Global Veterinary Microbiology and Veterinary Medicine Summit October 17-19, 2016 Chicago Illinois, USA
(5 Plenary Forum-1 Event)

Chair

Natascia Bruni

Istituto Farmaceutico Candioli, Italy

Co-Chair

Sue Hudson Duran

Auburn University, USA

Session Introduction

Mahdi Saeed

Michigan State University, USA

Title: Egg-derived antibody for prevention and treatment of E. coli –induced diarrhea

Biography:

Mahdi Saeed is the Professor of Epidemiology and Biostatistics in Michigan State University. He has completed his DVM degree in 1973 from University of Baghdad and

PhD from Washington State University

Abstract:

Antimicrobials are still the major tools for treating infectious diseases. However, the emergence of multidrug resistant bacterial

pathogens and the observation that antibiotic treatment might actually increase the production of some bacterial-derived toxins,

limit the usefulness of this approach, Enterotoxigenic Escherichia coli (ETEC) strains that produce heat stable enterotoxin are the

leading cause of traveler’s diarrhea and a major cause of diarrheal disease in developing countries, especially among children. They

are transmitted by contaminated food and water and cause profuse watery diarrhea which results in rapid dehydration and death

in human neonates. The objective of this project is to produce antibodies against STa that can serve as a therapeutic preparation

against diarrheal disease caused by ETEC strains. In order to produce large amounts of high avidity antibodies, a novel approach

was pursued which consists in purification of ETEC-STa, conjugation with bovine serum albumin and immunization of egg-laying

hens to produce STa-specific egg yolk-derived antibodies. Chickens were selected due to the well-known capacity of birds to generate

strong immune responses to protein antigens exceeding those generated in rabbits and other mammalian species, pooled egg-yolk

samples from 24 different birds immunized with STa were produced. These samples were transferred and characterized in terms

of levels of antibodies and antibody avidity by the Immunology Research Unit (IRU) at the University of Maryland. ELISAs to

measure STa antibody titers and avidity were set up and validated using the following reagents: Lyophilized E. coli STa, lyophilized

STa hyper-immune rabbit serum, STa-containing egg yolk extract from an immunized bird (positive control) and purified IgY. ELISA

checker board titrations assessing different experimental conditions and reagents were performed which allowed establishing the

adequate experimental conditions for measurements of STa-specific rabbit IgG and chicken IgY antibodies with good sensitivity and

reproducibility. The STa antigen purified at MSU was potent and produced very high absorbance values with low background. The

rabbit antiserum prepared at MSU was also highly reactive and yielded elevated antibody titers, above 1×100 EU/ml. The optimal

conditions for the measurement of chicken IgY STa antibodies included: Coating with STa at 1 pg/ml in PBS buffer overnight at 4

oC, washing plates after each incubation 6 times with PBS Tween 0.05%, soaking plates for 2 min in between washes; blocking plates

with PBS 0.102 Tween, incubating samples for that 37 oC, using as corlugate: HRP-conjugated AffiniPure Rabbit anti-chicken IgY, Fc

Fragment Specific diluted 1/5000. A standard curve with known concentration of IgY (a commercial reagent) was used to interpolate

absorbance values and to report antibody titers in mass per volume (concentrations). To measure avidity, an ELISA was performed

as described above adding a 10 min overlay with 6M urea after incubation with the experimental samples with the purpose of

dissociating low affinity antibodies. An avidity index was calculated for each sample as the residual titer measured in the presence vs.

absence of urea. The STa IgY titers in the 24 egg yolk samples ranged between 470 and 6182 EU/ml or 390 and 5399 ng/ml (expressed

in antibody concentrations). The range of avidity indices (AI) were between 54 and 100% with a mean AI of 77.2. The avidity of STa

IgY antibodies was higher than the avidity of the rabbit antisera despite the higher antibody levels produced in rabbits.

Akdoucche Leila

Higher National Veterinary School, Algeria

Title: Importance of yeasts in the mammary infection of the cattle in the region of SidiM’Hammed Ben Ali, Wilaya of Relizane, Algeria

Biography:

Akdouche Leila is currently a Research Scholar in Higher National Veterinary School, Algeria. Her research interest includes microbiology and antimicrobial effect study.

