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Multiplex PCR for simultaneous detection of 3 major fish pathogens incriminated in bacterial septicemic syndrome Full text
2018
Mortada M. A. Hussein | Walid H. Hassan | Aya M. A. El-Wkeel
Fish with bacterial septicemic syndrome (BSS) exhibit very similar clinical signs regardless of the etiological agents. Members of Aeromonas, Pseudomonas, Vibrio, Edwardsiella, Streptococcus and Lactococcus species are considered the most reported bacterial pathogens incriminated in such syndrome. Aeromonas hydrophila, Edwardsiella tarda and Streptococcus iniae are 3 major pathogens share in the BSS associated losses in aquaculture and considered problematic for growth of tilapia and catfish production in Egypt. Therefore, rapid and accurate diagnosis is highly needed for controlling their disease outbreaks, particularly, in mixed infections. In an attempt to elucidate the main causative pathogen, a novel multiplex PCR (m-PCR) was newly designed in this study. The developed m-PCR involves amplifying the three multiple genes in single reaction based upon primers deduced from the regions carrying 16S rRNA, etfA and 16S RNA genes of A. hydrophila (Aeromonas spp.), E. tarda and S. iniae, respectively. Prior to perform m-PCR, individual PCR assays were carried out to adapt suitable laboratory and m-PCR assays conditions. The specificity of the developed m-PCR was confirmed by the fact that only specific fragments were amplified equivalent for 953, 415 and 300 bp corresponding to A.hydrophila, E. tarda and S. iniae, respectively, and that was evident with both extracted DNAs and the bacterial cells. More specifically, these specific bands were obtained also when either the extracted DNAs or the bacterial cells of the three pathogens mixed together in the reaction. The developed m-PCR is accurate, sensitive, fast and simple technique for the simultaneous detection of A. hydrophila (Aeromonas spp.), E.tarda, and S. iniae, three major bacterial pathogens involved in BSS incidence in Egypt.
Show more [+] Less [-]Effect of liver diseases on hormonal and biochemical parameters in Wistar albino rat Full text
2018
Amira, H. Mohamed | Kamal, H.H. | Walaa, M.S. Ahmed | Hanan, E. Saeed
The current study was performed to evaluate the effect of acute and chronic hepatotoxicity induced by paracetamol and thioacetamide respectively on serum hormonal levels and biochemical parameters. Female Wistar albino rats were divided into 3 equal groups (C), (P) and (T). Group (C) were kept as control, group (P) were received paracetamol orally (500 mg/kg b.wt) daily for 15 days and those of group (T) were injected thioacetamide (200 mg/kg b.wt) intraperitonialy twice/ week for 90 days. In P group, results revealed significant elevation in liver enzyme activities (ALT, AST and ALP), T4, insulin (7th day), estrogen (7th and 15th days), triglycerides (7th day) and cholesterol levels throughout the experiment while serum proteins and T4 (15th day) showed significant decreased values. Whereas, at 90th days of chronic intoxicated group (T) resulted in significant elevation in liver enzyme activities (ALT, AST and ALP), bilirubin, estrogen, T4, triglycerides (60th and 90th days) and T3 (120th day). While the levels of T4 and cortisol (60th day), serum total protein, albumin, globulin (90th day) and insulin (120th day) showed significant decreased values when compared to control group. In conclusion, both paracetamol and thioacetamide cause different degrees of damage in liver of rats leading to clear changes in their hormonal and biochemical profiles.
Show more [+] Less [-]Beneficial roles of boron supplementation in the ration of rams on productive activity and semenquality Full text
2018
Taha B. Ibrahim | Ahmed Abdel-Wahab | Ahmed H. El-Anwar | Shawky S. Ibrahim
The aim of this study is to determine the effects of boron supplementation in ration of rams on their productive activity and semen quality. For that purpose, twelve rams were divided into two groups (6 rams/ each). Rams of the first control group were fed the basal ration. Concerning the second group (boron group), the rams were fed the basal ration plus 400 mg boric acid (BA) / kg diet as a source of boron. The rams in both groups fed their corresponding ration for 4 months ad libitum. Blood samples were collected monthly throughout the experimental period. Semen samples were collected at the 12th and 16th weeks of the experiment. The results clarified that the serum levels of tetraiodotyrosine (T4) were observed to be elevated significantly (P<0.05) with growing of age in the rams fed B, but they didn't change in the control group. In addition, the semen quality was maintained normal with boronsupplementation as the individual sperm motilities and live/dead sperm cells percentages were similar to control. Also, boronsupplementation didn't induce any adverse effects on the serum levels of Mg when compared to control group. Thus, boronis considered a potential and useful supplement in ration of rams to improve the thyroid activities and didn't induce any adverse effects on either semen quality or serum magnesium level in rams.
