The use of polyethylene intramammary device in protection of the lactating bovine udder against experimental infection with Staphylococcus aureus or Streptococcus agalactiae.

The susceptibility of lactating bovine udder quarters fitted with a polyethylene intramammary device to infection was investigated. Following experimental challenge with Streptococcus agalactiae or Staphylococcus aureus, the incidence of infection was significantly (p less than 0.05) lower in intramammary device-fitted quarters compared to control quarters. In general, total foremilk and strippings milk somatic cell counts for intramammary device-fitted and control quarters were not significantly (p less than 0.05) different. Differential foremilk and strippings milk somatic cell counts were significantly (p less than 0.05) higher in samples from intramammary device-fitted quarters compared to control quarters.     

Effect of intramammary devices on the outcome of induced Escherichia coli infection of bovine mammary quarters

Eighteen Holstein cows, free of intramammary infection, were fitted with smooth (n = 9) or abraded (n = 9) intramammary devices (IMD) in 2 diagonally opposed quarters within 4 weeks after calving. The 2 other quarters of each cow were used as controls. Three to 6 weeks after IMD insertion, depending on when milk somatic cell counts returned to a base-line value of less than 4 X 10(5)/ml, all cows were subjected to bacterial challenge exposure in the front or rear quarters by intracisternal injection of about 30 colony-forming units of Escherichia coli/quarter. Challenge exposure was done immediately after milking. Three weeks after the initial bacterial exposure, the other quarter pairs were similarly challenge exposed. Quarter bacteriologic status, concentration of milk somatic cells, and clinical observations (rectal temperature, milk appearance, udder palpation, and general condition of the cow) were monitored. Infection developed in 14 of 16 (88%) quarters with smooth IMD vs 16 of 16 (100%) control quarters and in 7 of 17 (41%) quarters with abraded IMD vs 17 of 17 (100%) control quarters. The difference in infection frequency between quarters with smooth IMD and quarters with abraded IMD was significant (P less than 0.05). Protection against establishment of infection was associated with somatic cell counts greater than 8.0 X 10(5)/ml in milk collected immediately after milking (7 of 12 quarters) or 4 hours later (11 of 12 quarters). In 10 quarters (59%) of cows fitted with abraded IMD, secretory abnormalities appeared before bacterial challenge inoculation. Abnormal milk or visible blood was observed over periods varying from 2 weeks after insertion through the entire lactation.     

Effect of teat dipping with a germicide barrier teat dip in late gestation on intramammary infection and clinical mastitis during the first 5 days post-partum in primiparous cows

The effect of teat dipping with a barrier teat dip prior to parturition on intramammary infection (IMI) and clinical mastitis during the first 5 days post-partum was investigated in a split udder trial in 149 Holstein-Frisian heifers. Their left front and right hind quarters were dipped three times weekly (i.e. Monday, Wednesday and Friday) with a barrier teat dip containing 0.1% polyvidon iodine from day 260 of gestation until parturition. The opposite quarters (right front and left hind quarter) served as untreated control. Bacteria were isolated from 52.2% of quarter milk samples collected immediately after parturition prior to first machine milking. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were predominantly found in the samples (29.2 and 35.6% of the positive samples, respectively). At parturition 6.7% of the heifers showed signs of clinical mastitis and another 27.5% developed signs of clinical mastitis during the first five days of lactation. No significant differences were found between treated and control quarters regarding IMI and incidence of clinical mastitis. Teat dipping prior to parturition in primigravid dairy heifers did not improve udder health in this trial.     

Removal of hair surrounding the teat and associated bacterial counts on teat skin surface, in milk, and intramammary infections

The effectiveness of monthly removal of hair surrounding teats on the reduction of teat skin surface bacteria, and the incidence of intramammary infection (IMI), was studied for 10 months in a dairy farm. A split udder design was used where hair was removed on one side, left or right, with the other side serving as a control. Controls and treatment sides were randomly applied in a systematic fashion to 218 cows. Standard milking time pre- and post-milking hygiene practices were applied to all udders during the trial. Collection of teat skin swab solutions preceded aseptic collection of milk samples, performed at monthly intervals, immediately prior to milking. Teat skin bacterial counts did not differ between control and treated teats. Incidences of IMI were similar for treatment when compared with control mammary quarters, as measured by total or by pathogen type. In a second study, the effect of hair removal on the bacterial content of milk was determined using 40 cows. Treatments and allocations were as described. Udder half milk, milk from both mammary quarters of each udder half, was combined and diverted into separate buckets. Buckets were thoroughly cleaned and sanitized between milkings. A portion of bucket milk was collected 24 h after removal of udder hair. The total milk bacterial counts, and counts of psychrotrophs and thermoduric organisms were not reduced by udder hair removal. Results do not suggest that removal of udder hair leads to an improvement in milk quality as determined by milk bacterial content in the herd studied.     

