7^th International Veterinary Congress September 04-05, 2017 Paris, France

Biography:

Abstract:

The biotechnological advancements have made new technologies and skills which are now available to help the identification of new potentialities of camel as an animal of unique attributes which would not only benefit the human and animal health but would be a favorite to researchers. Agranian society and traditional breeders will remain in scenario all the time as they produce and manage most of the camels. Research direction would be therefore more relevant when Agrarian society adopts modern breeding, management and health practices and scientists suggest value addition to this unique species. A focus on nutraceutical value of camel meat and milk may create a big consumer base which would be viewed as unique human health advantage. The camel racing industry must upkeep and modernize their breeding programs to preserve the germ plasm of best racing breeds. A large-scale vaccination program should be on priority in addition to increase in nutrition scale to camels. The diagnostic laboratories and veterinary hospital facilities should be functional in thick camel population zones to prevent morbidity and mortality. The scientific papers published in the Journal of Camel Practice and Research indicated a genuine trend of camel science and current research. The missing links of research were drawn out and were enlisted as a future vision of camel science.

Biography:

Abstract:

Esophageal obstruction is frequently reported in camels. They are caused by ingestion of food or other foreign bodies such as rags and polyethylene bags. In adult animal’s obstruction may be caused by bezoars migrating from the rumen or pressure against the esophagus from the neighboring tissues such as abscess, lymph node or cysts. Diagnosis was based on clinical findings, passing stomach tube through the mouth and radiography. Endoscopy was conducted in some instances. Esophagostomy was successful to remove foreign bodies in the cervical region or even in the mediastinal region however rumenotomy was achieved to remove foreign bodies in the cardiac region. The operations were fraught with high success rate but complications were in the form of esophageal fistula.

Biography:

Abstract:

Currently, several analgesic drugs such as opioids and NSAIDs have been used effectively in veterinary medicine. However, the clinical efficacy of these drugs for use in camels is still unknown. The aims of this study were to apply thermal and mechanical nociceptive threshold testing techniques in camels and evaluate them with reference to their applications, limitations and the factors which can influence both the testing procedure itself and the animal’s responses. Seven healthy camels of different breeds were brought into individual stables a minimum of three days before each experiment to allow re-acclimatization to the study environment. The camels were equipped with a wireless thermal and mechanical threshold testing system (Topcat Metrology Ltd). The display unit was mounted on the humps of the camels with the help of a surcingle and Velcro strips. Mechanical threshold was measured by using a pneumatic actuator, and thermal threshold was measured by using a thermal probe, both attached to the skin of metacarpus. Each camel went through 18 mechanical thermal and thresholds stimulation on 3 occasions at one-week interval. All camels responded to the stimulations, and type of reaction to the stimulus with a clear-cut end-point of stimulation was recorded. In conclusion, nociceptive threshold testing in camel using mechanical and thermal stimuli will be accepted as standard tests in preclinical studies for development of analgesics for camel use.

Biography:

Abstract:

There are many diseases of camels in Saudi Arabia which are estimated to be 810 000. Eastern province is the largest one of area composing one third of camels in the country. The camels in Veterinary Teaching Hospital of KFU those treated for non-infectious diseases were (763) and infectious diseases (539). In infectious diseases, as following: respiratory 101, enteritis 86, urinary 19, nervous 31, mouth 42, eyes 111 and skin 149 camels were treated. The camel’s diseases have been diagnosed by bacteriological, serological, molecular, histopathological and necropsy methods by different laboratories. Mostly reasons for diseases and death have been attributed to climatic changes, different breeding/grazing methods and self-improper measurements. Availability of vaccination program and traditional treatments are carried out. Middle East Respiratory Syndrome Coronavirus (MERS-CoV) has been confirmed in Saudi Arabia with 977 cases and odd case from time to time. Transmission of MERS-CoV has been attributed for different routes behind incontinous occurrence in Saudi Araba (Hemida et al, 2015). Middle East Respiratory Syndrome (MERS) first reported in Saudi Arabia in 2012 is a viral respiratory illness that is new to humans caused by MERS Coronavirus (MERS-CoV). Coronaviruses are common viruses that most people get some time in their life. MERS-CoV likely came from an animal source in the Arabian Peninsula. Researchers have found MERS-CoV in camels from several countries. Studies continue to provide evidence that camel infections may play a role in human infection with MERS-CoV. Molecular tests are used to diagnose active infection (presence of MERS-CoV) in people who are thought to be infected with MERS-CoV based on their clinical symptoms and having links to places where MERS has been reported. Real-time reverse-transcription polymerase chain reaction (rRT-PCR) assays are molecular tests that can be used to detect viral RNA in clinical samples. Saudi Arabia has warned its citizens to wear masks and gloves when dealing with camels as health experts said the animal was the likely source of the Middle East Respiratory Syndrome (MERS) coronavirus. Many camels were destroyed in endemic areas.

Biography:

Julian A Skidmore has completed her Graduation from University of Cambridge (UK) with a PhD in “Reproduction of the dromedary camel” in 1994. After completing her PhD in 1994 she has remained at the Camel Reproduction Centre in Dubai, United Arab Emirates, as Scientific Director leading a team of scientists covering all aspects of camel reproduction physiology with particular emphasis on embryo transfer of fresh, cooled and frozen embryos, artificial insemination of fresh and frozen semen and hybridization of New and Old-World camelids.

Abstract:

As the reproductive efficiency of camels is low under natural pastoral conditions the use of embryo transfer is becoming much more popular to increase the reproductive potential of female camels. Good pregnancy rates of 65-75% are achieved after transfer of fresh day 7 embryos into day 5 or 6 recipients. Synchronizing ovulation between donors and recipients, however, poses problems as camels lack the cyclical corpus luteum of spontaneous ovulators. This means that conventional methods used in cattle that involve giving two injections of prostaglandin 11 days apart are unsuitable for use in camels. More recent studies have shown that pregnancy rates of between 50-70% can be achieved in recipients that ovulate too early (i.e. before the donor is mated) if they are treated with meclofenamic acid to maintain the CL, and in recipients that ovulate too late if they are maintained on progesterone from two days before transfer to five days post transfer. However, pregnancy rates are dramatically reduced to <40% after transfer of frozen/thawed embryos. Several factors could contribute to this reduced embryo viability post-thaw, including size of embryo, physical injuries caused by intracellular and extracellular ice formation, cryoprotectant toxicity, osmotic stress and chilling injuries. In addition, freezing and thawing can irreversibly disrupt the organization of an embryo’s cytoskeleton and thereby reduce its ability to subsequently develop, so further work is required to improve slow-freezing and vitrification techniques so that pregnancy rates increase after transfer of frozen/thawed embryos.

Biography:

Abstract:

Biography:

J L Chaudhary is working as Professor and the Head at Department of Animal Production, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, India. His experience includes various programs, contributions and participation in different events for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.

Abstract:

An experiment was conducted on nine draught camels (7-9 years old and 574 ± 10.7 kg BW) to study the effect of energy supplementation fed Moth Straw (Phaseolus acontifolius Jacq.) based diets along with various levels of energy in concentrate mixtures. The camels were randomly divided into 3 groups of 3 animals each and allotted 3 dietary treatments i.e. T₁: 65% TDN in concentrate mixture, T₂: 70% TDN in concentrate mixture and T₃: 75% TDN in concentrate mixture along with dry moth straw fed ad libitum as sole roughage. The concentrate mixture was fed as per requirement of draught camels. The camels were subject to payload of 2.8 kg/kg body weight (18% BW) on a 2-wheeled camel cart. The camels covered 25.5 km distance in 2.65±0.02 to 3.19±0.03 hr at an average speed of 1.71±0.09 m/sec in a continuous work during winter season. The total dry matter intake (TDMI) was significantly (P<0.05) higher in T₃ as compared to T₂ and T₁ groups while non-significant different was observed between T₂ and T₁ groups. The DCP and TDN intake was significantly (P<0.05) higher in T₃ and T₂ groups as compared to T₁ group. The digestibility of DM, CP and NFE was higher (P<0.05) in draught camels fed on higher energy concentrate mixture but there was non-significant difference between the treatment for digestibility of OM, CF and EE. The DE and ME contents did not differ significantly among the treatment groups. All draught camels were trained to pull the multipurpose tool carrier two wheeled cart. The draught was varied by varying payload on the two-wheeled camel cart and hydraulic dynamometer was used for measuring the draught. The draught (kgf) and power output (hp) was highest (P<0.05) in T₃ as compared to other treatment groups. The maximum variation in physiological responses was noted at 2.8 kg/kg body weight (18% BW) on a two-wheeled camel cart. However, increase in rectal temperature, respiration and pulse rate was 36.42±0.14, 37.73±0.13, 15.72±0.40, 18.92±0.13, 45.9±0.32, 49±0.02 before and after carting. The blood serum was collected and analyzed to change in the biochemical attributes before and after carting. The serum glucose, lactate, cholesterol and aspartate transaminase activity changed significantly (P<0.05) during draught of camels. The results indicated that the nutrient utilization and draught performance was better in camels fed higher energy levels in concentrate mixture and covered distance without any hurdle and tolerate the work stress efficiently without any apparent ill effect on the health.

Biography:

Shawaf T is working at Department of Clinical Studies, College of Veterinary Medicine, King Faisal University, Saudi Arabia. His experience includes various programs, contributions and participation in different events for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.

Abstract:

Analysis of cerebrospinal fluid (CSF) is a valuable aid in the diagnosis of neurological disorders in man and animals. Previous reports documented the normal constituents of CSF in camel cadavers after slaughter. The purpose of the present paper was to establish a safe technique of sampling CSF from live camels and compare the obtained values with their serum constituents. An experimental design was performed in seven apparently healthy camels. There were 4 males and 3 females of 4-9 years old. The animals were clinically examined then blood samples were obtained from the jugular vein for serum biochemical analysis. Each camel was then sedated with an intravenous injection using 2% xylazine hydrochloride. The caudal part of the neck was aseptically prepared. CFS sample was withdrawn from the Atlanto-occipital articulation. Lateral radiographs of the neck were obtained while the needle was in situ. Eighteen biochemical parameters were determined from the serum and CSF of each camel. Serum had most of CSF tested parameters and was extremely significant in comparison to serum parameters. CSF parameters had lower concentration of all the studied parameters than the serum, except for the sodium and chloride. The mean value of sodium (150.3±1.70 mmol/l) and chloride (114.5±1.49 mmol/l) in CSF were significantly (P<0.03) higher than their mean values in serum; 154.5±0.92 mmol/l and 131.3±2.5 mmol/l respectively. Gender had no relevant effect on the most CSF measured parameters.

Biography:

Surong Hasi is currently working as a Professor in College of Veterinary Medicine, Inner Mongolia Agricultural University, and is a Director of Camel Protection Association of Inner Mongolia. His research interests are mainly focused on the pharmacokinetic characteristics of veterinary drugs in different species, drug-drug interactions in animals, pharmacological activities of camel milk and Bactrian camel CYP enzymes.

Abstract:

The aim of the research is to study the in vivo activities of Bactrian camels' CYP3A enzymes by investigating the pharmacokinetic characteristics of CYP3A specific probe drug Midazolam in Bactrian camel, and the effect of Itraconazole on the pharmacokinetic behavior of Midazolam was studied simultaneously. Firstly, five healthy adult Bactrian camels were intramuscularly injected with the single dose of 0.1 mg/kg Midazolam, and then blood samples were collected from the jugular vein at different time following the administration. Secondly, after 7 days drug clearance period, these five experimental camels were injected intramuscularly with the single dose of 0.1 mg/kg Itraconazole for 4 consecutive days, and following 2 h of last injection, Bactrian camels were administered intramuscularly with the single dose of 0.1 mg/kg Midazolam again. Blood samples were collected by same route and same intervals as previous, and the plasma was separated by centrifugation. The plasma concentration of Midazolam was determined high performance liquid chromatography-UV detection, and the pharmacokinetic parameters of Midazolam were analyzed by WinNonLin 7.0 with non-compartmental model. The pharmacokinetic parameters of Midazolam in group probe drug only and in group enzyme inhibitor plus probe drug were as follows: the T_1/2 was 2.5±0.073 h and 3.674±0.29 h, T_max was 0.85±0.09 h and 0.54±0.06 h, C_max was 0.62±0.12 µg/mL and 0.80±0.06 µg/mL, AUC_0-t was 1.47±0.35 h·µg/mL and 2.15±0.15 h·µg/mL, V_d was 259.17±41.29 mL/kg and 152.09±22.49 mL/kg, CL was 53.46±14.25 mL/h/kg and 34.3±5.13 mL/h/kg, and MRT was 3.71±0.16 h and 4.60±0.52 h, respectively. Therefore, all the T_1/2, T_max, C_max and MRT of Midazolam in Bactrian camels were relatively low which indicated that Bactrian camels' CYP3A enzyme possess high activity on metabolism of Midazolam. Furthermore, the CYP3A enzyme was significantly inhibited by Itraconazole which can increase the T_1/2, C_max, AUC and MRT, and can reduce the T_max of Midazolam in Bactrian camel

Biography:

Guleng Amu is currently working as a Professor in College of Science, Inner Mongolia Agricultural University. Her research interests are mainly focused on Biomedical Engineering and Biological Physics.