Show more [+] Less [-]Prevalence of Salmonella Spp in Some broiler farmsin different Egyptian Governorates Full text
2018
H.A. Kaoud | M. A. El-Babbly | K. G. El-Iraqi | M.M. Khalil
This study was conducted to evaluatethe degree of biosecurity level with especial reference to Salmonella Spp as an example to explain the expected causes and risk factors that leads to spread them in poultry flocks in Egypt then studied farms evaluated for the sensitivity of Salmonella isolates to the most common disinfectants used in Egypt. About 300 samples (100 cloacal swabs, 100 liver and intestinal samples, 100 litter samples) were collected from 10 broiler farms with different age (at 0 old day, one week, 2,4 and 6 weeks of age) then the samples were investigated for Salmonella Sppand subsequently identified based on biochemical and serological tests.The obtained results showed that 35 Salmonella species were isolated from 10 broiler poultry houses (25%);(6%) and (4%) from cloacal swab;liver and litter, respectively. Average prevalence of Salmonella spp. was 11.33 % in open broiler houses whether raised Cobb, Ross or Sasso breeds. Salmonella Typhimurium, S. enteritidis and S.Kentucky were the most serovars out of the 35 detected isolates. There was great statistical significant difference in the sensitivity of Salmonella isolates to the most common disinfectants(P<0.05)asVerkon- S® achieved 3 log reduction, after 5-minute,Formalin and Phenique were achieved 3 log reductions against S. entriditisafter one-minute Aldekol Des- Gda® achieved one log reduction after one minute, TH4®,Biosentry® 904 and Iodophoreachieved 2 log reductions after 5 minutes.
Show more [+] Less [-]Genetic analysis of multidrug resistant Salmonella isolated from broiler chickens Full text
2018
Walid H. Hassan | Ahmed H. Abed | Abd El Rady Thabet | Eman A.M. El Nady
Salmonellosis is a major problem for the poultry industry, and this problem represents a critical food safety hazard. Resistance to antimicrobial agents within nontyphoidal Salmonellae is a serious problem. The present study aimed to analyze genetically some β-lactamase resistance genes and some virulence associated genes in Salmonella isolates from broiler chicken. Five hundred samples were collected from diseased broiler chickens of different ages (3-6 weeks) from different farms in Assiut Governorate in Egypt during the period from January 2015 to December 2015. Bacteriological examination showed that 26 Salmonella isolates were recovered with a prevalence rate of 5.2% Serotyping of Salmonella isolates showed that S. Enteritidis S. Typhimurium, and S. Kentuky were identified at rates of 50%, 30.8% and 19.2%, respectively. Results of antibiogram showed that 18 Salmonella isolates (92.3%) were multidrug resistant. All isolates were screened for the presence of 2 β-lactamase resistance genes (blaCTX and blaCMY) as well as 3 virulence genes (stn, invA and hilA) using multiplex PCR. The overall prevalences were 53.9% for blaCTX and 34.6% for blaCMY. Meanwhile, stn, invA and hilA genes were found in 96.2%, 100% and 84.7% of isolates, respectively.
Show more [+] Less [-]Evaluation of bivalent inactivated oil adjuvant African Horse Sickness vaccine in Guinea pigs and mice Full text
2018
Nashwa K. Madkour | S. K. Rofaiil.
In this study, neutralizing antibody response in Guinea pigs and mice inoculated by African horse sickness (AHS) bivalent inactivated oil adjuvant vaccine was similar when compared serologically with that in horse. Therefore, using G. pigs and mice may be a method of great value in potency test for routine immunization. Potency test of the prepared vaccine using an immunization challenge protocol in adult mice was studied, using challenge virus with 100, 50 and 25 LD50. The highest protection level (95%) was with the dilution of 50 LD50/0.05 ml
Show more [+] Less [-]Some virulence factors of Escherichia coli isolated from diarrheic calves Full text
2018
Ghada Sayed Ibrahim | Walid H. Hassan | Ragab Azzam | Abeer Ahmed El -Sayed Shehata
Escherichia coliare the most common cause of diarrhea in calves. Diarrhea in calvesremains one of the most important problems faced by livestock, causing great economic losses. Some strains ofE. coli characterized by the presence of specific virulence factors including haemolysin production, resistance to bactericidal effects of serum and Congo red binding activity. In this studyfecal samples were collected from 115 diarrheic calves aged from 3 days to one year and from different localities in Egypt along the period from February to August 2015.The prevalence of E. coli in diarrheic calves was 72.2%. 39.8% of isolated E. coli were haemolytic to sheep blood agar, 68.7% were serum resistant, 100% showed Congo red binding activity.
Show more [+] Less [-]Efficacy of application of an alcohol-based antiseptic hand rub or a 2% chlorhexidine gluconate scrub for immediate reduction of the bacterial population on the skin of dogs Full text
2018
Maxwell, Elizabeth A. | Bennett, Avery | Mitchell, Mark A.
