Relationship between oxidative stress parameters and lipid status of cows with subclinical mastitis caused by bacteria Staphylococcus aureus

Iako su mastitisi veoma aktuelna tema istraživanja, oni su i dalje jedno od najvažnijih oboljenja mlečnih krava koje prouzrokuje velike ekonomske gubitke u govedarskoj proizvodnji širom sveta. Tokom mastitisa smanjena je produkcija mleka, visoki su troškovi lečenja, mleko se odbacuje zbog upotrebe antimikrobnih preparata, a krave se često prevremeno isključuju iz proizvodnje. Zbog svojih fenotipskih i genotipskih osobina, Staphylococcus aureus ima veliki značaj u etiologiji oboljenja mlečne žlezde. Infekcije mlečne žlezde bakterijom S. aureus uglavnom su po toku supkliničke, sa povećanim brojem somatskih ćelija (eng. somatic cell count - SCC), bez vidljivih promena u mleku i na vimenu, teško se otkrivaju i imaju veoma nisku stopu izlečenja. Jednom inficirana životinja može izlučivati uzročnika infekcije tokom više laktacija, a i nakon oporavka četvrti inficiranih bakterijom S. aureus postoji velika mogućnost reinfekcije. Nakon što patogeni mikroorganizmi prodru u mlečnu žlezdu, bakterijski toksini, enzimi i komponente ćelijskog zida direktno utiču na funkciju epitelnih ćelija mlečne žlezde, ali takođe stimulišu brojne medijatore inflamacije. Zahvaljujući promenama u vaskularnom endotelu, polimorfonuklearni leukociti, kao što su neutrofili i makrofage, migriraju ka žarištu infekcije i infiltriraju mesto trauma, čime se značajno povećava SCC u mleku. Smatra se da neutrofilni granulociti doprinose eliminaciji bakterija, pre svega fagocitozom i intracelularnim ubijanjem, kao i oslobađanjem reaktivnih kiseoničnih vrsta (eng. reactive oxygen species - ROS) i antimikrobnih polipeptida. Nastale ROS oštećuju bakterije, ali ukoliko njihov nastanak prevaziđe mogućnost antioksidativne zaštite organizma da ih neutrališe nastaje oksidativni stres i, u zavisnosti od dužine i težine zapaljenskog procesa, manja ili veća oštećenje ćelija i tkiva domaćina. Detaljnije razumevanje veze između oksido-redukcionog statusa organizma i pojave mastitisa krava moglo bi dovesti do razvoja efikasnijih strategija u prevenciji ovog oboljenja. Ispitivanje parametara oksidativnog statusa krava sa mastitisom može doprineti boljem poznavanju patofiziologije mastitisa i ranom otkrivanju bolesti kako bi se blagovremeno započela terapija, smanjilo oštećenje vimena i pad proizvodnje mleka, a samim tim i ekonomski gubitci..Despite being frequently targeted in research, mastitis in cows has globally been contributing to high economic losses in dairy production. During mastitis, milk production is reduced, there are huge costs of therapy, milk discharge due to antibiotic use and premature culling of cows. Due to its phenotypic and genotypic characteristics, Staphylococcus aureus is of great importance in the etiology of mammary gland diseases. S. aureus infections of the mammary gland are mostly subclinical, with an increased somatic cell count (SCC), without visible changes in the milk and udder. They are difficult to detect and have a very low cure rate. Infected animals can excrete the pathogen during multiple lactations, and even after the recovery, there is a high possibility of reinfection with S. aureus. On the penetration of pathogenic microorganisms into the mammary gland, bacterial toxins, enzymes and cell wall components have a direct impact on the function of mammary gland epithelial cells, but they also stimulate numerous mediators of inflammation. Owing to the changes in the vascular endothelium polymorphonuclear leukocytes such as neutrophils and macrophages migrate to the site of infection and infiltrate the site of trauma, significantly increasing SCC in milk. Neutrophils are thought to contribute to the elimination of bacteria, primarily by phagocytosis and intracellular killing, as well as the release of reactive oxygen species (ROS) and antimicrobial polypeptides. The resulting ROS are harmful to bacteria, but if their occurrence exceeds the possibility of neutralization by the body's antioxidant protection, oxidative stress occurs and depending on the length and severity of the inflammatory process, damage to host cells and tissues. A more detailed understanding of the connection between the oxidation-reduction status of the organism and the occurrence of mastitis could lead to the development of more effective strategies in the prevention of this disease. Evaluation of the oxidative status parameters of cows with mastitis can contribute to a better knowledge of the pathophysiology of mastitis and early detection of the disease, enable the beginning of therapy in time, reduce udder damage and decline in milk production, and thus economic losses..