不孕育成牛人工诱导泌乳试验

选用5头（20-34月龄）无子宫和卵巢疾病的黑白花育成牛,研究用诱乳激素配合地塞米松和乳房按摩诱导泌乳的效果。试验牛每天每100kg体重注射0．5mL诱乳激素,连续注射8d,隔1d后每头每天再注射地塞米松磷酸钠20mg,连续注射3d,在注射激素药物期间每天按摩乳房三次,观察诱乳情况。结果5头育成牛全部诱乳成功,成功率100%。试验牛305d平均日产奶量8．32kg;同时对试验牛随后的繁殖和泌乳情况进行了跟踪观察,结果5头育成牛诱乳当年全部受胎并于次年产犊,泌乳期平均日产奶量11．86kg,产奶量比诱乳当年相对提高42．55％。     

Effect of anthelmintic treatment of second-year cattle on growth performance during winter housing and first lactation yield

The effect of nematode infections on growth performance during winter housing and on first lactation yield was investigated in heifers on 69 farms. Half of each herd was treated with albendazole after housing, which resulted in significantly reduced egg output, antibody titres against Ostertagia and Cooperia, and pepsinogen values compared with the untreated control groups. Albendazole-treated groups gained on average 0.007 cm day-1 heart girth (i.e. 0.05 kg day-1) more than control groups during the housing period (P less than or equal to 0.10), varying from -0.060 to +0.137 cm day-1 among herds. Albendazole-treated heifers produced 195.4 kg milk per 305-day lactation more than untreated heifers (P less than 0.01). Per herd, the mean milk yield response to treatment was +313.8 kg milk (P less than 0.01), varying from -876 to +1950 kg. Milk production of untreated heifers increased by 10.5 kg milk kg-1 extra body weight, adjusted for age, measured after the second grazing season (P less than 0.05). This suggests that nematode infections occurring in the first 2 years of life negatively influence milk production by reducing weight gains and hence body weight at calving.     

The Use of Induced Mammary Infections for Evaluating Dry Cow Treatment Products II. Trial of a Proposed Method to Compare Three Levels of Novobiocin

Infections were induced at the end of lactation in all udder quarters of 19 cows by the infusion of 0.2 mL of a 10^-4 dilution in milk of a six hour milk culture of Staphylococcus aureus, strain 305 (A.T.C.C No. 29740). Two right or two left udder quarters were infected at 15 days and the opposite two five days before the last milking of lactation. Following the last milking all four udder quarters of eight cows were treated with 400 mg novobiocin in 10 mL of 2% aluminum monosterate in peanut oil, gelled. All udder quarters of eight other cows were treated with 50 mg novobiocin in the same vehicle and the udder quarters of three cows were treated with the vehicle only.

At calving, eight of 32 quarters treated with 400 mg novobiocin were still infected, as were 18 of 32 treated with 50 mg of novobiocin and all those quarters treated with vehicle only. Results were identical from udder quarters infected 15 and five days before drying off.

No significant differences were found between quarters in milk yield on the last day of lactation, nor the length of the dry period. An increasing number of udder quarters were infected at calving with increase in lactation age of the cow, although the small number of cows would not allow a firm conclusion. A significant difference in results was found between front and hind udder quarters, only five of 32 front quarters were infected at calving as compared to 21 to 32 hind quarters.

The method proposed was found to give essentially the same results as those from a large field trial using the same antibiotic. It should therefore be useful in evaluation trials of new antibiotic products for dry cow treatment.

