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3. Evaluation of the General Organization of Veterinary Services control program of animal brucellosis in Egypt: An outbreak investigation of brucellosis in buffalo
H. I. Hosein, Hoda Mohamed Zaki, Nesreen Mohamed Safwat, Ahmed M. S. Menshawy, Sherin Rouby, Ayman Mahrous and Bahaa El-deen Madkour
Veterinary World, 11(6): 748-757
Background and Aim: Brucellosis is a major constraint to livestock production in Egypt as well as many developing countries worldwide. Bovine brucellosis is an economically important disease with reproductive failure as a principal manifestation resulting in abortion, premature birth and decreased milk production in females, and orchitis and epididymitis in males. In spite of the efforts of Egyptian veterinary services to overcome brucellosis, the disease is still prevalent in both animals and humans and represents one of the most important public health hazards in Egypt. The aim of the present work was to investigate the efficacy of the control program implemented by the General Organization of Veterinary Services in Brucella infected buffalo farm on serological, molecular, cultural, and histopathological basis. Brucella melitensis biovar 3 was recovered from 6 buffalo-cows.
Materials and Methods: Blood samples were collected from a total of 750 non-vaccinated lactating buffalo-cows. These animals were proved positive for Brucella by the Egyptian brucellosis national program. Sera were tested using buffered acidified plate antigen test and rose Bengal test as screening tests and complement fixation test as a confirmatory test. Positive animals were separated for slaughtering under the supervision of the Egyptian veterinary authorities. Remaining animals were tested every 3 weeks with slaughtering of positive cases and this continued until the remaining animals revealed three successive negative serological tests. Different lymph nodes (prescapular, prefemoral, mediastinal, retropharyngeal, and supramammary) were collected from 11 Brucella seropositive buffalo-cows slaughtered after being confirmed serologically as Brucella infected cases. Samples were collected and processed for bacterial isolation and nucleic acid detection using polymerase chain reaction (PCR). Parts of these specimens were fixed in 10% neutral buffered formalin for 48 h then processed by paraffin embedding technique.
Results: "Test and slaughter" policy was applied on Brucella infected dairy buffalo farm. The program continued for 6 months with slaughtering of positive cases until the herd was proved Brucella free. B. melitensisbiovar 3 could be recovered from six buffalo-cows. Universal PCR confirmed Brucella on genus level and Bruce-ladder multiplex, PCR confirmed the presence of B. melitensis on the species level. Histopathological examination of Brucella-infected lymph nodes revealed massive rarified and depleted lymphoid areas of both sub-capsular and deep cortical lymphoid follicles, macrophage cells granulomatous reaction, as well as fat, infiltrates, and chronic vasculitis. The chronic nature of Brucella lesions has been confirmed in this study as indicated by the chronic vasculitis and collagen deposition.
Conclusion: Freedom status from brucellosis in this study required 6 months which are considered long time allowing the spread of infection to other localities especially under unhygienic conditions, husbandry system favoring mixed populations of different ages, sex, aborted and pregnant, and lack of controlled movement of animals. Therefore, effective control of animal brucellosis requires surveillance to identify infected animal herds, elimination of the reservoirs, and vaccination of young heifers. B. melitensis biovar 3 is the cause of the Brucella outbreak in buffalo which still remains the prevalent type of Brucella in Egypt. The disease runs a chronic course allowing further spread of infection.
2. Scrotal circumference: A predictor of testosterone concentration and certain attributes of seminal vesicles influencing buffalo male fertility
S. Mahmood, A. Kumar, R. Singh, M. Sarkar, G. Singh, M. R. Verma and G. V. P. P. S. R. Kumar
Veterinary World, 11(6): 739-747
Aim: The aim of this study was to evaluate the relationship of scrotal circumference (SC) with plasma testosterone, seminal vesicles (SVs) weight, and its secretion as measurable indicators of fertility and also to sequence and establish phylogenetic relatedness of certain SV protein genes with other species as such integrated approach is lacking.
Materials and Methods: Altogether, 59 apparently healthy male buffaloes sacrificed at slaughterhouse were selected (irrespective of breed) for measuring SC and collecting blood and paired SVs. The SC was measured at greater curvature using soft thread. In the present study, blood plasma testosterone, cholesterol, protein, and glucose in addition to SV fructose, citric acid and proteins in SV fluid were also estimated. The SV tissue was fixed in RNAlater for RNA extraction.Male buffaloes were categorized as per total SV weight into Group I (<5.0 g), Group II (5.0-7.84 g), and Group III (>8.0 g) and dentitions-I (≤18 months), II (18-24 months), and III (≥24 months) to assess the effect of weight and dentition age on SC, SV weight, and its certain secretions. Data were analyzed using linear model procedure including Tukey HSD test and Pearson's correlation coefficient. Variance inflation and condition index were also used to assess multicollinearity.
