def2, and β-def3 expression levels within the 3 days of sampling, found in the milk of animals with sub- and clinical mastitis, indicates the loss of antiinflammatory protection of the affected cow’s udder. In contrast, the stable increase of IL-2 and TGF-β1 positive cells observed in the milk of mastitis-affected cows, and the similar expression of these factors in the milk of healthy animals, indicate the possible lack of involvement of these cytokines at an early stage of udder inflammation.
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Authors and Affiliations

K. Šerstņova
1
M. Pilmane
1
Z. Vitenberga-Verza
1
I. Melderis
1
Ł. Gontar
2
M. Kochański
2
A. Drutowska
2
G. Maróti
3 4
B. Prieto-Simón
5 6

  1. The Institute of Anatomy and Anthropology, RSU, 9 Kronvalda bulvaris, Riga, Latvia
  2. Research and Innovation Centre Pro-Akademia, Innowacyjna 9/11, 95-050 Konstantynow Lodzki, Poland
  3. Seqomics Biotechnology Ltd., Morahalom, Vallalkozak utja 7, Hungary
  4. Biological Research Center, Plant Biology Institute, Szeged, Temesvári krt. 62, Hungary
  5. Department of Electronic Engineering, Universitat Rovira i Virgili, C. de l’Escorxador, 43007 Tarragona, Spain
  6. ICREA, Pg. Lluís Companys 23, Barcelona, Spain
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Abstract

The aim of the present study was to investigate inline lactate dehydrogenase (LDH) dynamic changes based on different cow factors – different number and stages of lactation, milk yield, and the status of reproduction in clinically healthy dairy cows.

In the Herd Navigator system, LDH activity levels (μmol/min per litre) were measured using dry-stick technology. A total of 378 cows were selected. According to their reproductive status, the cows were classified as belonging to the following groups: Fresh (1 – 44 days after calving); Open (45 – 65 days after calving); Inseminated (1 – 35 days after insemination); Pregnant (35 – 60 days after insemination and pregnant). According to their productivity, the cows were classified into the following groups: <15 kg/day, 15 – 25 kg/day, 25 – 35 kg/day and >35 kg/day. The cows were milked with a DeLaval milking robot (DeLaval Inc. Tumba Sweden) in combination with a Herd Navigator analyser (Lattec I/S. Hillerød Denmark).

In conclusion inline dynamic changes in the milk LDH concentration may increase together with the rise in the lactation period frequency. The highest LDH level determinated in the group of the fresh cows ranged from 5 to 10 DIM, while the highest LDH concentration level was found in the fresh cow milk. Thus, there was a positive relationship between the milk concentration of LDH and the milk yield.

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Authors and Affiliations

R. Antanaitis
D. Malašauskienė
M. Televičius
V. Juozaitienė
A. Rutkauskas
G. Palubinskas
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Abstract

The aim of this study was to compare the effect of controlled-release monensin on the automatic registered body condition score (BCS), and biomarkers registered using a fully automated inline analyzer, such as milk β-hydroxybutyrate (BHB), milk yield (MY) and milk lactate dehydrogenase (LDH).
Two experimental groups were formed: (1) monensin group (GK) supplemented with monensin (a monensin controlled release capsule (MCRC) of 32.4 g, n = 42) and (2) control group (GO) (capsule containing no monensin, n = 42). Treatment began 21 days before calving, and the experiment was finished one month after calving. In order to gather data about MY, BHB, and LDH, Herd Navigator a real-time analyzer (Lattec I/S, Hillerød, Denmark) was used together with a DeLaval milking robot (DeLaval Inc., Tumba, Sweden). BCS was measured using 3D BCS cameras (DeLaval, DeLaval International AB). All data were registered at one, 15 and 30 days after calving. The statistical analysis was performed using SPSS 26.0 (SPSS Inc., Chicago, USA) package. It was concluded that in the group of cows with monensin supplement (a monensin controlled release capsule of 32.4 g,), the body condition score was statistically significantly higher at the 15th (+0.24, p=0.003) and 30th (+0.52, p<0.001) days after calving, the productivity of cows in this group increased by 10.25% from the 1st to the 15th day and by 22.49% from the beginning of the experiment to the 30th day (p<0.001), lactate dehydrogenase activities at the 15th and 30th days after calving in this group were lower (p<0.001), and also in this group, the number of cows with a value of β-hydroxybutyrate of 0.06 mmol/L decreased from the beginning of the experiment to 30 days after calving by 4.70% (from 19.00% to 14.30%) compared with the control group.
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Authors and Affiliations