Abstract:

The mastitis is one of the principal pathologies in the dairy bovine exploitation. The majority of the cases are caused by bacteria

but there are also cases caused by fungi. The objective of our study was to evaluate the occurrence of these fungi in mammary

glands of 39 cows (mastitic cows and clinically healthy cows) belonging to two types of farms (4 exploitations using manual milking

and 3 exploitations with milking machine) in the area of SidiM’Hammed Ben Ali, Wilaya of Relizane and to assess some risk factors

(the tubes of drug, animal excretions, goblets-milkers, the milker hands and the litter). For this purpose, 150 sample of milk and 94

swabs were carried out. Our results revealed the presence of a heavy load of fungi cells in healthy and in the mastitis milks with a

strong frequency of the Trichosporon sp. (43, 58%) followed by the Candida sp. (30.76%). The same yeasts were isolated from swabs.

Ahasan Mohammad Shamim

James Cook University, Australia

Title: Detection and antibiotic resistance of Gram negative enteric isolates from green sea turtles (Chelonia mydas) in Great Barrier Reef, Australia

Biography:

Mohammad Shamim Ahasan is currently pursuing his PhD at James Cook University, Australia. He has completed his MS in Tropical Animal Health from Institute of Tropical

Medicine, Belgium. He is working as an Assistant Professor (in deputation) in the Faculty of Veterinary and Animal Sciences under Hajee Mohammad Danesh Science and

Technology University, Bangladesh. He has published more than 15 articles in reputed journals.

Abstract:

Gastrointestinal disorders are one of the identified threats for mortality of endangered green turtle populations in rehabilitation

centres. In most cases, accurate diagnosis is difficult and broad-spectrum antibiotics are used for treatment. This study identified

and measured the antibiotic resistance of intestinal Enterobacteriaceae that were isolated from green sea turtles in northern Great

Barrier Reef, Australia. Deep cloacal swabs were taken aseptically from 76 green turtles ranging from juvenile to adult from different

locations including rehabilitation centers between June 2015 and January 2016. A total of 173 out of 371 Gram negative bacterial

isolates were identified as Enterobacteriaceae using culture dependent phenotypic, biochemical and molecular techniques. The

resistance against 12 antimicrobial agents belonging to 5 different antibiotic classes was determined using the broth microdilution

inhibition (MIC) technique. 18 different species were identified that represent 13 different genera of Enterobacteriaceae. The dominant

isolates were Citrobacter (18.5%), Edwardsiella (17.92%) and Escherichia (13.29%). The other Enterobacteriaceae members include

Salmonella, Proteus, Klebsiella, Enterobacter, Serratia, Morganella, Providencia, Pantoea, Cronobacter and Raoultella. The isolates

showed highest resistance to beta lactam antibiotics (86.13%) followed by quinolone derivatives (49.13%), aminoglycosides (47.98%),

chloramphenicol (18.50%) and potentiated sulphonamides (7.51%) respectively. 27.17% isolates were identified exhibited multidrug

resistance. These included Escherichia, Klebsiella, Citrobacter and Proteus. Isolates obtained from rehabilitation centers were

significantly resistant (p<0.05) to the antibiotics except gentamicin, streptomycin and potentiated sulfonamides. The environmental

isolates recovered from the turtles of Cockle Bay and Ollera Beach showed significant (P<0.05) resistance to ceftiofur. The findings of

this study demonstrate that enteric bacterial flora of green sea turtles is composed of a wide spectrum of Enterobacteriaceae including

several potential zoonotic pathogens with multiple drug resistance. These findings indicate that in the future, it may be important to

investigate ways to reduce the level of multi-resistant Enterobacteriaceae in rehabilitated turtles before they are released back into the

Ocean.

Chair

Sue Hudson Duran

Auburn University, USA

Co-Chair

Natascia Bruni

Istituto Farmaceutico Candioli, Italy

Session Introduction

Natascia Bruni

Istituto Farmaceutico Candioli, Italy

Title: In vitro bactericidal activity of lactoferricin and other enzymes on bacteria selected from dogs with pyoderma

Biography:

Natascia Bruni has completed her PhD from University of Turin and Postdoctoral studies from High Synthesis School of Gargnano, Italy. She is the Director of Research

and Development in Candioli Pharma. She has published more than 10 papers in reputed journals.

