Impact of a Novel Homeopathic Complex Medicine on the Management of Multiple Antibiotic-Resistant Bovine Mastitis: An Open-Label, Non-Randomized, Placebo-Controlled Trial.

BACKGROUND: Bovine mastitis is characterized by an inflammatory process in the mammary gland and represents one of the main diseases affecting a dairy herd. Management of mastitis is most commonly via antibiotics, but the rising incidence of multiple antibiotic resistance (MAR) means that additional options are needed. Homeopathic products can be administered in dairy farming for a range of clinical reasons and may be preferential due to the absence of residues. OBJECTIVES: The aim of this study was to assess the potential of a novel homeopathic complex medicine in managing bovine mastitis. METHODS: Twenty-four lactating Holstein cows with mastitis were divided into two groups: the homeopathic complex group received a homeopathic complex daily for 60 days at a dose of 20 g/d; the placebo group received the calcium carbonate vehicle without homeopathic medicines at the same dose and repetition. The main outcome measure was somatic cell count (SCC; cells/mL), with additional outcome measures including milk production (kg/d), milk constituents (percentage of protein, fat, lactose and total milk solids), and serum levels of cortisol, glucose, ammonia and lactic acid. All outcomes were measured at the beginning of the study and after 30 and 60 days. Milk samples were also collected from all animals at the beginning of the study, confirming a high (>0.2) MAR index for isolated bacterial cultures. RESULTS: Assessment of SCC showed a statistically significant difference favoring the homeopathic complex versus placebo group at day 60. A reduction in serum cortisol levels and an increase in fat, lactose and total milk solids in animals treated with the homeopathic complex at day 60 were also seen. Other outcome measures did not show statistically significant inter-group differences. CONCLUSION: The results of this non-randomized, open-label, placebo-controlled trial suggest the potential for a novel homeopathic complex medicine in management of multiple antibiotic-resistant bovine mastitis, thus offering dairy farmers an additional option to antibiotics and making dairy products safer for consumer health and milk production more sustainable.

Symposium review: Environmental effects on mammary immunity and health.

Environmental effects on pathogen abundance and access are precursors to mastitis. Indeed, high heat and humidity, and unsanitary housing and equipment, are associated with greater pathogen load and exposure. Although less is known about effects of environment on a cow's ability to resist infection, several indicators suggest that it can affect pathogen responses. Mastitis incidence and bulk tank somatic cell count vary with season, typically peaking in summer. Recent controlled studies have revealed that heat stress exposure results in changes in the microbiome of the cow and her environment, which may relate to negative effects on milk quality and cow health. Alternatively, specific pathogen loads may vary based on housing dynamics rather than associations with physical environment. Indeed, housing-related stressors, such as overcrowding and social group challenge, influence secretion of glucocorticoids, thus affecting pathogen resistance in the cow. Two key seasonal variables are photoperiod and temperature, specifically the heat stress consequent to elevated temperature and humidity. Shifts in light duration regulate immune function in other species, but apparently have limited effect on udder health of lactating cows. In contrast, in dry cows, short days increase peripheral blood mononuclear cell number and are associated with lower somatic cell count in the next lactation, compared with long days. With heat stress, elevated body temperature directly affects expression of immune-related genes in mammary tissue. Responses depend on duration of exposure and feature acute upregulation of immune-signaling pathways, followed by enrichment of other immune-related pathways after prolonged exposure. Most responses are transient and recover within 1 wk. Functionally, heat stress impairs some aspects of acquired immunity in dry cows, including antigen responses and lymphocyte proliferation, but apparently not innate immune function. However, heat stress in late gestation reduces neutrophil phagocytosis and killing in vitro, and neutrophils in circulation are reduced in vivo as are responses to pathogen challenge in the subsequent lactation. A holistic understanding of the complex interplay of environment, pathogens, and host is needed to inform advances in this area.

The effect of pre-calving injection of trace mineral supplements on periparturient disease incidence in pasture based dairy cows.

Trace minerals, have a role in immune function and a trace mineral supplement (TMS) can improve animal health in dairy herds. This prospective randomised clinical study assessed whether subcutaneous injection of 5.5 mL of TMS (40 mg zinc, 10 mg manganese, 5 mg selenium, 15 mg copper per mL), 14-28 days before planned start of calving (PSC) reduced clinical mastitis (CM), subclinical mastitis (SCM) and purulent vaginal discharge (PVD). From four farms, half of 1700 cows stratified on somatic cell count, age and breed were randomly allocated to treatment or no treatment. Occurrence of CM from - 7 to PSC + 100 days, SCM at PSC + 60 days and PVD at PSC + 24 days was analysed using survival analysis and Bayesian generalised mixed multivariable models. From -7 to PSC +30 days, TMS reduced the adjusted hazard ratio (HR) for CM at quarter and cow level (P < 0.001), with no evidence for an effect beyond 30 days. The adjusted OR (and 95% highest density interval, HDI) for the effect of TMS on CM from -7 to PSC +30 days was 0.40 (95% HDI, 0.26-0.63) at quarter level, 0.51 (95% HDI, 0.38-0.69) at cow level and for SCM, 0.72 (95% HDI, 0.54-0.95). The difference in CM incidence from TMS at the cow level was -2.0% (95% HDI, -3.4 to -1.1%) and -1.2% (95% HDI, -3.2 to - 0.6%) at quarter level. No clear effect was identified of TMS on cumulative incidence of PVD.