Results: Gross and histomorphological evaluation of SVs did not show any abnormality. Macronutrients (plasma protein, glucose, and cholesterol) showed non-significant (p>0.05) variation between groups. The SC and SV weight varied significantly (p<0.05) with a significant positive relationship with plasma testosterone, SV protein, fructose, and citric acid. In addition, testosterone concentration also showed increasing trend from Groups I to III but increased significantly (p<0.05) from Group II to III with positive and significant correlations with SV protein, fructose, and citric acid similar to SV weight and SC. Binders of sperm protein (BSP1, 3, and 5) genes (full length) were sequenced and established an evolutionary relationship which is lacking in buffalo.
Conclusion: The present findings established a significant positive correlation of SC with that of other fertility parameters related to SVs weight and its secretions: Fructose, citric acid, and protein (inclusive of BSPs sequenced full length), and testosterone. Therefore, the present integrated approach along with certain semen quality attributes reflecting epididymis function could be used as a predictive fertility marker for grading and selection of breeding bulls and their progenies to develop outstanding bull mother farm.
Keywords: male buffalo, morphology, scrotal circumference, seminal vesicles, sequencing, testosterone.
1. Calculate of withdrawal times of clenbuterol in goats to obtain safe times of slaughter
Lazuardi Mochamad, Bambang Hermanto and T. I. Restiadi
Veterinary World, 11(6): 731-738
Background and Aim: Clenbuterol as a β2-agonist drug was investigated according to the concentration of the drug available in the bodies of goats and according to the level of sensitivity of the instruments used for detection. The objective of the current study was to determine withdrawal times after giving a therapeutic dose that resulted in safe slaughters.
Materials and Methods: Five healthy male goats with a mean body weight of 20.64 kg were treated with a single dose of 5.10-3 mg/kg in the BW onto jugular vein. Whole blood samples of approximately 5 mL were taken in a time series at 5, 30, 60, 90, 150, 210, 270, 390, 510, 630, and 750 min. At 24 h posttreatment, all subjects were sacrificed, and 300 g samples of the liver were obtained. The plasma concentration and liver residue of the drug were observed by reverse-phase high-performance liquid chromatography.
Results: The drug reached a maximum concentration of 19.233±0.331 μg/mL at 5 min, and the elimination half-life was at 173.25 min. The limit detection was obtained at 0.053 μg/mL. A one-way analysis of variance between all goats showed that elimination of the clenbuterol in their bodies was similar (p=1.00), with a withdrawal time of 1,479.326 min and no residues in the liver (p<0.05).
Conclusion: Safe times for slaughter were determined to be at 2 days, 13 h, and 12 min as the 2nd safety factor (SF) time and 3 days, 1 h, and 58 min as the 3rd SF time with the liver organ free from residue.
Keywords: elimination half-life, new method for calculating withdrawal time, prescriptions for obtained β2-agonist, residues in liver.
23. Probiotic white cheese production using coculture with Lactobacillus species isolated from traditional cheeses
A. Ehsani, M. Hashemi, A. Afshari and M. Aminzare
Veterinary World, 11(5): 726-730
Aim: The aim of the present study was to investigate the viability of lactic acid bacteria isolated from traditional cheeses and cocultured in Iranian white cheese during ripening.
Materials and Methods: A total of 24 samples were isolated from 8 types of traditional cheeses in West Azerbaijan, Iran. Isolated species were cocultured with starter bacteria during the production of Iranian white cheese, and their viability was investigated up to 60 days of the refrigerated storage.
Results: Of 118 isolates of Lactobacillus, 73 isolates (62%) were confirmed as facultative heterofermentative and 45 isolates (38%) as obligate homofermentative. Of the facultative heterofermentatives, 28 isolates (24%) were Lactobacillus plantarum, 24 isolates (20%) were Lactobacillus casei, and 21 isolates (18%) were Lactobacillus agilis. Obligate homofermentatives were Lactobacillus delbrueckii (21%), Lactobacillus helveticus (14%), and Lactobacillus salivarius (3%). L. plantarum, L. casei and L. helveticus were found in high enough levels (106 CFU/g).
Conclusion: According to the obtained data, it is recommended that complex starters such as L. helveticus, L. plantarum, and L. casei can be used in industrial productions of cheese to obtain exclusive properties of traditional cheeses.
Keywords: heterofermentative, Lactobacillus, probiotic, starter, traditional cheeses.
22. The improvement of eggs quality of Mojosari duck (Anas javanica) with soybean husk fermentation using cellulolytic bacteria of Spodoptera litura
Sri Hidanah, Dady Soegianto Nazar and Erma Safitri
Veterinary World, 11(5): 720-725
Aim: This study was aimed to improve the quality of the eggs of Mojosari duck (Anas javanica) through complete feeding containing soybean husk was fermented using cellulolytic bacteria of Spodoptera litura.