     

重组溶葡萄球菌酶对奶牛乳房炎的疗效研究

以85头患乳房炎奶牛共104个乳区为研究对象,随机分为4组,每组26个,分别用重组溶葡萄球菌酶粉低（200U／乳区）、中（400U／乳区）、高（800U／乳区）三个剂量,对照组为青霉素G钠（160万IU／乳区）,每天早晚挤奶后乳池灌注给药,共4d,研究不同剂量重组溶葡萄球菌酶对奶牛乳房炎的治疗效果。试验结果显示,低、中、高剂量重组溶葡萄球菌酶均能有效清除感染乳区的链球菌、葡萄球菌、化脓隐秘杆菌等G^＋菌,大幅降低牛奶中的白细胞数,提高日产奶量。其中低剂量组对隐性乳房炎、临床型乳房炎的有效率和治愈率略优于青霉素G钠（P〉0．05）;中、高剂量对隐性乳房炎、临床型乳房炎的有效率和治愈率显著优于青霉素G钠（P〈0．05）;中、高剂量的疗效相当（P〉0．05）。重组溶葡萄球菌酶是一种很好的治疗奶牛隐性乳房炎和临床乳房炎备选药物。     

Impact of intramammary infections in dairy heifers on future udder health, milk production, and culling

Dairy heifers represent the future of a dairy herd, and are expected to freshen with a healthy and well-developed udder, capable of producing an optimal amount of high quality milk. A high proportion of heifers have infected mammary quarters at calving, with coagulase-negative staphylococci (CNS) being the most common cause. Staphylococcus aureus and environmental pathogens are also found. The aim of this paper is to summarize how intramammary infections during (late) gestation and early lactation impair the development of the mammary gland and negatively affect future udder health and milk production. Heifers calving with either subclinical or clinical mastitis are also at a higher risk to be culled in first lactation. The magnitude of the effect is most likely related to the virulence of the causative pathogen, the persistence of the infection when milk production has started, and the time of onset of infection. Histological changes in udder tissue from quarters infected with S. aureus are more pronounced than those in udder tissue from CNS-infected quarters. The longer the infections exist and the longer they persist into lactation, the larger the impact on heifers' future udder health and milk production will be. In general, CNS infections are cleared early in lactation and some studies show that CNS do not have a large impact on future milk production and udder health. Future research should elucidate to what extent pathogen-specific as well as host-related factors affect the persistence of IMI in early lactating heifers.     

Effect of continuous milking and prostaglandin E2 on milk production and mammary epithelial cell turnover, ultrastructure, and gene expression

Mammary epithelial cell (MEC) growth is reduced in continuously milked (CM) mammary glands, and administration of a mammogenic compound such as prostaglandin E(2) (PGE(2)) at parturition might improve MEC growth in CM tissue. The objectives were to 1) compare MEC turnover, ultrastructure, and gene expression in CM and involuting mammary tissue, and 2) evaluate the effects of CM and intramammary infusion of PGE(2) on early lactation MEC turnover, ultrastructure, mammary gene expression, milk yield, and composition. First- and second-lactation cows (n = 8) were used in a half-udder model, in which one-half was dry for 60 d (CTL) and the other was CM. Udder halves (n = 16) were assigned to a postpartum (PP) treatment of PGE(2) (+PGE(2); 875 mug/10 mL of medium-chain triglyceride oil) or no PGE(2) (-PGE(2)) treatment at parturition and at 72 h PP. Biopsies of CM and CTL quarters were obtained during milk stasis (MS) of the CTL half at 3 and 7 d after dry-off of the CTL half (3d-MS; 7d-MS) and postpartum (PP) at 2 and 4 d (2d-PP; 4d-PP). Milk yield was reduced (P < 0.01) in CM udder halves compared with CTL halves (13.2 vs. 22.1 kg/d), but reductions were less in second-lactation cows. The apoptotic index was greater (P < 0.05) in CTL glands than in CM glands (3d-MS, 0.52 vs. 0.11% and 7d-MS, 0.24 vs. 0.12, respectively). Proliferation of MEC was unchanged at 3d-MS, but was increased (P = 0.01) in CTL halves at 7d-MS compared with CM halves (3.10 vs. 0.93%). At 2d-PP, MEC proliferation was increased (P = 0.05) in CM halves compared with CTL halves (1.3 vs. 0.6%), but was unaffected by PGE(2) (P > 0.2). Apoptosis was elevated in early lactation regardless of treatment. Ultrastructure was unchanged by dry period length or PGE(2). In prepartum tissue, involution in CTL halves increased (P < 0.05) the expression of the proapoptotic genes Bcl-2-associated x protein (bax) and IGFBP5 and decreased (P < 0.05) alpha-lactalbumin expression compared with CM tissue. In PP mammary tissue, CTL halves expressed greater (P < 0.05) levels of ATP-binding cassette 1 (ABC1) and IGFBP5. Treatment with PGE(2) did not alter (P > 0.1) gene expression. The results confirm that CM reduced milk yield of cows with a mammary growth requirement. Reduced MEC turnover and milk yield were not alleviated by IMI of PGE(2), which indicates that peripartum PGE(2) concentrations in CM glands are not limiting mammary growth or milk synthesis.     