Subclinical mastitis in dairy cows in south-Asian countries: a review of risk factors and etiology to prioritize control measures.

Mastitis is a major production disease, causing significant economic losses for dairy farmers in South-Asian countries, as well as other parts of the world. Udder health control programs (UHCP) have been established in developed countries as an effective strategy for mastitis control but have not yet been introduced in South-Asian low-income countries like Bangladesh, India, Pakistan, and Sri Lanka. To launch UHCP successfully in dairy herds in South-Asia, it is important to know the current prevalence and risk factors for subclinical mastitis (SCM). Therefore, a narrative literature review was conducted with the aim to describe the dairy sector, the prevalence of SCM and its causal agents, risk factors for mastitis occurrence and the control measures suggested by different studies conducted in the selected countries. The literature revealed that India had the highest cattle population. Milking was mainly done by hand in all of the studied countries. Stall feeding was done in Bangladesh and Sri Lanka and limited access to grazing was also reported in some farms in India and Pakistan. There was substantial variation in the prevalence of SCM between studies in all 4 countries, ranging from about 20% to about 80%, but the average prevalence across all studies was high (50%). The most common causal agents for SCM were non-aureus staphylococci (NAS), Staphylococcus (S.) aureus, Streptococcus spp. and Escherichia (E.) coli. The management related risk factors reported for SCM were stall feeding of cows, a higher stock density, cracked floors, open drains, the presence of flies, poor drainage, peri-parturient diseases, infrequent dung removal and earth floors. The control measures suggested in these studies were to improve the hygiene and sanitation of cows, to improve the cleanliness of farms and milker's hands, to apply dry cow therapy, supplementing micronutrients and routine screening for SCM combined with taking intervention measures like isolation of cows or milking infected cows last, and proper treatment. Also, full hand milking, complete milking, machine milking, and providing feed and water immediately after milking have been recommended. Finally, we show that current literature often studies the same set of (non-manageable) risk factors, so more research is needed to obtain a comprehensive picture of the determinants of SCM. Randomized controlled trials are needed to truly quantify the effect of intervention under field conditions. Altogether, our work gives an overview of the udder health situation in South-Asia and provides the basis for the design of UHCP in this region.

Evaluation of fully oxidised ß-carotene as a feed ingredient to reduce bacterial infection and somatic cell counts in pasture-fed cows with subclinical mastitis.

AIMS: To assess the effect of oral supplementation with fully oxidised ß-carotene (OxBC) on bacteriological cure, the incidence of clinical mastitis, and somatic cell counts (SCC) in pasture-fed cows with subclinical intramammary infection. METHODS: Cows from four dairy herds were enrolled in early lactation if they had quarter-level SCC >200,000 cells/mL and they had a recognised bacterial intramammary pathogen in one or more quarters. They were randomly assigned to be individually fed from Day 0, for a mean of 40 days, with 0.5 kg of a cereal-based supplementary feed that either contained 300 mg of OxBC (treatment; n = 129 quarters) or did not (control; n = 135 quarters). Quarter-milk samples were collected on Days 21 and 42 for microbiology and SCC assessment. Bacteriological cure was defined as having occurred when the bacteria present on Day 0 were not isolated from samples collected on Days 21 or 42. A new intramammary infection was defined as a bacterial species isolated either on Day 21 or 42 differing from that isolated on Day 0. Clinical mastitis was diagnosed and recorded by herdowners up to Day 42. RESULTS: The bacteriological cure rate was greater for quarters from cows in the treatment group (13.9 (95% CI = 4.1-23.7)%) than for cows in the control group (6.9 (95% CI = 4.8-9.1)%; p = 0.02). The percentage of quarters that developed a new intramammary infection at Day 21 or 42 was greater for cows in the treatment group (17.9 (95% CI = 6.7-29.1)%) than for cows in the control group (13.0 (95% CI = 4.3-21.8)%; p < 0.01). The prevalence of quarters that were infected on Day 42 was less in cows in the treatment group (79.9 (95% CI = 62.3-97.6)%) than the control group (88.2 (95% CI = 78.4-97.9)%; p = 0.009). The incidence of quarters diagnosed with clinical mastitis by Day 42 was lower in cows from the treatment group (1/129 (0.78 (95% CI = 0.02-4.24)%)) than in cows from the control group (6/135 (4.44 (95% CI = 1.65-9.42)%; p = 0.03)). Mean quarter-level SCC was not statistically different between treatment groups (p = 0.34). CONCLUSIONS AND CLINICAL RELEVANCE: Feeding 300 mg/cow/day of OxBC resulted in a higher bacteriological cure rate, a lower prevalence of intramammary infection, and a lower incidence of clinical mastitis compared to untreated controls. However new intramammary infections increased in treated cows, and the magnitude of the increased bacteriological cure was low, resulting in 80% of cows remaining infected at Day 42. Therefore treatment with OxBC should be considered as an adjunct to other mastitis control measures.

Dynamics of the within-herd prevalence of Mycoplasma bovis intramammary infection in endemically infected dairy herds.