Abstract:

Acetaminophen is a medication used to treat pain and fever, also is a specific probe substrate of CYP1A enzyme. The pharmacokinetic characteristics of Acetaminophen in Bactrian camels were studied in this paper. The experimental Bactrian camels were randomly divided into two groups: group probe drug only and group enzyme inhibitor plus probe drug, respectively. A crossover design was carried out in two experimental periods following 15 days of drug clearance period. Acetaminophen was intramuscularly injected to 6 female camels by 4 mg/kg in group probe drug only, and equal number of female camels was intramuscularly administered by 4 mg/kg of Acetaminophen following 4 consecutive days of intramuscular administration of lomefloxacin by 0.4 mg/kg in group enzyme inhibitor plus probe drug. And then the blood samples were collected at different time intervals after administration of Acetaminophen, and the plasma was separated by centrifugation. The plasma concentration of Acetaminophen was determined by high-performance liquid chromatography (HPLC) after the samples' protein was precipitated by methanol directly, and the pharmacokinetic parameters of Acetaminophen were calculated by WinNonLin 7.0. The pharmacokinetic parameters of Acetaminophen in group probe drug only and in group enzyme inhibitor plus probe drug were as follow: the elimination half-life (T_1/2) was 7.34±0.57 h and 8.98±0.31 h, the time to peak concentration (T_max) was 1.70±0.51 h and 0.833±0.31 h, the maximum plasma concentration of (C_max) was 1.27±0.83 µg/mL and 1.53±0.46 µg/mL, the area under the curve (AUC_0-t) was 7.60±0.45 µg·h/mL and 10.71±0.25 µg·h/mL, the apparent volume of distribution (V_d) was 3787.81±236.37 mL/kg and 2885.98 ±73.11 mL/kg, the clearance (CL) was 359.35±33.49 mL/h/kg and 222.75±8.79 mL/h/kg, and the mean residence time（MRT）was 10.35±0.84 h and 13.04±0.55 h, respectively. Therefore, the Acetaminophen was rapidly absorbed and slowly eliminated by Bactrian camel, and the Bactrian camels' CYP1A enzyme was significantly inhibited by lomefloxacin which can increase the T_1/2, C_max, AUC and MRT of Acetaminophen and reduce the T_max of Acetaminophen in Bactrian camel of China.

Biography:

Abstract:

Computed radiography and computed tomographic scanning of normal stifle joint of camel obtained from cadaver was studied and compared. The radiography revealed only the bony structures involved in the joint whereas CT scan revealed both bony and soft tissues. Lateral, cranio-caudal and caudo-cranial radiographs were taken which revealed all bones and articular surfaces of stifle joint, CT scan studies were done on 3D, transverse and sagittal sections. Medial patellar ligament was not evidenced, however the medial femoropatellar ligament was distinguishable. Other important anatomical structures evidenced were cranial and caudal cruciate ligament, medial collateral ligament, lateral patellar retinaculum, middle patellar ligament, attachments of lateral and medial menisci and most of the associated muscles around stifle joint. CT scan also revealed the bony structures of the joint i.e. femoral trochlea, femoral condyles, tibial condyles, intercondylar tubercles, patella, tibial tuberosity etc. the purpose of this study was to provide a detailed computed tomographic anatomic reference for the dromedary stifle joint.

Biography:

Aristide Maggiolino is a Research Fellow in Animal Science at the Department of Veterinary Medicine of Bari University “A. Moro” (Italy). In the last years his researches focused on equid production, both for milk and meat from horses and donkeys. Several draught horses, such as donkey’s breeds risk extinction are nowadays endangered for the lack of an economically sustainable use. The main aim of his research is the evaluation of catecholamine plasma levels in donkeys because of their involvement as indicator of acute stress, considering innovation that this species is going to live for new techniques and technologies introduced in farm management.

Abstract:

Statement of the Problem: Catecholamines are among the most frequently investigated parameters for studying short-term welfare problems providing information regarding sympathoadrenal activity in response to acute stress conditions. The aim of the study was to evaluate levels of basal plasma catecholamines (adrenaline, noradrenaline and dopamine) in healthy donkeys.

Methodology & Theoretical Orientation: A total of 962 Martina Franca donkeys were used for this study: 684 females and 278 males, aged from 4 months to 24 years. Animals were subdivided into four age categories: under 12 months old, from 13 to 36 months, from 37 to 120 months and over 120 months. Blood samples were collected from jugular vein. At sampling, donkeys’ reaction was evaluated and subdivided into 3 different categories: no reaction; donkeys move the head; and donkeys tend to flee. The dataset of animals with ‘no reactions’ was used to calculate confidence intervals and was subjected to two-way ANOVA considering age and as independent variables to describe basal catecholamines values. The whole dataset was subjected to one-way ANOVA considering reaction as independent variable. Moreover, Pearson’s correlation coefficients between the 3 catecholamines were also evaluated.

Findings: Confidence intervals (CI) for noradrenaline concentration ranged between 239.98 ng/L and 255.07 ng/L, for adrenaline between 129.27 ng/L and 137.90 ng/L, dopamine concentrations between 149.62 ng/L and 160.80 ng/L and noradrenaline/adrenaline ratio between 1.91 and 2.05. Age and reaction to sampling affect catecholamine concentrations.

Conclusion & Significance: Catecholamines are particularly sensible and respond to minimal acute stress. Younger animals were probably more sensitive to contact with humans, considering that older animals are more used to being near man. However, when catecholamines are measured, it is necessary to observe also animal reaction to blood collection because could be itself cause of level alteration.

Biography:

Abdur Rahman is Lecturer in Animal Nutrition section at College of Veterinary and Animal Sciences, Jhang (Sub Campus UVAS, Lahore) and holds DVM Degree, MPhil and PhD in Animal Nutrition. He has more than seven years teaching experience at UVAS, Lahore and CVAS, Jhang. He is actively engaged in teaching and research. He has worked in many research projects at Pakistan and Turkey. He has published more than 15 research papers in international journals. He is providing services to different livestock farmers as well.

Abstract:

Statement of the Problem: To prevent the egg quality losses during storage times, the use of herbal products could be a useful tool in the industry. Much focus has been given on the use of herbs and herbal products to improve performance and to some extent on the quality in freshly laid eggs but limited research data are available for the impact of herbs on the storage quality of eggs. The present study was designed to evaluate the effect of Mentha piperita oil and mentha juice in feed and drinking water respectively, egg quality traits in laying hens at different storage time.

Methodology & Theoretical Orientation: A total of 252 Babcock laying hens were divided into 7 groups and each group was divided into 4 subgroups having 9 hens in each. Group A served as a control. Group A was fed basal diet without any supplementation. Group B, C and D were offered diets supplemented with mentha extract at 50, 100 and 200 mg/kg of feed while groups E, F and G diets were having same doses of mentha juice in drinking water. At the end of the study (56 days), a total of 252 eggs (36 eggs from each group) were collected randomly. 84 eggs were analyzed at zero day of storage while other eggs were stored at 4°C temperatures. Among these eggs, 84 were analyzed after 15 days and remaining 84 after 30 days of storage.

Findings: The results revealed that egg quality traits like egg shell breaking strength (ESBS), yolk color (YC), haugh unit (HU) and egg weight showed non-significant difference (P>0.05) among all the groups at different storage time periods.

Conclusion & Significance: Supplementation of mentha extract and its juice has no impact on egg quality parameters during various storage conditions.

Biography:

Asma Waheed Qureshi is working at Department of Zoology, Abdul Wali Khan University, Mardan, Pakistan. Her experience includes various programs, contributions and participation in different countries for diverse fields of study. Her research interests reflect in her wide range of publications in various national and international journals.

Abstract:

In present study Nigella sativa (seeds) and Fumaria parviflora (aerial), were used to treat buffaloes naturally infected with fasciolosis and their efficacy (%) was compared. In vivo, pre-and post-treatment (50 mg, 100 mg and 150 mg/kg body weight of the two medicinal plants) fecal egg-counts were determined following standard fecal egg count reduction in buffaloes (Babulus sp.) of Nili Ravi breed naturally parasitized with Fasciola sp. The infected buffaloes were randomly divided into 3 main groups i.e., A, B and C. Animals in groups A and B were further sub divided into three sub groups i.e., A1, A2, A3; B1, B2, B3. Sub-groups A1 and B1 were given dose level of 50 mg/kg body weight, A2, and B2, 100 mg while A3 and B3, 150 mg/kg body weight of Nigella sativa (seeds) and Fumaria parviflora (aerial), respectively. The group C having infected animals served as control. The two medicinal plants were found significantly (P<0.05) effective (42-100%) and safe to use against fasciolosis.

Biography:

Gholamreza Ghorbani Kharaji is working at Isfahan University of Technology, Iran. His experience includes various programs, contributions and participation in different countries for diverse fields of study. His research interests reflect in her wide range of publications in various national and international journals.

Abstract:

Lactating dairy cows (n=78, days in milk=103±26.5; body weight=630±76.8 kg) were fed a high-concentrate diet consisting of 35% forage and 65% concentrate. Cows were adapted for 14 d and then were sampled for 10 d. Although all cows were fed the same diet, rumenocentesis pH and minimum reticuloruminal pH values ranged from 5.05 to 6.98 and 4.78 to 6.08, respectively, and the acidosis index ranged from 0 to 28 pH×min/kg of dry matter intake (DMI). Cows were classified according to rumenocentesis pH as tolerant (pH≥ 6.0; n=26), marginal (5.8≤pH<6; n=21), and susceptible (pH<5.8; n=31). Cows were also classified according to reticuloruminal pH as susceptible if duration of an acidotic condition (pH<5.8) exceeded 330 min/d (an average of 920 min/d; n=9) and tolerant if the acidotic condition persisted <330 min/d (an average of 78 min/d; n=5). The classification based on rumenocentesis pH revealed that DMI during first 2 h after morning feeding decreased with increasing SARA (Subacute Ruminal Acidosis) susceptibility. This result was associated with greater daily DMI of tolerant cows which also were of greater body weight. Sorting against long particles and in favor of fine particles during the morning feeding (0 to 6 h) also increased with increasing SARA susceptibility. However, when cows were classified according to reticuloruminal pH, DMI and sorting activity were not significant. Milk fat percentage was greater in tolerant cows, whereas total yield and yield of other components was not significant. The proportions of isovalerate and valerate were correlated with increased SARA susceptibility in cows classified according to rumenocentesis pH. These results indicate that substantial variation exists in SARA susceptibility among high producing, mid-lactation dairy cows, and that cows characterized as tolerant to the high-grain diet exhibit greater DMI and milk fat percentage, but decreased sorting behavior and proportion of isovalerate in the rumen.

Biography:

Belgin Sırıken is an expert in Food Microbiology, Safety and Chemical Properties of Particularly Animal Origin Foods. She has completed her PhD at Ankara University, and now she is working as Prof. Dr. at Ondokuz Mayis University, Samsun, Turkey. Her focus is on Molecular Food Microbiology.

Abstract:

Statement of the Problem: Nowadays, millions of people have died because of the foodborne diseases. Cattle origin meats have also important role in human salmonellosis. The aim of this study was to determine Salmonella spp. in ground beef and raw meatball (cattle origin) samples consumed in Amasya province, Turkey.

Methodology & Theoretical Orientation: In the study, a total 100 samples (50 ground beef and 50 meatball samples) randomly collected from supermarkets and butchers in Amasya province were analyzed. Two enrichment step classic culture technique was applied for the microbiologic isolation. For the isolation, Buffered Peptone Water (BPW) was used for pre-enrichment step and Rappaport Vassiliadis Broth (RV-Broth) was applied for selective enrichment step and Xylose Lysine Tergitol 4 (XLT4 with supplement) was used for the selective agar. For the confirmation of the isolates in molecular levels, single target PCR assay was used. For this purpose, invA and oriC genes were determined in the isolates.

