Mechanisms by which mastitis affects reproduction in dairy cow: A review.

Reproductive performance is a key factor in determining the profitability of dairy farm, which is affected by many factors such as environment and diseases. Mastitis is a common and important disease, which has caused huge economic losses to the dairy industries worldwide. Mammary gland infection causes immune responses, resulting in the abnormal secretion of cytokines and hormones and abnormal function of the reproductive system such as the ovary, corpus luteum, uterus and embryo. Cows with mastitis have delayed oestrus, decreased pregnancy rate and increased risk of abortion. The adverse effects of mastitis on reproductive performance are affected by many factors, such as occurrence time, pathogen and cow factors. This paper primarily reviews the progress in the effects and mechanisms of mastitis on reproductive performance, with emphasis on maternal transcriptome, genomic analysis, epigenetic modification, microbiota, inflammatory regulation and immune evasion mechanism of mastitis, aiming to provide directions for the prevention and control of mastitis in the future.

Proposal of an iELISA for Mycoplasma bovis diagnosis in dairy cattle and associated risk factors / [Proposta de um iELISA para diagnóstico de Mycoplasma bovis em bovinos leiteiros e fatores de risco associados]

Mycoplasma bovis is a highly contagious agent associated with several pathologies in cattle. The detection of reactive antibodies to M. bovis by Indirect Enzyme-Linked Immunosorbent Assay (iELISA) identifies if there was an exposure to the microorganism. The current study aimed to optimize an iELISA from M. bovis total cell antigen, applying it to bovine serum samples, and to evaluate risk factors. Serum samples were obtained from 400 cows from 17 herds from Southeast Brazil. In the optimization of iELISA, the following was established: 2 µg/mL of antigen, sera dilution 1:300, and conjugate dilution 1:15000. The frequency was 62.3% (249/400) of reactive animals and 100% (17/17) of reactive herds. Risk factors were: herds with more than 100 animals (OR= 3.1; CI= 95%); Holstein breed (OR= 72.5; CI= 95%); cows (OR= 29.7; CI= 95%); intensive breeding system (OR= 3.3; CI= 95%); associated small ruminant production (OR= 4.4; CI= 95%); milk production above 500L (OR= 2.9; CI= 95%); no quarantine (OR= 1.5; CI= 95%); mechanical milking (OR= 5.5; CI= 95%) and cases of mastitis (OR= 5.5; CI= 95%). The proposed iELISA was able to detect antibodies reactive to M. bovis in bovine serum. Knowledge of these risk factors can assist in the implementation of prophylactic measures.(AU)

Mycoplasma bovis é um agente altamente contagioso relacionado a várias patologias em bovinos. A detecção de anticorpos reativos a M. bovis por Ensaio de Imunoadsorção Enzimática Indireto (iELISA) identifica se houve exposição ao microrganismo. O presente estudo teve como objetivo otimizar um iELISA de antígeno celular total de M. bovis, aplicando-o a amostras de soro bovino, bem como avaliar fatores de risco. Amostras de soro foram obtidas de 400 vacas de 17 rebanhos da Região Sudeste do Brasil. Na otimização do iELISA foram obtidos: 2µg/mL de antígeno, diluição dos soros 1:300 e do conjugado 1:15000. A frequência de animais reativos foi de 62,3% (249/400) e de 100% (17/17) para os rebanhos. Os fatores de risco foram: rebanhos com mais de 100 animais (OR= 3,1; IC= 95%); raça Holandesa (OR= 72,5;IC= 95%); vacas (OR= 29,7;IC= 95%); sistema intensivo (OR= 3,3; C= 95%); produção de pequenos ruminantes (OR= 4,4;IC=95%); produção de leite acima de 500L (OR= 2,9;IC= 95%); sem quarentena (OR= 1,5;IC= 95%); ordenha mecânica (OR= 5,5;IC= 95%) e casos de mastite (OR= 5,5;IC= 95%). O iELISA proposto foi capaz de detectar anticorpos reativos a M. bovis no soro bovino. O conhecimento desses fatores de risco pode auxiliar na implementação de medidas profiláticas.(AU)

Combined effect of mastitis and parity on pregnancy loss in lactating Holstein cows.

