Susan Sanchez

Publications of Susan Sanchez

• Bacterial contamination of commercial ear cleaners following routine home use.

  Authors: Sarah J Bartlett, Wayne S Rosenkrantz, Susan Sanchez

  Veterinary dermatology. 06/2011; 22(6):546-53.

  Ear cleaning solutions are designed for repeated use, which raises the possibility for bacterial contamination leading to recurrent or persistent infectious otitis. The purpose of this study was toEar cleaning solutions are designed for repeated use, which raises the possibility for bacterial contamination leading to recurrent or persistent infectious otitis. The purpose of this study was to investigate the prevalence of bacterial contamination of commercial ear cleaners following routine home use in dogs and to describe the characteristics that are associated with contamination. Used ear cleaner bottles and information regarding their use were obtained from canine owners visiting veterinary dermatologists. Both the bottle applicator tips and the solution contents were cultured for aerobic bacteria. Bacterial contamination was present on 10% of the bottle tips and in 2% of the solutions. Isolated bacteria included Staphylococcus pseudintermedius, Bacillus spp., coagulase-negative Staphylococcus spp., Micrococcus spp. and Burkholderia cepacia. The contamination rate was significantly higher on the applicator tips than in the solutions (P = 0.0076). The applicator tip contamination rate was significantly higher in expired samples (17%) than in-date samples (4%; P = 0.0277). The bottle sizes were significantly larger for the samples with contaminated applicator tips compared with noncontaminated tips (P = 0.0455). The contamination rate was significantly higher when Tris-EDTA was an ingredient. Cleanliness of the bottle, contact with the ear canal and infection status of the ear at time of culture had no bearing on the contamination rate. In summary, with routine home use of commercial ear cleaners, pathogenic bacterial contamination is of minor concern. This concern may increase when expired products or larger bottles of ear cleaner are used and when Tris-EDTA is an ingredient.

• Pathology in practice. Cutaneous mycobacteriosis.

  Authors: Victoria B Wilson, Raquel R Rech, Michaela G Austel, Cynthia L Bauer, Kenneth S Latimer, Susan Sanchez, Elizabeth W Howerth

  Journal of the American Veterinary Medical Association. 01/2011; 238(2):171-3.

• Panniculitis attributable to Mycobacterium goodii in an immunocompetent dog in Georgia.

  Authors: Paula M Krimer, Kelly M Phillips, Doris M Miller, Susan Sanchez

  Journal of the American Veterinary Medical Association. 11/2010; 237(9):1056-9.

  A 5-year-old 38.3-kg (84.5-lb) mixed-breed dog was examined because of acute onset of lethargy and anorexia. Four days later, a raised, firm, warm 15 × 10-cm lesion was detected in the right caudalA 5-year-old 38.3-kg (84.5-lb) mixed-breed dog was examined because of acute onset of lethargy and anorexia. Four days later, a raised, firm, warm 15 × 10-cm lesion was detected in the right caudal paralumbar area. Cephalexin treatment yielded a poor response. Formalin-fixed tissue and fluid samples from the cystic areas of the lesion were submitted for cytologic and histologic examinations, routine bacterial and mycobacterial culture, and genus identification and 16S partial sequencing via PCR assays. Cytologic examination revealed chronic pyogranulomatous inflammation. Histologic examination by use of routine, Giemsa, silver, acid-fast, and modified acid-fast stains revealed multifocal nodular granulomatous panniculitis without identifiable organisms. Mycobacteria were initially identified via PCR assay and mycobacterial culture within 3 days. Mycobaterium goodii was speciated by use of partial 16S RNA sequence analysis. The lesion resolved after long-term treatment with a combination of rifampin and clarithromycin and insertion of a Penrose drain. There has been no recurrence of the condition. M goodii is an environmental rapidly growing mycobacterium and is a zoonotic pathogen. Infections have not been previously reported in domestic animals in North America, although there are rare reports of infection in humans associated with surgery, especially surgical implants. Domestic animals are a potential sentinel for this non-tuberculous mycobacterial infection in humans, although lack of speciation in infections of domestic animals likely underestimates the potential public health importance of this pathogenic organism. Current microbiological molecular methods allow for a rapid and inexpensive diagnosis.

• Investigation of shell disease in map turtles (graptemys spp.).