Mukometra kao posledica ozbiljne stručne greške

An apathetic 14-year-old Hungarian Puli bitch was presented with a seven day history of vaginal discharge and reported ovariohysterectomy (OHE) conducted six years ago. Clinical examination showed elevated body temperature (39.5°C), enlarged abdomen, and vulvar swelling. Despite a reported OHE in the anamnesis, ultrasound examination of the abdomen demonstrated the presence of both ovaries and a large anechogenic zone resembling the uterus. Vaginal cytologic examination and hormonal analysis indicated that the bitch was in the oestrus phase of the cycle. After laparotomy, both intact ovaries and an enlarged uterus with marked adhesions and filled with liquid content were visualised. The cervix was situated in the caudal part of the abdominal cavity and connected to the uterus by only an adhesion stump. After extirpation, uterus and ovarian tissue samples were sent for histological examination. Histological findings of uterus and ovarian tissue samples indicated a diagnosis of mucometra. Obviously, OHE was not conducted professionally, and the procedure performed only prevented conception, but ovaries and uterus continued to be active and caused a serious health disorder after six years. When only a part of the ovaries or uterus is left after OHE, health complications can appear up to ten years later. Our case testifies that even if both ovaries and the entire uterus are left after OHE without communication with cervix and vagina, a bitch can live without noticeable health disorders up to six years. This underlines the importance of lymphatic drainage and resorption processes in the uterus as well as evacuation of uterus content through the vagina.Četrnaest godina stara kuja rase mađarski puli je dovedena u apatičnom stanju i sa anamnezom iscedka iz vagine koji je prisutan 7 dana. Prema rečima vlasnika ovariohisterektomija je urađena pre 6 godina. Kliničkim nalazom je utvrđena povišena telesna temperatura (39,5˚C) uvećan abdomen i otok vulve. Uprkos podacima iz anamneze ultrazvučni pregled je ukazivao na prisustvo oba jajnika i velike anehogene zone koja je ukazivala na prisustvo materice. Citološki nalaz brisa vagine i hormonska analiza su ukazivali na to da je kuja bila u estralnoj fazi ciklusa. Nakon laparotomije potvrđeno je prisustvo oba jajnika i uvećane materice koja je bila ispunjena tečnim sadržajem i sa velikim brojem adhezija. Cerviks je bio lociran u kaudalnom delu abdomena i pričvršćen za uterus samo adhezionim patrljkom. Nakon ekstirpacije uterus i ovarijumi su poslati na histološku analizu, kojom je utvrđena dijagnoza mukometre. Očigledno da prethodno urađena ovariohisterektomija nije bila obavljena stručno i da je urađeni postupak sprečio koncepciju ali su ovarijumi i materica nastavili da budu aktivni što je prouzrokovalo ozbiljan zdravstveni problem 6 godina kasnije. Kada nakon ovariohisterektomije ostanu samo delovi tkiva ovarijuma i uterusa zdravstvene komplikacije se mogu javiti i do 10 godina nakon operacije. Prikazani slučaj svedoči o tome da je, čak i u slučaju kada su nakon operacije ostavljeni oba ovarijuma i uterus bez komunikacije sa cerviksom i vaginom, kuja 6 godina živela bez ozbiljnijih zdravstvenih problema. Ovo naglašava značaj limfne drenaže i procesa resorpcije sadržaja iz materice, pored pražnjenja kroz cerviks i vaginu