Materials and Methods: This study consisted of three stages: The first stages, isolation and identification of cellulolytic bacteria from S. litura; the second stage, the fermentation of soybean husk through the application of bacterial cellulolytic isolate from the first stage; and the third stage, the application of the best complete feed formulation from the second stage to Mojosari duck.
Results: There are four dominant bacteria: Bacillus sp., Cellulomonas sp., Pseudomonas sp., and Cytophaga sp. Furthermore, the best reduction of the crude fiber of soybean husks is the use of Cellulomonas sp. bacteria. The final of the study, the quality of the eggs of Anas javanica, was improved, as indicated by cholesterol decrease from the yolk without the decrease of egg weight and eggshell thickness, although the decrease in egg yolk color was inevitable.
Conclusion: Soy husk fermentation using cellulolytic bacteria of S. litura was added to complete feeding can be performed to improve the quality of the eggs of Mojosari duck.
21. Effect of increasing levels of wasted date palm in concentrate diet on reproductive performance of Ouled Djellal breeding rams during flushing period
A. Allaoui, B. Safsaf, M. Tlidjane, I. Djaalab and H. Djaalab Mansour
Veterinary World, 11(5): 712-719
Aim: The aim of the study was to assess the effect of two levels of wasted date (WD) by replacing commercial concentrate on the reproductive performance of Ouled Djellal (OD) rams.
Materials and Methods: Eighteen mature (2-year-old) OD rams were equally allocated to three groups and fed during 11 weeks with one of three different experimental diets, that contained 0% (0 WD), 50% (50 WD), or 75% (75 WD) of WDs in concentrate diet. Live body weight (LBW), body condition scoring (BCS), scrotal circumference (SC), testicular weight (TW), sperm production and quality, plasma testosterone concentration (T), and sexual behavior (reaction time and number of mounts with ejaculation) were regularly recorded from every ram.
Results: LBW, SC, and TW changed significantly among diet groups and during the experimental period (p<0.001), the highest averages were recorded in (0 WD) group. LBW, BCS, SC, TW, semen volume, and percentage of the positive hypo-osmotic swelling test were (p<0.001) positively influenced by flushing period. Nevertheless, sperm concentration showed a significant (p<0.001) decrease at day 30, followed by a return to the initial values afterward. There were no differences (p>0.05) between diet groups for plasma testosterone concentration and semen attributes, except that (50 WD) group expressed the lowest overall value of semen concentration. Furthermore, neither time nor diet affected (p>0.05) sperm motility and reproductive behavior parameters.
Conclusion: It is possible to introduce WD as unconventional local feeding resources in flushing diet of breeding rams without disturbing their reproductive performance.
Keywords: body weight, flushing period, rams, semen, wasted date.
20. The study of effect of didecyl dimethyl ammonium bromide on bacterial and viral decontamination for biosecurity in the animal farm
Tippawan Jantafong, Sakchai Ruenphet, Darsaniya Punyadarsaniya and Kazuaki Takehara
Veterinary World, 11(5): 706-711
Aim: The aim of this study was to determine the effectiveness of the fourth-generation quaternary ammonium compounds, didecyl dimethyl ammonium bromide (DDAB), on the efficacy of bacterial and viral decontamination against pathogens commonly found in livestock industry including Salmonella infantis (SI), Escherichia coli, and avian influenza virus (AIV).
Materials and Methods: The DDAB was prepared at 500, 250, and 125 parts per million (ppm) for absent and present organic material. Meanwhile, 5% of fetal bovine serum in DDAB solution sample was used to mimic the presence of organic material contamination. 400 μl of each DDAB concentration was mixed with 100 μl of each pathogen (SI, E. coli, and AIV) and then incubated at room temperature or 4°C at various time points (5 s, 30 s, 1 min, 5 min, 10 min, 15 min, and 30 min). The activity of DDAB treatment was stopped using 500 μl of FBS. Each treatment sample was titrated on either deoxycholate hydrogen sulfide lactose agar plates or Madin-Darby canine kidney cells for bacteria and AIV, respectively. Each treatment was conducted in triplicates, and the pathogen inactivation was considered effective when the reduction factor was ≥ 3 log10.
Results: Our current study revealed that the DDAB inactivated SI, E. coli, and AIV under the various concentrations of DDAB, organic material conditions, exposure temperature, and exposure timing. In addition, the comparison of bactericidal and virucidal efficacy indicated that bacteria were more susceptible to be inactivated by DDAB as compared to viruses. However, DDAB showed marked inactivated differences in the absence or presence of organic materials.
Conclusion: The DDAB may be a potential disinfectant for inactivating bacteria and viruses, especially enveloped viruses, in livestock farms. It can be useful as a disinfectant for biosecurity enhancement on and around animal farm.