  Authors: Stephen J Hernandez-Divers, Patrick Hensel, Juliet Gladden, Sonia M Hernandez-Divers, Kurt A Buhlmann, Chris Hagen, Susan Sanchez, Kenneth S Latimer, Mary Ard, Alvin C Camus

  Journal of wildlife diseases. 08/2009; 45(3):637-52.

  Nineteen map turtles (Graptemys spp.) maintained under natural conditions were investigated because of chronic shell abnormalities. Animals were evaluated using a novel shell scoring system thatNineteen map turtles (Graptemys spp.) maintained under natural conditions were investigated because of chronic shell abnormalities. Animals were evaluated using a novel shell scoring system that divided the 54 scutes into six regions, with each region scored for lesion extent and severity, and summated to produce a total shell disease score (TSDS). Complete blood counts and various biochemistry analytes (total protein, albumin, globulin, urea, uric acid, 25-hydroxycholecalciferol, phosphorus, and ionized and total calcium) were measured. Under ketamine-medetomidine-morphine anesthesia, cytology tape strips and full thickness shell biopsies were collected aseptically for microbiologic, histologic (including scoring of biopsy quality), and ultrastructural evaluations. The TSDSs were low and ranged from 4 to 22 (median = 9) out of a possible score of 54. There were no correlations between TSDS and any hematologic or biochemistry parameter. The histologic quality of shell biopsies was good, and normal shell structure, by both light and electron microscopy, is described. Small clefts and pitting lesions were noted in 8/19 sections. There was no evidence of erosion, ulceration, inflammation, or infectious agents, but algae and diatoms were observed. Six biopsies yielded aerobic isolates (Chryseobacterium indologenes, Aeromonas hydrophila, Ralstonia pickettii, and Morganella morganii), whereas 11 shell samples grew various clostridial anerobes. No fungal organisms were cultured. Although the etiology of the lesions described remains unknown, the use of a scoring system in conjunction with full thickness biopsies is suggested to help standardize investigations into chelonian shell disease in the future.

• Backyard chicken flocks pose a disease risk for neotropic birds in Costa Rica.

  Authors: Sonia M Hernandez-Divers, Pedro Villegas, Carlos Jimenez, Stephen J Hernandez-Divers, Maricarmen Garcia, Sylva M Riblet, C Ron Carroll, Barry M O'Connor, Julie L Webb, Michael J Yabsley, Susan M Williams, Susan Sanchez

  Avian diseases. 01/2009; 52(4):558-66.

  Pathogens of free-ranging chickens create a risk of disease for wild birds, some of which migrate to the United States, as well as potential economic losses for resource-poor farmers. Free-roamingPathogens of free-ranging chickens create a risk of disease for wild birds, some of which migrate to the United States, as well as potential economic losses for resource-poor farmers. Free-roaming backyard chickens are commonly kept in shade-grown coffee plantations, habitats that attract large numbers of wild birds. The husbandry and pathogen prevalence of backyard chicken flocks in San Luis, Costa Rica, were investigated. Based on serologic evidence, Newcastle disease virus, infectious laryngotracheitis virus, infectious bronchitis virus, chicken anemia virus, and infectious bursal disease virus, as well as both Mycoplasma gallisepticum and Mycoplasma synoviae, appear to be significant diseases of this population, and thus, we consider these backyard chickens potential reservoirs for these diseases. There was no evidence of avian influenza. Interviews, clinical examinations, and microscopic examination of tissues led us to believe that poxvirus is also a significant cause of morbidity and mortality in these chickens. We found that Escherichia coli isolates were resistant against tilmicosin, tetracycline, ampicillin, amoxicillin with clavulanic acid, ticarcillin, and cephalothin, and contained genes considered responsible for conferring tetracycline resistance. Additionally, although production was not measured, we suspect that husbandry and lack of preventative medicine are directly related to the diseases reported, all of which negatively affect production.

• Molecular characterization reveals Salmonella enterica serovar 4,[5],12:i:- from poultry is a variant Typhimurium serovar.

  Authors: Katherine Zamperini, Vivek Soni, Douglas Waltman, Susan Sanchez, Elizabeth C Theriault, Jordan Bray, John J Maurer

  Avian diseases. 01/2008; 51(4):958-64.