M. Urbutis
1
V. Juozaitienė
2
G. Palubinskas
3
K. Džermeikaitė
1
D. Bačėninaitė
1
R. Bilskis
4
W. Baumgartner
5
R. Antanaitis
1

  1. Large Animal Clinic, Veterinary Academy, Lithuanian University of Health Sciences, Tilžės St. 18, LT-47181 Kaunas, Lithuania
  2. Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, K. Donelaičio St. 58, LT-47181, Kaunas, Lithuania
  3. Department of Animal Breeding, Veterinary Academy, Lithuanian University of Health Sciences,Tilžės 18, LT-47181 Kaunas, Lithuania
  4. Animal Husbandry Selections, Breeding Values and Dissemination Center, Agriculture Academy, Vytautas Magnus University, Universiteto St. 10A, Akademija, Lt-53361 Kaunas Distr., Lithuania
  5. University Clinic for Ruminants, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria
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Abstract

The aim of this work was to evaluate the relative gene expression levels of the cytokines IL- 1B, IL-8, IL-12, IFN-γ, IL-4, IL-10 and TGF-β in somatic milk cells of French Alpine breed, anestrous goats that were experimentally infected in the left mammary gland with Staphylococcus chromogenes during the lactation peak. Milk samples were obtained from both glands for 21 consecutive days post infection. Total RNA was extracted, and real-time PCR was conducted using primers specific to each cytokine. The relative RNA expression of the evaluated cytokines was determined by the comparative method 2-ΔΔCT, using milk from the right gland of the goats as a reference (control) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous control. According to the Wilcoxon test results, IL-1B and IL-12 expression levels showed significant differences compared to those in the control group (p<0.05) from 24 hours post infection until the end of lactation; on day three, IL1β, IL8, IL12 and TGF-β had a statistically significant change in expression with respect to those in the control group (p<0.05); closer to the end of the lactation period, there is no overexpression of the anti-inflammatory interleukins (IL-4 and TGF-β) which may reflect the effort of the host immune system to eradicate the microorganism from the mammary gland.

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Authors and Affiliations

R.A. Ruiz-Romero
D. Martínez-Gómez
R.A. Cervantes-Olivares
E. Díaz-Aparicio
A.E. Ducoing-Watty
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Abstract

This study aimed to identify bacterial pathogens in milk samples from dairy cows with subclinical and clinical mastitis as well as to assess the concentrations of oxidant-antioxidant parameters [malondialdehyde (MDA), reduced glutathione (GSH), and total GSH levels] in both blood and milk samples. From a total of 200 dairy cows in 8 farms, 800 quarter milk samples obtained from each udder were tested in the laboratory for the presence of udder pathogens. Cultivated bacteria causing intramammary infection from milk samples were identified by Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF). In addition, from tested animals 60 cows were selected includıng 20 healthy cows that were CMT negative, 20 cows with subclinical mastitis (SM), and 20 cows with clinical mastitis (CM) for detection of MDA, GSH, and total GSH levels in blood and milk samples. Three hundred and eighty (47.5%; 380/800), 300 (37.5%; 300/800), and 120 (15%; 120/800) of milk samples, respectively were CMT positive or SM and CM, and those positives were cows from different farms. We observed that 87.4% (332/380), 25.3% (76/300), and 34.2% (41/120) of cows with CMT positive, CMT negative, and CM had bacterial growth. The most predominantly identified bacteria were Staphylococcus chromogenes (18.7%) obtained mainly from SM and Staphylococcus aureus (16.7%) as the most frequent cause of CM. According to our results, dairy cows with CM had the highest MDA levels, the lowest GSH, and total GSH levels in both blood and milk samples however, high MDA levels and low GSH levels in milk samples with SM were observed. Based on our results, lipid oxidant MDA and antioxidant GSH could be excellent biomarkers of cow’s milk for developing inflammation of the mammary gland. In addition, there was no link between nutrition and MDA and GSH levels.
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Authors and Affiliations