Prevalence of udder bacteria in milk samples from four dairy goat herds

The prevalence of udder bacterial infections in 4 commercial goat herds in New South Wales was examined. Coagulase negative staphylococci were the predominant bacteria isolated being cultured from 13.3% of the 896 halves tested. Other bacteria isolated were Staphylococcus aureus (less than 1% of halves), streptococci (0.6%) and coliforms (2.0%). The low prevalence of udder bacteria contributing to the contamination of the milk was attributed to the milking management practices carried out in these herds, in particular pre-milking and post-milking antisepsis.     

Effect of setting a maximum milking time,from peak lactation,on production,milking time and udder health

Abstract

AIM: To examine the effect of setting a maximum milking time, from peak lactation until drying-off, on production, duration of milking, and udder health of dairy cows.

METHODS: Forty cows were assigned in twin-pairs to be either milked until cups were removed at a milk flow-rate threshold of 0.35 kg/minute (Control), or until cups were removed at a milk flow-rate threshold of 0.35 kg/minute, or maximum time, whichever came first (MaxT). The maximum time was set by determining the milking time of the 70th percentile cow when ranked from fastest to slowest, irrespective of yield. The milking routine was typical of that practised on dairy farms in New Zealand, and involved no pre-milking preparation. The study began at peak lactation (68 (SD 7) days in milk; DIM) and continued for 26 weeks. Duration of milking and milk yield were measured for each milking. Composition of milk was determined from weekly herd tests, and milk quality from fortnightly somatic cell counts (SCC). Completeness of milking and teat condition were assessed during the study. The bacterial status of quarter milk samples was determined at the beginning and end of the study, and all treated cases of clinical mastitis recorded. ANOVA was used to examine the effect of treatment group on variables of interest.

RESULTS: Total milk, fat and protein yields during the study period did not differ between treatments. On average, 30.3% of the morning and 27.6% of the afternoon milkings of MaxT cows reached the maximum time at which cups were removed, and were therefore shortened. While the average milking time of the slowest-milking cow was longer for the Control compared with MaxT group in Weeks 1–18, the average milking time did not differ between treatments. There was no difference in overall SCC, and the incidence of clinical mastitis, or the percentage of infected quarters at drying-off, was similar for the MaxT and Control cows.

CONCLUSION: The results show that setting a maximum milking time can reduce the milking time of slower-milking cows in a herd without compromising overall herd production and udder health.

CLINICAL RELEVANCE: Although the numbers of cows in the study were small there was no evidence of a major increase in SCC, or subclinical or clinical mastitis when a maximum milking time was set for slower-milking cows.     

Association of CXCR1 polymorphisms with apoptosis,necrosis and concentration of milk neutrophils in early lactating dairy heifers

Associations between polymorphisms in the candidate gene CXCR1, encoding the chemokine (C-X-C motif) receptor 1, and udder health have been identified before. In the present study, associations between the CXCR1 genotype (whole coding region) and apoptosis, necrosis, and concentration of milk polymorphonuclear neutrophilic leukocyte (PMNL) of 292 quarters belonging to 73 early lactating dairy heifers were studied. In uninfected quarters, % milk PMNL apoptosis was higher in c.980GG heifers [least squares means (LSM) 27%] compared to c.980AG heifers (LSM 16%), whereas in infected quarters, % milk PMNL apoptosis was higher in c.642GG heifers (LSM 29%) compared to c.642AG heifers (LSM 18%). Differences in milk PMNL concentration between infected and uninfected quarters were smaller in c.980AG heifers than in c.980GG heifers. An association between the CXCR1 genotype and necrosis of milk PMNL could not be demonstrated. Results indicate that CXCR1 polymorphisms influence viability and concentration of milk PMNL and provide a foundation for future research.     

