Intramammary infections in a dairy herd with a low incidence of Streptococcus agalactiae and Staphylococcus aureus infections.

In a dairy herd with a low incidence of intrammary infections due to Streptococcus agalactiae and Staphylococcus aureus, clinical mastitis remained a serious problem despite good control of nonclinical mastitis through postmilking teat disinfection and antibiotic therapy of known infected quarters at the end of lactation. During the 2-year study, the incidence of clinical mastitis was 0.88 cases/cow-year; 32.2% were caused by streptococcal species other than Str agalactiae and 33.5% by gram-negative organisms. Among all new infections detected, 54.1% were caused by streptococcal species other than Str agalactiae and 25.7% by gram-negative bacteria. Among new infections, 41.6% occurred during the nonlactating period or within a few days of calving. Incidence of clinical mastitis was highest in the 1st month of lactation. Among 84 gram-negative infections, 42.8% were caused by Klebsiella pneumoniae, 20.2% by Escherichia coli, and 23.8% by Enterobacter spp. Among the many serotypes of K pneumoniae and E coli, none was predominant.     

Prevention of clinical coliform mastitis in dairy cows by a mutant Escherichia coli vaccine.

A prospective cohort study was undertaken in two commercial California dairies. The treatment group, 246 cows, received three doses of a whole cell bacterin of J5 Escherichia coli (mutant of E. coli O111:B4) plus Freund's incomplete adjuvant vaccine (two in the dry period and one after calving) while 240 unvaccinated cows served as controls. Thirty-five cases of clinical coliform mastitis were diagnosed, six in vaccinated cows and 29 in unvaccinated cows. Bacteria isolated from the clinical cases included 15 E. coli five Klebsiella pneumoniae, three K. oxytoca, three K. ozaenae, five Enterobacter aerogenes, three Serratia marcescens and one Serratia spp. Four control cows were culled, three of them because of chronic coliform mastitis and one because of postcoliform infection agalactia. Incidence rate of clinical gram-negative mastitis was 2.57% in vaccinated cows and 12.77% in unvaccinated cows. The estimated risk ratio, the measure of risk of having clinical gram-negative mastitis for vaccinated cows to unvaccinated cows, was 0.20 (p less than 0.005), indicating a strong relationship between vaccination and lack of clinical gram-negative mastitis. The results of this trial indicate that the administration of the E. coli J5 vaccine is protective against natural challenge to gram-negative bacteria, and reduces the incidence of clinical gram-negative mastitis in dairy cows during the first three months of lactation.     

Risk factors associated with mastitis in dairy goats

Prevalence of intramammary infection in healthy goats was determined from 4,662 composite udder samples taken over a 9-month period. For each doe, a colostral sample and 2 milk samples were collected. Breed, age, number of days not lactating before kidding, number of lactation days, and kidding date were recorded. Coagulase-negative Staphylococcus spp were isolated from 17.5% of does, Staphylococcus aureus from 3.1%, Mycoplasma spp from 1.2%, Streptococcus spp from 0.3%, and gram-negative bacteria from 2.0%. Gram-negative organisms were associated with intermittent infections, whereas coagulase-negative staphylococci were associated with persistent infections. Intramammary infection was related to breed, number of days not lactating, and number of lactation days, as determined by log-linear analysis. Does of the Nubian breed, does with nonlactating periods of greater than 60 days, and does in the first and last third of a standard 305-day lactation appeared to be at higher risk for intramammary infection.     

Experimental intramammary infection of the dairy cow with Escherichia coli during the nonlactating period

The nonlactating mammary gland was experimentally inoculated with Escherichia coli. During the first half of the nonlactating period, 32% of 34 inoculated glands were temporarily infected. All intramammary infections were eradicated by the cow without therapy and no signs of mastitis were observed. During the 30 days before parturition occurred, 88% of 42 inoculated glands in the cows became infected. Twenty-three intramammary infections were eradicated by the cow and infection in 14 glands persisted after parturition occurred. Peracute toxic mastitis occurred in those cows with infected glands.     