The main objective of this study was to examine the combined effect of mastitis and parity on pregnancy loss (PL) in lactating Holstein cows. A secondary objective was to estimate the cost of mastitis including that of PL attributable to mastitis. A total of 1,774 lactation periods from 1,047 Holstein cows with different parities from one dairy farm were included in a matched case-control study. All study cows were diagnosed pregnant by transrectal ultrasonography on day 33 after timed artificial insemination (TAI). Case cows (n = 222 lactations) were those later diagnosed non-pregnant by transrectal palpation on day 47 or 75 after TAI. Control cows (n = 1,552 lactations) were those confirmed pregnant by transrectal palpation on day 75 after TAI. Case cows were matched with eligible controls according to year of calving and calving-to-conception interval (CCI) ± 3 days. Cows with different parities were classified as exposed to subclinical mastitis (somatic cell score (SCS) > 4.5 in at least one Dairy Herd Improvement Association (DHIA) test day) or clinical mastitis (with or without evidence of subclinical mastitis) during two exposure periods: 1-42 days before breeding or 1-75 days during gestation (1 to PL diagnosis day in case cows, or 1-75 day in control cows). Conditional logistic regression was used to model the odds of PL as a function of previous exposure to mastitis in different parities. Cost of PL attributable to mastitis ($/case) among cows with mastitis was estimated based on attributable risk calculated in the epidemiologic analysis. We observed a higher than expected combined effect between exposure to mastitis (subclinical or clinical) before breeding and parity 3 or ≥ 4, and during gestation and parity ≥ 4 on PL. The cost of PL attributable to mastitis was highest ($196/case) in cows in parity ≥ 4 affected with clinical mastitis during gestation. Overall, study results indicate the impact of mastitis on PL is higher in older cows (parity ≥ 3). Dairy farmers and attending veterinarians can consider the combined effect of mastitis and parity when evaluating causes for PL and strategies for optimizing reproductive performance in dairy cows.

Somatic cell count and type of intramammary infection impacts fertility from in vitro produced embryo transfer.

The objective of this study was to assess the impact of mastitis-causing bacteria and somatic cell count (SCC) on pregnancy per embryo transfer (P/ET) in Holstein-Gir crossbred (Girolando) lactating dairy cows. Cows (n = 1397) were subjected to a timed-embryo transfer protocol. Milk samples were collected two days before embryo transfer for SCC and bacteriological culture analyses. Pregnancy diagnosis was performed on days 31 and 66 after timed-embryo transfer. The animals were grouped according to the National Mastitis Council recommendations: Gram-positive environmental (EV+), Gram-negative environmental (EV-), Gram-positive contagious (C+), coagulase-negative staphylococci (CNS) and control (no bacterial growth). Additional analysis was made by categorizing bacteria based on degree of pathogenicity (Major or Minor). Bacterial growth reduced P/ET (P < .01) at both 31 and 66 days of gestation. The P/ET was lower (P < .05) at 31 days in EV- (30.1%) and EV+ (29.9%) groups and tended (P = .09) to be lower in the C+ group (36.6%) than the control group (44.0%). The P/ET from the Major group at 31 days of gestation was lower (P = .03) compared with the Minor and control groups (32.1 vs 41.1 vs 43.2%, respectively). Cows with SCC > 400,000 cells/mL had lower P/ET (P < .01) than animals with SCC < 200,000 cells/mL at both 31 (30.4% vs 40.8%) and 66 days (24.7% vs 32.2%) of gestation. Pregnancy loss was not different between bacterial isolates and SCC categories. Elevated SCC significantly reduced P/ET, whereas environmental agents and those with Major pathogenicity yielded the greatest reduction in P/ET.