Risk factors for udder edema and its association with lactation performance on primiparous Holstein cows in a large Florida herd, U.S.A

The objective of this study was to determine risk factors for udder edema (case–control study) and to evaluate the association of udder edema in primiparous Holstein cows with their lactation performance (cohort study). Values for the first 10 test day for milk yield, fat, protein, and somatic cell counts (SCC) linear score as well as mature equivalent (ME) 305 days milk yield, fat, protein, SCC linear score, and incidences of periparturient diseases from a computerized farm recording system were compared between primiparous cows with and without udder edema. Data consisted of 118 dairy heifers with udder edema, induced for parturition, and treated with diuretics and 889 control heifers that received no treatment. Primiparous cows giving birth to male calves, were 1.72 (1.01, 3.24; 95% CI) times more likely to develop udder edema than cows giving birth to female calves. Primiparous cows calving in winter season were 3.68 (1.09, 12.5; 95% CI) times more likely to develop udder edema than cows calving in summer. For each extra 10 cm in height the odds of udder edemas was 1.23 (1.03, 1.47; 95% CI). The first test day DHIA milk yield was lower in cows that developed udder edema (3.6 kg/day) than in normal cows. However, the subsequent test days and ME measurements were not statistically different between groups. Cows with udder edema were 1.62 (1.18, 2.14; 95% CI) times more likely to develop udder edema in the second lactation than control animals. It is concluded that the gender of the offspring, calving season and height at parturition were significant risk factors for udder edema. Only milk yield on the first DHIA test day was lower in cows with udder edema than in cows without udder edema.     

Effects of creep feeding, zeranol implants and breed type on beef production: II. Reproductive development and fat deposition in heifers

Effects of preweaning creep feeding and zeranol implants on reproductive tract development and udder and s.c. fat deposition were studied in 24 weanling heifers (7 mo of age) with Brahman and Romana Red sires and Angus and Angus x Brown Swiss F1 reciprocal crossbred dams. Creep treatment did not affect (P greater than .19) ovarian weight, ovarian size, uterine horn diameter or follicle number. Heifers from the three creep treatments did not differ (P greater than .25) in udder weight, total lipid or percentage of lipid in the udder. Noncreep-fed (NC) heifers had a greater (P less than .02) number of adipocytes/gram of udder tissue than did the long-term creep-fed (LC) and short-term creep-fed (SC) heifers. The LC heifers had larger udder (166.0 vs 152.7 m) and s.c. adipocytes (166.7 vs 148.8 m) than NC heifers. Heifers implanted with zeranol at 56 and 146 d of age had a greater (P less than .03) uterine horn diameter and heavier (P less than .02) uterine weight than unimplanted heifers. Percentage of lipid in the udder was lower (P less than .02) in heifers implanted with zeranol. Implanted heifers had smaller (P less than .10) s.c. adipocytes than unimplanted heifers. Breed of dam did not effect (P greater than .17) development of the reproductive tract of weanling heifers. Heifers from Angus dams had smaller (P less than .08) udders and less (P less than .10) total fat in the udder than those from F1 dams, and the heifers from F1 dams tended (P less than .12) to have larger udder adipocytes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of precalving intramammary treatment with pirlimycin in nulliparous Holstein heifers

A clinical trial was conducted to determine whether prepartum intramammary pirlimycin reduces the proportion of nulliparous heifers with intramammary infection (IMI) during early lactation and improves milk production. Quarter milk samples were collected from 428 heifers, systematically allocated to treatment and control groups, 6 to 12 d before the expected calving date and 2 to 8 d after calving. At the prepartum visit, heifers in the treatment group (n = 219) received an infusion of pirlimycin hydrochloride in all 4 quarters; the control heifers (n = 209) received no infusions. Intramammary infection was detected in 69% of the heifers and 33% of the quarters before calving. After calving, the proportion of treated heifers with IMI was significantly lower than the proportion of control heifers (31% versus 45%). Staphylococcus aureus was isolated from 10% of the heifers and 3% of the quarters before calving. After calving, the proportion of IMIs due to S. aureus was significantly lower in the treated heifers than in the control heifers (5.6% versus 10%). Antibiotic treatment increased the percentage of cures and prevented new IMIs caused by gram-positive bacteria after calving. The incidence of new IMIs caused by gram-negative bacteria and yeast was higher among treated heifers than among control heifers. There was no overall effect of treatment on milk production, but there was a significant interaction effect of treatment and the interval between treatment and calving. An increase of 302 kg of milk was observed when antibiotic treatment was applied more than 1 wk before calving. Treatment did not affect the milk somatic cell count on the 1st 3 test days after calving.     