We aimed to identify the dynamics of the within-herd prevalence of Mycoplasma (M.) bovis intramammary infection (IMI) in four dairy herds, estimate prevalence of M. bovis in colostrum and clinical mastitis cases and compare M. bovis strains from calves' respiratory and cow clinical mastitis samples. Within a six-month study period, cow composite milk samples (CMS) were collected three times during routine milk recording, first milking colostrum samples from all calving cows and udder quarter milk samples from clinical mastitis cases. Calf respiratory samples were collected from calves with respiratory disease. Pooled milk samples were analysed for M. bovis with the Mastitis 4B polymerase chain reaction (PCR) test kit (DNA Diagnostic A/S). Prevalence estimates were calculated with Bayesian framework in R statistical programme. cg-MLST was used for M. bovis genotyping. In Herd I and II first testing M. bovis IMI within-herd prevalence (95 % credibility interval (CI)) was 4.7 % (2.9; 6.8) and 3.4 % (2.3; 4.6), changing to 1.0 % (0.1; 1.7) and 0.8 % (0.1; 1.4) in Herd I and 0.4 % (0.0; 0.7) in Herd II at the next samplings. In Herd III and IV first testing M. bovis IMI within-herd prevalence was 12.3 % (9.7; 15.2) and 7.8 % (6.2; 9.5), changing to 4.6 % (3.0; 6.4) and 3.2 % (1.9; 4.8) in Herd III and to 2.8 % (1.9; 3.8) and 4.9 % (3.6; 6.4) in Herd IV at the next samplings. The estimated prevalence of M. bovis in colostrum ranged between 1.7 % (0.2; 2.8) and 4.7 % (2.7; 7.1) and in clinical mastitis cases between 3.7 % (1.7; 6.4) and 11.0 % (7.5; 15.2) in the study herds. M. bovis strains isolated from cows and calves clustered within herds indicating possible transmission of M. bovis between dairy cows and calves. Prevalence of M. bovis in colostrum and clinical mastitis cases as well as the within-herd prevalence of M. bovis IMI was low in endemically infected dairy herds.

Transfer of passive immunity in dairy calves: the effectiveness of providing a supplementary colostrum meal in addition to nursing from the dam.

Failed transfer of passive immunity (FTPI) in dairy calves - which is often due to the low amount of colostrum provided within a few hours after birth - remains a crucial issue. Enabling dairy calves to nurse colostrum from their dams could be useful in increasing intake and thus avoiding FTPI, but further potential effects on the health and welfare of both calves and dams should also be considered. In this study, 107 calf-dam pairs from two Italian dairy farms were alternately assigned to one of the following colostrum provision methods (CPMs): 'hand-fed method' (HFM) - the calf was separated from the dam immediately after birth and colostrum was provided by nipple-bottle (n = 50); 'nursing method' (NM) - the calf nursed colostrum from the dam for the first 12 h of life without farmer assistance (n = 30); and 'mixed method' (MM) - the nursing calf received a supplementary colostrum meal by nipple-bottle (n = 27). Serum of calves (1 to 5 days of age) and samples of their first colostrum meal were analysed by electrophoresis to assess immunoglobulin (Ig) concentration. Additionally, behavioural indicators of separation distress (calf and dam vocalisations; calf refusal of the first meal after separation; undesirable dam behaviour at milking) in the following 24 h were recorded as binary variables (Yes/No), and the health status of calves (disease occurrence and mortality) and dams (postpartum disorders and mastitis occurrence) were monitored for the first 3 months of life and 7 days after parturition, respectively. The lowest FTPI occurrence (calf serum Ig concentration <10.0 g/l) was found in the MM (11.1%) and the HFM (22.0%) compared with the NM (60.0%) (P<0.05), and the highest percentage of calves with optimal transfer of passive immunity (serum Ig concentration ≥16.0 g/l) was observed in the MM (55.6%). The lowest calf-dam separation distress was observed in the HFM (P<0.05). The highest calf disease occurrence was recorded in the HFM (64.0%) and the lowest in the NM (33.3%), with an intermediate value for the MM (44.4%) (P<0.05). No effect of the CPM was observed on dam health or calf mortality (P>0.05). The results of this study indicated that providing calves with a supplementary colostrum meal in addition to nursing from the dam (MM) is truly effective in maximizing passive immunity transfer. Anyway, specific strategies should be studied to minimise calf-dam separation distress.

Effects of an immunomodulatory feed additive on intramammary infection prevalence and somatic cell counts in a dairy herd experiencing major health issues.

A trial was conducted to determine if feeding OmniGen-AF® (OG) to 22 late lactation cows 60 days prior to and during the early dry period, a time of increased susceptibility to mastitis, could reduce disease incidence in a dairy herd experiencing major health issues. Treated cows (n = 11) consumed a ration containing OG [9 g/100 kg of body weight/day] beginning 60 days before dry-off, during the dry period, and through 30 days in milk (DIM). Control cows received the same ration during the dry period through 30 DIM only. Body weights, body condition scores (BCS), intramammary infection (IMI) prevalence, new IMI rates, somatic cell counts (SCC), milk yield, and adverse health events were measured. No differences were found between treatments for body weight or BCS. Adverse health event data at calving showed no differences between treatments except for percentage of cows with hyperketonemia, which was lower among treated cows (63.6% vs 100%). Prevalence of IMI from calving through 30 DIM for treated cows (6.1%) was lower than controls (11.05%); likewise, new IMI rate during this time for treated cows (0.61%) was lower than controls (5.81%). The SCC from calving through 30 DIM for treated cows (215,000/ml) was lower than controls (493,000/ml). Average production/day at the first DHIA test (~33 DIM) showed that treated cows produced more milk (39.9 kg) than controls (35.34 kg). In conclusion, feeding OG 60 days prior to dry-off reduced hyperketonemia and mastitis, lowered SCC, and numerically increased milk yield in a dairy herd experiencing major health issues.

