Temos presenciado um crescimento expressivo da ovinocultura de corte nos últimos anos. Algumas doenças de grande importância econômica tem sido motivo de estudos, não apenas pelo impacto econômico que representam, mas também para a saúde do animal e do homem. Com a crescente preocupação com resíduos de medicamentos pesticidas nos produtos de origem animal, perda da qualidade do leite e conseqüente baixo ganho de peso dos borregos de corte, os sistemas orgânicos de produção vem ganhando espaço. Sabe-se que a mastite subclínica é uma das responsáveis pelo baixo rendimento de carcaça na ovinocultura de corte. Neste experimento, optou-se por realizar tratamento com medicamento homeopático (Phytolaca decandra) das ovelhas com tetos diagnosticados com CMT 2+ e 3+, sem sinais de mastite clínica. Dois lotes de ovelhas com mastite subclínica foram usados. Um lote controle que recebeu placebo e o lote tratado que recebeu remédio homeopático duas vezes ao dia junto ao concentrado a partir da quarta semana do parto (de lactação). Foram colhidas duas amostras de cada tetos com mastite subclínica no início do experimento (30 dias do parto) e a cada 15 dias até o desmame quando os borregos tinham aproximadamente 60 - 65 dias de vida. Das amostras colhidas, uma seguiu para identificação do agente microbiológico e outra para a contagem de células somáticas (CCS). Ao final do estudo não houve diferença estatisticamente significativa entre as CCSs da secreção láctea quando comparadas antes e depois do tratamento homeopático no mesmo grupo, bem como quando comparou-se ambos os grupos (tratados e placebo). Porém observou- se que o grupo tratados apresentou redução significativa (P<0,05) nos isolados das amostras com Staphylococcus spp. e aumento significativo de amostras sem crescimento bacterianos (negativas) e ganho de peso estatisticamente significante (P<0,05) quando comparado ao grupo placebo. / We have seen an expressive growth of lamb production in the last years. Some diseases of economic importance have been studied not just by its economic impact but also for the animal and human health. With worry about pesticides and drugs\' remaining (left over) in animal origin products, the loss of milk quality and consequent lamb\'s low gain weight, the organic production systems are growing up. It\'s known that subclinical mastitis is one of the most dangerous and one of those held responsible for the decrease of weight-gain in lamb breeders. In this experiment, it was chosen to use homeopathic medication for ewes whose half udder was diagnosed 2+ and 3+ on CMT (California Mastite Test) without signs of clínical mastitis. Two groups of subclinical mastitis ewes were used. The first one received placebo BID (no treated group) and the second one received homeopathic medication (Phytolaca decandra) BID (treated group) in concentrated food since the 4Th week of lambing. Two milk samples were taken from each injured gland 30 days after lambing and each 15 days until 60-65 days after lambing, when the lambs were weaning. One of the milk samples went to microbiological identification and the other one to SCC (somatic cell count). By the end of the research, neither was it found statistical difference between samples of milk before and after homeopathic treatment from the three milk sampling in the same group nor did it happens when the groups treated and placebo were compared. However, it was observed that the treated group showed significant decrease (P<0,05) samples with Staphylococcus spp. and significant increase of samples without bacteriological growth as well as gain of weight statistically significant (P<0,05) when compared with the placebo groups.

