Homeopathic treatment as an alternative prophylactic to minimize bacterial infection and prevent neonatal diarrhea in calves.

Bovine neonatal diarrhea is common due low immunity in newborn calves, poor management (or absence) of sanitary barriers, and other factors. Newborn calves with diarrhea in the first days of life suffer failure to thrive and may die if left untreated. The aim of this study was to evaluate whether prophylactic administration of a homeopathic product (Dia 100®) can control bovine neonatal diarrhea in calves born on a farm with substantial sanitary challenges. We counted total bacteria and protozoan parasites in fecal samples. We measured serum glucose, total protein, globulin, albumin, cholesterol and triglycerides on days 1, 7 and 14 of life. Twenty newborn calves were maintained in individual stalls, and were divided in two groups: ten untreated animals (control) and ten animals treated with Dia 100®. Fecal consistency was evaluated daily. We diagnosed diarrhea in five animals in the treated group, and in all animals from the control group. Infections with Escherichia coli and Giardia duodenalis were identified as the responsible organisms. The E. coli count was low in the treatment group on day 7 of life compared with the control group. Antibiotics were given to eight animals in the control group, and to two animals in the treatment group. On day of life 7, serum levels of total protein and globulins were higher in the control group, but were lower on day 14. Serum levels of glucose and triglycerides were greater in treated animals on days 7 and 14, suggesting that the homeopathic product contributes to improvement of intestinal health and absorption and nutrients. We conclude that Dia 100® controls diarrhea with 50% of efficacy, and reduces antibiotic utilization.

Chronic endometritis in an Asian elephant (Elephas maximus).

A 48-yr-old female Asian elephant with a history of pododermatitis developed recurrent hematuria beginning in 2002. Transrectal ultrasonography and endoscopic examination in 2004 identified the uterus as the source of hematuria and excluded hemorrhagic cystitis. Treatment with Desloreline implants, antibiotics, and homeopathic drugs led to an improved general condition of the elephant. In July 2005, the elephant was suddenly found dead. During necropsy, the severely enlarged uterus contained about 250 L of purulent fluid, and histopathology revealed ulcerative suppurative endometritis with high numbers of Streptococcus equi ssp. zooepidemicus and Escherichia coli identified on aerobic culture. Additional findings at necropsy included: multifocal severe pododermatitis, uterine leiomyoma, and numerous large calcified areas of abdominal fat necrosis. Microbiologic culture of the pododermatitis lesion revealed the presence of Streptococcus agalactiae, Streptococcus equi ssp. zooepidemicus, Staphylococcus sp., Corynebacterium sp., and Entercoccus sp.

Use of simulated blood cultures for antibiotic effect on time to detection of the two blood culture systems BacT/ALERT and BACTEC 9240.

To avoid the influence of pre-analytical steps, this study was performed using sterile blood spiked with defined loads of microorganisms as inoculum. Time-to-Detection (TTD) was evaluated for the most frequently encountered bacteria comparing two commercially available blood culture systems, BD BACTEC 9240 (Becton Dickinson) and BacT/ALERT (Organon Teknika). The effect of the most widely used antibiotics on TTD was evaluated on both systems. TTD was measured with antibiotics at their trough and at increasing concentrations. The results show that the BACTEC PLUS system recovers more pathogens with shorter time to detection than the BacT/ALERT FAN system when beta-lactam antibiotics (Ampicillin, Cefotaxime) are present at their respective trough concentration corresponding to parenteral therapy. The two systems seem to be equally efficient when Gentamicin, Ciprofloxacin and Trimethoprim/sulfamethoxazole are used; in the case of Vancomycin, BACTEC seems more effective than BacT/ALERT.

Detection of Escherichia coli O157:H7 in meat by an enzyme-linked immunosorbent assay, EHEC-Tek.