Findings: Salmonella spp. were determined in 6 (6%) of samples. Distribution of 6 samples; 4 (n=50, 8%) of 6 was determined in ground beef samples and 2 (n=50, 4%) of 6 was determined in meatball samples.

Conclusion & Significance: Global incidence of Salmonella spp. infections in humans has shown a significant increase. There have been various Salmonella isolation ratio of ground beef and meatball samples reported from different part of the world, and the results are changing from 0.0% to 26.7%. In the Turkey, Salmonella spp. contamination ratio is changing from 0.0% to 18.0%. Our study results are between these ratios. To our knowledge, there has not been a study on this matter. As a result, ground beef and meatballs were contaminated with the most widespread foodborne bacteria, Salmonella spp. These kinds of samples may be a potential vehicle of transmission of Salmonella spp. to humans. Therefore, it is determined that Salmonella could be threat to public health via consumed ground beef or meatball samples.

Biography:

Abstract:

Statement of the Problem: In this study, the isolation of Lactobacillus spp. and Enterococcus faecium and investigation of usage potential of isolated bacteria as probiotic were aimed.

Methodology & Theoretical Orientation: In a study, 50 chicken intestines were investigated which were taken from a commercial chicken slaughter house in Samsun region. The samples were taken from intestine mucosa and inoculated to the selective mediums for the isolation of Lactobacillus spp. and E. faecium. Suspected colonies were identified by PCR. The isolated bacteria were investigated for bile (0.5-1%) and pH (3-5) resistance by bile and pH tolerance tests, respectively. The hydrophobicity’s of the isolates were tested by using 0.03% Congo Red Agar. The antibiotic resistances of the isolates were determined by Agar Gel Disc Diffusion Test with using 9 antibiotic discs. The Radial Diffusion Method was used for determining the antagonistic effects of the isolates against Escherichia coli.

Findings: Twenty E. faecium, 21 Lactobacillus spp. (9 of them were L. acidophilus) isolated from samples. All the isolates were found as resistant to tested bile and pH conditions. All the isolates were hydrophobic, but none of them had an antagonistic effect against Escherichia coli. Eight of E. faecium isolates were found as resistant to 8 antibiotics. One Lactobacillus spp. (other than L. acidophilus) and 1 L. acidophilus isolates were resistant to 5 and 7 antibiotics, respectively. These isolates were evaluated as multi-antibiotic resistant strains.

Conclusion & Significance: In conclusion, we evaluated that all the isolates were hydrophobic, resistant to bile and low pH conditions; but none of them had an antagonistic effect against Escherichia coli in tested conditions. These results indicated that the multi-antibiotic resistant strains of E. faecium, L. acidophilus and Lactobacillus spp. isolates had a potential of using as a probiotic and further in vivo studies had to be essential for these strains.

Biography:

Abstract:

Statement of the Problem: The investigation of the effects of the administration of Lactobacillus acidophilus to rats fed with 2% cholesterol on the changes in total protein (TP), albumin (Alb), total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), oxidized low-density lipoprotein (Ox-LDL) and high-density lipoprotein (HDL) was aimed.

Methodology & Theoretical Orientation: The animal material comprised of 30 adult-male-Sprague-Dawley-rats. Rats were divided into three groups. Control group (C) was fed with standard rat food for 8 weeks. Hypercholesterolemic group (HC) was fed with a ration comprising of the food with 2% cholesterol for 8 weeks. Hypercholesterolemic and probiotic-administrated group (HCL) was fed with a ration comprising of food with 2% cholesterol for 8 weeks and fed orally with 2x10⁸ cfu/ml/day L.acidophilus probiotic for the last 4 weeks of the trial. At the end of 8 weeks, the serum levels of TP, Alb, TC, TG, LDL and HDL were detected by auto-analyzer. The level of Ox-LDL was measured by ELISA in brain supernatants.

Findings: The slightly increase of TP level (p>0.05) in HC group was determined compared to control group. In HCL group, this level slightly decreased (p>0.05). The changes in albumin level among the groups were not significant (p>0.05). It was detected that TC level in HC group increased (p<0.05); but in HCL group, it decreased and approached to the level of control group (p<0.05). The TG level in HC group increased compared to control group (p<0.05); but in HCL group, it decreased compared to HC group (p<0.05). It was determined that the level of HDL in HC group decreased compared to control group (p<0.05), but increased in HCL group (p>0.05). The changes in Ox-LDL level in HC group was determined as increased compared to control group (p<0.05); but in HCL group, it decreased compared to HC group (p<0.05).

Conclusion & Significance: It was thought that the administration of Lactobacillus acidophilus as a probiotic might be useful in reducing the cholesterol level.

Biography:

Hussein. A. Abd El-aal working at Department of Food Science, Minia University, Minia, Egypt. His experience includes various programs, contributions and participation in different events for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.  

Abstract:

Pepper is an important crop in the world, because of its economic importance, also due to the nutritional and medicinal value of its fruits and spices used as food flavorings. Their fruits are considered a good source of antioxidant and biologically active compounds, such as carotenoids, flavonoids, vitamins, capsaicinoids and mineral elements.

The objective of this study was to evaluate the antioxidant activity and vitamin C content of different color pepper fruits.  Three sweet pepper cultivars (Capsicum annuum L.), blocky Red, blocky Yellow and green, and one hot (Capsicum frutescens) tabasco green and ripped tabasco (red).  Proximate composition, total soluble solids, vitamin C content and color (L, a, b) were determined.  Ether extracts were studied to evaluate total phenolic compounds, total flavonoids, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Tabasco red had the highest value of total solids (10.83%), protein (2.16%), ash (1.06%) and vitamin C (215.05mg/100g) fresh matter.  Total phenolic contents ranged from 214.3to 489.7mg as garlic acid/100g, and total  flavonoids ranged from 186.1  to 467.2 mg as quarestin/100g fresh sample, respectively. Ripped tabasco red had significantly (P≤ 0.05) higher value of DPPH scavenging activity (78.65 %) than other samples. Green had lower content of total phenolic compounds , vitamin C, and antioxidant activity than yellow and red samples.  

These results illustrate that red pepper had high content of total phenolic, flavonoids and vitamin C and exhibit strong antioxidant activity.    

Biography:

Alessandro Di Cerbo has obtained his Bachelor’s degree in Medical and Pharmaceutical Biotechnologies at University Vita-salute San Raffaele (Milan, Italy) in 2005. In 2007 he has achieved his Master’s degree in Medical Biotechnology at University of Modena (Italy), in 2011 he has obtained the title of PhD in Nanoscience and Nanotechnology at the same University and in 2016 he got the Specialization in Clinical Biochemistry at University “G. d’Annunzio” of Chieti (Italy). His scientific activities are highly interdisciplinary, ranging from nanotechnology to nanomedicine, microbiology, nutrition and translational medicine. He has published more than 50 papers in reputed journals.

Abstract:

Food allergies and food intolerances are clinically difficult to discriminate and are classified as adverse food reactions, whose causes are numerous. We evaluated the effect of a nutraceutical diet in relieving evident clinical symptoms related to cutaneous adverse food reactions such as drooling, back and neck intense itching, neck eczema, chronic conjunctivitis and stomatitis and skin lesions in 18 indoor-housed clients-owned cats. Cytological evaluations of ear, skin and gingival swabs revealed an increased turnover of keratinocytes while the oxytetracycline ELISA determination showed an unexpected high amount of oxytetracycline in all cats at the first visit. All cats were then randomly assigned to receive a standard (SD group) or a nutraceutical diet (ND group) for 60 days. We observed a significant reduction of the mean serum concentration of oxytetracycline, pruritus intensity and skin lesion severity (p<0.01, p <0.001, and p<0.001, respectively) in the ND group as well as a significant improvement in the clinical picture. Although a direct correlation between oxytetracycline presence within cat sera and CAFR-related symptoms has never been described, this study highlights the benefit of a specific nutraceutical diet supplementation in improving clinical symptoms and skin lesions in cats with CARF.

Biography:

Abstract:

In this study, tendinopathies and other soft tissue injuries were assessed in horses, in Lahore, Pakistan. Forty-eight horses were randomly selected and divided into three equal groups (n=16) i.e. Group A (Race), Group B (Polo) and Group C (Draft-purpose); each group was further subdivided into two equal sub-groups i.e. sound (n=8) and unsound (n=8). Soft tissue structures from various joints including elbow, carpus, fetlock, stifle, hock, and flexor tendons were scanned based on various sonographic parameters, including echogenicity, fiber alignment, severity of injury, soundness via ultrasound and the degree of prognosis, respectively. The results showed that echogenicity of lesions was highly significant (P≤0.014) in Groups A and B. Fiber alignment was also highly significant for Group B (Polo), followed by Group A (Race Horses), and significant for Group C (Draft-purpose). Severity of injury was highly significant (P≤0.003) for both Groups A and C. Soundness via ultrasound was highly significant for all three groups (P≤0.002 for Race and Polo, and (P≤0.012 for Draft Purpose); whereas status of prognosis was highly significant for Group A (P≤0.001), followed by Groups B and C (P≤0.006). Conclusively, injuries were more pronounced in Race horses, followed by Polo horses and subsequently Draft purpose horses.

A race horse with bilateral fetlock joint swelling in fore legs. The horse was suffering from chronic pain and marked lameness, with a decreased range of motion of both fetlock joints. The sonogram reveals an abnormal, coarse hyperechoic CDE with black areas marking fiber disruption and filling in by granulation tissue and debris. A race horse at Sardar Noor Hussain Farm (Raiwind), suffering from an acute non-painful swelling on the craniodorsal aspect of the right stifle joint. The sonogram reveals the presence of an organized hematoma with a large, soft cavity filled with an echogenic fluid separated into geometric cavities by thin, echogenic trabeculae.

Biography:

Abstract:

In Pakistan, sonographic findings regarding liver problems in pets have not yet been reported. This study characterized the various stages of liver disease for precise prognosis and treatment prospects in dogs in Lahore city. Liver size was sonographically ascertained and correlated with laboratory findings in 21 client-owned dogs divided into three groups (A, B, C), n=7. Group A comprised of healthy dogs which served as control. Group B comprised of dogs scanned for acute hepatitis, while Group C comprised of dogs suffering from chronic hepatitis. Based on presenting clinical findings, liver was scanned in B-mode using 5.0 MHz transducer. Hematology and biochemistry profile were assessed and body surface area calculated. Hypoproteinemia and bleeding diathesis supervened in hepatitis-afflicted dogs (Groups B and C), manifested by a significant decrease in albumin and platelet counts (P<0.00), and prolonged prothrombin time (PT) and APTT (P<0.00). ALT showed a significant increase (P<0.028) while increase in bilirubin was highly significant (P<0.00) for both Groups B and C. Correlation analysis between sonographically determined liver size and body surface area (BSA) revealed a strong correlation (0.9) between liver size and BSA in the acute hepatitis group, moderate (0.6) correlation in the Control Group; while, the Chronic Hepatitis Group C depicted a negative correlation (-0.5). Conclusively, sonographic liver size correlated well with BSA and with laboratory findings and the clinical picture. Conclusively, sonographic findings correlate well with clinical picture in acute and chronic hepatitis in dogs.

Biography:

Riaz Shah has completed his Graduation in Veterinary Sciences and Master’s/PhD degree in Animal Biotechnology with specialization in Livestock Reproductive Biotechnology and Post-doctoral experience from AgResearch Ltd., Ruakura Research Centre, Hamilton, Newzealand in area of goat cloning. His current areas of research are the Application of Advanced Reproductive Technologies like IVF, SCNT and Stem Cell Production for augmenting production of Livestock. The recent successes in cloning of livestock (Buffalo and Pashmina goat) first time in India through Handmade Cloning technique during his research endeavors at National Dairy Research Institute, Haryana and SKUAST-K, Kashmir, India, has opened opportunities to achieve excellence in the application of such techniques for transgenic animal production and stem cell research. He is currently working as a Professor and Head, Division of Animal Biotechnology at Faculty of Veterinary Sciences in SK Agricultural University, Srinagar, J&K, India.