Bovine mastitis remains one of the costliest diseases affecting the dairy industry. Individual susceptibility to mastitis and severity of infection varies between animals and can only be partially explained by genetics. As such, understanding how genetic predisposition coordinately interacts with epigenetic modifications and environmental exposures is necessary to bridge the gap in missing heritability. The role of DNA methylation in regulating the response to bacterial lipopolysaccharide (LPS) was first determined by performing reduced representation bisulfite sequencing on fibroblasts isolated from heifers at 5- and 16-months of age that exhibit an age-dependent up-regulation in LPS-responsiveness. More than 14,000 differentially methylated sites were identified between the two sets of cultures with a trend towards decreased methylation with age. Young cultures were also hyper-methylated in gene promoters regulated by NF-κB and exhibited lower expression in genes that regulate the innate immune response, suggesting that methylation contributes to gene regulation in fibroblast innate response. Previously, TLR4 expression was shown to differ in the age-dependent fibroblast model, however, it was not known if variation in TLR4 expression would affect mastitis severity. Therefore, fibroblasts were isolated from sixty lactating, adult Holstein cows and their expression of TLR4, along with LPS-induced production of IL-8 and IL-6, was used to rank the animals from high to low. Six high responders and six low responders were then experimentally infected in one mammary gland with E. coli. Overall, severity of mastitis was quite variable, with a few notable differences between high and low responders. High responding animals had an earlier increase in somatic cell count and febrile response that coincided with more efficient bacterial clearance. However, tissue damage and milk production did not differ between the two groups, indicating that while rapid up-regulation of the innate response addresses bacterial clearance, subsequent down-regulation is required to alleviate damage within the mammary gland. Finally, one-week old bull calves were subjected to treatment with either saline or LPS to determine if neonatal exposure to endotoxin would make calves less responsive to a second LPS challenge at 32-days of age. The initial treatment showed a large effect of LPS as measured by higher plasma IL-6 and TNF-α concentrations in calves treated with LPS over saline. Subsequent treatment of all 10 calves with LPS showed a very similar response between the two treatment groups and significant inter-animal variability in clinical response. Fibroblasts and monocyte-derived-macrophages (MDMs) were also isolated following initial treatment to determine if any changes occurred at the cellular level as a result of LPS exposure. Fibroblasts isolated from calves at 20-days of age had a very low response to LPS that did not differ between the early life treatments. MDMs isolated from calves at 28-days of age were more responsive to LPS, but again no differences were detected between the early life treatments. In summary, our results suggest that DNA methylation likely plays a role in the cellular response to LPS and may partially contribute to differences between animals in severity of E. coli mastitis, however, the appropriate in vitro phenotype to detect susceptible animals still needs to be characterized before epigenetic biomarkers can be identified, and perhaps modified by environmental interventions.

Mastitis, the inflammation of the mammary gland by bacterial infection, is one of the costliest diseases to the dairy industry primarily due to a loss in milk production. The aim of this study was to investigate the mechanisms underlying reduced milk production during mastitis. We hypothesized that bacterial endotoxin induces cell apoptosis, oxidative stress and increases hypoxia while inhibiting milk gene expression in the mammary gland. To test this hypothesis, mice were bred to pregnancy, and 3 days post-partum the left and right sides of the 4th pair of mammary glands were alternately injected with either the endotoxin liposaccharide (LPS, E. coli 055:B5, 100 ul of 0.2 mg/ml) or sterile PBS through the teat meatus. At 10.5 and 22.5 h post-injection, pimonidazole HCl, a hypoxyprobe, was injected intraperitoneally. At 12 or 24 h after the LPS injection, the fourth glands were individually collected (n=8 pairs) and analyzed for hypoxia, gene expression and oxidative stress. LPS treatment induced mammary gland inflammation as shown by increases in inflammatory cytokine expression (P < 0.001) and neutrophil recruitment at 12 and 24 h. LPS promoted cell apoptosis in a transient manner; an abundance of cleaved caspase 3 was evident only at 12 h after LPS challenge (P = 0.02). Increased H2O2 content was seen at 12 h (P < 0.001) but decreased dramatically after 24 h of LPS treatment (P < 0.001). Total antioxidative capacity tended to decrease at both 12 and 24 h (P = 0.067 and 0.061, respectively). In agreement with these findings, LPS activated Nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidative signaling in the mammary gland, demonstrated by increased expression of its target gene Nqo1 at 12 h (P = 0.05) and xCT at 24 h (P = 0.076). Hypoxyprobe staining, indicative of hypoxia, was greater in the alveoli of PBS-treated glands than LPS-treated glands at both 12 and 24 h. This suggests oxygen tension rises in response to LPS treatment. Conversely, milk expression genes, β-casein gene (CSN2) and α-lactalbumin (LALBA), were inhibited by LPS treatment across time. Expression of α-S1 casein (CSN1S1) mRNA increased with LPS treatment at 24 h, but protein expression was reduced at this same time point (P < 0.05). In summary, intramammary LPS challenge incurs inflammation, augments cell apoptosis, induces oxidative stress and activation of the Nrf2 antioxidation pathway, increases oxygen tension, and inhibits milk protein expression in the mammary gland. This study provides functional insight into mechanisms of reduced milk production during mastitis and provides possible approaches to combat reduction in milk production, such as enhancing the Nrf2-antioxidative signaling pathway and reducing inhibition of milk protein expression.