Udder health of dairy cows fed different dietary energy levels after a short or no dry period without use of dry cow antibiotics.

Reports on the effects of length of dry period (DP) on udder health of cows that were not treated with dry cow antibiotics are scarce. Additionally, the effects of a reduced dietary energy level for cows with a 0-d DP on udder health have not yet been studied. The aims of this study were (1) to compare effects of a 0-d or 30-d DP without use of dry cow antibiotics on udder health across the DP and subsequent lactation in dairy cows fed different dietary energy levels and (2) to evaluate associations between udder health and metabolic status of dairy cows. Five weeks before the expected calving date, Holstein-Friesian dairy cows (n = 115) were blocked for parity, expected calving date, and milk yield and somatic cell count (SCC) at their 2 last test days and were randomly assigned to 2 DP lengths: 0-d DP (n = 77) or 30-d DP (n = 38). Quarter milk samples were taken in wk 5 prepartum and in wk 1 and 5 postpartum. Proportion of quarters with elevated SCC (SCC ≥200,000 cells/mL) and proportion of udder pathogens in quarter milk samples did not differ between DP lengths among weeks. After calving, 102 of these cows were randomly assigned to 3 treatments: a 30-d DP with a standard energy level required for expected milk yield (30-d DP SEL; n = 36), a 0-d DP with the same energy level as cows with a 30-d DP (0-d DP SEL; n = 33), and a 0-d DP with a low energy level (0-d DP LEL, n = 33). From wk 8 of lactation onward, cows received either a glucogenic ration consisting of corn silage and grass silage or a lipogenic ration consisting of grass silage and sugar beet pulp at a standard or low energy level. During wk 1 to 7 postpartum, treatment did not affect SCC or SCC corrected for milk yield. During wk 8 to 44 of lactation, 0-d DP SEL cows had a greater SCC than 0-d DP LEL or 30-d DP SEL cows and had a greater SCC corrected for milk yield than 0-d DP LEL cows. During wk 1 to 44 of lactation, occurrence of at least 1 elevation of SCC (SCC ≥200,000 cells/mL after 2 wk of SCC <200,000 cells/mL) was not different among treatments. The 0-d DP SEL cows but not the 0-d DP LEL cows tended to have a 2.17 times greater hazard of having a case of clinical mastitis at any time in lactation than 30-d DP SEL cows. In wk 1 to 44 of lactation, lower fat- and protein- corrected milk yield and energy intake, greater energy balance, and greater plasma insulin concentration were associated with greater SCC. In conclusion, DP length did not affect udder health in the DP and in early lactation but seemed to decrease udder health for 0-d DP SEL cows in later lactation compared with 30-d DP SEL or 0-d DP LEL cows.

Effects of injectable trace mineral supplementation in lactating dairy cows with elevated somatic cell counts.

Objectives of this clinical trial were to evaluate the effects of injectable trace mineral supplementation (ITMS) on somatic cell count (SCC), linear score (LS), milk yield, milk fat and protein contents, subclinical mastitis cure, and incidence of clinical mastitis in cows with elevated SCC. Holstein cows from a commercial dairy farm in New York were evaluated for subclinical mastitis, defined as SCC ≥200×10(3) cells/mL on the test day preceding enrollment. Cows with a history of treatment for clinical mastitis in the current lactation and those pregnant for more than 150d were not eligible for enrollment. Cows fitting inclusion criteria were randomly allocated to 1 of 2 treatment groups. Cows assigned to ITMS (n=306) received 1 subcutaneous injection containing zinc (300mg), manganese (50mg), selenium (25mg), and copper (75mg) at enrollment (d 0). Control cows (CTRL; n=314) received 1 subcutaneous injection of sterile saline solution. Following treatment, visual assessment of milk was performed daily, and cows with abnormal milk (i.e., presence of flakes, clots, or serous milk) were diagnosed with clinical mastitis (CM). Chronic clinical mastitis was defined as cows with 3 or more cases of CM. Milk yield, milk fat and protein contents, SCC, and LS were evaluated once monthly. Additionally, randomly selected animals were sampled to test serum concentrations of selected minerals on d0 and 30 (n=30 cows/treatment). Treatment did not affect serum concentrations of calcium, magnesium, phosphorus, potassium, copper, iron, manganese, selenium, and zinc on d30. Injectable supplementation with trace minerals did not improve overall cure of subclinical mastitis (CTRL=42.8 vs. ITMS=46.5%), although a tendency was observed in cows with 3 or more lactations (CTRL=27.1 vs. ITMS=40.0%). Supplementation did not reduce treatment incidence of CM (CTRL=48.2 vs. ITMS=41.7%); however, it tended to reduce the proportion of cows diagnosed with chronic CM (CTRL=16.9 vs. ITMS=12.0%), particularly among first-lactation cows (CTRL=18.4 vs. ITMS=7.6%). Cure of subclinical mastitis was associated with higher serum concentrations of phosphorus and selenium on d30. Supplementing trace minerals to cows with elevated SCC had no effect on milk yield, milk fat and protein contents, SCC, and LS.

The influence of the rearing period on intramammary infections in Swiss dairy heifers: A cross-sectional study.