  Although Salmonella remains one of the leading causes of foodborne illnesses in the United States, the Salmonella enterica serovars and genetic types associated with most infections appear toAlthough Salmonella remains one of the leading causes of foodborne illnesses in the United States, the Salmonella enterica serovars and genetic types associated with most infections appear to fluctuate over time. Recently, the Center for Disease Control and Prevention (CDC) has reported an increase in cases of salmonellosis caused by Salmonella 4,[5],12:i:-. Similarly, this unusual Salmonella serovar has been isolated from cattle and poultry in the state of Georgia. We examined the genetic relatedness of Salmonella 4,[5],12:i:-, isolated from several different poultry companies and dairy farms in Georgia, by pulsed-field gel electrophoresis (PFGE). Several Salmonella 4,[5],12:i:- isolates had PFGE patterns identical or similar to PFGE patterns of Salmonella Typhimurium isolated from numerous animal sources. We identified distinct PFGE patterns for Salmonella 4,[5],12:i:- and matching Salmonella Typhimurium PFGE patterns, identifying four "distinct" strains. We focused a more specific analysis on the poultry Salmonella 4,[5],12:i:- and Salmonella Typhimurium isolates and found that of these Salmonella 4,[5],12:i:- isolates, 32% lacked the entire phase 2 antigen gene, fljB; 61% contained partial deletion(s); and 4% had partial deletion(s) in fljB and an adjacent gene hin, 5' to fljB. Thirteen percent contained smaller deletions or point mutations not identified by our DNA probes. The Salmonella 4,[5],12:i:- isolates were positive for several genes present in the Salmonella Typhimurium, including lpfE (100%), sseI(96%), and spvC (93%). Genetic analysis indicates independent, spontaneous mutations in fljB in at least four distinct Salmonella Typhimurium strains of animal origin circulating in nature.

• Occurrence of canine parvovirus type 2c in the United States.

  Authors: Charles Hong, Nicola Decaro, Costantina Desario, Patrick Tanner, M Camila Pardo, Susan Sanchez, Canio Buonavoglia, Jeremiah T Saliki

  Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 10/2007; 19(5):535-9.

  Canine parvovirus (CPV) type 2 (CPV-2) emerged around 1978 as a major pathogen of dogs worldwide. In the mid-1980s, the original CPV-2 had evolved and was completely replaced by 2 variants, CPV-2aCanine parvovirus (CPV) type 2 (CPV-2) emerged around 1978 as a major pathogen of dogs worldwide. In the mid-1980s, the original CPV-2 had evolved and was completely replaced by 2 variants, CPV-2a and CPV-2b. In 2000, a new variant of CPV (named CPV-2c) was detected in Italy and now cocirculates with types 2a and 2b in that country. The CPV-2c has also been reported from single outbreaks in Vietnam and Spain. This study was conducted to determine if CPV-2c occurs in the United States. Thirty-three fecal samples were collected from dogs in 16 states between April 2006 and April 2007 and were tested for CPV using real-time polymerase chain reaction (PCR). Positive samples were further tested using conventional PCR and minor-groove binding TaqMan PCR assays to determine the viral type and to differentiate vaccine strains from field strains. Twenty-seven samples were positive for CPV, 7 of which were CPV-2c from 5 states: Arizona, California, Georgia, Oklahoma, and Texas. Of the 7 isolates, 4 differed from European CPV-2c isolates by 2 additional single-nucleotide mutations at positions 4076 and 4104, the latter of which produces a ThrAla change at residue 440 located near a major antigenic site. The coast-to-coast geographic distribution of the states in which CPV-2c was detected strongly suggests that this new CPV variant is probably widespread in the United States. The continuous evolution of CPV requires that monoclonal antibody-based and nucleic acid-based diagnostic assays should be periodically checked for sensitivity on prevalent CPV strains.

• Prevalence of Mycoplasma bovis in backgrounding and stocker cattle operations.

  Authors: Monique C Wiggins, Amelia R Woolums, Susan Sanchez, David J Hurley, Dana J Cole, Douglas T Ensley, Mel E Pence

  Journal of the American Veterinary Medical Association. 06/2007; 230(10):1514-8.