Mastitis, or mammary gland inflammation, causes the greatest loss in profit for dairy producers. Mastitis susceptibility differs among cows due to environmental, physiological, and genetic factors. Prior research identified a genetic marker in a chemokine receptor, CXCR1, associated with mastitis susceptibility and decreased neutrophil migration. Current research seeks to identify reasons behind mastitis susceptibility by validating this model through in vivo challenge with Streptococcus uberis and studying specific mechanisms causing impaired neutrophil migration. Holstein cows with GG (n=19), GC (n=28), and CC (n=20) genotypes at CXCR1+777 were challenged intramammarily with S. uberis strain UT888. After challenge 68% of quarters from GG genotype, 74% from CC genotype and only 47% from GC genotype cows had ≥10 colony forming units/ml S. uberis for at least two sampling time points (P<0.05). However, among infected cows, number of S. uberis, somatic cell count, rectal temperature, milk scores and mammary scores were comparable among genotypes throughout infection. These findings suggest that cows with GC genotypes may be more resistant to S. uberis mastitis, but have similar responses if infected. To better understand the mechanisms associated with disease resistance, migration patterns in neutrophils from cows with different CXCR1+777 genotypes were evaluated. Neutrophils from cows with GG (n=11) and CC (n=11) genotypes were isolated and stimulated with zymosan activated sera (ZAS). Cells were fixed and stained for F-actin and evaluated for F-actin content, distribution, and cell morphology. Neutrophils from CC cows had significantly lower average F-actin polymerization than GG cows v (P=0.05). Directed migration of neutrophils from GG (n=10) and CC (n=10) genotypes was imaged and tracking data was analyzed for individual cells. Cells from GG genotype traveled further on an X axis and had higher X/Y movement towards IL8 compared to CC genotype, meaning they moved more directly towards IL8. Our findings suggest lower F-actin polymerization in combination with lower ability to directly move towards IL8 could impair neutrophil response to infection in cows with a CC genotype and may contribute to increased mastitis susceptibility. Finding what makes certain cows more susceptible to mastitis could lead to strategies aimed at improved prevention and treatment of mastitis.

La mammite bovine à S. aureus se caractérise, dune part, par la persistance dun nombre élevé de cellules inflammatoires (principalement des neutrophiles) dans le lait, et, dautre part, par la persistance de la bactérie dans la glande mammaire. La guérison dune mammite à S. aureus repose donc sur deux facteurs étroitement liés : le retour à une concentration normale de cellules dans le lait et léradication de lorganisme pathogène. Or, laccumulation et lactivation des neutrophiles au site de linfection requiert lexpression de cytokines pro-inflammatoires, telles que lIL-8 et le GM-CSF, et la croissance intra- et extracellulaire de S. aureus est augmentée en présence de cytokines inflammatoires, telles que lIL-1b et le TNF-a. La persistance de cytokines pro-inflammatoires semble donc être défavorable à la guérison dune mammite à S. aureus. Lexpression de la plupart des cytokines pro-inflammatoires est sous la dépendance du facteur nucléaire kB (NF kB). Par conséquent, NF kB pourrait être une cible thérapeutique intéressante pour améliorer ou soigner la mammite à S. aureus. Au début de ce travail aucune étude navait été dédiée au rôle de NF kB dans la mammite. Nous avons par conséquent tenté de clarifier ce rôle, ainsi que le potentiel de NF-kB en tant que cible thérapeutique dans la mammite bovine.