Evidence that mastitis can cause pregnancy loss in dairy cows: A systematic review of observational studies.

The objective of this study was to conduct a systematic review to identify and assess evidence and knowledge gaps in published observational studies that have investigated the relationship between mastitis and pregnancy loss (PL) in dairy cows. PubMed and ScienceDirect were used to search pertinent peer-reviewed research reports of interest. Screening of research reports was conducted at 3 levels: titles, abstracts, and full-text articles. The search identified 651 records for initial screening. The final screening process identified 8 qualified articles for review after removing 10 duplicate records, 582 titles, 31 abstracts, and 20 full-text articles. Two studies produced strong epidemiologic evidence indicating that (1) exposure to clinical mastitis during early gestation (first 45 d of gestation) is associated with subsequent PL during the following 90 d; and (2) subclinical mastitis 1 to 30 d before artificial insemination (AI) is associated with subsequent PL at 35 to 41 d of gestation. An additional study showed that exposure to clinical mastitis during early lactation in combination with low body condition can increase the risk of PL in dairy cows; however, the interaction effect between clinical mastitis and low body condition on PL was considered weak. Four other studies produced inconclusive evidence indicating that mastitis is a predisposing factor for PL in dairy cows, as the exposure risk period for mastitis overlapped with the follow-up period for diagnosis of PL in dairy cows. Finally, one study failed to identify a relationship between mastitis and PL in dairy cows. Further research is needed to (1) support the hypothesis that mastitis in combination with low body condition score (or other exposure factors) can increase the risk of PL, (2) compare the effect of clinical versus subclinical mastitis on PL, (3) compare the effect of mastitis before breeding and during gestation on PL, and (4) compare the effect of mastitis on PL in dairy cows during different lactations.

Effects of ovarian structures identified at Ovsynch™ enrolment, disease history and lactation variables on odds of pregnancy to a fixed-time artificial insemination after Ovsynch™.

OBJECTIVE: To determine the effects of ovarian structures present at Ovsynch™ enrolment, disease history, lactation variables and times bred on pregnancy per fixed-time artificial insemination (FTAI) following fixed-time insemination after Ovsynch and to determine the relationship between the size of ovarian follicular structures and concurrent presence of a corpus luteum (CL). METHODS: The study was conducted in a 3000-cow Holstein dairy herd. Over a 6-month period, 886 non-pregnant cows were examined by transrectal ultrasound and enrolled into 1132 Ovsynch events. Enrolled cows were synchronised using an Ovsynch-56 protocol. At enrolment, both ovaries were scanned by transrectal ultrasound to identify and record the presence and size of ovarian structures. Cows that did not return to oestrus were pregnancy tested 32-38 days post-insemination. Univariable and multivariable generalised linear mixed models (GLMMs) were used to determine the effect of each variable on pregnancy per FTAI. A univariable GLMM was also used to analyse the relationship between size of the dominant follicular structure and concurrent presence of a CL. RESULTS: CL size (P = 0.039) and presence of a luteal cyst (P = 0.002) at Ovsynch enrolment significantly increased the odds of pregnancy. Occurrence of lameness (P = 0.035) or mastitis (P = 0.008) between calving and enrolment significantly decreased the odds of pregnancy. Neither the presence nor size of a follicular structure significantly affected the odds of pregnancy, although cows with larger follicular structures were less likely to have a concurrent CL (P < 0.001). CONCLUSION: Presence of luteal tissue at Ovsynch enrolment and recent disease events affected pregnancy per FTAI in cows inseminated after Ovsynch.

Association between disease occurrence and fertility of dairy cows in three geographic regions of Chile.