Healthy replacement heifers are one of the foundations of a healthy dairy herd. Farm management and rearing systems in Switzerland provide a wide variety of factors that could potentially be associated with intramammary infections (IMI) in early lactating dairy heifers. In this study, IMI with minor mastitis pathogens such as coagulase-negative staphylococci (CNS), contagious pathogens, and environmental major pathogens were identified. Fifty-four dairy farms were enrolled in the study. A questionnaire was used to collect herd level data on housing, management and welfare of young stock during farm visits and interviews with the farmers. Cow-level data such as breed, age at first calving, udder condition and swelling, and calving ease were also recorded. Data was also collected about young stock that spent a period of at least 3 months on an external rearing farm or on a seasonal alpine farm. At the quarter level, teat conditions such as teat lesions, teat dysfunction, presence of a papilloma and teat length were recorded. Within 24h after parturition, samples of colostral milk from 1564 quarters (391 heifers) were collected aseptically for bacterial culture. Positive bacteriological culture results were found in 49% of quarter samples. Potential risk factors for IMI were identified at the quarter, animal and herd level using multivariable and multilevel logistic regression analysis. At the herd level tie-stalls, and at cow-level the breed category "Brown cattle" were risk factors for IMI caused by contagious major pathogens such as Staphylococcus aureus (S. aureus). At the quarter-level, teat swelling and teat lesions were highly associated with IMI caused by environmental major pathogens. At the herd level heifer rearing at external farms was associated with less IMI caused by major environmental pathogens. Keeping pregnant heifers in a separate group was negatively associated with IMI caused by CNS. The odds of IMI with coagulase-negative staphylococci increased if weaning age was less than 4 months and if concentrates were fed to calves younger than 2 weeks. This study identified herd, cow- and quarter-level risk factors that may be important for IMI prevention in the future.

Lack of efficacy of homeopathic therapy against post-calving clinical mastitis in dairy herds in the Waikato region of New Zealand.

AIM: To compare clinical and bacteriological cure rates of clinical mastitis following treatment with either antimicrobials or homeopathic preparations. METHODS: Seven spring-calving herds from the Waikato region of New Zealand were used to source cases of clinical mastitis (n = 263 glands) during the first 90 days following calving. Duplicate milk samples were collected for bacteriology from each clinically infected gland at diagnosis and 25 (SD 5.3) days after initial treatment. Affected glands were treated with either an antimicrobial formulation or a homeopathic remedy. Generalised linear models with binomial error distribution and logit link were used to analyse the proportion of cows that were clinical treatment cures and the proportion of glands that were classified as bacteriological cures, based on initial and post-treatment milk samples. RESULTS: Mean cumulative incidence of clinical mastitis was 7% (range 2-13% across herds) of cows. Streptococcus uberis was the most common pathogen isolated from culture-positive samples from affected glands (140/209; 67%). The clinical cure rate was higher for cows treated with antimicrobials (107/113; 95%) than for cows treated with homeopathic remedies (72/114; 63%) (p < 0.001) based on the observance of clinical signs following initial treatment. Across all pathogen types bacteriological cure rate at gland level was higher for those cows treated with antimicrobials (75/102; 74%) than for those treated with a homeopathic preparation (39/107; 36%) (p < 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: Using herds located in the Waikato region of New Zealand, homeopathic remedies had significantly lower clinical and bacteriological cure rates compared with antimicrobials when used to treat post-calving clinical mastitis where S. uberis was the most common pathogen. The proportion of cows that needed retreatment was significantly higher for the homeopathic treated cows. This, combined with lower bacteriological cure rates, has implications for duration of infection, individual cow somatic cell count, costs associated with treatment and animal welfare.

[Effectiveness of a homeopathic prophylaxis for bovine mastitis during dry-off and parturition]. / Zur Wirksamkeit einer homöopathischen Mastitisprophylaxe zum Trockenstellen und Abkalben beim Rind.

The present study examines the efficacy of a homeopathic dry cow prophylaxis in a randomized, placebo controlled case-control field trial. The study was conducted in 24 Brown Swiss farms in the Engadine (Swiss mountain region). The effect of the used homeopathic substances were combined with antibiotics in justified cases. At drying off and in the 3rd and 5th week of lactation the udders were clinically examined and quarter milk samples were taken for bacteriological and cytological analysis. In addition, milk recording data of the first 6 milk testing were included in the evaluations. The used homeopathic prophylaxis at drying off did not show any effect in the incidence of dry cow mastitis and mastitis in the first 120 days of lactation. However, at day 21 post partum significantly fewer animals in the verum group showed a bacteriological finding of a major pathogen, but more animals in this group suffered from a secretion disorder. It has been shown that at the 6th milk test pp significantly more animals of the verum group had a somatic cell count below 100'000 cells/ml than the control group.

Dans cette étude, on contrôle l'efficacité d'une prévention homéopathique lors du tarissement dans le cadre d'une étude randomisée versus placebo. L'étude a été réalisée dans 24 exploitations en Engadine, élevant de la race brune. L'efficacité des produits homéopathiques utilisés a également été considérée en tenant compte des antibiotiques appliquées dans les cas fondés. Chez tous les animaux on a recueilli un échantillon de chaque quartier lors du tarissement ainsi qu'en 3ème et 5ème semaine de lactation. Ces échantillons ont été examinés du point de vu bactériologique et quant au nombre de cellules. En outre, le taux de cellules des 6 premiers contrôles laitier a été intégré dans le calcul des résultats. La prévention homéopathique lors des tarissement n'a pas montré d'effet quant à l'apparition de mammites durant la période de tarissement ou durant les 120 premiers jours de la lactation. On a toutefois pu montrer que, 21 jours postpartum, le nombre d'animaux présentant des résultats bactériologiques avec des germes importants pour les mammites était significativement plus bas mais qu'un plus grand nombres d'animaux souffraient des troubles de la sécrétion. Lors du 6ème contrôle laitier postpartum, il y avait dans le groupe test plus d'animaux avec un taux de cellule inférieur à 100'000/ml que dans le groupe de contrôle.