L'ampia letteratura sul periodo di transizione delle bovine richiama sempre più ai rapporti negativi fra problemi di salute e performance delle bovine. Dimostrazioni sempre più numerose confermano l'instaurarsi, soprattutto nel primo mese di lattazione, di una condizione di “stress da malattia”, non sempre associata a manifestazioni cliniche, ma con le stesse conseguenze: i fenomeni infiammatori. I principali obiettivi dei nostri studi, sono stati quelli di meglio caratterizzare gli stati infiammatori puntando l'attenzione su due aspetti: 1°) le cause di variazione della risposta infiammatoria alla infezione microbica e i suoi rapporti con la produzione di latte, 2°) le conseguenze che i fenomeni infiammatori hanno a livello di ingestione degli alimenti, delle riserve corporee, della produzione quanti-qualitativa e quindi dell'efficienza alimentare. relativamente al primo aspetto, nelle bovine sottoposte ad analoga infezione si è osservata una risposta infiammatoria di intensità diversa e che pare in buona relazione con precedenti fenomeni infiammatori in atto o pregressi; inoltre la risposta più intensa ha portato ad un maggior calo produttivo. Le conseguenze sulla efficienza energetica sono state studiate comparando due gruppi di bovine caratterizzate da minori (Lo-lfi) o maggiori (UP-lfi) valori di un indice di funzionalità epatica (lfi). Da ciò è emerso che, nel primo mese di lattazione, specie dopo avvenuta correzione per l'energia mobilizzata delle riserve corporee, l'efficienza è minore nei soggetti con basso lfi per il probabile aumento dei costi del sistema immunitario, aumento che pare prolungarsi ben oltre il fenomeno infiammatorio. / The extensive literature on transition period of the cows suggests the growing frequency of negative relationship between health problems and performance. Many evidences confirm during the first month of lactation, the presence of a particular condition named “stress disease”, which is not always associated to clinical symptoms but has the same consequences: the inflammatory processes. The principal objective of this study was to better characterize the inflammatory processes with particular attention to: 1) cause of the changes in inflammatory response to a bacterial infection and its relationship with the milk production, 2) consequences of the inflammatory processes on feed intake, body reserves, quantity and quality of milk production and then on feed efficiency. relatively to the first, we have observed a different intensity of the inflammatory response (in bovine submitted at the same infection), and this response seems correlated to previous inflammatory processes, probably not resolved; moreover, the higher intensity in the inflammations response can be responsible of the milk production decline. The consequences on feed efficiency have been studied through the comparison between two groups of cows characterized by higher (up-lfi) or low (lo-lfi) values of the liver functionality index; from that was demonstrated that during the first month of lactation, the cows with low lfi had lower efficiency values, probably caused increased charge of the immunitary system and prolonged beyond the inflammatory status itself.

Thesis (M.S.)--Mississippi State University. Department of Animal and Dairy Sciences. / Title from title screen. Includes bibliographical references.

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 “OrbeScal” was evaluated for drying cows. A comparison of Candida genus commercial identification systems diagnostic accuracy was via classical methods.

Mastitis causes largest economic losses in dairy production. Sub-clinical mastitis is detected in 50% of lactating cows in Lithuania. Curing effect is transient if major causes and factors of mastitis are not eliminated. Prophylactic measures are directed to strengthen the immune system of the cow and to define and to eliminate all factors causing mastitis. Bacteria are present on skin of the teat in the surroundings. Use of antiseptics of cow teats before and after milking is the most effective preventive procedure that can prevent from new mastitis cases. Antiseptic solutions help to fight most opportunistic and contagious bacterial infections causing mastitis. Teat antiseptics can reduce mastitis incidence by 50 to 75 %. It is recommended to use antiseptic solutions for udder and teats, as they improve physical condition of the udder and milk quality. Agents for udder and teat antiseptics contain bactericidal, skin protecting components in aqueous solution. Bactericidal activity is defined to kill germs during chemical and biological reactions. Active ingredients are iodoforms, chlorhexidins, sodium hypochloride, sodium chloride, lactic acid, hypochloric acid, antimicrobial proteins and fatty acids. Teat dipping reduces bacterial milk contamination, and spreading of pathogens on the teats during the milking. For the present moment this procedure is also used in Europe. Spraying teats with two different concentrations of iodine solution before and after the milking... [to full text]