The objective was to analyze the association between disease occurrence during early lactation and reproductive performance and survival of dairy cows in high-producing herds, under different management practices in three geographic regions of Chile. Data included 30,757 lactation records of cows calving from January 2013 to June 2014 in three different locations: Central (C) area (n = 6198 cows in eight herds), Central-South (CS) area(n = 17,234 cows in 12 herds), and South (S) area (n = 7325 cows in six herds). Data were analyzed using logistic regression and ANOVA, considering cow as the experimental unit. Covariables offered to the models included parity number, season of calving, cow and herd relative milk yield, geographic location, and management system. Average milk yield (305 ME) per cow were 12,091, 11,783, and 6852 kg for C, CS, and S regions, respectively. Time from calving to first service and time to conception were consistently greater for cows with at least one disease event within 50 days in milk (DIM), for cows that were reported lame, or for cows that had mastitis or metritis. The odds (95% confidence interval) of pregnancy at 150 DIM (P150) and the odds of survival until 150 DIM (S150) for cows that had at least one disease event within 50 DIM were 0.84 (0.79-0.91) times the odds of pregnancy and 0.25 (0.22-0.28) times the odds of survival for healthy cows. The odds of P150 for cows located in the C and CS areas were 1.56 (1.36-1.80) and 1.16 (1.04-1.30) times the odds of P150 for cows in the S area. The odds of S150 for cows located in the C and CS areas were 0.48 (0.37-0.62) and 0.54 (0.42-0.67) times the odds of S150 for cows in the S area. These data suggested that cow health status and geographic location are significantly associated with reproductive performance and survival in this population of Chilean dairy cows.

Impact of subclinical and clinical mastitis on sensitivity to pain of dairy cows.

A total of 90 cows from three commercial farms were used to evaluate the relationship between subclinical mastitis and clinical mastitis and thermal nociceptive threshold. Milk strips from all udder quarters were tested for clinical mastitis with visual inspection of milk and udder alterations and for subclinical mastitis using California Mastitis Test. Milk yield was recorded, milk was sampled and further analyzed for somatic cells count (SCC). Cows were considered healthy when SCC200 000 cells/ml and no visual alterations in milk and/or udder, with moderate subclinical mastitis when SCC>500 000 cells/ml and no visual alterations in milk and/or udder and with clinical mastitis when visual alterations in milk and/or udder were detected. Nociceptive threshold was evaluated with the thermal threshold meter apparatus applied to the rear legs. Thermal threshold (TT) decreased when we compared healthy cows with cows presenting clinical mastitis and tended to decrease when we compare healthy cows with those with moderate subclinical mastitis. TT was lower at the ipsilateral rear leg compared with the contralateral leg to the infected mammary gland. TT linearly decreases as log10SCC increased and it showed sharp decrease as log10SCC exceed the value of 6.4. Increase in one unit of log10SCC increased the odds of low thermal threshold (lower than 55.8°C). Subclinical mastitis might be a welfare issue as it tended to decrease nociceptive thermal threshold.

Métodos automatizados de diagnóstico de mastite em vacas leiteiras: uma revisão / Automated diagnosis methods of mastitis in dairy milk cattle: a review

A mastite é a infecção mais frequente dos animais destinados à produção de leite e que mais onera a pecuária leiteira. As perdas econômicas são causadas por diversos fatores: medicamentos, morte ou descarte precoce de animais, bem como em laticínios, pela queda na qualidade do produto final. Rotineiramente, quando se pretende verificar a incidência de mastite em vacas leiteiras utilizam-se os métodos convencionais e subjetivos, como a caneca telada e o California Mastitis Test. A automação na pecuária leiteira é uma inovação tecnológica que visa a minimizar os erros de manejo e otimizar os processos dentro da atividade, dentre os quais o diagnóstico precoce de casos subclínicos de mastite. O uso de sensores que utilizam parâmetros objetivos de mensuração tais como: condutividade elétrica, contagem de células somáticas, presença da enzima lactato desidrogenase, mensuração da cor e a homogeneidade podem ser a alternativa para detecção rápida e precisa de mastite em bovinos, especialmente quando se trata de sistemas automatizados, pois o diagnóstico precoce minimiza as perdas econômicas. Diante do exposto, objetivou-se realizar uma revisão bibliográfica sobre os métodos automatizados de detecção de mastite existentes atualmente...

