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Multi-drug resistance (MDR) Escherichia coli have become a major threat due to their ability to overcome different types of antibiotics. However, Extended Spectrum β-lactamase E. coli (ESBLE) imposes an additional threat due to their ability to resist the 3rd generation cephalosporins. Accordingly, our study aimed to investigate the antibiogram profile of ESBLE isolates obtained from sheep. A Total of 40 ESBLE isolates were included in this study which represents sheep fecal and milk samples. Twelve antibiotics were selected to perform antibiotic sensitivity tests following standard microbiological methods. The results of the study showed that the highest resistance percentages were recorded for tetracycline 97.5%, ciprofloxacin 80%, trimethoprim/sulfamethoxazole 65%, and streptomycin 57.5%. While other antibiotics recorded lesser values. On the other hand, all isolates were susceptible to gentamycin and tobramycin each at 92.5%, followed by chloramphenicol and levofloxacin each at 82.5% and nitrofurantoin 72.5%. While fewer values of sensitivity were recorded for streptomycin, trimethoprim/sulfamethoxazole, azithromycin, nalidixic acid, ciprofloxacin and, tetracycline. The study concluded that ESBLE of sheep origin that have additional resistance features to other antibiotics could be a major threat for spreading resistance and contaminating the environment and finally impose negative impact for response to antibiotic treatment in humans.

Shepherd dogs have been implemented in the transmission and distribution of many threatening pathogens. The presence of extended-spectrum-cephalosporin resistant Escherichia coli (ESCR E. coli) in dog feces can constitute a significance risk to human health due to transmission of antibiotics resistance from dogs to humans, other animals and the surrounding environment. Therefore, in this study, phenotypic and molecular characterization of fecal ESCR E. coli were investigated in shepherd dogs accompanied sheepherders in urban areas. Sixty-seven fresh fecal samples were collected from shepherd dogs from different regions of Mosul city. Bacteriological examination of ESCR E. coli was done using MacConkey agar with cefotaxime followed by subsequent PCR confirmation of the CTX-M gene using specific primers and molecular characterization using specific primers directed to CTX-M-1, 2 and 9 groups. The results of bacterial examination showed successful confirmation of ESCR E. coli which has been isolated from fecal samples of shepherd dogs 58.2% (39/67). In addition, detection of CTX-M gene was confirmed in 53.7% (36/67) of E. coli isolates. Furthermore, molecular characterization of CTX-M gene revealed the presence of only one genotype belongs to CTX-M-1. However, both of CTX-M-2 and CTX-M-9 genotypes were not detected in this study. This study concluded that shepherd dogs have an essential role in carrying and spreading of ESCR E. coli especially in urban regions.

Paratuberculosis or Johne’s disease is a chronic debilitating disease mainly infects ruminants and caused by Mycobacterium paratuberculosis. Previous serological studies in Mosul city confirm the presence of positive reactants for paratuberculosis in cattle. However, culture methods to confirm the disease need a long incubation period and also special media. Raw cow’s milk is considered as potential source for transmission of M. paratuberculosis in cows’ herds. Accordingly, this study aimed to detect the presence of M. paratuberculosis specifically in the raw cow’s milk using polymerase chain reaction (PCR) technique as a rapid, sensitive and reliable method. A total of 50 samples of raw cow’s milk were collected from cows suffering from emaciation and unresponsive to antibiotic treatment. All the samples were subjected to DNA extraction and direct amplification PCR. The results showed that 3 (6%) out of 50 milk samples were positive for M. paratuberculosis. This is the first study in Mosul city that confirms the presence of M. paratuberculosis in raw cow’s milk using PCR technique. In conclusion, raw cow’s milk could be an important source for M. paratuberculosis infection in dairy cows, and also PCR technique could be helpful in rapid diagnosis of paratuberculosis.

Lipopolysaccharide (LPS) of P. multocida B:2, a causative agent of haemorrhagic septicaemia (HS) in cattle and buffaloes, is considered as the main virulence factor and contribute in the pathogenesis of the disease. Recent studies provided evidences about the involvement of the nervous system in pathogenesis of HS. However, the role of P. multocida B:2 immunogens, especially the LPS is still uncovered. Therefore, this study was designed to investigate the role of P. multocida B:2 LPS to induce pathological changes in the nervous system. Nine eight-month-old, clinically healthy buffalo calves were used and distributed into three groups. Calves of Group 1 and 2 were inoculated orally and intravenously with 10 ml of LPS broth extract represent 1 × 1012 cfu/ml of P. multocida B:2, respectively, while calves of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. Significant differences were found in the mean scores for clinical signs, post mortem and histopathological changes especially in Group 2, which mainly affect different anatomic regions of the nervous system, mainly the brain. On the other hand, lower scores have been recorded for clinical signs, gross and histopathological changes in Group 1. These results provide for the first time strong evidence about the ability of P. multocida B:2 LPS to cross the blood brain barrier and induce pathological changes in the nervous system of the affected buffalo calves.