G. Ozbey
1
Z. Cambay
1
S. Yilmaz
2
O. Aytekin
1
F. Zigo
3
M. Ozçelik
1
B. Otlu
4

  1. Department of Medical Services and Techniques, Vocational School of Health Services, Firat University, Rectorate Campus, 23119, Elazig, Turkey
  2. Department of Biochemistry, Faculty of Veterinary Medicine, Firat University, Rectorate Campus, 23119, Elazig, Turkey
  3. Department of Nutrition and Animal Husbandry, University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, Košice, 04001, Slovakia
  4. Department of Medical Microbiology, Faculty of Medicine, Inonu University, Main Campus, 44280, Battalgazi, Malatya, Turkey
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Abstract

In a dairy cow herd consisted of herd of 200 lactating Holstein-Friesian cows and heifers, clinical signs of mastitis in 40 out of 170 animals were observed. Treatments with antibiotics were proved ineffective. Milk bacterial cultures from 15 affected animals revealed Pseudomonas aeruginosa. An autogenous vaccine was administered subcutaneously, twice in a month period, to all adults. Cases of clinical mastitis declined significantly (p<0.0001) during next 3 months.
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Authors and Affiliations

E.J. Petridou
1
I.A. Fragkou
2
S.Q. Lafi
3
N.D. Giadinis
4

  1. Laboratory of Microbiology and Infectious Diseases, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Greece
  2. Veterinary Service, Ministry of Rural Development and Food, Athens, Greece
  3. Department of Pathology and Animal Health, Faculty of Veterinary Medicine, Jordan University of Science and Technology, P.O. Box 3030, Irbid, Jordan
  4. Clinic of Farm Animals, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Greece
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Abstract

This study aimed to determine the levels of milk cell total protein (TP), reduced nicotinamide adenine dinucleotide phosphate (NADPH), total glutathione (tGSH), activities of glucose-6-phos- phate dehydrogenase (G6PD) and glutathione peroxidase (GPx) in subclinical mastitic cows. Milk from each udder was collected and grouped by the California Mastitis Test. Then, a somatic cell count (SCC) was performed, and the groups were re-scored as control (5–87 × 103 cells), 1st group (154–381 × 103 cells), 2nd group (418–851 × 103 cells), 3rd group (914–1958 × 103 cells), and 4th group (2275–8528 × 103 cells). Milk cell TP, NADPH, tGSH levels, G6PD, and GPx ac- tivities were assessed. Microbiological diagnosis and aerobic mesophyle general organism (AMG, cfu/g) were also conducted. In mastitic milk, TP, NADPH, and tGSH levels, and G6PD and GPx activities were significantly reduced per cell (in samples of 106 cells). In addition, milk SCC was positively correlated with AMG (r=0.561, p<0.001), NADPH (r=0.380, p<0.01), TP (r=0.347, p<0.01) and G6PD (r=0.540, p<0.001). There was also positive correlation between NADPH (r=0.428, p<0.01), TP (r=0.638, p<0.001) and AMG. NADPH was positively correlated with TP (r=0.239, p<0.05), GPx (r=0.265, p<0.05) and G6PD (r=0.248, p=0.056). Total protein was positively correlated with tGSH (r=0.354, p<0.01) and G6PD (r=0.643, p<0.001). There was a negative correlation between tGSH and GPx activity (r=-0.306, p<0.05). The microbiological analysis showed the following ratio of pathogens: Coagulase-Negative Staphylococci 66.6%, Streptococcus spp 9.5%, Bacillus spp 9.5%, yeast 4.8%, and mixed infections 9.5%.

As a conclusion, when evaluating the enzyme and oxidative stress parameters in milk, it is more suitable to assign values based on cell count rather than ml of milk. The linear correlation between the SCC and AMG, milk cell NADPH, TP and G6PD suggests that these parameters could be used as markers of mastitis.