Abstract:

Statement of the Problem: Pashmina (cashmere) goat is among the few species of livestock which can survive in their inhabitant harsh cold arid climate. This goat produces world’s finest fibre. Due to high risk of genetic loss via inbreeding, reproductive technologies need to be implemented. Cloning is the fastest way to multiply the limited superior germplasm. Handmade cloning (HMC) is an alternative method of cloning which eliminates the use of costly sophisticated micromanipulator tools demanding greater degree of skill, in comparison to SCNT. The present study was designed to optimize in vitro continuous culture system for development of zona free handmade cloned pashmina goat blastocysts and they’re in-vivo development after laparoscope aided intra uterine transfer into synchronized recipients.

Methodology & Theoretical Orientation: Skin derived fibroblasts at 5^th to 9^th passage was used as nucleus donor cells for HMC experiments. Cumulus–oocyte complexes (COCs) were in vitro matured and stripped of their cumulus investment and zona pellucida. Protrusion cone-guided bisection was performed for enucleation.

Findings: The cleavage and blastocyst rates were determined at day 7 of embryo culture. G1. G2 medium gave the best cleavage percentage (86.84±2.26) while as RVCL (commercial medium from Cook^TM, Australia) gave the best blastocyst percentage (15.01±4.58). WID (well in drop) culture system was found to be most efficient with highest cleavage and blastocyst percentages i.e., 84.34±4.15 and 21.65±1.69 respectively. On day 45 post embryo transfer into 19 receipients, 3 pregnancies were detected out of which only one carried to term.

Conclusion & Significance: Using cost effective HMC technique, we successfully report the live birth of first handmade cloned cashmere goat. The birth weight of the cloned kid was 2.4 kg, like female kids from naturally bred Pashmina goats during the same period. No significant differences in growth rate between cloned goat and naturally bred goats (1.2 kg/month) were observed till it reached sexual maturity. Also, estrous cycle of the cloned goat was observed to be normal and bred normally. In our study, the embryos were cultured in a serum free media which could explain the normal birth weight of the cloned kid. Microsatellite analysis confirmed that the cloned kid was genetically identical to the fibroblast cell donor dam. In conclusion, this study elucidated the production of hand-made cloned blastocysts using a continuous culture system and birth of healthy cloned kid from Pashmina goat.

Biography:

Abdur Rahman is Assistant professor in animal Nutrition section at College of Veterinary and Animal Sciences, Jhang (Sub Campus UVAS, Lahore) and holds DVM Degree, M.Phil and PhD in Animal Nutrition. He has more than seven years teaching experience at University of Veterinary and Animal Sciences, Lahore and CVAS, Jhang. He is actively engaged in teaching and research. He has worked in many research projects at Pakistan and Turkey. He has published more than 20 research papers in international journals. He has attended many international and national scientific conferences and he is providing services to different livestock farmers as well

Abstract:

Statement of the Problem: Egg quality losses during storage period from laying to transportation pose hazardous damage to farm economics. Sustainability and maintenance of internal egg quality can be achieved by alternative therapy like herbal and plants products. Current study was designed to investigate the effects of dietary supplementation of Pistacia terebinthus seed meal in laying hens on egg quality traits during different storage time. Methodology & Theoretical Orientation: For this purpose a total of 192 Babcock white laying hens were divided into 6 groups with 4 subgroups in each and fed diets containing 0, 10, 20, 30, 40 and 50 g kg-1 terebinth (Pistacia terebinthus) to each group accordingly. At the end of experiment, 48 eggs were collected from each group at 2 consecutive days. From the first day collection, 12 eggs from each group was analysed on the same day without storage. Remaining eggs were stored at +4°C. Then after 10, 20 and 30 days of storage, 12 eggs from each group were analyzed for internal egg quality parameters. Findings: Results revealed that terebinthus has shown significantly positive effect (P<0.05) on haugh unit values at 20 g kg-1 and 40 g kg-1 inclusion rate at days 30 of storage. Similarly, yolk color index value was also increased (P<0.05) at 3% inclusion level of terebinthus at days 20 of storage while egg shell breaking strength and egg weight remained unaffected (P>0.05) at various dose levels during different storage duration.  Conclusion & Significance: It is concluded that terebinthus seed meal could be used to extend the storage time of eggs without adverse effects on quality of eggs.

Biography:

Alper Ciftci is an expert about Molecular Microbiology and Vaccine Development. He finished his PhD at Ankara University, and now he works as Associate Professor at Ondokuz Mayis University, Samsun, Turkey. He focuses on working development and validation of commercial products such as vaccine and diagnostic kits.

Abstract:

Statement of the Problem: In this study, the investigation of the levels of some biochemical parameters in the blood of sheep due to the vaccination for the prevention of brucellosis was aimed.

Methodology & Theoretical Orientation: 35 months-old and female sheep were used as an animal material. The vaccine was given in single dose against Brucella melitensis and every month blood was drawned from Vena jugularis for 4 months. The vaccine, vaccinated 1 dose via conjunctiva, contains Brucella melitensis Rev1 (0.5-2×10⁹ cfu). Some biochemical parameters in sheep such as ALP, ALT, AST, LDH, glucose, total protein and albumin measured using autoanalyzer.

Findings: In consequence of evaluation of the Rose Bengal Plate agglutination tests, the sera belonging to bloods taken before vaccination were negative and 120 serums belonging to experimental group showed a positive reaction. The SAT results of the serums which were evaluated after RBPT showed that antibody titers (P<0.001) increased significantly at first month according to the pre-vaccine, but it was determined that while from the second month began to fall. It was determined that there was no statistically significant changes in glucose, AST, ALT activity after vaccination (P>0.005), a significant amount of total protein and ALP decreased after vaccination (P<0.005), while the LDH levels were significantly increased (P<0.005).

Conclusion & Significance: It was determined that in first month of increased antibody of conjunctival Brucella vaccine which was carried out in low doses to protect against Brucellosis affected the amount of LDH, ALP, total protein and albumin in subsequent months, it was approaching the level of the pre-vaccine group and conjunctival Brucella vaccine was considered to be used as a safe house to protect them from Brucellosis.

Biography:

Christelle Navarro is the Medical Director – Companion Animals - for Virbac. She manages a team of veterinarians involved in phase IV trials and collaboration with Universities and specialists across the World. She graduated from Alfort Veterinary School and is certified from Veterinary Dermatology and Pain management.

Abstract:

Objectives: The objective of this study was to assess the efficacy and the onset of activity of two ear preparations containing two different corticosteroids: hydrocortisone aceponate (Easotic, Virbac, France) or betamethasone valerate (Otomax, MSD, France) in dogs with OE.

Materials & Method: 120 privately owned dogs with nonparasitic OE were selected from two different Chinese veterinary hospitals. Dogs were divided into 2 groups (60 dogs each) treated with Easotic (1ml pump, once daily for 5 days) or Otomax (5-8 drops twice daily for 7 days). Clinical efficacy was assessed every day up to Day 7, via a bacterial and fungal cytology score (0 to 5), the presence of an auricular discharge, the number of ear scratches per ten minutes and a medical efficacy score (-1 to +3). Scores between groups in each site were analysed by one-way ANOVA.

Results: Altogether, 73% of dogs had a mixed bacterial and Malassezia OE, 21% had only bacteria and 6% Malassezia yeast. On Day 7, bacteria and yeasts were absent in both treatment groups. A significantly faster decrease in bacterial score (from D2 or D5, in site 1 or 2 respectively) and fungal score (from D2 or D1) was seen with Easotic than with Otomax (p < 0.05). Auricular discharge and pruritus disappeared before Day 7 in both groups with a quicker improvement for Easotic than with Otomax (as from D1 for pruritus, p<0.05) (Fig. 1). Both medications led to a complete OE recovery (medical score = 3). There were no adverse reaction observed with both products.

Conclusion: In this comparative field study, both ear treatments were effective to cure nonparasitic otitis externa in dogs. Dogs treated with Easotic had faster improvement of clinical signs than dogs treated with Otomax, and both products were equally effective on day 7.

Biography:

Francesco Sgarangella is Professor of Public health, Faculty of Veterinary Medicine of Sassari, Director of Prevention Service of ASSL of Sassari and regional manager about African swine fever in both domestic pig in wild boar. He is also component of the Project Unit for the eradication of African swine fever in Sardinia. This regional organization began a series of measures to eradicate the disease that afflicts the pig sector of Sardinia. Among the various measures put in place the depopulation of feral pigs activities and the identification of the surveillance and protection zones.

Abstract:

Statement of the problem:

The 2017 bovine tubercolosis Control Program (CP) for the Province of Sassari aims to research and remove residual sources of TB infection to ensure the achievement of the Officially tuberculosis-free qualification (Leg.Dec.no.196/1999). Active surveillance is based on pre-movement tuberculin test (IDT) to monitor all bovine transport, passive surveillance rely on post-mortem inspection at the slaughterhouses and, in addition, ASSL of Sassari performs a thorough check of all animals involved in road accidents. The purpose of this work is to describe a case of tuberculosis in a calf urgently slaughtered after a road accident in an area named Goceano.

Methodology & Theoretical Orientation:

In the event of road accidents, where emergency slaughter is needed, ASSL intervenes with official veterinary surgeons of animal health and food hygiene services. At the abattoir, in case of suspect of tuberculosis, after the histological exam, the lymph nodes are subjected to colony examination.

Findings:

An 8-month calf, urgently slaughtered after a car accident, at the anatomopathological exam showed periportal lymphadenopathy with chronic granulomatous lymphadenitis, the consequent lab test confirmed a positivity for M.Bovis. Out of 29 animals tested in the herd of origin, only the calf's mother showed positivity to IDT, while at the anatomopathologic exam resulted negative. Finally, the histological and cultural examination on tonsillary lymph-nodes was evaluated also negative.

Conclusion:

The evidence shows that tuberculosis is a problematic disease that requires surveillance activity beyond the actions ordinarily provided by control plans. In fact, given that no routine herd control was due on short-term, considered that the calf was not destined to slaughter in a short time, and that the mother sent to abattoir did not show suspicious lesions, failure to detect the presence of infection in cattle constitutes an important risk factor for spreading the disease.

Biography:

Francesco Sgarangella is Professor of Public health, Faculty of Veterinary Medicine of Sassari, Director of Prevention Service of ASSL of Sassari and regional manager about African swine fever in both domestic pig in wild boar. He is also component of the Project Unit for the eradication of African swine fever in Sardinia. This regional organization began a series of measures to eradicate the disease that afflicts the pig sector of Sardinia. Among the various measures put in place the depopulation of feral pigs activities and the identification of the surveillance and protection zones

Abstract:

Statement of the Problem:

The goal of the 2015-2017 Extraordinary Program (EP), approved by the European Commission, had the objective of eradicating African swine fever from the regional territory. According to risk maps elaborated by the Regional Epidemiological Observatory, in 2015, the EP presupposed an official check on pig farms regarding: registrations, biosecurity and animal welfare measures. The aim of this work is to illustrate the results of non-compliance evidenced during official controls under EP, classify their typology, connect them with different territorial risks in order to define intervention priorities and to reduce the risk of spread of ASF virus .

Methodology & Theoretical Orientation:

Data were entered weekly on the Animal Health Service information system (E-VET), a computerized database based on pig farm registry. The use of farm codes and geographic coordinates has allowed us to categorize non-conformities and contextualize them across the regional territory through multi-stratum maps (GIS technology).

Findings:

Out of 2,950 controlled farms, 1864 non-conformities were detected; 44.6% were about biosecurity, 26% regarding pigs registration, the remaining part of non-compliance concerned reproductive parameters.

Conclusion & Significance:

The described model, based on the employment of a local software implemented with the non-compliance data ascertained during official monitoring activities and the subsequent data mapping on thematic maps (GIS), highlighted the major criticalities on pig farm registry management and biosecurity measures, particularly in wild pigs ASF infected areas. By associating the different types of non-conformities to the territorial context, which is diverse from an epidemiological point of view, it was possible to model the timing of official controls during the 2016 activity, prioritizing official controls to high risk farms.

Biography:

Christelle Fontaine is a Medical Manager, Companion Animals, Virbac. She is involved in phase IV trials and collaboration with universities and specialists across the world. She graduated from the French Veterinary School of Maison Alfort, in Paris in 2007. 

Abstract:

Introduction: In order to benefit from the widest protection while keeping on a limited number and volume of vaccines administered, mixing of vaccines from different laboratories are commonly performed by veterinarians. The study aim was to confirm the safety and immunogenicity of the vaccines association Canigen^TM DHPPi (Virbac) and Nobivac^TM L4 (MSD) in dogs.