Resistance to gentamicin and amikacin of gram-negative organisms isolated from horses

Resistance of gram-negative bacteria to gentamicin has become an increasingly common problem among clinical isolates from human beings. Susceptibility of isolates from horses to gentamicin and amikacin was evaluated for the period from July, 1983 to June, 1985. All isolates of Escherichia coli, and species of Enterobacter, Klebsiella, Proteus, and Pseudomonas examined were susceptible to amikacin, except 2 of the 46 Pseudomonas isolates. In contrast, 13 to 50% of isolates were resistant to gentamicin. Escherichia coli, and Klebsiella, Proteus, and Enterobacter species isolates were highly significantly more susceptible to amikacin (P less than 0.01) than to gentamicin. Pseudomonas spp (P = 0.13) were not significantly different in susceptibility to the 2 drugs. There was significant variation among genera in their susceptibility to gentamicin (P = 0.002), primarily because of the frequency of resistance in isolates of Klebsiella spp and Proteus spp, compared with the other 3 organisms (E coli, Enterobacter spp, and Pseudomonas spp). There was no significant difference of susceptibility to amikacin among the genera studied (P = 0.06).     

Comparison of bacteriologic culture of blood and necropsy specimens for determining the cause of foal septicemia: 47 cases (1978-1987)

Diagnosis of bacterial septicemia was confirmed by results of bacteriologic culture of antemortem blood samples and/or necropsy specimens obtained from 47 foals up to 8 days old. Gram-negative bacteria were isolated from all 47 foals, and mixed infections with more than one organism were involved in 26 (55%). Escherichia coli, Actinobacillus spp, and Klebsiella pneumoniae were the most frequent isolates, infecting 55, 34, and 23% of foals, respectively. Gram-positive bacteria and anaerobic bacteria were isolated only from foals with mixed infections with gram-negative organisms. Clostridium perfringens was the only anaerobe isolated. In 38 (81%) of 47 foals with confirmed septicemia, blood samples were culture-positive. Thirty-two septicemic foals subsequently died, allowing a comparison to be made between the species of bacteria isolated by culture of blood with those recovered by culture of internal organs at necropsy. Blood failed to yield any gram-negative organisms in 12 (37.5%) of 32 foals from which a gram-negative pathogen was isolated at necropsy. Forty-three percent of the gram-negative bacteria, including 59% of the E coli, and 10% of the gram-positive bacteria found in septicemic foals at necropsy were not detected earlier by results of bacteriologic culture of blood.     

Occurrence of gram-negative bacteria in hens' eggs depending on their source and storage conditions

The aim of this study was to analyse the qualitative composition of Gram-negative microbes, mainly of the family Enterobacteriaceae, including pathogenic bacteria such as Salmonella, in the albumens and yolks and on the shells of hens' eggs, depending on their source and on the temperature and duration of their storage. A total of 375 table eggs were studied, from a large-scale poultry farm, a small-scale poultry farm and a supermarket. Each group was divided into 5 subgroups according to the temperature and duration of their storage during the study. Two serotypes of bacteria of the genus Salmonella were identified: S. Enteritidis and S. Arizonae. Strains of Salmonella spp. were also isolated. Apart from Salmonella and Escherichia coli, among the most frequently isolated bacteria of the family Enterobacteriaceae were Enterobacter spp., Klebsiella spp. and Citrobacter freundii. Qualitative analysis of the bacterial microflora of the eggs also showed the presence of other Gram negative bacteria, including Acinetobacter spp., Pseudomonas spp., Tatumella ptyseos, Providencia stuartii, Serratia liquefaciens, Flavimonas oryzihabitans, Vibrio metschnikovii, Leclercia adecarboxylata, Kluyvera spp., Rahnella aquatilis, Proteus mirabilis, and Achromobacter spp. The study demonstrated that the conditions applied, i.e., the temperature and duration of storage, did not significantly influence the prevalence of particular species of Gram-negative bacteria in the eggs. However, based on the analysis of contamination of eggs with Salmonella depending on their source, it can be concluded that the system in which the hens are housed affects the risk of contamination of eggs with these pathogens.     

Experimental infection of bovine mammary glands with Streptococcus agalactiae during the nonlactating period

Two experiments were done to determine the rate of intramammary infection (IMI) in dairy cows during the nonlactating period. In experiment 1, all glands were equally exposed to Klebsiella pneumoniae and Streptococcus agalactiae for the first 3 weeks after the start of the nonlactating period. Nearly all new IMI was caused by S agalactiae. In experiment 2, the susceptibility of nonlactating mammary glands to new IMI by S agalactiae after direct inoculation into the lactiferous sinus was determined. During the 1st month of the nonlactating period, 30% of inoculated glands became infected. During the last month of the nonlactating period, 90% of inoculated glands became infected.     