Mūsų tyrimų duomenimis analizuotoje fermoje yra pakankamai aukštas išmilžis, kuris už 2007 metus sudarė 6234 kg per laktaciją. Karvių bandoje per mažą procentą sudaro pimaveršės (11,3%). Vyresnės karvės bandoje pasiskirsto beveik vienodai : antraveršės – 24,7 %, trečiaveršės- 39,1 % ir ketvirtos laktacijos- 24,7 %. Karvių sergamumas slaptuoju mastitu priklauso nuo produktyvumo. Mūsų tyrimai rodo, karvės, kurių produkcija per laktaciją buvo 3000 kg pieno - iki 200 tūkst./ml somatinių ląstelių turėjo 20,93 %, 3001 – 5000 kg – 55,8 % ir virš 5001 kg – 23,26 % karvių. Antroje bandomoje grupėje, karvių somatinių ląstelių skaičius buvo 201 – 400 tūkst./ml pirmaveršių buvo 20,0 %, 3001 – 5000 kg- 43,3% ir virš 5001 kg – 36,6 %. Trečioje bandomojoje grupėje ( 24 karvės) 401 tūkst./ml SLS turėjo 25 %, 3001 – 5000 kg – 16,7% ir virš 5001 kg – 58,3 %. Tyrimai rodo, kad didėjant karvių produktyvumui jos dažniau serga slaptuoju mastitu. Tuo mūsų tyrimų duomenys sutampa su J. Rudejevienės (2007), E Aniulio (2007), Sederevičiaus (2004) tyrimų duomenimis. Analizuojant pažeistus karvių tešmens ketvirčius pastebėjome, kad vienas ketvirtis buvo pažeistas pas 40,0 %, du ketvirčiai - 30 %, trys ketvirčiai - 16,7 % ir visi ketvirčiai 13,3 %. Šio tyrimo duomenys nevisiškai sutampa su J. Klimaitės (2005) duomenimis. Manome, kad pažeistų ketvirčių skaičius priklauso nuo karvių laikymo būdo, sanitarinių ir kitų priežasčių. Pieno sanitarinė būklė priklauso nuo tvarto higienos, spenių paruošimo melžimui... [toliau žr. visą tekstą] / According to the data of our studies, there is quite high milk yield in analyzed farm. This milk yield composed 6234 kg per lactation during 2007. In the herd of cows there is a small percent of first-calf cows (11,3%). Older cows are spread in the herd almost equally: second- calf cows – 24,7 %, third-calf cows- 39,1 %, and cows of fourth lactation- 24,7 %. Cows’ morbidity with subclinical mastitis depends on productivity. Our study shows that cows whose production during lactation was 3000 kg- till 200 thousands/ml of somatic cells had 20,93 %, 3001 – 5000 kg – 55,8 % and over 5001 kg – 23,26 % of cows. In the second experimental group the somatic cells count 201 – 400 thousands/ml had 20,0 % of first-calf cows, 3001 – 5000 kg- 43,3% and over 5001 kg – 36,6 %. In the third experimental group ( 24 cows) 401 thousands/ml of SCC (somatic cells count) had 25 %, 3001 – 5000 kg – 16,7% and over 5001 kg – 58,3 %. Studies show that during the rise of productivity of cows they more often suffer from subclinical mastitis. According to this aspect our studies data are compatible with research data of J. Rudejevienė (2007), E Aniulis (2007), Sederevičius (2004). While analysing damaged udder quarters we noticed that one quarter was damaged at 40,0 %, two quarters - 30 %, three quarters - 16,7 % and all four quarters- 13,3 %. The data of this study are only partially compatible with data of J. Klimaitė (2005). It is clear that the number of damaged quarters depends on the way of cows’... [to full text]