Methodology: Fifteen SPF male and female Beagle puppies of 9-10 weeks of age on Day 0 participated in this controlled randomized trial. Puppies, received two subcutaneous injections twenty-one days apart for the primary vaccination course. In group 1 (n=5), puppies received one dose of Canigen^TM DHPPi vaccine rehydrated with one dose of Nobivac^TM L4 vaccine. Puppies belonging to group 2 (n=5) and group 3 (n=5) served as controlled and were vaccinated with respectively Canigen^TM DHPPi/L and Nobivac^TM DHPPi (MSD) reconstituted with Nobivac^TM L4. Puppies were followed for forty-nine days for both, the appearance of any adverse reaction and seroconversion, assessed by either seroneutralisation assays or micro-agglutination tests. Statistical analysis were performed using Unistat 6.0 software. P<0.05 was considered as significant.

Results: On Day 0, none of the puppies (n=0/15) had any antibody against the vaccines strains. After vaccination, no significant difference could be demonstrated between the groups regarding the antibody titres against the core vaccinal strains of Canigen^TM vaccine and the Leptospira serovars of Nobivac^TM vaccine at the times of onset of immunity (Mann-Whitney U test, p>0.05). No general health parameter alteration or local injection site side effect was observed.

Conclusion: Canigen^TM DHPPi and Nobivac^TM L4 vaccines can be mixed and administered simultaneously to dogs without any major negative impact on the safety and immunogenicity of each respective vaccine.

Biography:

Christelle Fontaine is a Medical Manager, Companion Animals, Virbac. She is involved in phase IV trials and collaboration with universities and specialists across the world. She graduated from the French Veterinary School of Maison Alfort, in Paris in 2007. 

Abstract:

Introduction: Bovine serum albumin (BSA), one component of culture media frequently used for vaccines manufacturing can be one of the allergens responsible for immediate-type allergic reactions post vaccination in dogs. Allergic reactions are the major pharmacovigilance reported signs in dogs given Leptospira vaccines. The study aim was to compare the quantity of BSA in various dog Leptospira vaccines.  

Methodology: Four commercial vaccines from different manufacturers were used. The vaccines tested contained two (Canigen^TM L, Virbac), three (Vaccine B) or four (Vaccine D and E) serovars of Leptospira. The concentration in BSA in each vaccine was determined by ELISA assay (Bovine Serum Albumin Assay, Cygnus Technologies). Statistical tests were performed using the non-parametric Dunn's test, Jmp v10 software (SAS Institute). P<0.05 was considered as significant.

Results: The lowest concentration in BSA was observed in Canigen^TM L vaccine, with values at the low detection limit of the method used, ie 1 µg/mL. Vaccine B had a concentration in BSA above the detection threshold, i.e., 64 µg/mL. For Vaccines C and D, mean concentrations in BSA were respectively 21.9±7.6 µg/mL and 6.0±0.2 µg/mL. Significant differences could be observed between Canigen^TM L vaccine and Vaccine B (p=0.01) and between Canigen^TM L vaccine and Vaccine C (p=0.04).

Conclusion: The commercial vaccine containing two serovars of Leptospira, Canigen^TM L, was the one with the lowest concentration in BSA, which is in line with the highest safety reported from the field. Interestingly, the concentration in BSA was not strictly correlated to the number of Leptospira serovars included in each of the vaccines, which could be explained by special filtration steps applied during the vaccines manufacturing processes. In conclusion, the vaccine containing two serovars of Leptospira, Canigen^TM L remains globally purer than vaccines containing additional serovars of Leptospira, which might explain its highest safety.

Biography:

Shigeru Sato earned MS degree in Pathology of Bovine Leukemia from Iwate University in 1980, thereafter he worked at the Miyagi Prefectural Federation of Agricultural Mutual Aid Association from 1980-2006. He earned PhD in ruminal bacteria and immune response of calves from Tohoku University in 1991, and also in immunity of periparturient cows from Kitasato University in 1998. Now he is a Faculty Member of Iwate University, and teaches a course on Food Animal Internal Medicine and Bovine Clinics.

Abstract:

Objectives: Japanese black beef cattle produces high quality meats, whereas clinical or subclinical ruminal acidosis (SARA) have been frequently occurred due to feeding a high-grain diet with small amount of forage. However, the characterization of reticulo-rumen fluid is unknown in Japanese black cattle for fattening. Therefore, the objective of this study is to characterize the rumen and reticulum fluid, especially pH and VFA, in Japanese black beef cattle.

Materials & Methods: Japanese black beef cattle equipped with fistula in the middle (n=3) and later (n=3) fattening stage were used. Cattle were fed much concentrates and hay twice a day by the routine manner, respectively. Ruminal and reticular pH measured continuously every 10 min using a radio transmission system (YCOW-S; DKK-TOA Yamagata, Japan). Rumen and reticulum fluids for VFA, NH₃-N, and lactic acid analysis were collected in the 4 hours after morning feeding. The 24-h mean pH, and VFA, NH₃-N, and lactic acid concentrations were analyzed using two-way repeated measures ANOVA with Bonferroni’s multiple-comparison post-test, and Pearson’s correlation coefficients were calculated between pH and other parameters in the rumen and reticulum.

Results: The 24-h mean ruminal and reticular pH were low in each of the middle (5.5-6.6 and 5.1-6.2) and later stage (5.3-6.1 and 5.5-6.1), and the ruminal pH was tended to be lower than the reticular pH. Furthermore, the ruminal and reticular pH values of the later stage showed a tendency to be lower than the middle stage. No significant difference was observed in the ruminal and reticular VFA, NH₃-N, and lactic acid concentrations. Although high variations among the individual cattle were observed, a positive correlation was identified between the ruminal and reticular pH. However, 24-h mean pH was not significantly correlated with VFA, NH₃-N, and lactic acid in the rumen and reticulum.

Conclusion: Low ruminal and reticular pH were identified in the middle and later fattening stage of Japanese black beef cattle, and the ruminal pH was slightly lower than reticular pH. Further research is required to reveal the pathophysiological evaluation of SARA in Japanese black beef cattle. 

Biography:

Aline Camargos has her expertise in reproduction of domestic animals. Her most recent projects aim to develop new technologies for animal reproduction. Her research group has developed a software to analyze bovine sperm, a Computer-Assisted Sperm Analysis named SlimCASA. Financial support by CNPq and IF Goiano.

Abstract:

Statement of the Problem: The objective was to test three methods of low cost image capture, in order to make possible the future use of the software in development. Methodology & Theoretical Orientation: Twenty frozen semen samples were used to make the images. One drop of 10μL of semen was added on a slide, and this was carried under a microscope attached to a microscope camera and with a digital camera and a camera phone positioned in the eyepieces of the microscope by means of a holder. A 20-second videos were made. From them, pictures were taken, which the technician used to do the sperm count to compare the range and accuracy of each image capture device. The results were submitted to analysis of variance and Tukey's test, at 5% of significance. Findings: The mean number of spermatozoa counted in the images differed, being 145.10 ± 98.72 for the microscope camera, 17.90 ± 11.25 for the digital camera and 407.80 ± 299.42 for the cellular camera (p <0.05). Conclusion & Significance: It was observed a great variation of the performance of the different types of image capture, being that the camera of the cell obtained the best performance.

Biography:

Aline Camargos has her expertise in reproduction of domestic animals. Her most recent projects aim to develop new technologies for animal reproduction. Her research group has developed a software to analyze animal sperm, a Computer-Assisted Sperm Analysis named SlimCASA. Financial support by CNPq, FAPEG and IF Goiano.

Abstract:

Statement of the Problem: This study aimed to develop a software for bovine sperm evaluation, from images taken under a microscopy. This program should be compatible with notebooks and PCs for home use, easy to use, in the Portuguese language and free. It is intended, with this software, standardize the sperm motility analysis carried out by veterinarians during andrological examinations at field, cheapening the cost of acquisition of specific equipment. Methodology & Theoretical Orientation: Seventy two images of bovine semen were made by microscopy from thawed semen doses. The 0.5 mL reed were thawed for 30 seconds in a water bath at 37 ° C. Semen drops were analyzed by CASA (Hamilton Thorne Research) and deposited on slides with cover slip for microscopy. The images were obtained from phase contrast microscope (Jenamed2) with 1.3 MP camera attached (Coleman). The software was developed from resources already available in an open source Java solution called ImageJ. Initially, the video images were converted into frames and subjected to some treatments, using only 8-bit color and segmenting grayscale so that the software could do the analysis of the image particles. The 72 semen samples were analyzed by CASA and by the software SlimCASA. For statistical analysis, the results of the counts were subjected to analysis of variance (SAS, 2012) and Tukey’s test, at a significance level of 5%. Findings: The average values of the sperm motility did not differ and were 64.69 ± 19.34 and 64.47 ± 9.15 by CASA and by the software SlimCASA, respectively (p> 0.05). Conclusion & Significance: In conclusion, the developed software showed the same efficiency as the motility evaluation carried out by Hamilton Thorne CASA.

Biography:

Mahmoud Khajeh is working at Paytakht Pet Hospital, Iran, Tehran. His experience includes participation in various programs, contributions and participation in different events for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.

Abstract:

A female Rottweiler 9-years old dog with anorexia, polydipsia, polyuria, vaginal discharge was referred to Paytakht Pet Hospital. After a precise clinical evaluation, the case was referred to perform abdominal radiography and sonography. A well circumscribed but non-encapsulated mass with 16×12 cm in size was detected in the cervix of the uterine. The mass had hemorrhagic and necrotic feature. The surgical procedure (OHE) was done and a mass was observed in the cervix. The mass was sent to pathology. Paraffin-embedded tissue sample was prepared and stained with H & E. The sections were surveyed by clinical pathologist and fibrosarcoma was diagnosed. Highly cellular fibroblastic proliferation in herringbone pattern was observed and cells had scant cytoplasm, tapering elongated dark nuclei with increased granular chromatin, variable nucleoli. Mitotic activity presents with normal forms. In our acknowledgement, this is first report of fibrosarcoma occurring in the cervix.

Biography:

Mohammadreza Ferdows is working as a Veterinary Specialist at Paytakht Pet Hospital, Iran. His experience includes various programs, contributions and participation in different events for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.

Abstract:

Genital system neoplasms are rare in dogs, although tumors of the testis in males are the most frequent. Hemangiomas are common in dogs, but rare in other domestic animals. A Great Dane dog with testicular mass was referred to Paytakht Pet hospital. All the scrotal masses were evaluated with ultrasonography. Serum biochemistries, CBC, urinalysis (UA), and thoracic radiographs and abdominal ultrasound were performed to rule out lung and visceral metastases. Cytology sample was prepared and stained with Giemsa and hemangioma was diagnosed. Surgical recommendation included orchiectomy with intrascrotal ablation although according to owners’ preference just surgical excision of the masses was supposed to perform. The surgical samples of the intrascrotal tissue were collected and fixed in buffered formalin solution, embedded in paraffin, and sectioned at 3 µm. The sections were stained routinely with hematoxylin and eosin (H & E). Each section was evaluated by two experienced clinical pathologists and hemangioma was diagnosed according to published criteria. After surgical excision of the tumor, the dog was followed-up. The dog was clinically evaluated every 5-6 months during a 1-year follow-up to detect the presence of local recurrences and/or distant metastases. Tumor recurrence was observed after 8 months and the case died after 12 months. The testicles are the second most common sites for tumors and hence castration has been recommended for prevention. The breeding dogs can be involved by a variety of the testicular and scrotal tumors, so increase of acknowledgement of the case owners in Iran is important issue. Although, orchiectomy with intrascrotal ablation is necessary for dogs with intrascrotal hemangioma.

Biography:

Ms Sima Nikkhah working as a student at Islamic Azd University, Research and science Branch, Veterinary Radiology Departement, IRAN. Her international experience includes various programs, contributions and participation in different countries for diverse fields of study. Her research interests reflect in his wide range of publications in various national and international journals.  

Abstract:

 Anecdotal evidence has suggested that guinea pigs have a high prevalence of ocular lesions. Hetotrophic bone formation (also known as osseous choristoma) is a condition unique to the guinea pig that result in new bone formation in the ciliary body that can be visible as a white, crystalline opacity at the limbus¹.

  A 13 months old Guinea pig refer to exotic and ophtalmology departments of Paytakht Pet Hospital because of ocular lesion. History taking shows the owner had information about role of  low level of  Vitamin C in diet. Depends on this thinking he use over taking of Vitamin C (food material and supplement) for himself Ginuea pig diet order.

