Brucellosis is a common zoonotic disease in Egypt. However, there are limited data available on the genetic diversity of brucellae circulating in Egypt and other Mediterranean areas. One hundred and nine Brucella (B.) strains were isolated from different animal species in thirteen Egyptian governorates. Multi-locus variable number tandem repeats (VNTRs) analysis (MLVA-16) was employed to determine the geographical relatedness and the genetic diversity of a panel of selected Egyptian strains (n = 69), with strains originating from Italy (n = 49), Portugal (n = 52), Greece (n = 63), and Tunisia (n = 4). Egyptian B. melitensis strains clustered into two main clusters containing 21 genotypes. Egyptian B. abortus strains clustered into three main clusters containing nine genotypes. The genotypes were irregularly distributed over time and space in the study area. Egyptian strains of B. melitensis showed MLVA-16 patterns closer to that of Italian strains. Egyptian B. abortus strains isolated from cattle share the same genotype with strains from Portugal and similar to strains from Italy with low genetic diversity. Strains with similar MLVA patterns isolated from different governorates highlight the movement of the pathogen among governorates. Hence, it may also reflect the long endemicity of brucellosis in Egypt with earlier dispersal of types and great local genetic diversity. Open markets may contribute to cross-species transmission and dissemination of the new types nationwide. The presence of West Mediterranean lineages of B. melitensis and relatedness of B. abortus strains from the studied countries is a result of the socio-historical connections among the Mediterranean countries. Transnational eradication of brucellosis in the Mediterranean basin is highly demanded.

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. melitensis biovar 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 popul...

Aeromonas species are increasingly recognized as enteric pathogens. Faecal samples from 20 cow, 45 sheep; 60 goat and 60 camels were examined for the presence of Aeromonas species, which was also sought in the available drinking water (55 well water and 52 drinking chlorinated tap water were also examined). Aeromonas species was isolated more frequently from goats (21.7%) than from other animal groups sampled and isolated more frequently from well water (38.2%) than chlorinated supplies (23.0%). A. hydrophilia was the most dominant species isolated from different kinds of samples (13.4%). Whereas A. sobria and A. caviae were isolated in much lower rates 4.7 and 2.1% respectively. There was significant association between the isolation of Aeromonas species from all animal faeces and its presence in drinking water. All isolated strains were examined for the characteristics that are reputed to have roles in pathogenicity. The data reported in this study indicates that the distributions of virulence factors, that regulate the pathogenicity of Aeromonads, are different in clinical and enviromental samples. Aeromonas isolates exhibited multi-drug resistanc amoxicillin, carbenicillin and ampicillin. The most potent antibiotics against Aeromonas species isolated in this study were ceftriaxone, ceftazidime, cefotaxime, cefepime. Essential oils have been tested for in vitro and in vivo antimicrobial activity. Clove, Olive and Peppermint oil exhibited a wide spectrum of antimicrobial activity against all strains used in this study, showed a zone of inhibition ranging from 10.00±0.8 to 14.82±0.41 mm in diameter. Minimum Inhibitory Concentration (MIC) for selected oils ranged from 12.8 to 25.6 mg mL −1 . Treatment of mice with essential oil for 15 days led to enhance antibody levels in all treated groups and significant clearance of A. hydrophilia from animals. The treated animals had minimal histopathological changes and lower bacterial loads in the organs examined. In conclusion, these findings indicate that aeromonads have the potential to cause human illness and confirm the role of water as vehicles for Aeromonas diseases. This study also demonstrated that the multi-factorial nature of the diseases and the influence of environmental conditions in the expression of the putative virulence properties. These results suggested the potential value of essential oils as an additional or supporting treatment in gastrointestinal inflammations.

Klebsiella pneumoniae is a Gram-negative enterobacterium that has historically been and currently remains, a significant cause of human disease and several kinds of infections in animals. In the present work, trials for the isolation of Klebsiella pneumoniae from diseased and apparently healthy farm animals (cows, sheep, goats and camels) were done for recognition of Klebsiella pneumoniae subspecies. It was noticed that there was a marked variation between incidences of Klebsiella pneumoniae subspecies in examined animals as regards to health condition. The frequency was greater among samples collected from diseased animals 25.2% as compared with apparently healthy one 5.5%. It was found that there was great difference between the prevalence of Klebsiella isolated from various animal origins. On biochemical identification Klebsiella pneumoniae subsp. pneumoniae was the most prevalent followed by Klebsiella pneumoniae subsp. ozaenae and Klebsiella pneumoniae subsp. Rhinoscleromatis. Klebsiella pneumoniae subsp. rhinoscleromatis was not isolated from apparently healthy animals. The in vitro sensitivity of isolates of Klebsiella pneumoniae subspecies recovered from different animal species to 23 antimicrobial agents was tested. It was found that were resistance to cefoxitin, cefotaxime, cefoperazone, ceftazidime, ceftriaxone, aztreonam, amoxicillin and ampicillin. The most potent antibiotics showing 100% activity against Klebsiella pneumoniae subsp. isolated in this study were imipenem, ciprofloxacin, norfloxacin, gentamicin and kanamycin. While 96.2% of all examined isolates were sensitive to amoxicillin/clavulanic acid and ticarcillin/clavulanic acid. SDS-PAGE analysis showed that CPSs of Klebsiella pneumoniae subspecies contained wide variety of different molecular weights which ranged from 15.52 kDa to106.29 kDa and gave 10-13 bands. Evaluation of humoral immune response of mice immunized with CPSs was done using ELISA. It was found that the highest immune response was obtained 14 days post 1st dose of immunization with extracted CPSs of the three Klebsiella pneumoniae subspecies. When mice were immunized with CPSs of Klebsiella pneumoniae subspecies, they were protected against virulent challenge with homologous or heterologous strains, as a result the mortality rates were reduced from 80, 75 and 65 to 5 to 15% of Klebsiella pneumoniae subsp. pneumoniae, Klebsiella pneumoniae subsp. ozaenae and Klebsiella pneumoniae subsp. rhinoscleromatis, respectively. Determination of the reisolation rates of Klebsiella pneumoniae subspecies from lung, liver and spleen were done on both immunized mice and challenged and on control mice. Histopathological studies have been done on both dead infected non-immunized and immunized mice. Lungs were the main organs that showed macroscopic and microscopic pathological changes. Finally, ELISA has been used for detection of CPSs of Klebsiella pneumoniae subspecies antibodies in sera of examined animals. The sensitivity of ELISA using CPSs extracted from Klebsiella pneumoniae ...

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