Investigation of the specificity of an enzyme-linked immunosorbent assay (ELISA) for detection of Escherichia coli O157:H7 in raw meats (EHEC-Tek; Organon Teknika Corp.) revealed that the target antigens of the detection reagent, monoclonal antibody 4E8C12, were present in numerous serotypes of E. coli and that their ELISA reactivity was influenced by bile salts, acriflavine, and heat. The specificity of the ELISA was improved by a modified test protocol incorporating immunocapture.

Evaluation of methods for the isolation and detection of Escherichia coli O157 in minced beef.

This study has evaluated enrichment and detection procedures for the isolation and detection of Escherichia coli O157 inoculated into minced beef. The use of a 24 h enrichment in modified EC broth containing novobiocin allowed low numbers of contaminating cells to multiply to levels detectable on culture media and by ELISA test kits. Total analysis time was reduced by the use of the Dynabead immunomagnetic separation system. The use of the Petrifilm Test Kit-HEC for E. coli O157:H7 and Organon Teknika EHEC-TEK system detected low numbers of contaminating cells following enrichment and reduced analysis time by 1 d. The incorporation of cefixime and tellurite into Sorbitol MacConkey Agar increased the rate and ease of isolation of E. coli O157 and its use is therefore recommended.

Comparison of beta-glucuronidase-based substrate systems for identification of Escherichia coli.

Methods based on the measurement of beta-glucuronidase have been shown to be specific and inexpensive for the identification of Escherichia coli from bacterial colonies within 1 h. Recently, commercial systems incorporating beta-glucuronidase substrates were introduced. Rapid Identification Method E. coli (Austin Biological Laboratories, Curtin Matheson Scientific, Inc., Houston, Tex.) and Rapid Detect E. coli (Organon Teknika, Morris Plains, N.J.) are single-tube test combinations to simultaneously measure beta-glucuronidase (fluorescence at 366 nm), o-nitrophenyl-beta-D-galactopyranoside (yellow), and indole (red). To determine the accuracy and utility of these two systems, we used them to test 169 E. coli and 150 non-E. coli and compared them with conventional substrate tests. The Rapid Detect test was more efficient than the Rapid Identification Method in demonstrating beta-glucuronidase activity, but the commercial systems were equal to each other and to the conventional tests for o-nitrophenyl-beta-D-galactopyranoside and indole. There were no false reactions by either system.

Evaluation of a new generation of plastic culture bottles with an automated microbial detection system for nine common contaminating organisms found in PLT components.

BACKGROUND: A microbial detection system (BacT/ALERT 3D, bioMérieux [formerly Organon Teknika]) has previously been validated with a variety of bacterial contaminants in PLTs. The recovery of nine organisms seeded into PLTs with new plastic culture bottles was studied in comparison to the current glass bottles. The use of plastic instead of glass would be expected to reduce the risk of injury. STUDY DESIGN AND METHODS: Isolates of Bacillus cereus, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Serratia marcescens, Streptococcus viridans, and Propionibacterium acnes were inoculated into Day 2 (>24 hr <48 hr) apheresis PLT units to 10 and 100 CFUs per mL. Replicate samples (4 mL) were inoculated into both current- and new-generation standard aerobic and anaerobic bottles. RESULTS: All organisms (with the exception of P. acnes) were detected in a mean time of 9.3 to 18.9 hours (10 CFUs/mL) or 8.7 to 18.2 hours (100 CFUs/mL). In aggregate (with the exception of P. acnes), the plastic and glass aerobic bottles had a mean difference in detection of 1.2 hours (p < 0.0001), and the plastic and glass anaerobic bottles had a mean difference of 3.3 hours (p < 0.0001). In all cases, the mean detection time was superior or clinically comparable (within 0.1 hr) with the new plastic bottles. P. acnes (an anaerobic organism) was detected with the new and current anaerobic bottles in a mean of 72.8 and 90.4 hours (10 CFUs/mL) or 64.0 and 80.8 hours (100 CFUs/mL), respectively. The narrower bottle neck and smaller inoculation septum present with the new-generation plastic bottles were inoculated with comparable ease to that of the glass bottles. CONCLUSIONS: These data demonstrate that the new plastic bottles are clinically comparable or superior to the current glass standard aerobic and anaerobic culture bottles.