Prevalence of periparturient diseases and effects on fertility of seasonally calving grazing dairy cows supplemented with concentrates.

The objectives were to characterize the prevalence of periparturient diseases and their effects on reproductive performance of dairy cows in seasonal grazing farms. A total of 957 multiparous cows in 2 farms (555 in farm A and 402 in farm B) were evaluated and diseases characterized. At calving, dystocia, twin birth, stillbirth, and retained fetal membranes were recorded and grouped as calving problems. On d 7±3 and 14±3 postpartum, cows were evaluated for metritis and on d 28±3 for clinical endometritis based on scoring of the vaginal discharge. From parturition to 30 d after artificial insemination (AI), prevalence of mastitis, lameness, and digestive and respiratory problems were recorded. For subclinical diseases, diagnosis was based on blood samples collected from 771 cows and analyzed for concentrations of Ca, nonesterified fatty acids (NEFA), and ß-hydroxybutyrate. Cows were considered as having elevated NEFA concentration if the concentration was ≥0.70 mM, subclinical ketosis if the ß-hydroxybutyrate concentration was ≥0.96 mM, and subclinical hypocalcemia if the Ca concentration was ≤2.14 mM. Ovaries were scanned on d 35±3 and 49±3 postpartum for determination of estrous cyclicity. All cows were enrolled in a timed AI program and inseminated on the first day of the breeding season: on average, 86 d postpartum. Overall, 37.5% (359/957) of the cows presented at least 1 clinical disease and 59.0% (455/771) had at least 1 subclinical health problem. Prevalence of individual diseases was 8.5% for calving problems, 5.3% for metritis, 15.0% for clinical endometritis, 13.4% for subclinical endometritis, 15.3% for mastitis, 2.5% for respiratory problems, 4.0% for digestive problems, 3.2% for lameness, 20.0% for elevated NEFA concentration, 35.4% for subclinical ketosis, and 43.3% for subclinical hypocalcemia. Clinical and subclinical diseases had additive negative effects on reproduction, delaying resumption of estrous cyclicity and reducing pregnancy per AI (P/AI). Occurrence of multiple diseases further reduced reproductive efficiency compared with a single disease. Individually, subclinical hypocalcemia, elevated NEFA concentration, metritis, and respiratory and digestive problems reduced estrous cyclicity by d 49 postpartum. Elevated NEFA concentration, calving problem, metritis, clinical and subclinical endometritis, and digestive problems reduced P/AI on d 65 after AI. Moreover, calving problems and clinical endometritis increased the risk of pregnancy loss between gestation d 30 and 65. Serum concentrations of Ca and NEFA were negatively correlated, and both were associated with prevalence of uterine diseases. In conclusion, periparturient diseases were highly prevalent in seasonally calving grazing dairies and affected cows had delayed resumption of estrous cyclicity, reduced P/AI, and increased risk of pregnancy loss.

Inferring relationships between clinical mastitis, productivity and fertility: a recursive model application including genetics, farm associated herd management, and cow-specific antibiotic treatments.

A dataset of test-day records, fertility traits, and one health trait including 1275 Brown Swiss cows kept in 46 small-scale organic farms was used to infer relationships among these traits based on recursive Gaussian-threshold models. Test-day records included milk yield (MY), protein percentage (PROT-%), fat percentage (FAT-%), somatic cell score (SCS), the ratio of FAT-% to PROT-% (FPR), lactose percentage (LAC-%), and milk urea nitrogen (MUN). Female fertility traits were defined as the interval from calving to first insemination (CTFS) and success of a first insemination (SFI), and the health trait was clinical mastitis (CM). First, a tri-trait model was used which postulated the recursive effect of a test-day observation in the early period of lactation on liability to CM (LCM), and further the recursive effect of LCM on the following test-day observation. For CM and female fertility traits, a bi-trait recursive Gaussian-threshold model was employed to estimate the effects from CM to CTFS and from CM on SFI. The recursive effects from CTFS and SFI onto CM were not relevant, because CM was recorded prior to the measurements for CTFS and SFI. Results show that the posterior heritability for LCM was 0.05, and for all other traits, heritability estimates were in reasonable ranges, each with a small posterior SD. Lowest heritability estimates were obtained for female reproduction traits, i.e. h(2)=0.02 for SFI, and h(2)≈0 for CTFS. Posterior estimates of genetic correlations between LCM and production traits (MY and MUN), and between LCM and somatic cell score (SCS), were large and positive (0.56-0.68). Results confirm the genetic antagonism between MY and LCM, and the suitability of SCS as an indicator trait for CM. Structural equation coefficients describe the impact of one trait on a second trait on the phenotypic pathway. Higher values for FAT-% and FPR were associated with a higher LCM. The rate of change in FAT-% and in FPR in the ongoing lactation with respect to the previous LCM was close to zero. Estimated recursive effects between SCS and CM were positive, implying strong phenotypic impacts between both traits. Structural equation coefficients explained a detrimental impact of CM on female fertility traits CTFS and SFI. The cow-specific CM treatment had no significant impact on performance traits in the ongoing lactation. For most treatments, beta-lactam-antibiotics were used, but test-day SCS and production traits after the beta-lactam-treatment were comparable to those after other antibiotic as well as homeopathic treatments.