Abstract:

Lactoferricin (LFcin) is a peptide with antimicrobial activity against microorganisms such as Gram negative and positive bacteria,

including some antibiotic-resistant pathogens. Others enzymes like dextrozyme, alcalase and amylase may be associated to the

LFcin to enhance its antimicrobial action. The aim of this study was to evaluate the in vitro activity of LFcin associations with

dextrozyme (LFD), alkalase (LFL) and amylase (LFM) against Staphylococcus pseudintermedius, Escherichia coli, Proteus vulgaris

and Pseudomonas aeruginosa strains isolated from dogs with pyoderma. The evaluation was performed using minimum inhibitory

concentrations with a microtiter plate dilution method starting by an LFcin/enzymes solution at 11%. The bacterial inoculums in

log-phase growth were prepared in brain heart infusion broth (BHI) with a turbidity of 0.5 McFarland, corresponding to 102 to 103

cells/ml. Dilutions in BHI at 2:1 (7.5%), 1:1 (5.5%), 1:2 (3.7%) and 1:5 (1.8%) were used for each bacteria. Negative and positive

controls were included. All the wells were incubated 10 μl on blood agar at 37 °C for 48 hours to confirm the bacterial inhibition. The

associations LFD and LFM showed bactericidal activity against all the isolates at 7.3%, at 5.5%, only for S. pseudintermedius at 3.7%.

The LFL showed inhibition at 3.7% for all strains and resistance at 1.8%. These results suggest that the associations of LFcin with other

enzymes improve its antimicrobial activity. The LFL exhibits in vitro bactericidal activity even against a strain of multidrug resistant

P. aeruginosa at low concentrations. LFcin and its associations should be a new topical treatment of skin infections.

Eugene

University of Calgary, Canada

Title: he changing clinical manifestations and pathological processes Histophilus somni infections in cattle

Biography:

Eugene has received his Doctor of Veterinary Medicine (DVM) in 1972 from the Western College of Veterinary Medicine, Saskatoon, Saskatchewan. He has spent three

years practicing in northeastern Alberta before returning to the Western College of Veterinary Medicine on Alberta-sponsored fellowship to complete Post-graduate degree.

He has received MVS degree from the University of Melbourne, Australia in 1977. In 1977, he accepted a position with the Ambulatory Clinic at the Western College of

Veterinary Medicine, Saskatoon, Saskatchewan and has spent his time working with a general interest in beef cattle medicine. He became associated with Feedlot Health

Management Services in 2003.

Abstract:

Histophilus somni, a small, pleomorphic, Gram-negative bacteria, causes an infection commonly referred to as “Histophilosis”,

implying that the infection is systemic even though the clinical manifestation may be specific to an organ system. It was first

identified in 1960 as the cause of infectious thromboembolic meningoencephalitis (ITEME) in cattle fed in confinement and this

encephalitic form of the infection was recognized to be the main manifestation. More recently, this manifestation is diagnosed less

commonly and infection with H. somni has been shown to be an important cause of sporadic “sudden death” in groups of recently

weaned, confined cattle and associated with diverse pathological processes involving the heart, thoracic space and lungs. Modern

microbiology has demonstrated and continues to demonstrate characteristics of the organism that contribute significantly to its’

virulence but may not be present in all isolates. Pathogenic and commensal strains have been shown to exist. The diagnosis of

Histophilosis historically was made by isolating the pathogen microbiologically which was often difficult because of prior treatment

with antimicrobials. However, confirmation currently is made using molecular techniques like immunohistochemical (IHC) staining

of H&E fixed tissues or polymerase chain reaction (PCR) of fresh swabs taken from the lesions. In vitro examination of the organism

suggests it is sensitive to most antibiotics, although the minimum inhibitory concentration (MIC) needed to prevent growth of H.

somni has apparently increased in recent years. The various clinical and pathological manifestations will be discussed with emphasis

on diagnosis and complications of clinical management.

Zeaur Rahim

International Centre for Diarrheal Disease Research, Bangladesh

Title: Mycobacterium orygis: Emerging causative agent of tuberculosis in cattle in Bangladesh

Biography:

Zeaur Rahim is a citizen of Bangladesh. He is a Microbiologist. After completion of Master degree in Biological Science, he became affiliated with icddr,b as Research Trainee for a period of three years since the year 1981. During his long career at icddr,b Dr. Rahim obtained training in the field of Water Bacteriology, Diarrhoeal Disease and finally Tuberculosis. In the year 2001, Dr. Rahim established Tuberculosis Laboratory at icddr,b to set up conventional culture and susceptibility testing of M. tuberculosis. Dr. Rahim published 75 papers in peer reviewed bio-medical journals and associated with different journals as reviewers manuscript. 

Abstract:

Bangladesh is one of the tuberculosis (TB) endemic countries of the world. National TB control program has proper attention on

human TB, but TB in cattle is neglected. In this study, vital organs of the dead cows were analyzed after postmortem to isolate

and identify Mycobacterium from the vital organs of the dead cows, perform drug susceptibility testing and genotyping of isolates.