Mastitis es la infección más frecuente en animales destinados a la producción de leche y más oneroso a la ganadería lechera. Las pérdidas económicas son causadas por diversos factores: medicamentos, muerte o descarte precoz de animales, así como en lacticinios, por la caída en la calidad del producto final. Rutinariamente, cuando se pretende verificar la incidencia de mastitis en vacas lecheras se utilizan los métodos convencionales y subjetivos, como la taza de red y el California Mastitis Test. La automatización en la ganadería lechera es una innovación tecnológica que busca minimizar los errores de manejo y optimizar los procesos dentro de la actividad, entre los cuales el diagnóstico precoz de casos subclínicos de mastitis. El uso de sensores que utilizan parámetros objetivos de mensuración tales como: conductividad eléctrica, conteo de células somáticas, presencia de la enzima lactato deshidrogenasa, mensuración del color y la homogeneidad que pueden ser la alternativa para detección rápida y precisa de mastitis en bovinos, principalmente cuando se trata de sistemas automatizados, pues el diagnóstico precoz minimiza las pérdidas económicas. Delante el expuesto, se ha tenido por objetivo realizar una revisión bibliográfica sobre los métodos automatizados de detección de mastitis existentes actualmente...

Rapid mastitis detection assay on porous nitrocellulose membrane slides.

We have developed a rapid mastitis detection test based on the immobilization of tag-specific antibody molecules, the binding of double-tagged amplicons, and as a secondary signal a conjugate of black carbon nanoparticles having molecules of a fusion protein of neutrAvidin and alkaline phosphatase at their surface. The antibodies were inkjet printed onto three different nitrocellulose membrane slides, Unisart (Sartorius), FAST (GE Whatman), and Oncyte-Avid (Grace-Biolabs), and the final assay signals on these slides were compared. The blackness of the spots was determined by flatbed scanning and assessment of the pixel gray volume using TotalLab image analysis software. The black spots could be easily read by the naked eye. We successfully demonstrated the detection of specific amplicons from mastitis-causing pathogens in less than 3 h. Using a similar protocol, we also showed that it was possible to detect specific amplicons from four different mastitis-causing pathogens (six strains) on the same pad. The influence of two different printing buffers, phosphate-buffered saline (pH 7.4) and carbonate buffer (pH 9.6), on the functionality of the primary antibodies was also compared.

[Herd-specific estimation of milk yield reduction due to recurrent clinical mastitis]. / Herdenspezifische Schätzung der Milchleistungsminderung durch wiederholte klinische Mastitis.

To decide about investments in preventive measures improving udder health it is important that the farmer knows the monetary loss due to mastitis on his specific farm. The Saxon dairy herd improvement association (Sächsischer LandeskontrolIverband e.V., LKV) is developing a computer program called "Daten- und Informationsplattform Tier (DIT)"which determines - among other analyses - the milk loss caused by mastitis for a specific herd based on individual cow's mastitis and daily milk yield data. The present article introduces the linear mixed model which is taken as a basis for calculating the reduction in performance through clinical mastitis. The short and long term decrease in daily milk yield is described by expanding the lactation curve model of Ali and Schaeffer (1987). For falculating the short term drop, the model includes the laps of time in days since the mastitis incident as secon-ddegree polynomial. The coefficients are estimaged specifically for the first respectively every following case of mastitis (class of episode). Classes of episode are also considered calculating the long term decrease by estimating lactation curves without mastitis as well as corresponding to the classes of episode. By integrating the statistic software R (R Development Core Team, 2012) into the processes of the DIT the estimation of the farm specific model parameters is largely automated on the servers of the LKV.Thereby, milk yield can be estimated for every day in milk according to episode number and laps of time since the incident or with no mastitis incident respectively for a particular period of time in a specific dairy herd. The loss resulting from reduced performance due to clinical mastitis is specified by adding up the differences and can serve as a valuable basis for management decisions.