Prevalence and antimicrobial susceptibility patterns of bacteria from milkmen and cows with clinical mastitis in and around Kampala, Uganda.

BACKGROUND: Identification of pathogens associated with bovine mastitis is helpful in treatment and management decisions. However, such data from sub-Saharan Africa is scarce. Here we describe the distribution and antimicrobial susceptibility patterns of bacteria from cows with clinical mastitis in Kampala, Uganda. Due to high concern of zoonotic infections, isolates from milkmen are also described. METHODOLOGY/PRINCIPAL FINDINGS: Ninety seven milk samples from cows with clinical mastitis and 31 nasal swabs from milkmen were collected (one sample per cow/human). Fifty eight (60%) Gram-positive isolates namely Staphylococci (21), Enterococci (16), Streptococci (13), Lactococci (5), Micrococci (2) and Arcanobacteria (1) were detected in cows; only one grew Staphylococcus aureus. Furthermore, 24 (25%) coliforms namely Escherichia coli (12), Klebsiella oxytoca (5), Proteus vulgaris (2), Serratia (2), Citrobacter (1), Cedecea (1) and Leclercia (1) were identified. From humans, 24 Gram-positive bacteria grew, of which 11 were Staphylococci (35%) including four Staphylococcus aureus. Upon susceptibility testing, methicillin-resistant coagulase-negative staphylococci (CoNS) were prevalent; 57%, 12/21 in cows and 64%, 7/11 in humans. However, methicillin-resistant Staphylococcus aureus was not detected. Furthermore, methicillin and vancomycin resistant CoNS were detected in cows (Staphylococcus hominis, Staphylococcus lugdunensis) and humans (Staphylococcus scuiri). Also, vancomycin and daptomycin resistant Enterococci (Enterococcus faecalis and Enterococcus faecium, respectively) were detected in cows. Coliforms were less resistant with three pan-susceptible isolates. However, multidrug resistant Klebsiella, Proteus, Serratia, Cedecea, and Citrobacter were detected. Lastly, similar species grew from human and bovine samples but on genotyping, the isolates were found to be different. Interestingly, human and bovine Staphylococcus aureus were genetically similar (spa-CC435, spa-type t645 corresponding to ST121) but with different susceptibility patterns. CONCLUSIONS/SIGNIFICANCE: CoNS, Enterococci, Streptococci, and Escherichia coli are the predominant pathogens associated with clinical bovine-mastitis in Kampala, Uganda. Multidrug resistant bacteria are also prevalent. While similar species occurred in humans and cows, transmission was not detected.

Reevaluation of vitamin E supplementation of dairy cows: bioavailability, animal health and milk quality.

Although vitamin E has been known as an essential nutrient for almost 80 years, we are far from a complete understanding of all the aspects related to bioavailability and its effects on health and milk quality in dairy cows. Vitamin E is a generic descriptor for two families of lipid-soluble compounds, the tocopherols and the tocotrienols, of which α-tocopherol has the highest biological activity. Commercially available α-tocopherol supplements for dairy cows contain either the natural RRR form or the synthetic (all-rac) form, which contains all the eight possible stereoisomers (four possessing the 2R and four possessing the 2S configuration) in equimolar amounts. Recent data clearly suggest that an almost complete discrimination against the 2S isomers occurs in dairy cows. Thus, 1 g of the all-rac form is essentially equivalent to 0.5 g of the RRR form. With respect to the effect of vitamin E supplementation of dairy cows on health and milk quality, the majority of published studies suggests that vitamin E supplementation at the level 1000 to 4000 IU/cow per day during the dry period reduces both the frequency of intramammary infection and that of clinical mastitis and improves milk quality, as shown by a reduction in the levels of somatic cell count (SCC)/ml in milk, decreased plasmin activity and increased oxidative stability of milk. However, a recent study from the Netherlands suggested that vitamin E supplementation at the 3000 IU/cow per day level during the dry period when combined with high levels of plasma vitamin E at dry-off (>14.5 µmol/l) increases the incidence of mastitis. Data from previously unpublished survey studies and those from published vitamin E feeding trials, in which high levels of blood vitamin E were observed, were reanalyzed. All farms selected for the analysis implemented oral administration of vitamin E at the 3000 IU/cow per day level throughout or during the late dry period (4 weeks before the expected day of parturition). Dairy cows were divided into three groups, depending on blood α-tocopherol levels at dry-off: high (>6.25 µg/ml), medium (between 6.25 and 4.25 µg/ml) and low (<4.25 µg/ml). Data indicate that there were no differences in the incidence of mastitis and in the level of SCC/ml of milk between the three groups. Thus, supplementation of 3000 IU vitamin E/cow per day in the late dry period remains recommended because it is generally associated with decreased risk of mastitis. Conditional or opposite effects have not been repeated and require further research before changing recommendations for vitamin E supplementation.

Pathogen group specific risk factors at herd, heifer and quarter levels for intramammary infections in early lactating dairy heifers.