After post-mortem, part of the tissue samples from vital organs was processed for histopathological examination. For culture, tissue

homogenate samples were inoculated on Lowenstein-Jensen slants. Characteristic colonies were confirmed Mycobacterium by acidfast

staining and colony morphology. Anti-TB drug sensitivity testing was performed following proportion method. For molecular

characterization, DNA was extracted from the culture of Mycobacterium for RD analysis, single nucleotide polymorphism for genes like;

gyrB, mmlp6, TbD1 and PPE55 genes. Genotyping was performed by standard spoligo and VNTR-MIRU typing. Histopathological

examination, demonstrated prominent granuloma, caseous necrosis and calcification as proof of TB. Mycobacterium grew from the

vital organs of 18 out 21 dead cattle. AFB staining, sensitivity to PNB indicated that the strains belong to M. tuberculosis complex.

Molecular characterization indicated that these strains belong to new species M. orygis. Isolated strains were sensitive to all first line

anti-TB drugs. M. orygis is an emerging species of Mycobacterium causing cattle TB. Detailed epidemiological study of M. orygis

needs to be conducted to control this disease. Sensitivity to all first line anti-TB drugs indicated that human infection with this TBbacillus

could be successfully treated with anti-TB drugs.

Indira T Kudva

United States Department of Agriculture-Agricultural Research Service, USA

Title: Shiga toxin-producing Escherichia coli and recto anal junction persistence in ruminants: A study of bacterial-epithelial interactions

Biography:

Indira T Kudva is a Research Microbiologist and Lead Scientist at the National Animal Disease Center, USDA, Ames, Iowa. She has received her BSc in Zoology and

MSc in Medical Microbiology from India, PhD in Microbiology, Molecular Biology and Biochemistry from the University of Idaho and trained as a Postdoctoral Fellow at

the University of Idaho, Massachusetts General Hospital and Harvard Medical School. She has over 25 years of experience in the field of microbiology, molecular biology

and infectious diseases. She has 29 peer-reviewed publications, 3 invited reviews, 27 meeting abstracts, 18 invited talks, 8 funded grants and novel inventions (4 patent

applications). She is also an adjunct Assistant Professor at the School of Veterinary Medicine, Iowa State University, the Executive Editor for the “Virulence Mechanisms

of Bacterial Pathogens” book, 5th Edition, ASM press and is on the Editorial Boards of the Applied and Environmental Microbiology (ASM press) and the SRL Proteomics

and Bioinformatics (Sci Res Literature) journals.

Abstract:

Escherichia coli O157:H7 (O157) was the first Shiga toxin-producing E. coli serotype to be associated with bloody diarrhea or

hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS) in humans. It has since been implicated in several outbreaks

in the U.S. and globally. Non-O157 STEC have not been associated with major outbreaks but still cause 64% of all STEC infections

in the U.S. Cattle are well documented ruminant reservoirs of STEC and the primary site of STEC persistence in these animals is the

rectoanal junction (RAJ). To investigate this persistence at the RAJ we have analyzed O157 adherence at a histological level, developed

a novel adherence assay using squamous epithelial cells at the RAJ, evaluated role of well characterized STEC adherence proteins,

in STEC attachment to the RAJ and explored the similarities in RAJ-STEC interactions with another ruminant animal. STEC show

distinct adherence patterns on the columnar epithelial and squamous epithelial (RSE) cells at the RAJ. The novel RSE cell adhesion

assay provides a convenient means of directly evaluating bacterial interactions with host-specific cells. We have determined that

proteins other than LEE and intimin-γ proteins are involved in STEC adherence to RSE cells. Such proteins with adhesin potential

have been shortlisted using proteomics for development of efficacious anti-adhesion modalities. We have also found that bison and

cattle RAJ share similar distribution of epithelial cell markers and O157 adheres to RSE cells from both animals in similar patterns,

supporting bison as likely ‘wildlife’ reservoirs for O157.

Beatriz Quinones

United States Department of Agriculture-Agricultural Research Service, USA

Title: Virulence and antimicrobial resistance profiles of Shiga toxin-producing Escherichia coli O157 and non-O157 recovered from domestic farm animals in rural communities in Northwestern Mexico

Biography:

Beatriz Quinones has completed her PhD in Molecular and Cell Biology and Postdoctoral studies in Plant and Microbial Biology from the University of California at Berkeley.

She is currently a Research Molecular Biologist in the Produce Safety and Microbiology Research Unit at the USDA/ARS-Western Regional Research Center. Her research

projects are aimed at the detection and genotyping of bacterial and viral food borne pathogens and have resulted in the publication of more than 35 papers in peer-reviewed

journals. Her ongoing collaborations with academic institutions in Mexico are focused on the detection of food borne pathogens in agricultural regions for export produce

Abstract:

Shiga toxin-producing Escherichia coli (STEC) are zoonotic enteric pathogens associated with human gastroenteritis worldwide.