Short communication: experimentally induced mastitis reduces weight shifting between the rear legs while standing in dairy cows.

The objectives of this study were to evaluate changes in weight shifting between legs while standing on a weighing platform in response to endotoxin-induced clinical mastitis, and to evaluate the effect of the nonsteroidal antiinflammatory drug flunixin meglumine on weight distribution between legs while standing in dairy cattle with endotoxin-induced clinical mastitis. Clinical mastitis was induced in 10 primiparous and 9 multiparous lactating dairy cows (days in milk=55 ± 12; mean ± standard deviation) by intramammary infusion of 100 µg of Escherichia coli lipopolysaccharide (LPS) into the right rear quarter. Four hours later, 10 animals were randomly assigned to receive flunixin meglumine intravenously (2.2mg/kg of body weight; treated group) and 9 received an equivalent volume of sterile isotonic saline solution (control group). Body temperature was monitored rectally 3d before LPS infusion, immediately before LPS infusion, and 4, 7, 10, 13, 16, and 28 h after LPS infusion. The weight applied to each leg was recorded while cows were standing on a weighing platform on the day before the challenge and 7, 10, 13, 16, and 28 h after LPS infusion. Two measures of weight shifting between the rear legs were calculated for each recording session: the standard deviation of the weight applied to the legs over time and the frequency of steps. The LPS infusion resulted in a consistent case of clinical mastitis approximately 4h after the LPS infusion, as assessed by the presence of visible swelling and elevated rectal temperature in all cows. However, control animals had a higher temperature 7h after LPS infusion compared with treated animals (40.8 vs. 39.0°C; standard error of the difference=0.2). Overall, weight shifting between the rear legs was decreased 7h after the LPS infusion compared with baseline, and this decrease was not affected by treatment with flunixin meglumine. It is likely that weight shifting increases friction between the swollen udder and the legs, increasing the pain experienced by the cow. Thus, cows with endotoxin-induced mastitis avoided shifting weight, particularly at the times when the most severe signs of inflammation occurred. Further research is needed to assess the efficacy of flunixin meglumine in mitigating udder pain and the accuracy of behavioral measures such as weight shifting in assessing analgesia in cows with mastitis.

Behavioral changes in dairy cows with mastitis.

Mastitis is a frequent and painful disease in dairy cows. However, pain detection and alleviation in mastitic cows has been overlooked. The objectives of this study were to measure behavioral changes in dairy cows with clinical mastitis and to investigate the effect of intramammary infusion of an antibiotic on lying behavior and behavior during milking. In experiment 1, 42 lactating cows were used: 14 mastitic cows and 28 control cows. Mastitic and control cows were subjected to an evaluation of pain responses on d 1 (mastitis detection day), and 2, 3, and 7d after the last antibiotic treatment (d 10+). The antibiotic treatment was administered to mastitic cows twice a day, starting on d 1, for at least 3 consecutive days. Behavioral changes were evaluated by measuring lying behavior, reactivity during milking (stepping, lifting, and kicking), weight distribution, and hock-to-hock distance. Overall, mastitic cows spent less time lying down on d 2 compared with control cows. The percentage of time lying on the mastitic quarter side did not differ significantly between mastitic and control cows. No differences were observed between control and mastitic cows on the number of steps per 24h on each day. Restless behavior during milking did not differ between treatments. Restless behaviors differed significantly within mastitic cows between days. Frequency of kicks per minute was higher on d 1 compared with d 2, frequency of lifts was higher on d 1 and 2 compared with d 10+, and frequency of steps was higher on d 2 compared with d 10+. The variability of weight that mastitic cows applied to the leg on the mastitic quarter side was higher on d 1 than on d 10+. For control cows, the variability in weight applied to the homologous leg of the mastitic quarter side leg was higher on d 1 compared with d 2 and 3. The hock-to-hock distance did not differ between treatments. Mild clinical mastitis might not cause sufficient pain to observe marked changes in behaviors. However, cows showed differences in lying time and reactivity during milking and slight differences in the laterality of lying. To further develop methodologies for assessing pain in mastitic cows, it is worth applying the methodologies used in this study to cows with moderate to severe mastitis, followed by their validation using analgesic treatment, to ensure that any change is a pain-specific behavior rather than a simple reflex. In experiment 2, no effect of intramammary infusion of the antibiotic was observed on lying behavior or behavior during milking. Cows with mild clinical mastitis present behavioral changes in lying behavior and at milking time, which could be associated with discomfort.