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Authors and Affiliations

P.P. Akalin
Y. Ergün
N. Başpinar
G. Doğruer
A. Küçükgül
Z. Cantekin
M. İşgör
M. Saribay
E. Koldaş
A. Baştan
S. Salar
S. Pehlivanlar
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Abstract

Mastitis is one of the most crucial diseases of dairy animals. Especially subclinical mastitis (SCM) has negative impacts on of dairy economy in term of reducing milk quality and quantity also premature culling and cost of therapy. Staphylococci are important etiological agents in SCM. The aim of the study was to investigate the biofilm production and antibiotic resistance profiles of Staphylococcus spp. other than S. aureus isolated from milks of Anatolian water buffalo with subclinical mastitis. Twenty-two coagulase negative staphylococci (CNS) identified phenotypically were also identified with PCR as Staphylococcus spp. other than S. aureus. Biofilm productions were investigated both by Congo Red Agar Method and PCR. The antibiotic resistance profiles of the isolates were determined by Disc Diffusion Method and they were antibiotyped. Only three (13.6%) isolates were biofilm positive both phenotypically and genotypically. All isolates except for two were resistant against at least two antibiotics. Multidrug-resistance among the isolates was low (13.6%). Antibiotyping results showed that the similarities among the strains were between 30-100%. Genotyping of the strains revealed that a genetic heterogeneity was found among CNS isolates and their similarities were between 43% and 93%. In conclusion, CNS isolates identified as subclinical mastitis agents in buffaloes showed a high antibiotic resistance profile especially against oxacillin and vancomycin. Further studies should be conducted to investigate new mechanisms and/or genes responsible for antibiotic resistance in buffaloes.
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Authors and Affiliations

H. Gurler
1
A. Findik
2
M.G. Sezener
2

  1. Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, University of Ondokuz Mayis, Samsun, Turkey
  2. Department of Microbiology, Faculty of Veterinary Medicine, University of Ondokuz Mayis, Samsun, Turkey
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Abstract

The aim of this study was to compare the effect of parenteral and oral supplementation of Selenium (Se) and vitamin E (VTE) on selected antioxidant parameters in blood and colostrum as well as their effect on the incidence of mastitis in dairy cows during the final phase of gravidity (6 weeks) and first two weeks after calving. For the practical part of the study 36 dairy cows of Slovak pied breed in the second to fourth lactation-gestation cycle were selected. The animals weredivided into three groups: the control (C) and two experimental groups (D1 and D2). The selected groups were treated as follows: in group D1 products containing Se (Selevit inj.) and vitamin E (Erevit sol. inj.) were administered intramuscularly twice, six and three weeks prior to parturition; in group D2 a vitamin-minerals supplement in the form of sodium selenite (Na2SeO3) and dl-α-tocopherol acetate were supplemented orally for six weeks calving. The blood samples were collected from the vena jugularis in dairy cows approximately 42 days before calving (control sampling), on parturition day, and the 14th day after calving. Higher concentrations of Se and VTE were found in the blood plasma samples of both experimental groups collected on the day of parturition. In addition, the orally supplemented group (D2) showed higher Se and α-tocopherol concentrations in blood plasma on the14th day after calving as well a reduction of occurrence of mastitis by about 25 % compared to the control group. The relationship between inflammatory response and oxidative stress was also confirmed. The concentrations of milk malondialdehyde indicating lipid peroxidation during mastitis were significantly higher in milk samples from infected cows than in milk samples from healthy animals in each monitored group. In order to prevent oxidative stress and moderate inflammatory response in dairy cows it is very important to optimally balance their nutritive needs with an appropriate ratio of Se and VTE supplements. Therefore we still recommend supplementation of the cows’ postpartum dietwith 0.5 mg of Se/kg dry matter (DM) and 102 mg of dl-α-tocopherol acetate/kg DM to stabilize their optimal blood levels, stimulate the activity of glutathione peroxidase and reduce the incidence of mastitis.
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Authors and Affiliations

M. Vasiľ
1
F. Zigo
1
Z. Farkašová
1
E. Pecka-Kielb
2
J. Bujok
2
J. Illek
3

  1. Department of Nutrition and Animal Husbandry, University of Veterinary Medicine and Pharmacy in Kosice, Komenského 73, Košice, 04001, Slovakia
  2. Department of Biostructure and Animal Physiology, Wroclaw University of Environmental and Life Sciences, Norwida 31, 50-375 Wroclaw, Poland
  3. Large Animal Clinical Laboratory, Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences, Palackého tř. 1946/1, 612 42 Brno, Czech Republic

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