Clinical pharmacokinetics of carbenicillin, carfecillin, ticarcillin and BL-P 1654 in dairy cows

The minimal inhibitory concentrations (MIC) of carbenicillin, ticarcillin and BL-P 1654 for gram-negative udder pathogens were determined using the agar plate dilution method. The MIC of the drugs for 50% and 90% of the isolates examined ranged for Escherichia coli and Aerobacter spp. from 1.56 to 25 micrograms/ml, and for Klebsiella spp. and Pseudomonas spp. from 3.12 to 50 micrograms/ml. The Serratia spp. were relatively non-susceptible for the drugs studied (MIC greater than 50 micrograms/ml). Each drug was administered intravenously at 5 g and 15 g per cow to different groups of cows with normal and inflamed quarters of the udder. Distribution and elimination kinetic parameters calculated from serum drug level data were very similar to those of other beta-lactam antibiotics. Although drug concentrations in milk from inflamed quarters were higher than in milk from normal quarters, they were considerably below the MIC for the majority of gram-negative udder pathogens. The data suggest that parenteral treatment of gram-negative udder infections with carbenicillin, carfecillin, ticarcillin and BL-P 1654 at the dose levels used in the present study is unlikely to result in a bacteriological cure and would probably be clinically ineffective.     

Changes in bacterial and fungal ocular flora of clinically normal horses following experimental application of topical antimicrobial or antimicrobial-corticosteroid ophthalmic preparations

OBJECTIVE: To determine effects of topical antimicrobial and antimicrobial-corticosteroid preparations on the ocular flora of horses. animals: 40 horses. PROCEDURE: One eye was treated 3 times daily for 2 weeks with one of the following ointments: (1) neomycin-bacitracin-polymyxin B, (2) 0.6% prednisolone-0.3% gentamicin, (3) neomycin-polymyxin B-0.05% dexamethasone, or (4) treated (artificial tears) control. Contralateral eyes of treated control eyes served as untreated control eyes. Corneal and conjunctival specimens for bacterial and fungal cultures were collected prior to initiation of treatment, after 1 and 2 weeks of treatment, and 2 weeks after concluding treatment. Changes in culture growth quantity scores of bacterial and fungal species were analyzed. RESULTS: The most common species before treatment were the following: gram-positive bacteria included Streptomyces spp (66%), Staphylococcus spp (46%), Bacillus spp (32%), and Streptococcus spp (32%); gram-negative bacteria included Moraxella spp (28%), Escherichia coli (24%), Acinetobacter spp (18%), and Enterobacter spp (14%); and fungi included Aspergillus nidulans (56%), Cladosporium spp (32%), and Aspergillus fumigatus (22%). In all groups, the percentage of positive bacterial culture results, growth quantity score of gram-positive bacteria, and number of bacterial species isolated decreased at week 1 and increased at week 2, whereas growth quantity score of gram-negative bacteria decreased throughout treatment. Differences were not significant among groups. Fungal growth quantity score decreased during treatment in all groups. Repopulation of bacterial and fungal species occurred. CONCLUSIONS AND CLINICAL RELEVANCE: All interventions decreased the number of microorganisms. Repopulation of normal flora occurred during and after treatment.     

Antimicrobial activity of bovine bactericidal permeability-increasing protein-derived peptides against gram-negative bacteria isolated from the milk of cows with clinical mastitis