Volume of blood required to detect common neonatal pathogens.

OBJECTIVE: To determine the minimum volume of blood and the absolute number of organisms required for detection of bacteremia and fungemia by an automated colorimetric blood culture system (BacT/Alert, Organon Teknika). DESIGN: Common neonatal pathogens, Escherichia coli, Streptococcus agalactiae (group B streptococcus (GBS): one American Type Culture Collection (ATCC) strain and one clinical isolate), Staphylococcus epidermidis, and Candida albicans, were seeded into blood to produce bacteremia or fungemia with low colony counts (1 to 3 colony-forming units (CFU) per milliliter) and ultra-low colony counts (<1 CFU/ml). For each organism, 96 culture bottles were inoculated with either 0.25, 0.5, 1.0, or 4.0 ml of the two seeded blood concentrations. Blood culture bottles were incubated in the BacT/Alert device for 5 days, and time to positivity was noted when applicable. All bottles were subcultured on plated media. DATA ANALYSIS: The Poisson statistic was used to calculate the probability of finding at least one viable CFU per inoculated culture bottle. The fraction of culture bottles with positive findings per group was divided by the probability of one or more organisms present to give the positivity index. RESULTS: Plated subculture identified no growth of organisms not detected by the colorimetric detection system. The false-positive rate for the automated device was less than 1%. The positivity index for the GBS clinical isolate was 1.13, for the GBS ATCC isolate 0.96, for S. epidermidis 0.94, for C. albicans 0.97, and for E. coli 0.95. There was a statistically significant difference with time to positivity and inocula volume (p <0.01), but the difference was not clinically important. CONCLUSIONS: If one or two viable colony-forming units are in the blood inoculated into culture media, the BacT/Alert system will detect growth rapidly. Because there appears to be a sizable subset of neonates who are at risk of sepsis with a colony count less than 4 CFU/ml, then a 0.5 ml inoculum of blood into the culture media is inadequate for sensitive and timely detection of bacteremia. One to two milliliters of blood should increase microorganism recovery in the face of low-colony-count sepsis.

Controlled evaluation of 5 versus 10 milliliters of blood cultured in aerobic BacT/Alert blood culture bottles.

Bottles developed for use in the BacT/Alert automated blood culture system (Organon Teknika Corp., Durham, N.C.) can accept up to 10 ml of blood without falling below a 1:5 ratio of blood to broth. We compared the yield and speed of detection of microorganisms in 13,128 adequately filled, paired, aerobic bottles inoculated with 5 versus 10 ml of blood at three university hospitals. A total of 798 microorganisms causing sepsis grew in one or both bottles. The overall recovery of microorganisms from 10-ml samples exceeded that from 5-ml samples (P < 0.001); the increased yield attributed to the additional 5 ml in the samples was 7.2%. The increased yield from 10-ml inocula was most marked for Escherichia coli (P < 0.01) and other members of the family Enterobacteriaceae (P < 0.001). Ten-milliliter samples did not yield more gram-positive bacteria, nonfermentative gram-negative rods, or yeasts. When both bottles were positive, the bottles inoculated with 10 ml of blood showed growth sooner (P < 0.001). Earlier detection with 10-ml inocula was especially notable for coagulase-negative staphylococci (P < 0.001), streptococci (P < 0.001), E. coli (P < 0.025), and other members of the family Enterobacteriaceae (P < 0.025). We conclude that an increase in the volume of blood inoculated into BacT/Alert aerobic blood culture bottles from 5 to 10 ml will increase the overall yield and the speed of detection of clinically important blood pathogens.