The Beth Israel synagogue - a work by architect Milan Kapetanović

Predmet članka je sefardska sinagoga Bet Jisrael, koja je od 1908. do 1944. postojala u Beogradu, u Ulici cara Uroša br. 20. Ovu građevinu u mavarskom stilu izveo je građevinski inženjer i preduzimač Viktor Azriel, prema projektu i pod nadzorom arhitekte Milana Kapetanovića. U tekstu je prikazan životni put Milana Kapetanovića, sa posebnim osvrtom na stručnu delatnost, ali je njegov najveći deo posvećen istoriji i arhitekturi sinagoge Bet Jisrael.The Beth Israel Synagogue was located at 20 Cara Urosa Street. Built in 1908, it was designed and its construction monitored by architect Milan Kapetanović (1859-1934, Belgrade). Civil engineer and contractor Victor Azriel (Belgrade, about 1875-1942, Belgrade) was responsible for the construction. Kapetanović finished high school and Technical School in Belgrade and graduated architecture from the Superior Technical School in Munich. Since 1887 until 1905, he taught descriptive geometry at the Technical School in Belgrade. Before World War 1, he was minister for the national economy and minister of civil works after the war. Kapetanović designed a number of private and public buildings in Belgrade, including the home of Jevrem Grujic, the Vukanovic residence, the Dorcol elementary school, the Class Lottery House (with engineer Miloš Savković), in the Neo-Renaissance style. Together with architect Milorad Ruvidić, he designed the Serbian Pavilion at the International Exhibition in Paris, in 1900, in Serbo-Byzantine style. The Beth Israel Synagogue has a special place in Kapetanović's body of work. Built in the Maori style, it followed the trend of the European synagogues of the time. The main characteristics of the Beth Israel Synagogue were picturesqueness and polychromy. Decorative elements, fitting harmoniously into the whole, dominated the bichrome facade. Two metal domes dominated the front side of the building. It was well proportioned and in harmony with its surroundings. The synagogue was heavily damaged during World War 2. Today's Gallery of Frescoes was built on its remains.Ovaj rad učestvovao je 2000. godine na 44. Nagradnom konkursu Saveza jevrejskih opština Jugoslavije

Morphological characteristics and expression of estrogen and progesterone receptors in the canine endometrium during the estrus cycle, cystic endometrial hyperplasia and pyometra

The estrus cycle of bitches is divided into four phases: proestrus, estrus, diestrus and anestrus, during which different morphological changes, and also cyclic changes of estrogen and progesterone receptors are present. Several pathological changes can be differentiated on the endometrium, but one of these is the most important - cystic endometrial hyperplasia, which frequently develops into pyometra. The aim of the present study was to describe morphological characteristics, and expression of estrogen and progesterone receptors on the endometrium of mixed-breed bitches during the different phases of the estrus cycle, cystic endometrial hyperplasia and pyometra. The uterus and ovaries of 36 mixed breed bitches in different phases of the estrus cycle and also with cystic endometrial hyperplasia (CEH) and chronic purulent endometritis - pyometra were examined macroscopically, histopathologically, and immunohistochemically for estrogen receptors (ER) and progesterone receptors (PR). During proestrus uterine cells showed a weak reaction for both estrogen and progesterone receptors, but during estrus a large number of uterine cells showed a strong reaction on estrogen receptors and moderate reaction on progesterone receptors. On the contrary, during diestrus the scores for the estrogen receptors decreased, while the progesterone receptors level increased - uterine cells expressed strong reaction for progesterone receptors, and moderate reaction for estrogen receptors. Uterine cells in cystic endometrial hyperplasia expressed a strong reaction for estrogen receptors, and moderate reaction for progesterone receptors, but on the other hand the uterine cells in the uterus with pyometra expressed a moderate to strong reaction for progesterone receptors, and a weak reaction for estrogen receptors. In further investigations it would be interesting to perform quantitative analysis for both estrogen and progesterone receptors during different phases of the estrus cycle and also in the uterus with cystic endometrial hyperplasia and pyometra

Analysis of lactoferin gene polymophism and its association to milk quality and mammary gland health in Holstein-Friesian cows