  OBJECTIVE: To characterize the prevalence of Mycoplasma bovis infection in backgrounding and stocker cattle operations and compare bacteriologic culture with PCR assay for detection of M bovis.OBJECTIVE: To characterize the prevalence of Mycoplasma bovis infection in backgrounding and stocker cattle operations and compare bacteriologic culture with PCR assay for detection of M bovis. DESIGN: Prospective descriptive study. ANIMALS: 432 calves, 3 to 9 months old, from 9 operations. PROCEDURES: 2 nasal swab specimens were collected from each calf. Swab specimens were evaluated via bacteriologic culture and PCR assay for organisms of the class Mollicutes and M bovis. Culture results were considered negative if no growth occurred within 21 days. Positive results were indicated by characteristic colony formation with PCR assay confirmation. Deoxyribonucleic acid was extracted from 1 swab specimen for direct PCR assay for Mollicutes and M bovis. RESULTS: Of 432 calves, 374 (87%) had positive results for Mollicutes via PCR assay and 63 (15%) via culture. Seven (2%) calves had positive results for M bovis via PCR assay and 10 (2%) via culture. Prevalence of Mollicutes at the farm level ranged from 54% to 100% via PCR assay and from 0% to 59% via culture. Prevalence of M bovis at the farm level ranged from 0% to 4% via PCR assay and from 0% to 6% via culture. Calves that shed M bovis were significantly more likely to have a fever than were calves that did not shed M bovis. CONCLUSIONS AND CLINICAL RELEVANCE: M bovis was detected at a low level in recently purchased backgrounded and stocker calves in Georgia. Although slightly more infected calves were detected via culture and PCR assay together, PCR assay appeared to accurately identify M bovis at the farm level.

• Prevalence and characterization of urinary tract infections in dogs with surgically treated type 1 thoracolumbar intervertebral disc extrusion.

  Authors: Kevin S Stiffler, M A McCrackin Stevenson, Susan Sanchez, Jeanne A Barsanti, Erik Hofmeister, Steven C Budsberg

  Veterinary surgery : VS : the official journal of the American College of Veterinary Surgeons. 07/2006; 35(4):330-6.

  OBJECTIVE: To determine the prevalence of urinary tract infections (UTI), factors that correlate positively with UTI, and whether identified UTI are most likely community- or hospital acquired inOBJECTIVE: To determine the prevalence of urinary tract infections (UTI), factors that correlate positively with UTI, and whether identified UTI are most likely community- or hospital acquired in dogs with surgically treated type 1 thoracolumbar intervertebral disc (IVD) extrusions. STUDY DESIGN: Prospective cross-sectional clinical study. SAMPLE POPULATION: Dogs (n=92) that were surgically treated for a thoracolumbar extradural compressive spinal cord lesion that was consistent with type 1 IVD extrusion. METHODS: Dogs were evaluated for bacterial lower UTI when possible by cystocentesis and urine culture before surgery, and 48-72, 96-120 hours, and 7 days after surgery while hospitalized. Paraparesis, confirmation of thoracolumbar extruded nucleus pulposus, and informed owner consent were required for study inclusion. Urine specimens (n=297) were cultured and both objective and subjective clinical data were obtained. RESULTS: Prevalence of UTI in dogs with surgically treated type 1 thoracolumbar IVD extrusion was 27% (25 dogs). Temporal prevalence of UTI was 15% (13/89) before surgery, 12% (11/91) at 2-3 days, 16% (12/76) at 4-5 days, and 20% (8/41) at 7 days after surgery. Statistically significant factors affecting UTI prevalence included neurologic and urinary status, sex, administration of perioperative antibiotics, and amount of time body temperature was <35 degrees C during anesthesia. CONCLUSION: UTI are common in dogs with surgically treated type 1 thoracolumbar IVD extrusion. Females, dogs that cannot ambulate or voluntarily urinate, dogs not administered perioperative cefazolin, and dogs whose body temperature falls <35 degrees C during anesthesia have a higher incidence of UTI. CLINICAL RELEVANCE: All dogs with surgically treated type 1 thoracolumbar IVD extrusion should be monitored for the presence of UTI; however, close attention should be paid to females and dogs that cannot ambulate or voluntarily urinate.

• Dissemination of fluoroquinolone-resistant Campylobacter spp. within an integrated commercial poultry production system.

  Authors: Umelaalim Idris, Jingrang Lu, Marie Maier, Susan Sanchez, Charles L Hofacre, Barry G Harmon, John J Maurer, Margie D Lee

  Applied and environmental microbiology. 06/2006; 72(5):3441-7.