OBJECTIVE: To evaluate antimicrobial activity of bovine bactericidal permeability-increasing protein (bBPI)-derived synthetic peptides against mastitis-causing gram-negative bacteria. SAMPLE POPULATION: Bacterial isolates from the milk of cows with clinical mastitis. PROCEDURES: 3 peptides were synthesized with sequences corresponding to amino acids 65 to 99 (bBPI(6,599)) or 142 to 169 (bBPI(142,169)) or the combination of amino acids 90 to 99 and 148 to 161 (bBPI(9,099, 148,161)) of bBPI. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of these peptides against bacterial isolates from cows with mastitis were determined by use of a standardized broth microdilution assay. The ability of these peptides to retain their antimicrobial activity in serum and milk was also evaluated. Finally, bacterial lipopolysaccharide (LPS)-neutralizing activity of these peptides was assayed with the Limulus amebocyte lysate test. RESULTS: Of the 3 peptides tested, bBPI(9,099, 148,161) had the widest spectrum of antimicrobial activity, with MIC and MBC values ranging from 16 to 64 Mg/mL against Escherichia coli, Klebsiella pneumoniae, and Enterobacter spp and from 64 to 128 Mg/mL against Pseudomonas aeruginosa. None of the peptides had any growth-inhibitory effect on Serratia marcescens. The antimicrobial activity of bBPI(9,099, 148,161) was inhibited in milk, but preserved in serum. Finally, bBPI(142,169) and bBPI(9,099, 148,161) completely neutralized LPS. CONCLUSIONS AND CLINICAL RELEVANCE: bBPI(9,099, 148,161) is a potent neutralizer of the highly proinflammatory molecule bacterial LPS and has antimicrobial activity against a variety of gram-negative bacteria. The ability of bBPI(9099,148161) to retain antimicrobial activity in serum suggests a potential therapeutic application for this peptide in the management of gram-negative septicemia.     

Host-response patterns of intramammary infections in dairy cows

Many different bacterial species have the ability to cause an infection of the bovine mammary gland and the host response to these infections is what we recognize as mastitis. In this review we evaluate the pathogen specific response to the three main bacterial species causing bovine mastitis: Escherichia coli, Streptococcus uberis and Staphylococcus aureus. In this paper we will review the bacterial growth patterns, host immune response and clinical response that results from the intramammary infections. Clear differences in bacterial growth pattern are shown between bacterial species. The dominant pattern in E. coli infections is a short duration high bacteria count infection, in S. aureus this is more commonly a persistent infection with relative low bacteria counts and in S. uberis a long duration high bacteria count infection is often observed. The host immune response differs significantly depending on the invading bacterial species. The underlying reasons for the differences and the resulting host response are described. Finally we discuss the clinical response pattern for each of the three bacterial species. The largest contrast is between E. coli and S. aureus where a larger proportion of E. coli infections cause potentially severe clinical symptoms, whereas the majority of S. aureus infections go clinically unnoticed. The relevance of fully understanding the bovine host response to intramammary infection is discussed, some major gaps in our knowledge are highlighted and directions for future research are indicated.     

Nosocomial infections and bacterial antibiotic resistance in a university equine hospital

A base-line study of bacteria isolated from horses admitted to the Veterinary Medical Teaching Hospital during a 6-month period was performed to determine the extent of multiresistant nosocomial infections caused by gram-negative aerobic bacteria other than Salmonella spp. Results of this study indicated that 21.9% of the 105 horses from which cultures and sensitivities were available had developed nosocomial gram-negative aerobic infections, with high rates of resistance to gentamicin, kanamycin, and trimethoprim sulfadiazine, three of the most often prescribed antibiotics in this hospital. In addition, a prospective study of antibiotic-resistant bacteria of fecal origin was performed to determine whether there was a change in the degree of antibiotic resistance of a horse's intestinal flora while the horse was hospitalized. Bacterial culturing for gram-negative lactose fermenting bacteria was done on fecal specimens collected directly from the rectum on day 1 and day 7 of a horse's hospitalization. Susceptibility testing was done on each isolant. Of the 24 paired fecal specimens obtained, Escherichia coli and Klebsiella sp isolated on day 7 were resistant to a significantly higher number of antibiotics than day 1 isolants (P = 0.003, P = 0.043, respectively).     

Efficacy of intramammary infusion of ceftiofur hydrochloride at drying off for treatment and prevention of bovine mastitis during the nonlactating period

This study evaluated the efficacy of intramammary infusion of ceftiofur hydrochloride for the treatment of intramammary infections present at the last milking of lactation and for prevention of new intramammary infections during the nonlactating period. Cows were randomly assigned to five treatments (untreated negative control, 125, 250, and 500 mg of ceftiofur, and a positive control group receiving 300 mg cephapirin benzathine). A dose of 125 mg of ceftiofur per mammary quarter was effective for treatment of existing infections present at the time of milk cessation, but only the 500-mg dose of ceftiofur per mammary quarter was effective for both treatment of existing intramammary infections at the time of milk cessation and for prevention of new intramammary infections during the nonlactating period.     