Lactoferrin (LTF) is a glycoprotein, a member of transferrin gene family which plays an important role in immune mechanisms in the mammary glands of cows. The amount of lactoferrin increases during inflammatory processes and viral infections. The aim of this investigation was to monitor the distribution of lactoferrin gene genotypes and its connection to milk quality and the occurrence of mammary gland diseases in 46 Holstein-Freisian cows of different age (2-7 years) on a farm near Belgrade. DNA was isolated from blood samples, and the polymorphism of lactoferrin gene was deterimined by PCR-RFLP method using the restriction enzyme Eco RI. We found two alelic forms of this gene in cows included in these experiments (A and B) and two genotypes (AA and AB) in a ratio 71.7% to 28.3%. The genotype BB was not found in this sample. In order to determine the degree of differences between genotypes we used discriminant analysis which has shown that there is a statistically significant difference between genotypes AA and with respect to productive parameters. When analyzed separately, the only parameter which differed significantly (p=0.021) between two genotypes was total milk production. Individuals with observed genotypes are most similar for the amount of milk fat (p=0.271). There is no statistically significant difference in the number of somatic cells in milk samples between the examined genotypes. [Projekat Ministarstva nauke Republike Srbije, br. III46002

Investigations of efficacy of intramammary applied antimicrobials and glucocorticosteroides in the treatment of subclinical and clinical mastitis in cows

Inflammation of the mammary gland, mastitis in cows, presents one of the most acute problems in intensive dairy production, inflicting huge economic losses. In the course of one year, 80 samples were taken at investigated farms from udder quarters of cows with clinical mastitis and 160 samples from udder quarters of cows with subclinical mastitis. The efficacy of three preparations, A, B, and C, was examined in the treatment of clinical and subclinical mastitis in cows. The investigations indicate that antibiotic preparation A (neomycin, polimixine B, oleandomycin and prednisolone) exhibited a greater efficacy in the treatment of clinical mastitis caused by Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Micrococcus sp., but a smaller efficacy in the treatment of subclinical mastitis caused by Staphylococcus aureus. Preparation B (amoxicillin, clavulanic acid and prednisolone) exhibited a higher efficacy in the treatment of clinical mastitis caused by Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Micrococcus, but a weaker effect in the treatment of subclinical mastitis caused by Staphylococcus aureus. Preparation C (procaine penicillin G, streptomycin, neomycin sulfate and prednisolone acetate) exihibited efficacy in the treatment of clinical and subclinical mastitis caused by Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Micrococcus, Staphylococcus aureus and Esherichie coli. [Projekat Ministarstva nauke Republike Srbije, br. III 46002: Molekularno-genetička i ekofiziološka istraživanja u zaštiti autohtonih animalnih genetičkih resursa, očuvanju dobrobiti, zdravlja i reprodukcije gajenih životinja i proizvodnji bezbedne hrane

Effects of chestnut tannins supplementation of prepartum moderate yielding dairy cows on metabolic health, antioxidant and colostrum indices

This study investigated the effects of dietary supplementation with chestnut tannins (CNT) on metabolic and antioxidant status of prepartum cows along with their colostrum quality. Pregnant multiparous Holstein cows were paired according to parity and body condition score, and assigned either to a diet supplemented with 20 g/d of commercially available product containing chestnut tannins (CNT, n=20) or to an unsupplemented control diet (CON, n=20) for the last 25±2 d of pregnancy. Serum metabolite, insulin and antioxidant capacity indices were measured in blood samples taken at d 25 and d 5 before expected parturition. Chemical composition and IgG concentration were determined in colostrum samples collected from the first milking postpartum. The addition of CNT led to lower BUN (P=0.02) and consequently higher serum glucose (P=0.02) and insulin (P<0.01) concentrations which were associated with lower circulating NEFA (P<0.01) and BHBA (P<0.01) in CNT group than those of CON. The serum paraoxonase 1 (PON 1) activity and total antioxidant capacity (T-AOC) were higher at-5 d in CNT than in CON (P<0.01, P=0.03; respectively). Close-up CNT improved lactose percentage and IgG concentration (P=0.03, P=0.04; respectively) and tended to improve percentage of protein and SNF (Solid Not Fat) in primary colostrum (P=0.06, respectively), without affecting colostrum fat and total solid (P=0.98, P=0.43; respectively). Supplementation of CNT in the diet during close-up period did not have adverse effects on metabolic profiles prepartum. Instead, this feeding regimen was more beneficial to antioxidant capacity and colostrum quality than feeding the control diet