  While characterizing the intestinal bacterial community of broiler chickens, we detected epsilon-proteobacterial DNA in the ilea of 3-day-old commercial broiler chicks (J. Lu, U. Idris, B. Harmon, C.While characterizing the intestinal bacterial community of broiler chickens, we detected epsilon-proteobacterial DNA in the ilea of 3-day-old commercial broiler chicks (J. Lu, U. Idris, B. Harmon, C. Hofacre, J. J. Maurer, and M. D. Lee, Appl. Environ. Microbiol. 69:6816-6824, 2003). The sequences exhibited high levels of similarity to Campylobacter jejuni and Campylobacter coli sequences, suggesting that chickens can carry Campylobacter at a very young age. Campylobacter sp. was detected by PCR in all samples collected from the ilea of chicks that were 3 to 49 days old; however, it was detected only in the cecal contents of chickens that were at least 21 days old. In order to determine whether the presence of Campylobacter DNA in young chicks was due to ingestion of the bacteria in food or water, we obtained commercial broiler hatching eggs, which were incubated in a research facility until the chicks hatched. DNA sequencing of the amplicons resulting from Campylobacter-specific 16S PCR performed with the ileal, cecal, and yolk contents of the day-of-hatching chicks revealed that Campylobacter DNA was present before the chicks consumed food or water. The 16S rRNA sequences exhibited 99% similarity to C. jejuni and C. coli sequences and 95 to 98% similarity to sequences of other thermophilic Campylobacter species, such as C. lari and C. upsaliensis. The presence of C. coli DNA was detected by specific PCR in the samples from chicks obtained from a commercial hatchery; however, no Campylobacter was detected by culturing. In order to determine whether the same strains of bacteria were present in multiple levels of the integrator, we cultured Campylobacter sp. from a flock of broiler breeders and their 6-week-old progeny that resided on a commercial broiler farm. The broiler breeders had been given fluoroquinolone antibiotics, and we sought to determine whether the same fluoroquinolone-resistant strain was present in their progeny. The isolates were typed by pulsed-field gel electrophoresis, which confirmed that the parental and progeny flocks contained the same strain of fluoroquinolone-resistant C. coli. These data indicate that resistant C. coli can be present in multiple levels of an integrated poultry system and demonstrated that molecular techniques or more sensitive culture methods may be necessary to detect early colonization by Campylobacter in broiler chicks.

• In vitro efficacy of a buffered chelating solution as an antimicrobial potentiator for antifungal drugs against fungal pathogens obtained from horses with mycotic keratitis.

  Authors: William L Weinstein, Phillip A Moore, Susan Sanchez, Ursula M Dietrich, Richard E Wooley, Branson W Ritchie

  American journal of veterinary research. 05/2006; 67(4):562-8.

  OBJECTIVE: To determine whether a novel third-generation chelating agent (8 mM disodium EDTA dehydrate and 20 mM 2-amino-2-hydroxymethyl-1, 3-propanediol) would act as an antimicrobial potentiator toOBJECTIVE: To determine whether a novel third-generation chelating agent (8 mM disodium EDTA dehydrate and 20 mM 2-amino-2-hydroxymethyl-1, 3-propanediol) would act as an antimicrobial potentiator to enhance in vitro activity of antifungal medications against fungal isolates obtained from horses with mycotic keratitis. SAMPLE POPULATION: Fungal isolates (3 Aspergillus isolates, 5 Fusarium isolates, 1 Penicillium isolate, 1 Cladosporium isolate, and 1 Curvularia isolate) obtained from horses with mycotic keratitis and 2 quality-control strains obtained from the American Type Culture Collection (ATCC; Candida albicans ATCC 90028 and Paecilomyces variotii ATCC 36257). PROCEDURE: Minimum inhibitory concentrations (MICs) against fungal isolates for 4 antifungal drugs (miconazole, ketoconazole, itraconazole, and natamycin) were compared with MICs against fungal isolates for the combinations of each of the 4 antifungal drugs and the chelating agent. The Clinical and Laboratory Standards Institute microdilution assay method was performed by use of reference-grade antifungal powders against the fungal isolates and quality-control strains of fungi. RESULTS: Values for the MIC at which the antifungal drugs decreased the growth of an organism by 50% (MIC50) and 90% (MIC90) were decreased for the control strains and ophthalmic fungal isolates by 50% to 100% when the drugs were used in combination with the chelating agent at a concentration of up to 540 microg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: The chelating agent increased in vitro activity of antifungal drugs against common fungal pathogens isolated from eyes of horses with mycotic keratitis.