Bacterial flora of the conjunctiva and nasal cavity in normal and diseased captive bustards.

A survey was carried out to describe the normal aerobic bacterial flora of the conjunctiva and nasal cavity of captive houbara bustards (Chlamydotis undulata), kori bustards (Ardeotis kori), and white-bellied bustards (Eupodotis senegalensis) maintained at the National Avian Research Center, Abu Dhabi, United Arab Emirates. A total of 58 samples were examined from the nasal cavity and 55 samples from the conjunctiva of healthy bustards. There was no bacterial growth in 45% of conjunctival samples. Bacteria isolated from the conjunctiva of healthy birds included Micrococcus spp., Staphylococcus auricularis, Staphylococcus xylosus, Staphylococcus capitis, Staphylococcus warneri, Bacillus spp., and Enterobacter amigenus. Bacteria isolated from the nasal cavity of healthy birds included Bacillus spp., Micrococcus spp., S. auricularis, S. xylosus, Staphylococcus simulans, Staphylococcus saprophyticus, Staphylococcus hyicus, Staphylococcus cohnii, Staphylococcus sciuri, Aerococcus spp., and Providencia rettgeri. These findings were compared with bacterial isolates from bustards with clinical signs of ocular or upper respiratory tract diseases. Mycoplasma spp., Pseudomonas aeruginosa, Pseudomonas stutzeri, Proteus mirabilis, Escherichia coli, Klebsiella spp., Aeromonas hydrophila, and Staphylococcus aureus were the pathogenic bacteria isolated from the conjunctiva of 34.3% bustards with ocular discharges. Mycoplasma spp., P. aeruginosa, Pseudomonas spp., P. mirabilis, E. coli, Klebsiella pneumoniae, and S. aureus were the pathogenic bacteria isolated from the nasal cavity of 74% bustards with upper respiratory tract diseases.     

Function of phagocytes obtained from lacteal secretions of lactating and nonlactating cows

Phagocytes, macrophages and neutrophils, were obtained from lacteal secretions of lactating (n = 13) and nonlactating cows (n = 14). Secretions from nonlactating cows were collected at 7 and 14 days after cessation of lactation. Phagocytes were incubated in vitro with Staphylococcus aureus or Escherichia coli, and function was assessed by fluorescent microscopy of cell suspensions stained with acridine orange and crystal violet. A greater percentage of macrophages from nonlactating cow secretions collected on day 14 phagocytized bacteria than did those collected on day 7. A greater percentage of macrophages from nonlactating cow secretions collected on days 7 and 14 phagocytized bacteria than did neutrophils obtained from the same secretions. A similar percentage of phagocytes from nonlactating cow secretions phagocytized bacteria, compared with phagocytes from lactating cow secretions. Results indicated that the intramammary macrophage may be most important in defense of the mammary gland during the early nonlactating period, because it was more phagocytic than the neutrophil and was more active at 14 days than at 7 days into the nonlactating period.     

Cause, occurrence, and clinical signs of mastitis and anorexia in cows in a Wisconsin study

Microbiologic culture revealed the following cause of mastitis and anorexia in 145 cows in Wisconsin to be Escherichia coli, 66 cows; Klebsiella spp, 3; Corynebacterium pyogenes, 27; streptococci, 21; staphylococci, 20; yeasts, 1; and no bacterial growth, 7. Mastitis was detected with approximately equal frequency throughout the year. Escherichia coli was isolated throughout the year, but was more common and was the predominant organism during the summer. Corynebacterium pyogenes was isolated most often in winter and spring; streptococci in fall, winter, and spring; and staphylococci throughout the year. Corynebacterium pyogenes caused most of the mastitis in nonlactating cows. Escherichia coli, C pyogenes, streptococci, and staphylococci were isolated with about equal frequency at parturition, whereas E coli was the predominant cause of mastitis in early and late lactation. Of cases of mastitis, 27% were seen 10 days before and after parturition. Local and systemic clinical signs of infection were similar for all causes, except that C pyogenes caused more (P less than 0.01) malodorous and purulent milk than did other organisms and was isolated more commonly from quarters with injured teats. Recovery was significantly (P less than 0.01) higher in cows with E coli infections, compared with recovery in cows with gram-positive organism infections. Cows with C pyogenes infections frequently had quarters that ultimately ceased lactation. A few cows were recumbent at initiation of antimicrobial therapy and a few were not eating 24 hours later; however, 50% of these cows recovered. Criteria such as season of year, stage of lactation, appearance of milk and udder, and appetite permitted the cause (gram-negative or gram-positive organisms) of the mastitis to be predicted with 77% accuracy.     