Risk factors for intramammary infections caused by coagulase-negative staphylococci, contagious major pathogens and environmental major pathogens in early lactating heifers were evaluated at the herd, heifer and quarter levels. In total, 764 quarters of 191 dairy heifers in 20 randomly selected farms in Flanders (Belgium) were sampled. Quarter milk samples were collected between 1 and 4 days in milk and between 5 and 8 days in milk for bacteriological culture. Data were analyzed using multivariable, multilevel logistic regression analysis. Higher average herd milk somatic cell count (>200,000 cells/mL), not having an effective fly control strategy, contact with lactating cows prior to calving and moderate to severe udder edema prior to calving increased the odds of intramammary infections caused by contagious major pathogens. Poor heifer hygiene and lack of mineral/vitamin supplementation prior to calving were risk factors for intramammary infection caused by environmental major pathogens. Teat apex colonization with coagulase-negative staphylococci prior to calving seemed to protect quarters against intramammary infections caused by major pathogens. Poor heifer hygiene before calving, a non-clipped udder and not practicing of teat dipping prior to calving increased the odds of intramammary infection with coagulase-negative staphylococci. Although management is important in the prevention and control of intramammary infections in early lactating heifers, most variation in the prevalence of intramammary infections resided at the heifer and quarter levels, indicating that the susceptibility for intramammary infections around calving is mainly determined by heifer and quarter characteristics.

Vitamin E supplementation during the dry period in dairy cattle. Part I: adverse effect on incidence of mastitis postpartum in a double-blind randomized field trial.

A randomized, controlled field trial with dairy cows demonstrated an adverse effect of vitamin E supplementation during the dry period on mastitis incidence in early lactation. This study was conducted on farms with historically high rates of mastitis to investigate the benefit of vitamin E supplementation on udder health; however, the outcome showed an adverse effect. The aim of the study was to evaluate whether daily supplementation of 3,000 IU of vitamin E to dairy cows during the dry period could improve udder health in commercial herds with a high incidence of mastitis. On 5 dairy farms, dry cows were randomly divided into 2 experimental groups: a high and a low group. Both groups received a dry cow mineral mix providing 3,000 or 135 IU of vitamin E/cow per day, respectively, between dry-off and calving for a mean period of 8 wk. Providing 3,000 IU of vitamin E exceeds NRC standards, but this amount has been used in previous studies. The experiment, as well as the majority of the statistical analysis, were carried out blinded. Blood was sampled 3 times before calving and on calving day. Serum was analyzed for vitamin E and cholesterol. Vitamin E and the vitamin E:cholesterol ratio were analyzed as dependent variables in mixed models and Student's t-tests to study trends in time and differences between groups. Relative risk calculation and survival analysis were used to study the effect of supplementation on mastitis incidence in the first 3 mo of lactation. The results showed that vitamin E supplements increased both absolute vitamin E and the ratio of vitamin E to cholesterol in blood. In the high group, significantly more subclinical and clinical cases occurred, showing the same trend on all farms. In this study, an initial vitamin E level at dry off above 14.5 µmol/L was a risk factor for clinical mastitis, suggesting that the vitamin E status at the start of the dry period is important. It is recommended to work out exactly at what threshold vitamin E is harmful for udder health before new trials with high dosages of vitamin E are started. Additionally, further research is required to investigate the mechanism by which vitamin E affects udder health.

Vitamin E supplementation during the dry period in dairy cattle. Part II: oxidative stress following vitamin E supplementation may increase clinical mastitis incidence postpartum.

The aim of this study was to evaluate, retrospectively, which physiological states influenced the effect of vitamin E supplements during the dry period on the level of oxidative stress at 2 wk antepartum. Furthermore the effect of oxidative stress at 2 wk antepartum on the risk of clinical mastitis in early lactation was investigated. Cows experience oxidative stress around calving. Vitamin E is able to decrease oxidative stress by scavenging free radicals. Normally, vitamin E radicals formed when vitamin E reacts with free radicals are regenerated by a network of other antioxidants, termed the "vitamin E regeneration system" (VERS). In case of vitamin E supplementation, VERS should be sufficient to regenerate formed vitamin E radicals; if not, oxidative stress might increase instead of decrease. Additionally, the level of oxidative stress and vitamin E might be important physiological states to evaluate before supplementation. In a clinical trial, 296 cows on 5 farms were randomly divided into 2 groups, supplemented with a mineral mix between dry off and calving that supplied 3,000 or 135 IU/d, respectively. Blood samples collected at dry off and 2 wk antepartum were analyzed for vitamin E, reactive oxygen metabolites, ferric-reducing ability of plasma, glutathione peroxidase, and malondialdehyde. Cows were allocated retrospectively into 8 subgroups based on the level of oxidative stress, vitamin E, and VERS status at dry off. To evaluate whether differences in physiological states at dry off influenced the effect of vitamin E supplementation on the level of oxidative stress, group effects (supplemented vs. control) were studied with Student's t-test for all 8 subgroup at 2 wk antepartum. Differences in physiological states at dry off influenced the effect of vitamin E supplements. In 2 insufficient VERS subgroups, the supplemented group had higher levels of free radicals at 2 wk antepartum compared with the control group. Relative risk calculation was used to study the effect of oxidative stress at 2 wk antepartum on the incidence of mastitis in the first 100 d of lactation. Higher levels of oxidative stress at 2 wk antepartum were related to higher risk of clinical mastitis. In conclusion, not every dry cow responded well to high vitamin E supplementation. This subgroup analysis provides a possible explanation for the unexpected adverse effects observed in the clinical trial.