• Microbiologic findings in feedlot cattle with acute interstitial pneumonia.

  Authors: Amelia R Woolums, Gary L Mason, Larry L Hawkins, Corrie C Brown, Shamita M Williams, Jared A Gould, Jocelyn J Fox, Scott D Sturgeon, Jenny L Anderson, Florence E Duggan, Susan Sanchez, Paula B Barrett, Sharon W Chitwood

  American journal of veterinary research. 12/2004; 65(11):1525-32.

  OBJECTIVE: To test the hypothesis that feedlot cattle with acute interstitial pneumonia (AIP) have bacterial infection of the lung or liver and concurrent bovine respiratory syncytial virus (BRSV)OBJECTIVE: To test the hypothesis that feedlot cattle with acute interstitial pneumonia (AIP) have bacterial infection of the lung or liver and concurrent bovine respiratory syncytial virus (BRSV) infection significantly more often than pen mates without AIP ANIMALS: 39 feedlot cattle with signs consistent with AIP and no history of treatment with antimicrobials and 32 healthy control cattle from the same pens. PROCEDURES: Lung and liver specimens were obtained postmortem for bacterial or mycoplasmal culture and histologic examination; lung tissue was assessed for BRSV infection immunohistochemically. RESULTS: Among affected cattle, 26 had AIP confirmed histologically. Lung tissue from 11 cattle with AIP yielded microbial respiratory tract pathogens on culture; tissues from control animals yielded no microbial growth. In 4 cattle with AIP and 2 control animals, liver abscesses were detected; bacteria were isolated from abscessed tissue in 3 and 1 of those animals, respectively. Immunohistochemically, 9 cattle with AIP and no control animals were BRSV-positive. Histologically, 9 AIP-affected cattle had only acute alveolar damage with exudation, and the other 17 had acute exudation with type II pneumocyte hyperplasia. No lesions of AIP were detected in control animals. Only 4 AIP-affected cattle had bacterial infection of the lung with concurrent BRSV infection. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that microbial respiratory tract pathogens are more common in cattle with AIP than in healthy pen mates. Control of bacterial pneumonia late in the feeding period may reduce the incidence of AIP at feedlots where AIP is a problem.

• Serological response to Pasteurella multocida NanH sialidase in persistently colonized rabbits.

  Authors: Susan Sanchez, Shaikh Mizan, Charlotte Quist, Patricia Schroder, Michelle Juneau, Donald Dawe, Branson Ritchie, Margie D Lee

  Clinical and diagnostic laboratory immunology. 10/2004; 11(5):825-34.

  Pasteurella multocida is a mucosal pathogen that colonizes the upper respiratory system of rabbits. Respiratory infections can result, but the bacteria can also invade the circulatory system,Pasteurella multocida is a mucosal pathogen that colonizes the upper respiratory system of rabbits. Respiratory infections can result, but the bacteria can also invade the circulatory system, producing abscesses or septicemia. P. multocida produces extracellular sialidase activity, which is believed to augment colonization of the respiratory tract and the production of lesions in an active infection. Previously, it was demonstrated that some isolates of P. multocida contain two unique sialidase genes, nanH and nanB, that encode enzymes with different substrate specificities (S. Mizan, A. D. Henk, A. Stallings, M. Meier, J. J. Maurer, and M. D. Lee, J. Bacteriol. 182:6874-6883, 2000). We developed a recombinant antigen enzyme-linked immunosorbent assay (ELISA) based on the NanH sialidase of P. multocida and demonstrated that rabbits that were experimentally colonized with P. multocida produce detectable anti-NanH immunoglobulin M (IgM) and IgG in serum, although they demonstrated no clinical signs of pasteurellosis. In addition, clinically ill pet rabbits infected with P. multocida possessed IgM and/or IgG antibody against NanH. The NanH ELISA may be useful for the diagnosis of P. multocida infections in sick rabbits as well as for screening for carriers in research rabbit colonies.

• Rapid detection of Campylobacter coli, C. jejuni, and Salmonella enterica on poultry carcasses by using PCR-enzyme-linked immunosorbent assay.