Bacterial colonization of intravenous catheters in young dogs suspected to have parvoviral enteritis

OBJECTIVE: To determine the prevalence of bacterial colonization of IV catheters among young dogs suspected to have parvoviral enteritis, to identify the organisms responsible for catheter colonization, and to determine the antimicrobial susceptibility of organisms that were obtained. DESIGN: Case series. ANIMALS: 100 dogs. PROCEDURE: Catheters were aseptically removed when fluid therapy was discontinued, the catheter was replaced, or the dog died. The distal tip of the catheter was cut off, split open, and vortexed with sterile saline (0.9% NaCl) solution. The saline solution was plated on culture plates, which were then incubated and examined for bacterial growth every 24 hours for 72 hours. All bacteria cultured were identified, and antimicrobial susceptibility was determined. RESULTS: Bacteria were isolated from 22 catheters. Most bacteria that were isolated were of gastrointestinal tract or environmental origin (Serratia odorifera, S. liquefaciens, S. marcescens, Acinobacter anitratus, Citrobacter freundii, Klebsiella pneumoniae, K. oxytoca, Escherichia coli, Enterobacter spp). Only 2 gram-positive organisms were isolated (Staphylococcus intermedius and Streptococcus spp). High percentages of organisms were resistant to penicillin, lincomycin, cloxacillin, erythromycin, and cephalexin. Percentages of organisms resistant to amikacin, enrofloxacin, chloramphenicol, potentiated sulfonamides, and amoxicillin-clavulanic acid were low. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that IV catheters may be colonized with bacteria in 22% of young dogs suspected to have parvovirus infection.     

Aerobic bacterial flora of nesting green turtles (Chelonia mydas) from Tortuguero National Park, Costa Rica.

Bacteriological examination of 70 nesting green turtles (Chelonia mydas) from Tortuguero National Park, Costa Rica was performed to investigate nasal and cloacal aerobic bacteria. A total of 325 bacterial isolates were obtained, including 10 Gram-negative and three Gram-positive genera. Two hundred thirty-nine were Gram-negative and 86 were Gram-positive isolates. Klebsiella pneumoniae was the most common microbe identified in turtle samples: 27/70 (38.5%) in cloacal, and 33/70 (47.1%) in nasal samples. The Enterobacteriaceae family, including Enterobacter agglomerans, E. cloacae, Escherichia coli, Klebsiella oxytoca, K. pneumoniae, and Serratia marcescens, was the largest Gram-negative group of bacteria recovered and comprised 127 of 239 (53.1%) of the Gram-negative isolates. Staphylococcus species was the largest Gram-positive bacteria group, including S. aureus, S. cromogenes, S. epidermis, and S. intermedius, and made up 63 of 86 (73.2%) of the Gram-positive isolates recovered. The results of this study demonstrate that the aerobic bacterial flora of nesting green turtles at Tortuguero National Park is composed of a very wide spectrum of bacteria, including several potential pathogens.     

The incidence of antibiotic resistance among coliform bacteria isolated from food.

Three-hundred-and-seventy-eight strains of coliform bacteria were isolated from specimens of commonly sold milk and food products. Klebsiella and Enterobacter spp. were predominating. Resistance to sulphonamides, streptomycin, and chloramphenicol occurred in only 5, 1, and 2 strains, respectively. No tetracycline-resistant strains were found. Two-hundred-and-two strains (54 %) were resistant to ampicillin. In genetic crosses with a sensitive strain of E. coli Κ 12 W 3132 transmissible R factors could not be demonstrated in any of the resistant coliform strains.It is concluded that food is not a significant source of antibiotic resistant enteric bacteria. It may, however, be suggested that food is a source of potentially pathogenic gram-negative bacteria which points out the importance of strict hygienic surveillance of food production.