Our study characterized the genotypic diversity and virulence profiles of O157 and non-O157 STEC strains, recovered from

domestic animals in the agricultural Culiacan Valley in Northwestern Mexico. By using a selective enrichment and isolation protocol,

serotype O157:H7 isolates were identified in 40% (26/65) of the recovered isolates from cattle, sheep and chicken feces. The clinicallyrelevant

non-O157 serotypes O8:H19, O75:H8, O111:H8 and O146:H21 represented 35.4% (23/65) of the isolates, mostly from sheep.

Analysis of the allelic diversity indicated that the O157:H7 isolates were highly related but a greater genotypic diversity was observed

in the non-O157 isolates. Genotyping assays revealed the presence of virulence genes coding for adhesins, cytotoxins, effectors and

Shiga toxin (Stx) subtypes in the tested strains. To examine the relative toxicities of the STEC strains, a fluorescent Vero cell based

assay was employed to measure the inhibition of protein synthesis by Stx. Non-O157 strains with serotypes O8:H19 O75:H8 and

O146:H8 were found to have an enhanced ability to inhibit protein synthesis in Vero cells while a reduced cytotoxicity was observed

for all O157:H7 strains. The STEC strains exhibited antimicrobial resistance to aminoglycosides, tetracyclines, cephalosporins and

penicillin which are commonly used in Mexico. In conclusion, zoonotic STEC with virulent genotypes are present in animals on

small farms in the Culiacan Valley and these findings emphasize the need for the development of control measures and surveillance

of antimicrobial resistance in an important agricultural region in Northwestern Mexico.

Eben Oosthuysen

New Mexico State University, USA

Title: Blood gas analysis as diagnostic tool for early detection of respiratory disease in cattle

Biography:

Eben Oosthuysen is currently pursuing his Doctoral degree in Ruminant Nutrition at New Mexico State University, USA. His research focus on alternative management

practices to improve feedlot receiving calf health in preparation for the soon anticipated restriction on antibiotic use in food producing animals. He has published his research

findings as proceedings and abstracts in more than 17 occasions.

Abstract:

Bovine respiratory disease is the most significant health problem facing in the U.S. beef industry. Blood oximetry is used in human

medicine to diagnose respiratory disease and has been correlated to arterial hypoxia in cattle. Therefore, we evaluated the possible

correlation between blood parameters and health and performance of immune-stimulated and hay-supplemented calves. Crossbred

heifers (n=705; 179±0.58 kg) were assigned to 48 pens and 4 treatments in a randomized complete block design. No metaphylaxis

(antibiotic) was used and calves received a corn gluten feed based ration (Ramp). Treatments were a factorial arrangement

of supplemental hay (+HAY vs. –HAY) and immune stimulation (+IMMUN vs. –IMMUN). Calves assigned to +HAY received

supplemental alfalfa hay for the first 14 days and +IMMUN calves received a DNA immunostimulant (Zelnate) on day 0. Calf average

daily gain was greater (P<0.01) for +HAY than –HAY during the first 14 days but lower from day 14 to 28. Immune stimulated calves

had lower (P<0.01) average daily gains from day 28 to 56 and from day 0 to 56. Treatments did not affect (P≥0.18) calf morbidity,

mortality or blood parameters (pH, glucose, lactate, sO2). Blood sO2 correlated (P<0.05) with mortality (R2=0.08) and blood glucose

correlated with first (R2=-0.22) and second (R2=-0.13) medical treatment. Lactate correlated (P<0.05) with first medical treatment

(R2=-0.12) and mortality (R2=-0.12). These correlations suggest possible application of blood parameters as diagnostic tool.

Chair

Sue Hudson Duran

Auburn University, USA

Co-Chair

Mahdi Saeed

Michigan State University, USA

Session Introduction

Juana Liz Vidal Arboleda

University of Antioquia, Colombia

Title: Characterization of the genetic variability of field strains of Brucella canis isolated in Antioquia, Colombia between 2005-2013

Biography:

Juana Liz Vidal Arboleda has completed her undergraduate studies in Microbiology at the University of Antioquia. She has participated as Colciencias Young

Researcher of the project in the Rio Grande II Reservoir. From 2009-20013, she did specialization in Veterinary Clinical Laboratory in the UDCA (University of

Applied and Environmental Sciences), currently pursuing her Master’s degree in Animal Science in University of Antioquia and serves as Microbiologist and

Bioanalyst in the Laboratory of Veterinary Microbiology, Faculty of Agricultural Sciences at the University of Antioquia.