  Authors: Yang Hong, Mark E Berrang, Tongrui Liu, Charles L Hofacre, Susan Sanchez, Lihua Wang, John J Maurer

  Applied and environmental microbiology. 07/2003; 69(6):3492-9.

  Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a seriesContamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 x 10(2) and 4 x 10(1) CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100- to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa = 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for SALMONELLA: With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry.

• Pharmacokinetics of imipenem in dogs.

  Authors: Corrie W Barker, Weijiang Zhang, Susan Sanchez, Steven C Budsberg, F Douglas Boudinot, M A McCrackin Stevenson

  American journal of veterinary research. 07/2003; 64(6):694-9.

  OBJECTIVE: To determine the plasma pharmacokinetics of imipenem (5 mg/kg) after single-dose IV, IM, and SC administrations in dogs and assess the ability of plasma samples to inhibit the growth ofOBJECTIVE: To determine the plasma pharmacokinetics of imipenem (5 mg/kg) after single-dose IV, IM, and SC administrations in dogs and assess the ability of plasma samples to inhibit the growth of Escherichia coli in vitro. ANIMALS: 6 adult dogs. PROCEDURE: A 3-way crossover design was used. Plasma concentrations of imipenem were measured after IV, IM, and SC administration by use of high-performance liquid chromatography. An agar well antimicrobial assay was performed with 3 E coli isolates that included a reference strain and 2 multidrug-resistant clinical isolates. RESULTS: Plasma concentrations of imipenem remained above the reported minimum inhibitory concentration for E coli (0.06 to 0.25 microg/mL) for a minimum of 4 hours after IV, IM, and SC injections. Harmonic mean and pseudo-standard deviation half-life of imipenem was 0.80 +/- 0.23, 0.92 +/- 0.33, and 1.54 +/- 1.02 hours after IV, IM, and SC administration, respectively. Maximum plasma concentrations (Cmax) of imipenem after IM and SC administration were 13.2 +/- 4.06 and 8.8 +/- 1.7 mg/L, respectively. Time elapsed from drug administration until Cmax was 0.50 +/- 0.16 hours after IM and 0.83 +/- 0.13 hours after SC injection. Growth of all 3 E coli isolates was inhibited in the agar well antimicrobial assay for 2 hours after imipenem administration by all routes. CONCLUSIONS AND CLINICAL RELEVANCE: Imipenem is rapidly and completely absorbed from intramuscular and subcutaneous tissues and effectively inhibits in vitro growth of certain multidrug-resistant clinical isolates of E coli.

• Evaluation of broiler litter with reference to the microbial composition as assessed by using 16S rRNA and functional gene markers.

  Authors: Jingrang Lu, Susan Sanchez, Charles Hofacre, John J Maurer, Barry G Harmon, Margie D Lee

  Applied and environmental microbiology. 03/2003; 69(2):901-8.

  Very little is known about the microbial composition of animal bedding wastes, including poultry litter, and what is known has been deduced from standard culture methods, by which some fastidiousVery little is known about the microbial composition of animal bedding wastes, including poultry litter, and what is known has been deduced from standard culture methods, by which some fastidious organisms that exist in the environment may not be detected. We evaluated the bacterial composition of poultry litter by using a combination of culture and molecular detection. Total aerobic bacteria in poultry litter were detected by culture at 10(9) CFU/g of material. Enteric bacteria such as Enterococcus spp. and coliforms composed 0.1 and 0.01%, respectively, of the total aerobic cultivatable bacteria in poultry litter; no Salmonella strains were detected by culture. In order to characterize the most abundant bacterial groups, we sequenced 16S ribosomal DNA (rDNA) genes amplified by PCR with microbial community DNA isolated from poultry litter as the template. From the 16S rDNA library, 31 genera were identified. Twelve families or groups were identified with lactobacilli and Salinococcus spp. forming the most abundant groups. In fact, 82% of the total sequences were identified as gram-positive bacteria with 62% of total belonging to low G+C gram-positive groups. In addition to detection of 16S rDNA sequences associated with the expected fecal bacteria present in manure, we detected many bacterial sequences for organisms, such as Globicatella sulfidofaciens, Corynebacterium ammoniagenes, Corynebacterium urealyticum, Clostridium aminovalericum, Arthrobacter sp., and Denitrobacter permanens, that may be involved in the degradation of wood and cycling of nitrogen and sulfur. Several sequences were identified in the library for bacteria associated with disease in humans and poultry such as clostridia, staphylococci, and Bordetella spp. However, specific PCR targeting other human and veterinary pathogens did not detect the presence of Salmonella, pathogenic Escherichia coli, Campylobacter spp., Yersinia spp., Listeria spp., or toxigenic staphylococci. PCR and DNA hybridization revealed the presence of class 1 integrons with gene cassettes that specify resistance to aminoglycosides and chloramphenicol. Only from understanding the microbial community of animal wastes such as poultry litter can we manage animal disease and limit the impact of animal waste on the environment and human and animal health.