  A full ophthalmic examination was performed direct and indirect ophthalmoscopy and with slit lamp biomicroscopy, intraocular pressure using the ICARE Tonovet tonometer after topical anesthesia (no evidence of glaucoma) without ocular discomfort, show white lesion at limbus protruding slightly into the cornea because of heterotrophic bone formation at  cilliary body in Guinea pig 

Treament: No treatment is recommended. role of ascorbic acid metabolism by guinea pig in disease pathogenesis of the ciliary body also remains unknown¹, But because of Vit C promotes calcium deposition when at high concentration in tissue²  recommended to owner for diet modification.

Biography:

Dr. Mahan Bitaraf working at Paytakht Pet Hospital,Iran. His international experience includes various programs, contributions and participation in different countries for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.  

Abstract:

This study was performed to evaluate adding of steroidal or non-steroidal anti-inflammatory agents or topical medroxyprogesterone acetate to tetracycline and Vit C regime for management of ocular alkali burns.

Alkali wounds were inflicted on the central cornea of guinea pigs by applying a round filter paper, 12mm in diameter, soaked in 1M NaOH for 30 sec. Only one eye in each guinea pig was used. A total of fifty guinea pigs were divided into five groups of ten animals. Ten guinea pigs after the alkali burn were designated as the control group (without treatment). In other groups, topical tetracycline and systemic Vitamin C were used. Besides them, in group A, topical medroxyprogesterone acetate 1%, in group B, topical prednisolone acetate 1% and in group C, topical diclofenac was used. In group D, corneal ulcers were treated just with tetracycline and Vitamin C. Clinical outcome was monitored daily by corneal opacity, duration of blepharospasm, corneal vascularization and duration of ocular discharge. After three weeks, corneas were excised for histopathologic analysis. Samples were monitored by evaluating of corneal thickness (μ), numbers of epithelial rows, keratocyte density, stromal vascularization, stromal inflammation and stromal collagen arrangement.

Comparative evaluation of groups showed that groups A, B and C had significantly lower discharge days and groups B and C had significantly shorter duration of blepharospasm than control group.

In microscopic evaluation of corneas, group B and C had significant lower degree of corneal vascularization and group B had significant lower degree of corneal inflammation than control group.

In conclusion, the regimen of topical prednisolone 1% combined with vitamin C and tetracycline may have therapeutically valuable in early (first three weeks) treatment of alkali burn cornea.

Biography:

Belgin Siriken is an expert about food microbiology, safety and chemical properties of particularly animal origin foods. She finished her PhD at Ankara University, and now she works as Prof. Dr. at Ondokuz Mayis University, Samsun, Turkey. She focuses on working molecular food microbiology.

Abstract:

Statement of the Problem: Ground beef is a raw food of animal origin that constitutes a signifcant portion of the Turkish diet. It is a good medium for the rapid growth of microorganisms. The bacteria normally found on the surface of meat are distributed throughout the entire product during the mincing and mixing process used to produce raw ground meat. It has been known as a vehicle for transmission of organisms such as E. coli, E. coli O157 or H7 and L. monocytogenes.  Methodology & Theoretical Orientation: In the study, a total 70 ground beef samples were tested for contamination by L. monocytogenes  and E. coli O157:H7. Isolation of E.coli O157:H7 was carried out FDA (2001). Fr this Tryptone Soya Broth with novobiocin and TC-SMAC (Tellurite Cefixime Sorbitol MacConkey) agar plates were used enrichment step and specific media, respectively. Isolation of. L. monocytogenes and other Listeria spp. Listeria Enrichment Broth and Fraser Broth were used pre-enrchment and enrichment step, respectively. For the isolation, Listeria selective agar was used. Findings: Listeria spp. was present in 27  (38.6%) of the samples. L. monocytogenes was detected 9 (12.9%), 7 (10.0%) as L. innocua, 5 (7.1%) as L. ivanovii, 4 (5.67%) as L. seeligeri and 2 (2.9%) as L. welshimeri.  However, E. coli O157:H7 was not detected in any of the samples. . Conclusion & Significance: The incidence of L. monocytogenes in ground beef indicates the need for improved equipment and facility, sanitation in slaughterhouse and in ground beef preparation and handling facilities. The presence of L. monocytogenes in ground beef samples is concerning public health risk. More sensitive methods should be used instead of conventional culture techniques in order to properly detect and characterize contamination by E. coli O157:H7.

Biography:

Alper Ciftci is an expert about Molecular Microbiology and Vaccine Development. He finished his PhD at Ankara University, and now he works as Associate Professor at Ondokuz Mayis University, Samsun, Turkey. He focuses on working development and validation of commercial products such as vaccine and diagnostic kits.

Abstract:

Statement of the Problem: In this study, the investigation of the levels of some biochemical parameters in the blood of sheep due to the vaccination for the prevention of brucellosis was aimed.

Methodology & Theoretical Orientation: 35 months-old and female sheep were used as an animal material. The vaccine was given in single dose against Brucella melitensis and every month blood was drawned from Vena jugularis for 4 months. The vaccine, vaccinated 1 dose via conjunctiva, contains Brucella melitensis Rev1 (0.5-2×10⁹ cfu). Some biochemical parameters in sheep such as ALP, ALT, AST, LDH, glucose, total protein and albumin measured using autoanalyzer.

Findings: In consequence of evaluation of the Rose Bengal Plate agglutination tests, the sera belonging to bloods taken before vaccination were negative and 120 serums belonging to experimental group showed a positive reaction. The SAT results of the serums which were evaluated after RBPT showed that antibody titers (P<0.001) increased significantly at first month according to the pre-vaccine, but it was determined that while from the second month began to fall. It was determined that there was no statistically significant changes in glucose, AST, ALT activity after vaccination (P>0.005), a significant amount of total protein and ALP decreased after vaccination (P<0.005), while the LDH levels were significantly increased (P<0.005).

Conclusion & Significance: It was determined that in first month of increased antibody of conjunctival Brucella vaccine which was carried out in low doses to protect against Brucellosis affected the amount of LDH, ALP, total protein and albumin in subsequent months, it was approaching the level of the pre-vaccine group and conjunctival Brucella vaccine was considered to be used as a safe house to protect them from Brucellosis.

Biography:

Pasquale Centoducati is a Full Professor of Animal Science at the Department of Veterinary Medicine of Bari University “A. Moro” (Italy). One of his main research topics in the last ten years has been the study of meat and milk production by equids (horses and donkeys). In particular, his researches focused on milk produced by these species, and on the possibility of marketing. The main aim of his research is the recognition of a good heating process for donkey milk and how to give the possibility of traceability of heat treatments.

Abstract:

Statement of the Problem: Donkey milk is characterized by high lactose together with low protein amount, low casein (CN)/whey protein (WP) ratio and high level of non-protein nitrogen (NPN) fraction. For this reasons it is the most similar to breast human milk and mare milk, although poor in lipid content and caloric value and so, inadequate as exclusive food in infants for the first year of life. Heat treatment represents an essential step for donkey milk commercialization, and heat treatment traceability had an important role in food control system.

Methodology & Theoretical Orientation: For the trial 270 milk samples were used, 70 donkey milk samples, 70 cows’ milk samples and 70 ewe milk samples. All samples were treated at 72°C for 15 seconds, in continuos. After this, they were suddenly refrigerated at 4°C. Raw and heated milk of all species was analyzed for alkaline phosphatase and peroxidase activity. Enzyme activity was measured by spectrophotometric methods.

Findings: Raw cow milk showed mean ALP values of about 800 × 10³ mU/L, ewe milk 1000 × 10³ mU/L and donkey milk 6 × 10³ mU/L. After heat treatment cow and ewe ALP activity were lower than 300 mU/L. Differently, ALP activity in heated donkey milk decreased of the 10% if compared to raw milk, and remained on values of about 5,5 × 10³ mU/L.

Conclusion & Significance: The ALP and LPO activity in donkey milk is about 100 times lower than what reported for cow and ewe milk. However, although this enzyme was studied and resulted useful for heat treatment traceability in cow milk, it is not the same for donkey milk. In fact, it is partially inactivated and results at high levels of activity after heat treatment. Moreover, LPO activity resulted different from what observed for cow and ewe milk.

Biography:

Alper Ciftci is an expert in Molecular Microbiology and Vaccine Development. He finished his PhD at Ankara University, and now he works as an Associate Professor at Ondokuz Mayis University, Samsun, Turkey. He focuses on working development and validation of commercial products such as vaccine and diagnostic kits.

Abstract:

Statement of the Problem: Staphylococcal mastitis is one of the major economic problems of cattle. The aim of this study was to identify Staphylococcus species that cause mastitis, to examine the virulence factors of these species and to determine the relation between these factors with the pathogenic and apathogenic species.

Methodology & Theoretical Orientation: In this study, 84 Staphylococcus strains isolated from milk samples of 120 cows with clinical and sub-clinical mastitis and 80 healthy cows in different herds in the Aydın region were examined. The species-specific identification of Staphylococcus spp. was carried out with a commercial identification kit (Api-Staph, Bio Merieux®, Lyon, France). The virulence factors of the Staphylococcus isolates were determined with regard to coagulase, protein A, DNase, TNase, capsule, slime and biofilm, hemolytic activity, staphylokinase, hemagglutination, methicillin resistance tests.

Findings: In the study, 37 S. aureus, 13 S. hyicus, 9 S. simulans, 8 S. chromogenes, 5 S. lentus, 5 S. epidermidis, 2 S. haemolyticus, 2 S. hominis, 1 S. auricularis, 1 S. warneri, 1 S. sciuri were isolated and identified. The 41.6% of strains were determined as coagulase positive. In the coagulase positive strains, the rate of protein A, DNase, TNase, capsule, hemolyse, staphylokinase, slime (in agar), biofilm (microdilution) and hemagglutination characteristics  were found 71.4%, 48.6%, 11.4%, 40%, 97.1%, 40%, 28.6%, 37.1% and 17.1%, respectively. In the coagulase negative strains, the rate of these characteristics were found 10.2%, 12.2%, 2%, 8.2%, 82%, 32.7%, 12.2%, 12.2% and 10.2%, respectively. The methicillin resistance rates in the coagulase positive and negative strains were determined as 2.9 and 16.3%.  

Conclusion & Significance: In conclusion, it was of the opinion that animals are potential carrier of Staphylococcus strains that are pathogen for human.

Biography:

Alper Ciftci is an expert in Molecular Microbiology and Vaccine Development. He finished his PhD at Ankara University, and now he works as Associate Professor at Ondokuz Mayis University, Samsun, Turkey. He focuses on working development and validation of commercial products such as vaccine and diagnostic kits.

Abstract:

Statement of the Problem: Enterococci are commensal organisms of human and animals, and may cause diseases in particular conditions. Several virulence factors are responsible in the production of diseases. The aim of this study was to isolate enterococci from fish and to determine virulence factors of the isolates.

Methodology & Theoretical Orientation: In this study, 200 fish samples that were collected from different commercial fish farms establishment in Aegean region from Turkey. For isolation, samples were inoculated to selective medium and for the identification at the level of the genus, Gram staining, catalase tests on slide, gas reproduction from mannitol and reproduction at 45°C were made to presumptive positive colonies. The strains that gave positive results in these tests and were denominated as Enterococcus spp. have been observed for their various phenotypic and biochemical properties to identify them at species level. For genomic identification of enterococci for being E. faecalis, ddl gene targeted PCR was performed. Measurement of the Aggregation Substance (AS) of the enterococci was performed by clumping assay. Gelatinase (Gel) activity of enterococci was tested in gelatine medium. The detection of cytolysin activity was made in blood agar.

Findings: A total of 26 (13%) E. faecalis strains were isolated from live, moribund and dead fish collected from fish farms in Aegian Region. Cytolysin production was not detected in any of E. faecalis strains. Of 26 strains tested, 27% was found to produce AS. Gel activity was found in 11.5% of strains.

Conclusion & Significance: The presence of strains with important virulence factors in enterococci from fish was established. It was suggested that these strains have the potential of producing disease in human and animals.

Biography:

Alper Ciftci is an Expert in Molecular Microbiology and Vaccine Development. He finished his PhD at Ankara University, and now he works as Associate Professor at Ondokuz Mayis University, Samsun, Turkey. He focuses on working development and validation of commercial products such as vaccine and diagnostic kits.

Abstract:

Statement of the Problem: Aeromonas hydrophila is known to be the causative agent of bacterial hemorrhagic septicemia in fresh water fish. Zoonotic properties of A.hydrophila increase the importance of infection in terms of public health. The aim of this study was to develop the ready-to-use-PCR-kit and evaluate the stability of kit for A. hydrophila.