Abstract:

Introduction: Brucella canis, a facultative intracellular pathogen is responsible for canine brucellosis, a zoonotic disease that

affects canines causing abortions and reproductive failures, human infection may cause mild, asymptomatic or serious medical

conditions. Since 2005 the presence of Brucella canis was demonstrated in Antioquia and the circulating strains were identified

as B. canis type-2, besides recognizing in them the operon virB. In order to learn more about the circulating strains the entire

genome of a wild strain of Brucella canis denoted Oliveri was sequenced, which showed insertions and deletions speciesspecific.

Objective: Characterize the genetic variability of field strains of Brucella canis isolated in Antioquia during 2005-2013.

Methods: A set of six pairs of primers that would allow identification of indels identified in the strain of B. canis str. Oliveri was

designed, after this conventional PCR and sequencing of the amplified products was performed on 30 strains of Brucella canis,

Brucella suis 1330 and bv. 1, Brucella melitensis 16M and vaccine strains of Brucella abortus S19 and RB51.

Results: 5 of 30 field strains studied share indels with the sequenced strain, the remaining 25 strains differ in a deletion exclusive

from the Oliveri strain. Inside the deletions a related one with a critical epimerase for the synthesis of carbohydrates from the

surfaces is present, which is related to the rough characteristics of the colonies like those presented by the field strains. A second

deletion is related to N-acetyltransferase, which prevents the action of aminoglycosides, its absence could facilitate the action

of these antibiotics in the bacteria. A third 6 bp deletions in chromosome II, is related to the ABC transporters superfamily,

which perform the transport of nutrients through the membrane and it is corresponded with a unique characteristic of the

species metabolism. A final deletion related to a glutamic acid tranference RNA is the only exclusive in Oliveri strain, this

deletion may result from the adaptation of the bacteria to intracellular lifestyle more secure and stable with a constant supply

of nutrients, additional we could talk about the effect of speciation and host specific adaptation or as in other Brucella species a

possible recombination process. Inside the insertions studied, one related with an anti prophage repressor which interacts with

prophage repressor genes facilitating horizontal gene transfer was presented; a second insertion evaluated as 60 bp is part of a

hypothetical protein with no specific function but that facilitates differentiation of Brucella canis from other species.

Conclusions: The field strains isolated in Antioquia present genotypic variations from the sequenced strain Oliveri, indicating

that this is not the only strain circulating in the region,which would be useful in future studies to

Juana Liz Vidal Arboleda

University of Antioquia, Colombia

Title: Prevalence of mastitis causing bacteria isolated in two diagnostic laboratories in Antioquia (Colombia), between the years 2013 and 2015

Biography:

Juana Vidal has completed her undergraduate studies in Microbiology at the University of Antioquia. She has participated as Colciencias Young Researcher

of the project in the Rio Grande II Reservoir. From 2009-20013, she did specialization in Veterinary Clinical Laboratory in the UDCA (University of Applied and

Environmental Sciences), currently pursuing her Master’s degree in Animal Science in University of Antioquia and serves as Microbiologist and Bioanalyst in the

Laboratory of Veterinary Microbiology, Faculty of Agricultural Sciences at the University of Antioquia.

Abstract:

Bovine mastitis continues to be the most economically important disease in dairy cattle worldwide and is caused by a

broad spectrum of infectious agents. In the last decade, the main bacteria isolated from mastitic cows in Colombia have

been Streptococcus agalactiae and Staphylococcus aureus. The state of Antioquia is one of the regions in Colombia with major

milk production. Almost 10% of the milking cows present subclinical mastitis diagnoses by somatic cell count or at list with

CMT. Therefore, the microbiological culture is an indispensable preventive and control tool, because it helps producers to

take appropriate decisions. This poster used reports from cultures of submitted milk samples to two veterinary diagnostic

laboratories located in Antioquia. Between the years 2013 and 2015, 7780 milk samples were processed for microbiological

culture at the ICMT-Colanta Laboratory and at Laboratory of Microbiology, School of Veterinary Medicine, University of

Antioquia, using a standard protocol. The most commonly isolated bacteria from all samples were coagulase-negative

Staphylococcus (20.7%), followed by S. aureus (17.9%), S. agalactiae (14.3%), S. uberis (9.1%), others Streptococcus (6.7%), S.

dysgalactiae (5.9%) and others bacteria (3.0%). Gram-negative bacilli were found in 10.0% of the samples; whereas no growth

occurred in 12.0% of cultures. In conclusion, the main bacterial genus isolated was Staphylococcus with a percentage of 38.6%.