• Class 1 integron-associated tobramycin-gentamicin resistance in Campylobacter jejuni isolated from the broiler chicken house environment.

  Authors: Margie D Lee, Susan Sanchez, Martha Zimmer, Umelaalim Idris, Mark E Berrang, Patrick F McDermott

  Antimicrobial agents and chemotherapy. 12/2002; 46(11):3660-4.

  Using PCR, we screened 105 isolates of poultry-associated Campylobacter jejuni for the presence of class 1 integrons. Of those isolates, 21% (22 of 105) possessed the integrase gene, but only 5Using PCR, we screened 105 isolates of poultry-associated Campylobacter jejuni for the presence of class 1 integrons. Of those isolates, 21% (22 of 105) possessed the integrase gene, but only 5 isolates produced an amplicon in a 5'-3' conserved sequence PCR directed toward amplification of the resistance cassettes. DNA sequencing demonstrated that all five isolates possessed the aminoglycoside resistance gene, aacA4.

• Characterization of multidrug-resistant Escherichia coli isolates associated with nosocomial infections in dogs.

  Authors: Susan Sanchez, M A McCrackin Stevenson, Charlene R Hudson, Marie Maier, Tameka Buffington, Quyen Dam, John J Maurer

  Journal of clinical microbiology. 11/2002; 40(10):3586-95.

  Multidrug-resistant opportunistic pathogens have become endemic to the veterinary hospital environment. Escherichia coli isolates resistant to 12 antibiotics were isolated from two dogs that wereMultidrug-resistant opportunistic pathogens have become endemic to the veterinary hospital environment. Escherichia coli isolates resistant to 12 antibiotics were isolated from two dogs that were housed in the intensive care unit at The University of Georgia Veterinary Teaching Hospital within 48 h of each other. Review of 21 retrospective and prospective hospital-acquired E. coli infections revealed that the isolates had similar antibiotic resistance profiles, characterized by resistance to most cephalosporins, beta-lactams, and the beta-lactamase inhibitor clavulanic acid as well as resistance to tetracycline, spectinomycin, sulfonamides, chloramphenicol, and gentamicin. E. coli isolates with similar resistance profiles were also isolated from the environment in the intensive care unit and surgery wards. Multiple E. coli genetic types were endemic to the hospital environment, with the pulsed-field gel electrophoresis fingerprint identified among E. coli isolates from diseased animals and the hospital environment matching. The extended-spectrum cephalosporin resistance in these nosocomial E. coli isolates was attributed to the cephamycinase-encoding gene, bla(CMY2). Chloramphenicol resistance was due in part to the dissemination of the florfenicol resistance gene, flo, among these isolates. Resistance encoded by both genes was self-transmissible. Although bla(CMY2) and flo were common to the polyclonal, nosocomial E. coli isolates, there was considerable diversity in the genetic compositions of class 1 integrons, especially among isolates belonging to the same genetic type. Two or more integrons were generally present in these isolates. The gene cassettes present within each integron ranged in size from 0.6 to 2.4 kb, although a 1.7-kb gene cassette was the most prevalent. The 1.7-kb gene cassette contained spectinomycin resistance gene aadA5 and trimethoprim resistance gene dfrA17.

• Animal issues associated with Escherichia coli O157:H7.

  Authors: Susan Sanchez, Margie D Lee, Barry G Harmon, John J Maurer, Michael P Doyle

  Journal of the American Veterinary Medical Association. 11/2002; 221(8):1122-6.

• Animal sources of salmonellosis in humans.

  Authors: Susan Sanchez, Charles L Hofacre, Margie D Lee, John J Maurer, Michael P Doyle

  Journal of the American Veterinary Medical Association. 09/2002; 221(4):492-7.