Methodology & Theoretical Orientation: The oligonucleotide primers which targeted phospholipase C gene were designated with using PUBMED-Blast. The optimization of PCR was conducted through combination of reactives. After optimization, the most suitable PCR conditions were performed for specifity. For this aim, related and unrelated bacterial strains were used. The sensitivity of PCR was investigated by the dilutions of DNAs of A.hydrophila strain. For the determination of stability of optimized PCR, 52 PCR mixtures without template DNA were lyophilized. These mixtures were diluted every week and A.hydrophila template DNA was added to the mixture. The mixtures were amplified with optimized PCR conditions.

Findings: The optimized PCR conditions with using designated primers were gave specific bands of 303 bp for A.hydrophila. When used, related and unrelated bacterial strains except for A.hydrophila, no band were shown in the PCR. These results showed us that the designated primers with optimized conditions were specific for A.hydrophila. The sensitivity of the PCR was determined as 70 pg of DNA. This concentration was used to determine the stability of lyophilized PCR mixtures. During 22 weeks period, the PCR gave positive band of 303 bp for A.hydrophila. After 23 weeks, no bands were shown.

Conclusion & Significance: For the development of ready-to-use-PCR-kit, the oligonucleotide primers were designated. After optimization, the kit gave results up to 22 months, that the primer used in this study was only A.hydrophila specific and was able to detect 70 pg of DNA.

Biography:

Anastasiya Varlamova is a Postgraduate Student of the Third Course of All-Russian Scientific Research Institute of Fundamental and Applied Parasitology of Animals and Plants named after K I Skryabin. The spheres of her scientific work are Veterinary Parasitology and Pharmacology. She studies new dosage forms of drugs, tests new methodics of researches with her Scientific Advisor. The aim of her work is decreasing the toxicity of drugs and reduction of drug residues in food.

Abstract:

Statement of the Problem: The supramolecular complexes of anthelmintics used to treat cestode infection with sodium salt of glycyrrhizic acid (Na₂GA) were created by mechanochemical technology to enhance the solubility, bioavailability of the drugs. In this regard the aim of our study was to examine the efficacy of them against hymenolepiosis of white mice.

Methodology & Theoretical Orientation: The studies were made on 48 mice experimentally infected with Hymenolepis nana at the dose of 200 eggs per head. The complexes of albendazole, fenbendazole, niclozamide and praziquantel with Na₂GA produced by mechanochemical modification of substance in activator of percussive-grating type were used in tests. Mice of different groups of 6 animals each on the 13 day after infection received per os complexes of albendazole, fenbendazole and praziquantel at the dose of 2,0 mg/kg of active substance (AS) on 1% starch gel and niclozamide at the dose of 20 mg/kg. The substances of albendazole, fenbendazole and praziquantel at the dose of 2.0 mg/kg and niclozamide at the dose of 20 mg/kg of AS were as the basic drugs. Control animals received starch gel. The efficacy of drugs was determined on the fourth day after drugs administration by the results of helminthological autopsy of mice intestine and calculation the amount of cestodes.

Findings: The complexes of praziquantel and niclozamide with Na₂GA showed the best efficacy: 100% and 83.4% respectively against hymenolepiosis. Complexes of albendazole and fenbendazole with Na₂GA were not effective enough. Basic drugs – substances of albendazole, fenbendazole, praziquantel showed 46.7; 46.7; 66.7% efficacy respectively at the dose of 2.0 mg/kg and niclosamide showed 40.0% at the dose of 20 mg/kg.

Conclusion & Significance: The efficacy of supramolecular complexes of praziquantel and niclozamide was approximately 1, 5-2 times higher than the efficacy of the basic drugs. The study was supported by RSF â„–14-16-00026.

Biography:

Emre Kaya is a PhD in Department of Biochemistry, Faculty of Veterinary Medicine, Firat University, Elazig, Turkey. He is continuing his scientific studies in various subjects in the same place. His work focuses more on oxidative stress and its prevention.

Abstract:

Statement of the Problem: Diethylnitrosamine (DEN) is a potent genotoxic nitroso carcinogen that induces hepatocellular carcinomas in rat liver.

Methodology & Theoretical Orientation: In this study, adult male Wistar-Albino rats were used. Rats were divided into five groups as follows; control group, lycopene group (lycopene was administered), DEN group (DEN was administered), lycopene+DEN group (lycopene administration was started 10 days before the DEN administration) and DEN+lycopene group (Lycopene administration was started together with the DEN administration). DEN was administered to rats at 200 mg/kg/bw, a single dose i.p. for 90 days. Lycopene was administered to rats every other day at 10 mg/kg/bw, gavage for 10 days. Plasma aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine kinase (CK) activities and cholesterol levels were determined.

Findings: It wasn’t found statistically significant difference in plasma AST, ALT, ALP, LDH, CK activities and cholesterol level in the lycopene group when compared with the control group. Plasma AST, ALT, ALP, LDH, CK activities and cholesterol levels were detected to significant increases in the DEN group when compared with the control group (p<0.001). Plasma AST, ALT, LDH activities and cholesterol level were detected to decrease in the lycopene+DEN and DEN+lycopene groups when compared with the DEN group (p<0.05). However, plasma ALP and CK activities were not changed in the lycopene+DEN and DEN+lycopene groups when compared with the DEN group.

Conclusion & Significance: It was observed that DEN causes a change to in some liver specific enzymes in plasma of DEN administered rats. These results indicate that lycopene might have a protective effect against DEN induced hepatotoxicity by regulating these enzymes.

Biography:

Gulay Ciftci is an expert in Molecular Biochemistry and Proteomics. She finished PhD at Ankara University, and now she works as an Associate Professor at Ondokuz Mayis University, Samsun, Turkey. Her work focuses on hormones, proteins and genes.

Abstract:

Obesity and its associated co-morbidities have become a global health concern. Peptides such as leptin, cholecystokinin, glucagon-like peptide 1 and acylated ghrelin have been shown to influence energy balance in peripheral tissues and brain. It was suggested that nesfatin-1 could play a role in the control of food intake and energy expenditure. Leptin resistance is a common phenomenon in obesity. Central and peripheral injection of nesfatin-1 exerts its food reducing effects via a leptin dependent mechanism. Nesfatin-1 is a recently discovered peptide derived from the precursor protein nucleobindin2 (NUCB2) and described to be expressed in the rat hypothalamus. In quest of novel appetite regulating molecules, NEFA/nucleobindin2, a peroxisome proliferator-γ receptor-activated gene was re-discovered in immortalized cell lines and later in the hypothalamus of rodents. Nesfatin-1 and its precursor NUCB2 possess anorexigenic properties. Nesfatin-1 reduces food intake following central injection in rats, mice or goldfish. Subsequent research showed a more widespread distribution of NUCB2/nesfatin-1 in the rat brain and a prominent expression in the rat stomach with 10-fold higher expression levels in the gastric oxyntic mucosa compared to the brain. A third ventricle injection of either NUCB2 or nesfatin-1 at similar doses reduces food intake in ad libitum fed rats during the dark phase. Moreover, the anorexigenic effect of nesfatin-1 is independent from leptin. Nesfatin-1’s anorexigenic effect was confirmed in further studies where nesfatin-1 was injected centrally in ad libitum fed rats and fasted animals. Whether species differences play a role remains to be further investigated. In summary, current data clearly point towards a role of central nesfatin-1 in the regulation of food intake, whereas peripheral nesfatin-1 seems to be rather involved in the reduction of gastric motility in dogs, increase of hepatic oxidation and glucose homeostasis with an increase of glucose-stimulated insulin release from the pancreas in rats and humans.

Biography:

Gulay Ciftci is an Expert in Molecular Biochemistry and Proteomics. She finished PhD at Ankara University, and now she works as an Associate Professor at Ondokuz Mayis University, Samsun, Turkey. She focuses on working hormones, proteins and genes.

Abstract:

Metabolic disorders present an increasing public health concern and can significantly undermine an individual’s quality of life. Over the last decade, dynamic changes in dietary nutrient ingestion have rapidly increased the prevalence of metabolic disorders. Recently, scientists have proposed that metabolic disorders might result from an alteration in gut microbiota composition. The term “probiotic” originates from the Greek word meaning “for life”. In 1989, Fuller defined the term probiotic as “a live microbial feed supplement which beneficially affects the host animal by improving its intestinal balance”. It is well established that five bacterial phyla, firmicutes, bacteroidetes, actinobacteria, proteobacteria, and verrucomicrobia, are dominant components of the gut microbiota. More than 90% of the bacterial populations are gram-negative anaerobes and include the predominant genera bacteroides, eubacterium, bifidobacterium, and fusobacterium. Diet controls the composition of these bacteria, which are crucial in the development of metabolic disorders. Accumulating evidence suggests that supplementation of probiotics and prebiotics could have preventative and therapeutic effects on CVD due to a reduction in total serum cholesterol, low-density lipoprotein (LDL-cholesterol), and inflammation. In particular, changes of gutmicrobiota composition are strongly associated with increased adiposity, cell dysfunction, metabolic endotoxemia, systemic inflammation]. The change in intestinal microbiota and the reduced bacterial diversity were also observed in obese conditions. The change of these bacteria compositions increases susceptibility to infections, immune disorders, inflammation, oxidative stress and insulin resistance, events that are mediated by metabolic endotoxemia, which involves exposure to noxious intestinal products, particularly lipopolysaccharides. Consequently, the present review aimed to provide an objective perspective of the current knowledge regarding the impacts of food constituents on gut microbiota and health.

Biography:

Sibel Gür is working at Afyon Kocatepe University, Turkey. His international experience includes various programs, contributions and participation in different countries for diverse fields of study. His research interests reflect in his wide range of publications in various national and international journals.  

Abstract:

Bovine viral diarrhea virus (BVDV) is among the most prominent infections in the aspect of economic consequences in dairy industry. The number of mild or big size dairy enterprises having been increasing in Turkey. However, majority of the farm animal production is still having been in small private enterprises in all over the Anatolia. Health problems in the cattle are mainly due to main viral infections rather than bacterial ones. Ruminant pestiviruses are the most prevalent disease beside Infectious Bovine Rhinotracheitis. In this study, pestivirus was investigated serologically in Afyonkarahisar province in the west Anatolia which takes place in 8^th order of cattle livestock in the Turkey with 342.601 cattle as 2015 data. Blood sera samples were collected from only family type small farms from boroughs of Afyonkarahisar province; BaÅŸmakçı, Bayat, Bolvadin, Çay, Çobanlar, Dazkırı, Dinar, EmirdaÄŸ, Evciler, Hocalar, Ä°hsaniye, Ä°scehisar, Kızılören, Sandıklı, SinanpaÅŸa, Sultandağı and Åžuhut. There were less than 40 animals (average 2-10 cattle) in every farm. Ages of the animals were 6 months old and above (av. nearly 3 years old). Sex was ignored but most of the animals were female. Breeding aim was generally dual. The all cattle were clinically healthy during sampling and not vaccinated for the infection in last two years. Blood serum samples were controlled using Serum Neutralisation and SN₅₀ test. Out of 17 boroughs, seropositivity was detected in all of them between 50.2% and 92.8%. In total, pestivirus antibodies were found to be in 958 of 1.279 cattle (74.9%). Average antibody titer values were the highest in 1/20 dilution while slight increase was observed from 1/80 point as a sign of recent infection. Incidence of viral infections is generally lower in the small farms comparing intensive breeding having been big scale enterprises. However, obtained results showed considerable risk potential even in small private farms. Proper technical support seems is an urgent needs to prevent losses due to wide range of clinical disorders by BVD.

Biography:

Harikrishna Pillai has completed his Master of Veterinary Science and Doctorate of Philosophy in Animal Biochemistry from Indian Veterinary Research Institute. He has done his Doctoral Thesis on the Protein Regucalcin. He has done purification, expression, structural and functional studies of Regucalcin of Water buffalo (Bubalus bubalis) in various cell types. His research is first to report the presence of regucalcin in stem cells and spermatozoa. He investigated the anti capacitatory effects and putative cryoprotective effect of regucalcin in spermatozoa using recombinant protein. His research so far has generated five first-author peer-reviewed journal articles. Currently he is working as Laboratory Manager in Almarai, Saudi Arabia. His career objective is to further his research capabilities by pursuing a Post-doctoral position in a reputed laboratory to learn, develop, and diversify himself.