Associations between udder health and reproductive performance in United Kingdom dairy cows.

The objective of this research was to evaluate the relationship between udder health and reproductive performance in UK dairy cows. Data from 80 herds were restructured such that each unit of data represented a 2-d period during lactation where a cow was at risk of becoming pregnant. Multilevel discrete-time survival models were then used within a Bayesian framework to explore associations between reproductive outcomes and a variety of potential explanatory variables. Separate models were constructed using 2 different univariate binary outcomes: a cow becoming pregnant during a risk period and a cow becoming pregnant as a result of a given service. Potential explanatory variables included occurrence of clinical mastitis and a categorical representation of individual cow somatic cell count (SCC), both at a variety of timings relative to the risk period. Posterior predictions were used to assess model fit and to check model building assumptions. These demonstrated that the model represented the data well. Within-sample Monte Carlo simulation (i.e., use of the model to predict outcomes for cases within the data set, repeated over a large number of iterations) was used to illustrate results as posterior predicted relative risks. A negative association was found between reproductive performance and cases of clinical mastitis over a wide time frame relative to the risk period (from 28 d before to 70 d after the risk period). A similar negative association with the probability of a service leading to a pregnancy (pregnancy rate) was observed over the same time frame. Higher SCC recordings (i.e., those more likely to be associated with an intramammary infection) were also associated with decreased reproductive performance, especially where an individual cow SCC of greater than 399,000/mL was recorded in the 30 d following a risk period or service. This research demonstrates that both clinical and subclinical mastitis are associated with a reduction in reproductive performance, and that this influence varies in magnitude but can be exerted over a prolonged period.

The association between teat end hyperkeratosis and teat canal microbial load in lactating dairy cattle.

Most pathogens that cause bovine mastitis invade the udder lumen through the teat canal. Amino acids and intercellular lipids may support microbial colonisation of the teat canal epithelium by pathogenic microorganisms. The aim of this study was to investigate the association between teat end hyperkeratosis, which is induced by machine milking, and teat canal microbial load. Contralateral teats, which differed in teat end hyperkeratosis scores, were identified in a split-udder experiment. The teat canal's microbial load was evaluated using the wet and dry swab technique. Staphylococcus (S.) aureus, Streptococcus (Sc.) uberis, Escherichia (E.) coli and other coliforms were detected by agar plate cultures. The positive detection of E. coli and the log(10)-transformed E. coli load of a teat canal were significantly associated with the teat end hyperkeratosis score (P<0.05). There were significant differences with respect to positive findings for E. coli, as well as the microbial load of E. coli and Sc. uberis, between the less-calloused and the more-calloused teat of a pair. For S. aureus, no significant associations between hyperkeratosis score and teat canal microbial load were detected. In general, a teat with a highly calloused teat end had an increased teat canal microbial load compared with the contralateral teat, characterised by a lower callosity. The results of the present study indicate that the environmental pathogen load is associated with teat end hyperkeratosis. Further research is needed to identify factors that may affect teat canal microbial load in lactating dairy cattle.

Chronic mastitis is associated with altered ovarian follicle development in dairy cattle.