The pattern of isolation of mastitis causing bacteria in Colombia has changed over last three years with an increase in the

percentage of isolates of CNS and a decrease in isolates of S. agalactiae.

Juana Liz Vidal Arboleda

University of Antioquia, Colombia

Title: Phylogenetic analysis of the Brucella canis str. Oliveri isolated in Antioquia, Colombia

Biography:

Juana Liz Vidal Arboleda has completed her undergraduate studies in Microbiology at the University of Antioquia. She has participated as Colciencias Young Researcher of

the project in the Rio Grande II Reservoir. From 2009-20013, she did specialization in Veterinary Clinical Laboratory in the UDCA (University of Applied and Environmental

Sciences), currently pursuing her Master’s degree in Animal Science in University of Antioquia and serves as Microbiologist and Bioanalyst in the Laboratory of Veterinary

Microbiology, Faculty of Agricultural Sciences at the University of Antioquia.

Abstract:

recognized and exhibit different host preference; comparative analysis of this genus have show a close genetic relationship between

species. To study the evolution of these species have developed multiple phylogenetic analyzes on which consensus has been separated

into a clade B. melitensis and B. abortus, in another clade marine species and one species consisting of B. suis and B. canis, recognizing

Brucella ovis as the ancestral species, suggesting that the initial contact to pigs, goats and cattles occurred from contact with infected

sheep.

Objective: To establish the phylogenetic relationships and the time of divergence of Brucella canis strain Oliveri.

Methods: Concatenated sequences of these genes were used: glK, trpE, cobQ, aroA, dnaK, rpoB, gap, gyrB, Omp2a, Omp2b, Omp25 and

Omp3 in 24 species of Brucella, including Brucella canis str. Oliveri. Using the Mega 6 program phylogenetic analysis was performed

by the method of neighbor joining with the substitution model Tamura-Nei and 10000 Bootstraps repetititons. The molecular clock

hypothesis among Brucella species was tested and the test of relative rates of Tajima between Brucella canis str. Oliveri and Brucella

canis ATCC was performed.

Results: Phylogenetic analysis including O. anthropi as out group indicate that Brucella ovis was the first lineage split is the most

basal species. The clade with greater genetic diversity is formed by B. suis where it will also find strains of B. canis suggesting a

recent divergence. B. abortus and B. melitensis appear as sister species, being each one monophyletic. Aquatic species B. ceti and

B. pinnipedialis are part of the same clade but separate from the terrestrial strain. Within B. canis all strains present a simultaneous

divergence since they split from their common ancestor, corroborating the result of the test of relative rates of Tajima with P=0.31731.

The molecular clock hypothesis is rejected between species indicating that the rate of evolution of all species of Brucella is not the

same.

Conclusions: The strain of Brucella canis str. Oliveri like others canis species diverged from Brucella suis. The Brucella canis species

had a similar rate of evolution and a genetic distance, so is not possible define which diverged first.

Benson C Iweriebor

University of Fort Hare, South Africa

Title: Climate change and tick-borne bacteria (Rickettsia spp., Anaplasma spp., and Ehrlichia spp.) in ticks collected in the Karoo regions of Eastern Cape, South Africa

Biography:

Benson C Iweriebor is working at AIDS Virus Research Laboratory, Department of Microbiology, University of Venda, South Africa. His research interest include AIDS and Microbiology.

Abstract:

Background: There has been a projected global increase in the distribution and prevalence of infectious diseases with climate change

thus suggesting a pending societal crisis. Global warming causes a wide spectrum of consequences on the human health, including

changes in the spread of tick borne pathogens. Ticks generally play an important role in the transmission and ecology of infectious

diseases. Climatic factors (temperature, rainfall and humidity) strongly influenced the ecology, development, behavior and survival

of ticks and the transmission dynamics of the diseases they transmit.

Objective: The objective of this study was to determine the prevalence of tick-borne bacterial pathogens of the genera Rickettsia spp.,

Ehrlichia spp., and Anaplasma spp. among ticks collected from domesticated ruminants in some selected localities in the Eastern Cape

Province of South Africa where there has been a decreased precipitation and slight increase in temperature recently.

Methods: Between February and May, 2016, a total of 1200 ticks were collected from domesticated ruminant heads in some selected

communities’ within the Nkonkonbe and Chris Hanne District Municipalities in the Eastern Cape, South Africa. Ticks were identified

based on morphological criteria. Genetic detection of tick-borne bacteria belonging to Rickettsia, Anaplasma and Ehrlichia genera

was performed by PCRs. Positive amplicons were sequenced and phylogenetically analyzed.