Abstract:

Statement of the problem: Regucalcin/Senescence Marker Protein-30 (RGN/SMP-30), a multifunctional calcium binding protein, has many pivotal roles in calcium homeostasis, ascorbic acid biosynthesis and having anti-prolific and anti-apoptotic functions. All these functions are very important in stem cells. However, there are no reports regarding the role of RGN in stem cells. The aim of the present study was to investigate the expression of RGN in buffalo derived Wharton’s jelly (BWJ) and fetal fibroblast (BFF).

Methodology: Gravid buffalo uteri of (~40- 50 days old) were procured. Mesenchymal stem cells (MSCs) were isolated from BFF and BWJ and cultured in DMEM with 15% FBS. The expression of RGN in BFF and BWJ was elucidated by qPCR, immunostaining and western blot analysis.

Results: The real-time PCR products of BWJ and BFF were confirmed on 2% agarose gel electrophoresis by the presence of 197bp product. An immunoreactive band of 34 kDa specific for RGN was detected in Western blot of BWJ and BFF. Cellular localization in both BWJ and BFF by immunocytochemistry revealed that RGN is located in both cytoplasm and nucleus.

Conclusion & Significance: This study is the first report on the expression of RGN in stem cells of any kind. It confirms the presence of RGN in fetal stem cells in mRNA and protein level. As this work provides initial information regarding the presence of RGN in MSCs, further investigation is needed to find out the role of RGN in important role in the differentiation, proliferation and aging of stem cells.

Biography:

Jobin Thomas has completed his Master of Veterinary Science in Veterinary Immunology from Indian Veterinary Research Institute, India. He investigated on the disease canine parvoviral enteritis. His research mainly includes antigenic identification, cell culture, recombinant protein expression, serological diagnosis using LAT and Dot blot of CPV-2. Currently he is doing his PhD in Bovine Tuberculosis at IREC, SaBio, UCLM, Spain.

Abstract:

Statement of the Problem: Parvoviral enteritis is a major cause of mortality in puppies, caused by canine parovirus-2 (CPV-2). Vaccination is the major prevention tool; however the emergence of new antigenic variants (CPV-2a, 2b, 2c) has raised a great challenge on the success of vaccination. The present study documents screening of fecal samples from suspected puppies for CPV-2 and sequencing of some of the positive samples to find out the locally prevalent antigenic types.

Methodology: Fecal samples were collected form puppies showing typical symptoms of parvoviral enteritis, presented at Veterinary Polyclinic, Indian Veterinary Research Institute, India and viral DNA was isolated. Polymerase chain reaction was done using newly designed primers and epidemiological analysis was carried out with respect to age, sex and breed. Oligonucleotide sequencing was done using the amplified product and phylogenetic analysis was performed with other sequences reported from India and abroad using Neighbor joining method based on p-distance model with the bootstrap test based on 2,000 resamplings in MEGA 6 software.

Results: Out of 44 samples, 23 were found to be positive for CPV-2 which was confirmed by 564 bp products on 1% agarose gel (Figure 1). Epidemiological analysis revealed that the disease was found to be more common in Labrador pups and 3-6 months male puppies were mostly affected. The positive cases represented 17.4% vaccinated puppies. Sequencing and BLAST analysis of 15 random positive samples (Gen bank accession numbers: KJ-364526, KM-003870 to KM003883) indicated 99% identity with CPV-2a. However, there were   mutations observed at positions 297 (Ser 297 Ala) and 440 (Thr 440 Ala) amino acid residue, and hence typed as new antigenic variant of CPV-2a. Moreover, another mutation at position 264 (Gly 264 Val) was present only in 10 sequenced samples (10/15) which was not reported previously from India.  Also there was a mutation at nucleotide position 3978 in two sequences in which cytosine was replaced by thymine. Phylogenetic analysis indicated that the genetic changes between the Indian sequences were much lesser when compared with foreign sequences. All the isolates in the present study showed more similarity with sequences from China.

Conclusion & Significance: CPV-2 is a major threat to puppies despite of regular vaccination, as the present study has shown that there is a significant percentage of infection in vaccinated puppies. The emergence of new antigenic variants makes the vaccine efficacy questionable. Further investigation is needed to determine the variation in the prevalence of antigenic types and evaluation the efficacy of current vaccines against new types of CPV-2

Biography:

Julio V Figueroa  is currently working as a Researcher and Head of the National Research Center for Veterinary Parasitology, INIFAP, in Jiutepec, Morelos, Mexico. He obtained his Veterinary Medicine Degree from The State of Mexico Autonomous University in Toluca, Mexico, and the MSc and PhD degrees in Veterinary Pathology and Microbiology at the University of Columbia-Missouri, in Columbia, MO, USA. He has conducted research on tick borne diseases of cattle during the past 30 years and has published over 70 research papers in peer reviewed international journals.

Abstract:

The high economic impact caused by one of the most important tick-borne disease in Mexico, bovine babesiosis, has led to the search and development of effective techniques that facilitate its diagnosis and control. The objective of this study was to compare the serological indirect fluorescent antibody test (IFAT) and the indirect enzyme-linked immunosorbent assay (iELISA) for the identification of cattle exposed to Babesia spp. Negative and positive sera controls, sera from experimentally infected animals and sera from animals under field conditions were evaluated. For the iELISA, the recombinant proteins RAP-1α (Babesia bigemina) and MSA-1 (B. bovis) were used as antigens. The technique iELISA demonstrated high sensitivity and diagnostic specificity rates using the RAP-1-α antigen. The sensitivity and diagnostic specificity rates for the indirect ELISA with the B. bigemina RAP-1-α recombinant antigen were 90% and 98.75%, respectively, whereas a sensitivity and diagnostic specificity of 73.33% and 81.25%, respectively, were obtained with the IFAT. The kappa index (κ) between the IFAT (in vitro culture-derived B. bigemina antigen) and iELISA (RAP-1-α) was 0.44. Although the serological tests do not have the capacity to discriminate cattle from three different geographic regions in Mexico that are exposed to either Babesia bovis and / or B. bigemina, it was concluded that the iELISA (RAP-1-α) is a tool with high sensitivity and diagnostic specificity, able to lead to a better knowledge on the epidemiology of bovine babesiosis, providing useful information for the design of control strategies and management of cattle babesiosis, one of the most important tick-borne disease in Mexico.

Biography:

Julio V Figueroa  is currently working as a Researcher and Head of the National Research Center for Veterinary Parasitology, INIFAP, in Jiutepec, Morelos, Mexico. He obtained his Veterinary Medicine Degree from the State of Mexico Autonomous University in Toluca, Mexico, and the MSc and PhD degrees in Veterinary Pathology and Microbiology at the University of Columbia-Missouri, in Columbia, MO, USA. He has conducted research on tick borne diseases of cattle during the past 30 years and has published over 70 research papers in peer reviewed international journals.

Abstract:

The Babesia bigemina RAP-1 antigen is a 58 kDa protein located in the merozoite roptries of the protozoan parasite, which participates in the invasion to the bovine erythrocyte. rap-1 sequences of B. bigemina isolates from Mexico, Brazil, Argentina, Uruguay and Puerto Rico are known. However, the degree of conservation of rap-1 sequence in different geographic isolates of Mexico is not known. The aim of this study was to analyze rap-1α1 sequences in 8 different isolates of B. bigemina from Mexico. The methodology included the extraction of genomic DNA from B. bigemina-infected erythrocytes; PCR amplification of rap-1α1 using  rap-1F and rap-1R as primers; Cloning the amplified fragment (1440 bp) into TOPO 10 vector and transforming competent E. coli cells; Selection of clones and purification of recombinant plasmids; Sequencing and analysis of rap-1α1 from the B. bigemina isolates, obtained from at least 3 recombinant clones per isolate, sequencing the inserts in both directions; and sequence alignments and assembling with the CLC application genomic workbench 4.8. The identity of the rap-1α1 sequences was determined by a homology search with BLAST tools. The results showed amplification of rap-1α1 in all the B. bigemina isolates. The comparative analysis of nucleotide sequences revealed a high degree of conservation (98-99% identity) between the rap-1α1 genes of the different Mexican isolates, compared to the reference rap-1α1 sequence. BLASTX analysis revealed identities up to 99% in the deduced amino acid sequences of RAP-1 compared to the sequence of the reference isolate. The high degree of conservation in rap-1α1 sequences among geographically distant Mexican isolates suggests that there is no strong bovine immune pressure that translates into genetic variation of this particular gene. The RAP-1 antigen is a viable candidate for inclusion in a diagnostic test for bovine babesiosis caused by B. bigemina in Mexico.

Biography:

Julio V Figueroa is currently working as a Researcher and Head of the National Research Center for Veterinary Parasitology, INIFAP, in Jiutepec, Morelos, Mexico. He obtained his Veterinary Medicine Degree from the State of Mexico Autonomous University in Toluca, Mexico, and the MSc and PhD degrees in Veterinary Pathology and Microbiology at the University of Columbia-Missouri, in Columbia, MO, USA. He has conducted research on tick borne diseases of cattle during the past 30 years and has published over 70 research papers in peer reviewed international journals.

Abstract:

Lack of information about Mexico for corroborating hemoparasitic infections to wild ruminants led to the present study which is done to determine the prevalence of babesiosis and anaplasmosis in water buffaloes that are raised in areas of high incidence of ticks. A total of 233 blood samples were collected in four buffalo production farms in the gulf coast of Mexico. For the detection of circulating antibodies against Babesia spp. and Anaplasma spp., the sera were evaluated using the indirect fluorescent antibody test (IFAT) and the enzyme-linked immunosorbent assay (ELISA) techniques, respectively. Molecular analysis of the samples was performed using the nested polymerase chain reaction (nPCR) using specific primers for each hemoparasite species. The overall seroprevalence rate (54.5%, 57.1% and 7.29% for Babesia bovis, Babesia bigemina and Anaplasma marginale, respectively) and molecular evidence (39.48%, 14.16% and 2.57% for B. bovis, B. bigemina and A. marginale, respectively) that were obtained as results in the present study allow to conclude that the water buffaloes, raised in the four production farms, not only demonstrate the immune system exposure of animals to these hemoparasites species, but also, that they are carriers of the infection caused by B. bovis, B. bigemina and A. marginale, as evidenced by the detection of specific circulating antibodies and hemoparasites DNA, respectively. 

Biography:

Márcia M G Jusi has a degree in Biological Sciences from the Faculty of Agrarian and Veterinary Sciences (FCAV), UNESP-Jaboticabal, SP, Brazil. She was a CNPq Fellow of Scientific Initiation in 2005, in the area of ​​Biological Control. She was also a FAPESP Fellow of Scientific Initiation, in 2007, on diagnosis of leishmaniasis in wild canids and procionids. She completed a Master's degree in 02/2011 at FCAV, in the area of ​​Molecular Biology with cloning and expression of Leishmania chagasi A2 protein. In March 2011, she started her PhD course at the same university and finished it in February 2015 with scholarship and research funding by FAPESP, she carried out a study on cellular and humoral immune response of mice immunized with the A2 recombinant Leishmania chagasi protein. She is currently a Post-doc Business Scholar (PIPE-FAPESP), developing an immunochromatographic test for canine ehrlichiosis using a recombinant protein in the company Imunodot.

Abstract:

Statement of the Problem: Canine monocytic ehrlichiosis (CME), is an infectious-contagious disease caused by the intracellular bacteria, Ehrlichia canis. The diagnosis of CME in the acute phase allows the early treatment and improves the prognosis. However, the clinical diagnosis is impaired by the non-specific clinical signs. Therefore, serological tests provide a more sensitive diagnosis. The search for specific antibodies anti-E. canis by the Fluorescent Antibody Test (IFAT) is considered the gold standard for CME diagnosis. The objective of this work is to develop and validate a rapid immunochromatographic diagnostic kit for detection IgG antibodies to E. canis recombinant proteins of naturally infected dogs serum samples. A kit originally brazilian, which would contribute to early of diagnosis, as well as the benefit and costs. Since it is quick and easy to perform, the veterinarians themselves in their clinics would perform it.

Methodology & Theorical Orientation: We produced the recombinant E. canis protein in large scale, as well, the colloidal gold, this later, using the tannic acid and sodium citrate method. We also produced the conjugate (Protein A conjugated to colloidal gold) for nitrocellulose membrane tests.

Findings: We detect reactivity for positive serum canine and not for the negative serum canine by immunochromatographic test in preliminar tests. 

Conclusion & Significance: The test on the nitrocellulose membrane demonstrated that as the recombinant protein as and protein A reacted very well with the dogs’s serum samples once establishing the optimal concentrations of antigen, conjugate and dilution of the sera.