Dietary Supplementation of Chestnut Tannins in Prepartum Dairy Cows Improves Antioxidant Defense Mechanisms Interacting with Thyroid Status

Cows in the peripartal period undergo changes in thyroid hormones and are susceptible to lipomobilization and/or oxidative stress. The addition of chestnut tannins as polyphenolic compounds in the diet may improve feed efficiency and prevent oxidative stress-related health disorders in transition cows. However, the relationship between chestnut tannin supplementation and thyroid function, which plays an important role in metabolic regulation, has not been investigated in dairy cows. This study was conducted to investigate the effects of chestnut tannin supplementation during the close-up period on thyroid status and to evaluate the interaction between thyroid hormones and oxidative stress biomarkers in prepartum dairy cows. Forty multiparous Holstein cows were fed either a diet containing chestnut tannins (CNTs, n = 20, 1.96 g chestnut tannins/kg feed, dry matter) or a non-supplemented diet (CON, n = 20) during the last 25 ± 2 days of gestation. Blood samples were collected on the first day of study (before chestnut tannin supplementation) and d 5 before parturition to measure hormonal and oxidative stress indices. Serum concentrations of T3 (p = 0.04) and T4 (p = 0.05) were higher in CNT cows than in the CON group on day 5 before parturition. Thyroid status of CNT cows was associated with higher serum total antioxidant capacity (T-AOC, p < 0.01), activities of superoxide dismutase (SOD, p = 0.03) and glutathione peroxidase (GPx, p = 0.01), and reduced glutathione concentration (GSH, p = 0.05). Serum thiobarbituric acid reactive substances (TBARS) were lower (p = 0.04) which was associated with lower aspartate aminotransferase (AST, p = 0.02), and lactate dehydrogenase (LDH, p = 0.01) activities in the CNT than in the CON group. Estradiol and progesterone did not differ between CNT and CON cows. Chestnut tannin supplementation improves antioxidant protection, prevents oxidation-reduction processes, reduces the degree of liver cell membrane damage, and protects thyroid tissue from damage, allowing higher T3 and T4 synthesis. Considering the importance of the thyroid hormone status before parturition, mechanisms of thyroid hormone regulation in CNT-supplemented dairy cows require more detailed investigations

Efficiency evaluation of a bivalent vaccine in the prophylaxis of mastitis in cows

Mastitis in cows represents one of the most actual problems in intensive dairy production. The longtime different approaches to the treatment of mastitis have not offered a suitable solution, and the problem of mastitis is still present and acute. Prevention of pathogen penetration into the mammary gland, its colonization and multiplication impose a constant need for regular inspections of milk, as well as preventive and therapeutic measures to reduce the incidence of mastitis. Studies in the field of vaccination of ruminants against mastitis pathogens suggest a limited success in obtaining significant results in immunoprophylaxis. Considering the results of other researchers and our own research, and bearing in mind the problems mastitis caused by Staphylococcus aureus and Streptococcus agalactiae represent, the aim of this study was the preparation and testing of an indigenous vaccine prepared with two stable strains, SAU 7 (S. aureus) and SAG 3 (Str.agalactiae). The dose was 5 mL/cow and consisted of inactivated bacterial S. aureus SAU 7 cells in a concentration of 1x1010 cfu/mL and Str. agalactiae SAG 3 in a concentration of 4 x 109 cfu/mL. The number of somatic cells in the milk samples during the whole study period was higher in vaccinated cows in both groups compared with the control, but this difference was not statistically significant. The concentration of immunoglobulin IgG in the milk of vaccinated cows was significantly higher than the concentration of this class of proteins in the milk of unvaccinated cows. [Projekat Ministarstva nauke Republike Srbije, br. III 46002

Izolacija Candide albicans i Citrobacter freundii iz duboko zamrznutog semena bika