Results: The ticks were identified as belonging to the genera Rhipicephalus (980) and Amblyomma (220) respectively. Genetic

screening for Rickettsia spp., Ehrlichia spp. and Anaplasma spp. revealed the presence of Rickettsia spp. and Ehrlichia spp. respectively

while none was positive for Anaplasma spp. in the tick samples collected. Ticks from cattle were highly infected with Rickettsia spp.

while Ehrlichia spp. was isolated mainly from ticks collected from sheep. No positive result was obtained from ticks collected from

goats.

Conclusion: The observation that these pathogens are present in ticks on animals within homesteads suggests that inhabitants of

these communities could be infected and tourists visiting the areas should be cautious of tick-biting. The findings of this study show

that zoonotic pathogens are present in ticks in the studied localities. This information will be helpful in the epidemiology of tickborne

zoonotic diseases in the country as well as in creating awareness about such diseases in the veterinary, medical and tourism

sectors, as they may be the mostly affected.

M A Abouelkhair

University of Tennessee, USA

Title: SpA(KKAA) as a candidate Staphylococcus pseudintermedius Vaccine

Biography:

Mohamed Adel Salaheldin Abouelkhair has completed his Master’s degree from University of Sadat City, Egypt. He is an Assistant Lecturer in Bacteriology,

Mycology and Immunology Department, University of Sadat City, Egypt. He is currently a graduate PhD student in Comparative and Experimental Medicine

Graduate Program, College of Veterinary Medicine, University of Tennessee, USA.

Abstract:

Staphylococcus pseudintermedius is an important opportunist bacterium that frequently causes diseases in dogs and

occasionally causes zoonotic infections to human beings. It is commonly associated with canine pyoderma and is also

frequently associated with urinary tract, wound and surgical site infections. The development of a staphylococcal vaccine

is a challenging and prior infection with S. pseudintermedius is not associated with protective immunity. The envelope of S.

pseudintermedius is decorated with staphylococcal protein A (SpA), which captures the Fcγ portion of immunoglobulins to

prevent opsonophagocytosis and associates with the Fab portion of V(H)3-type of B cell receptors to trigger B cell superantigen

activity and consequently ablating adaptive immune responses. We show that mutation of Ig-binding domains of SpA with

amino acid substitutions Glutamine (Q) with Lysine (K) and aspartate (D) with Alanine (A) abolished the ability of the

resulting variant SpA(KKAA) to bind Fc gamma or Fab V(H)3 of B cells which may pave the way for the prevention and therapy

of staphylococcal disease in dogs.

 

Marita Vedovelli Cardozo

Sao Paulo State University , Brazil

Title: Klebsiella pneumoniae and Escherichia coli producing extended spectrum β-lactamase in broilers production chain

Biography:

Marita Vedovelli Cardozo is a Biologist and Master in Microbiology from Universidade Estadual Paulista (UNESP). Presently, she is a Doctoral student at the same

institution and her work is focused in microbiology with an emphasis on food safety, infectious diseases and public health.

 

Abstract:

The extended spectrum β-lactamases (ESBL) are enzymes that hydrolyze the β-lactam ring of penicillins, cephalosporins

and aztreonam, conferring resistance to these antimicrobials. There are an increasing number of infections caused by

Gram-negative bacteria producing extended spectrum β-lactamases and the emergence of new therapeutic resources does

not follow the evolution of resistance mechanisms, making them a serious public health problem. Brazil is the largest exporter

of chicken means that the presence of ESBL and it becomes also a potential risk in the agribusiness sector. Based on this, the

present study aims to characterize ESBL production in Enterobacteriaceae isolated from broilers production chain, including

animals, food and consumers. Of the 300 samples collected from cloacal swab, chicken meat and human feces, 36 were resistant

to cefotaxime (CTX) in antimicrobial phenotypic screening. After, by microarray molecular detection, it was found positive

isolates to CTX-M gene. Each positive CTX-M sample was subjected to PCR to confirm the presence of CTX-M-1 and

CTX-M-2 genes and the sequencing showed that all positive samples from CTX-M-1 group belonged to CTX-M-15 gene and

all positive samples from CTX-M-2 group belonged to CTX-M-2 gene. Moreover, identification by MALDI-TOF showed that

36 isolates, 15 were classified as Klebsiella pneumoniae and 21 isolates belong to Escherichia coli. These results evidence of the

correlation of resistance genes between animals and humans show the need to reduce the use of antibiotics in the production

of broilers in the country to combat the spread of antibiotic resistance.