Prevention of intramammary infections in dairy cows by the use of a premilking teat dip method with a foaming iodophor dip agent

In this study we investigated the efficacy of premilking teat dipping with a foaming iodophor teat dip in a negative controlled field study. Incidence of new intramammary infections (IMI), incidence of clinical mastitis, influence on somatic cell count (SCC) and the characteristics of udder tissue and teats were used as parameters to evaluate clinical efficacy. Predipping was compared with a negative control using a split-udder experimental design. Right teats were predipped with a foaming disinfectant containing 0.27% iodine while left teats served as controls. The latter were conventionally cleaned with damp cloth towels and dried manually with disposable paper towels ("best cleaning practice"). All teats were dipped after milking with the same dip. There were no differences between treated and control quarters with respect to incidence of new IMI during the study period (treated quarters: 6.6% vs. untreated: 6.95%), incidence of clinical mastitis (30 cases in the treatment group vs. 39 cases in the control group) and geometric mean of SCC of quarter milk samples. Spectrum of detected pathogens was also comparable. Condition of udder tissue and teat ducts did not differ between treated and control quarters.     

Pharmacokinetics of polymyxin B administered via the bovine mammary gland

Polymyxin B was infused into normal, chronically inflamed, and acutely inflamed quarters of the mammary gland of lactating cows at dosages ranging between 1 and 2 million units (100–200 mg) per quarter. Samples of milk from treated and non-treated quarters, jugular venous blood, subcutaneous abdominal (mammary) venous blood, and urine were collected at intervals after treatment and were assayed using microbiological test methods for polymyxin B concentrations. The drug was not absorbed from normal and chronically inflamed quarters; more than 90% of the infused dose was recovered in milk within 24 h after treatment, and drug residues were detected up to the ninth milking. Drug concentrations in milk from acutely inflamed quarters were significantly lower than in milk from normal quarters; 55% of the infused dose was recovered in the milk within 24 h after treatment. The drug was detected in milk from non-treated quarters, in blood from the subcutaneous abdominal vein, and in the urine during 36–48 h after acutely inflamed quarters were infused with the drug. These data indicate that polymyxin B is well distributed throughout, and is absorbed to a significant degree into the systemic circulation from the acutely inflamed udder.     

Field trials on the prophylaxis of intramammary infections in pregnant heifers

The study was carried out in 5 farms on 174 pregnant heifers. Clinical examination of the udder and bacteriological tests of quarter secretion were performed between the 8th and 3rd week before parturition, and then the animals were divided into a control group (64 heifers) and 3 experimental groups and immediately treated. A group of 32 experimental heifers was injected once with antioxidants (Vitamin A--600,000 i.u.; Vitamin D3--200,000 i.u.; Vitamin E--1.5 mg/kg b.w., Selenium--0.022 mg/kg b.w., i.m.). The next group (26 heads) was intramammary infused with antibiotic DC product (cloxacillin). Heifers from last experimental group (52) were injected with lysosyme dimer in a single dose of 0.02 mg/kg b.w. Clinical and bacteriological examinations were made during the first week after calving. The presence of bacteria was found in secretion of 22.6-38.9% udder quarters in 56.2-71.2% of pregnant heifers. The number of infected quarters (cows) did not change distinctly in the first week after calving except the lysozyme dimer group, where a decrease by 30% was noted. The percentage of quarters with elevated somatic cell count was higher in antibiotic DC group and closely similar in the other groups. None of examined methods showed an acceptable prophylactic effect. Clinical mastitis cases during first week after parturition were mostly caused by Escherichia coli, Staph. chromogenes, Staph. simulans, Staph. aureus, Staph. hyicus, Str. uberis, Str. acidominimus and Enterococcus faecalis.