Microbiological examination of semen as part of health control of bulls is of great importance for the success cattle reproduction. Deep-frozen semen ofbulls should be free of pathogenic and conditionally pathogenic microorganisms and controlled production of seed should allow out. According to the OIE recommendations unfrozen dose of semen should not contain more than 5000 cfu / ml of saprophytic microorganisms. According to the Law of Veterinary Medicine of Serbia, Article 115 (Traffic reproductive material) it is prohibited to trade, import and export of seeds for artificial insemination, ova and fertilized eggs containing pathogens of animals or more bacteria than allowed, or that their biochemical, biophysical and morphological properties are not eligible for reproduction. Clear limits of number and types of allowed saprophytic microorganisms in deep- frozen semen is not listed. Microbiological analysis included a sample of 351 semen ofbulls in pure culture was isolated Candida albicans in 9 samples, Citrobacter freundii in 5 samples, while other samples were free of microorganisms. Infection with Candida albicans are well described in the literature, and it is known that can cause mastitis, endometritis and abortion in cows. Citrobacter freundii is known as a cause of nosocomial infection of respiratory, urinary tract and blood in humans. It was stated that 29% of opportunistic infections in humans are caused by these microorganisms. In veterinary medicine, infection with Citrobacter freundii have been described in dogs, laboratory animals, fish and turtles. It is known that Citrobacter species can cause abortions and mastitis in cows, but literature data on the effect of pathogen Citrobacter freundii on cows reproductive health are low. Isolation of this organism from the semen ofbulls has not been described in the available literature. On the basis of our recommendations and OIE Veterinary Act, and the little available literature on infections (especially C freundii) it is recommended that the deep-frozen bull semen is unusable for arteficial insemination.Mikrobiološki pregled semena kao deo zdravstvene kontrole bikova od velikog je zna;aja za uspeh reprodukcije goveda. Duboko zamrznuto seme bikova treba da bude slobodno od patogenih, kao i od uslovno patogenih mikroorganizama, što kontrolisana proizvodnja semena treba da omogući. Prema OIE preporukama otopljena doza semena ne treba da sadrži više od 5000 cfu/ml saprofitskih mikroorganizama. Prema Zakonu o veterinarstvu Republike Srbije, članu 115 (Promet reproduktivnog materijala) “Zabranjen je promet, uvoz ili izvoz semena za veštačko osemenjavanje, jajnih ćelija i oplođenih jajnih ćelija koji sadrži uzročnike bolesti životinja ili veći broj bakterija od dozvoljenog ili koji svojim biohemijskim, biofizičkim i morfološkim svojstvima ne ispunjavaju uslove za reprodukciju”. Jasna granica broja i vrste dozvoljenih saprofitskih mikroorganizama u duboko zamrznutom semenu nije navedena. Mikrobiološkom analizom obuhvaćen je 351 uzorak duboko zamrznutog semena bikova, u čistoj kulturi je izolovana Candida albicans u 9 uzorka, Citrobacter freundii u 5 uzoraka, dok su ostali uzorci bili slobodni od mikroorganizama. Infekcije krava sa Candida albicans su dosta opisane u literaturi, i poznato je da može da izazove mastitise, endometritise i pobačaje kod krava. Citrobacter freundii je poznat kao uzročnik nozokomijalnih infekcija respiratornog, urinarnog trakta i krvi ljudi. Navedeno je da je 29% oportunističkih infekcija kod ljudi izazvano ovim mikroorganizmom. U veterinarskoj medicini infekcije sa Citrobacter freundii su opisane kod pasa, laboratorijskih životinja, riba i kornjača. Poznato je da Citrobacter vrste mogu izazvati mastitise i pobačaje krava, ali literaturni podaci o patogenom delovanju Citrobacter freundii na reproduktivno zdravlje krava su oskudni.Izolacija ovog mikroorganizma iz duboko zamrznutog semena bikova nije opisana u nama dostupnoj literaturi. Na osnovu preporuke OIE i našeg Zakona o veterinarstvu, a zbog malo dostupne literature o infekcijama (posebno C. freundii) data je preporuka, da je duboko zamrznuto seme bika neupotrebljivo za veštačko osemenjavanje