Connection between mastitis and fertility is multifaceted; therefore, several aspects need more elucidation. In particular, the aim was to investigate if naturally occurring chronic mastitis has an effect on ovarian function. At the time of slaughter, a milk sample and both ovaries were collected from 68 cows. The presence and intensity of chronic mastitis was diagnosed by the combined evaluation of bacteriological examination and somatic cell count of the milk of each individual quarter according to the measures of the National Mastitis Council. Animals were divided into 4 groups characterized by a low (n=15), mild (n=14), intense (n=19), or severe (n=16) degree of infection. A count of visible follicles on each ovary was followed by a quantitative analysis of microscopic traits on a selected group of animals (n=16). The latter included the classification and count of the entire preantral follicle population, and the morphometric analysis of the vascular bed extension and connective stroma in the cortical region. Finally, the expression of growth and differentiation factor-9 (GDF-9) was studied. The number of follicles with diameters ranging from 1 to 3 mm and 4 to 7 mm was not affected by the degree of infection. A significant effect of the degree of udder infection was observed on the number of follicles with a diameter larger than 8 mm. Furthermore, the intensity of mastitis had no effect on the number of primordial and primary follicles, but severely affected cows showed a lower number of secondary follicles (0.5±0.1 vs. 0.2±0.03). Quantitative analysis demonstrated a decrease in the density of blood vessels (6.30±1.08 vs. 4.68±0.28) expressed as ratio of vascular bed/total area) and a higher incidence of fibrous stroma (1.60±0.99 vs. 6.04±3.08 expressed as ratio of connective tissue/total area) in the cortical area of the most affected animals. Finally, the level of GDF-9 protein within the oocytes of different follicle size was lower in the animals with the severe form of chronic mastitis (1.34±0.05 vs. 0.78±0.21 expressed as arbitrary units). In conclusion, decreased fertility of cows with chronic mastitis takes place through an effect on the ovary altering the dynamics of folliculogenesis. Within the ovary, this implies a reduction of the vascular bed and an increase in the fibrotic tissue together with a direct effect on oocyte-specific factors as GDF-9, all of which are essential regulatory elements of folliculogenesis.

Observed and expected combined effects of clinical mastitis and low body condition on pregnancy loss in dairy cows.

The objective was to compare the observed and expected combined effects of clinical mastitis before timed artificial insemination (TAI) and low body condition at 70 d postpartum (dpp) on pregnancy loss in dairy cows. Cows were examined for pregnancy by ultrasonographic examination 28-32 d after TAI; the presence of an embryo with a heartbeat was the criterion used to determine pregnancy. Cows diagnosed pregnant were re-examined by transrectal palpation of the uterus and its contents 28 d later to confirm pregnancy status and to identify pregnancy loss. Eighty-eight (17%) of 512 cows were diagnosed with pregnancy loss. Cows affected with clinical mastitis before insemination and a body condition score (BCS)≤2.75 at 70 dpp were 2.03 times more likely to experience pregnancy loss, compared to cows without clinical mastitis and with a BCS>2.75 (RR=2.03; 95% CI=1.15, 3.60; P=0.01). This observed combined effect for pregnancy loss (RR=2.03) was higher than the expected combined effect based on adding (RR=1.39) or multiplying (RR=1.42) absolute independent excesses due to clinical mastitis or low body condition.

Mastitis and fertility disorders in cows.

Udder inflammations, independently of significant losses in milk yield and alteration of its quality, can negatively affect cows' reproduction efficiency. Mastitis causes changes in many active constituents, both in milk and blood. Pathogenic changes in existing constituents and new active chemical compounds, generated during disease, can affect other organs, particularly the reproductive system and its mechanisms. Fertility disorders in mastitic cows are mostly connected with the activity of cytokines (especially TNF-alpha), cortisol, prostaglandin F2-alpha, reactive oxygen species (ROS), and classic inflammatory mediators. The successful treatment of mastitis as well as protection against udder infections should be considered important methods for prophylaxis of fertility disorders in cows. The first Polish trials indicated that injection of supportive drugs (antioxidants or lysozyme dimer or flunixin meglumine) to intramammarily treated cows can increase fertility in cows with mastitis.