OBJECTIVE To compare the efficacy of application of an alcohol-based antiseptic (80% ethyl alcohol) hand rub (ABAHR) with that of a 2% chlorhexidine gluconate scrub (CGS2) for immediate reduction of the bacterial population on the skin of dogs. ANIMALS 50 client-owned dogs with no evidence of skin disease. PROCEDURES On each dog, 2 areas of hair on the ventral aspect of the abdomen were clipped with a No. 40 blade and cleared of debris. A direct contact plate holding tryptic soy agar with polysorbate 80 and lecithin was gently pressed (for 2 seconds) on each skin site (preapplication sample). The CGS2 and ABAHR were each aseptically applied to 1 skin site on each dog. A direct contact plate was subsequently applied to each site in a similar manner (postapplication sample). All plates were cultured, and bacterial isolates were identified and quantified by the number of CFUs per plate. RESULTS Application of the CGS2 and ABAHR significantly decreased skin bacterial colony counts, compared with findings for preapplication samples. The number of CFUs per plate or postapplication percentage reduction in CFUs per plate did not differ between treatments. There were no adverse skin reactions associated with either application. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that applications of ABAHR and CGS2 were equally effective at immediately reducing the bacterial population on the skin of dogs, and there was no significant difference in percentage reduction in colony counts between the 2 applications.
Show more [+] Less [-]Impact of a trap-neuter-return event on the size of free-roaming cat colonies around barns and stables in Quebec: A randomized controlled trial Full text
2018
Bissonnette, V. | Lussier, B. | Doize, B. | Arsenault, J.
The objective of this study was to evaluate the impact of a trap-neuter-return (TNR) event on the size of free-roaming rural cat colonies in Quebec. This prospective randomized, controlled study included 18 cat colonies around barns and stables that were randomly assigned to either a TNR group (10 colonies of 7 to 27 cats; 14.3 cats on average) or a control group (8 colonies of 7 to 26 cats; 14.5 cats on average). The number of cats in each colony was calculated from the images obtained by camera-trapping at: baseline (T0), 7.5 mo (T7), and 12 mo (T12). At baseline, the TNR group was subjected to a TNR event. When taking into account adults only, a significant growth difference was observed in the number of cats between the TNR group and the control group at T7 (P = 0.03). When including kittens as well as adults, a trend towards a lower growth of the TNR group compared to the control group was noted at T7 (P = 0.06). There was no difference in the number of kittens between the 2 groups at T7 (P = 0.49) or at T12 (P = 0.36). There was a trend towards more emigration in the control group at T12 (P = 0.095). Isolated TNR events have a low and temporary impact on colony size in Quebec's rural cat colonies.
Show more [+] Less [-]Effect of a syringe aspiration technique versus a mechanical suction technique and use of N-butylscopolammonium bromide on the quantity and quality of bronchoalveolar lavage fluid samples obtained from horses with the summer pasture endophenotype of equine asthma Full text
2018
Bowser, Jacquelyn E. | Costa, Lais R. R. | Rodil, Alba U. | Lopp, Christine T. | Johnson, Melanie E. | Wills, Robert W. | Swiderski, Cyprianna E.
OBJECTIVE To evaluate the effect of 2 bronchoalveolar lavage (BAL) sampling techniques and the use of N-butylscopolammonium bromide (NBB) on the quantity and quality of BAL fluid (BALF) samples obtained from horses with the summer pasture endophenotype of equine asthma. ANIMALS 8 horses with the summer pasture endophenotype of equine asthma. PROCEDURES BAL was performed bilaterally (right and left lung sites) with a flexible videoendoscope passed through the left or right nasal passage. During lavage of the first lung site, a BALF sample was collected by means of either gentle syringe aspiration or mechanical suction with a pressure-regulated wall-mounted suction pump. The endoscope was then maneuvered into the contralateral lung site, and lavage was performed with the alternate fluid retrieval technique. For each horse, BAL was performed bilaterally once with and once without premedication with NBB (21-day interval). The BALF samples retrieved were evaluated for volume, total cell count, differential cell count, RBC count, and total protein concentration. RESULTS Use of syringe aspiration significantly increased total BALF volume (mean volume increase, 40 mL [approx 7.5% yield]) and decreased total RBC count (mean decrease, 142 cells/μL), compared with use of mechanical suction. The BALF nucleated cell count and differential cell count did not differ between BAL procedures. Use of NBB had no effect on BALF retrieval. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that retrieval of BALF by syringe aspiration may increase yield and reduce barotrauma in horses at increased risk of bronchoconstriction and bronchiolar collapse. Further studies to determine the usefulness of NBB and other bronchodilators during BAL procedures in horses are warranted.
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