Haemorrhagic septicaemia (HS) is an acute, fatal, septicaemic disease of cattle and buffaloes caused by one of two specific serotypes of Pasteurella multocida B:2 and E:2 in Asian and African, respectively. It is well known that HS affect mainly the respiratory and digestive tracts. However, involvement of the nervous system in pathogenesis of HS has been reported in previous studies without details. In this study, nine buffalo calves of 8 months old were distributed into three groups. Animals of Group 1 and 2 were inoculated orally and subcutaneously with 10 ml of 1 × 10(12) cfu/ml of P. multocida B:2, respectively, while animals of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. All calves in Group 1 and Group 3 were euthanised after 504 h (21 day) post-infection, while calves in Group 2 had to euthanise after 12 h post-infection as they develop sever clinical signs of HS. Significant differences were found in Group 2 in the mean scores of clinical signs, gross and histopathological changes which mainly affect different anatomic regions of the nervous system. In addition, successful bacterial isolation of P. multocida B:2 were obtained from different sites of the nervous system. On the other hand, less sever, clinical, gross and histopathological changes were found in Group 1. These results provide for the first time strong evidence of involving of the nervous system in pathogenesis of HS, especially in the peracute stage of the disease.

Pasteurella multocida is responsible for one of the major animal diseases with economic importance in both developed and developing countries. P. multocida B:2 causes haemorrhagic septicemia (HS) in cattle and buffaloes, which result in heavy economic losses through direct effect, resulting in high mortality rates and indirect effects through impairment of the animal productivity. It is well known that HS affects mainly the respiratory and digestive tracts of host animals. However, involvement of the reproductive system in the pathogenesis of HS has not been reported previously. This study was designed to present detailed detection and histopathological changes of the reproductive system and mammary glands of buffaloes experimentally infected with P. multocida B:2. Briefly, nine healthy buffalo calves aged 8-month-old were divided into three groups of three calves each. Calves from groups 1 and 2 were inoculated subcutaneously and orally with 10 mL of 1 × 1012 cfu/mL of P. multocida B:2, respectively, while calves of group 3 were inoculated orally with 10 mL of phosphate buffer saline as a sham control. All the calves in group 1 had to be euthanized after 12 h post-infection, while calves in group 2 were euthanized after day 21 post-infection. Successful isolation and PCR confirmation of P. multocida B:2 was achieved from different parts of the reproductive system, including ovary, oviduct, uterine horn, uterine body and vagina as well as mammary glands and supramammary lymph nodes of the buffaloes in group 1 after 12 h post-infection. However, P. multocida B:2 was not detected in the reproductive organs of buffaloes in group 2 and group 3 after 21 days post-infection. Significant differences (P

Background Brucella melitensis is the most important pathogenic species of Brucella spp. which affects goats and sheep and causes caprine and ovine brucellosis, respectively. Serological tests for diagnosis of brucellosis such as Rose Bengal plate test (RBPT) and enzyme-linked immunosorbent assay (ELISA) usually utilize smooth lipopolysaccharides (S-LPS) as a diagnostic antigen which could give false positive serological reactions. Outer membrane proteins (OMP) of B. melitensis have been used as alternative diagnostic antigens rather than S-LPS for differential serological diagnosis of brucellosis, mainly in ELISA with single recombinant OMP (rOMP) as a diagnostic antigen. Nevertheless, the use of single format mainly showed lack of sensitivity against the desired rOMP. Therefore, this study aimed to determine whether a newly developed rOMPs indirect ELISA (rOMPs I-ELISA), based on combination of rOMP25, rOMP28 and rOMP31of B. melitensis, has a potential benefit for use in the serodiagnosis of brucellosis. Methods In this study, omp25, omp28 and omp31 of B. melitensis were cloned and expressed using prokaryotic pET-32 Ek/LIC system and their respective rOMPs were combined as one coating antigen to develop rOMPs I-ELISA. Three groups of BALB/c mice were used to elicit antibody response. Group 1, infected with B. melitensis strain 0331 field strain; group 2, injected with B. melitensis Rev.1 vaccine strain and group 3, infected with Yersinia enterocolitica O:9. Antibody responses in three groups of mice were investigated using Rose Bengal plate test (RBPT) and rOMPs I-ELISA. Results The production of rOMP25, rOMP28 and rOMP31 of B. melitensis were achieved and Western immunoblotting analysis demonstrated their reactivity. The RBPT was unable to differentiate the vaccinated mice (group 2) and mice infected with Y. enterocolitica O:9 (group 3) and categorized them wrongly as positive for brucellosis. In contrast, the rOMPs I-ELISA was able to differentiate the mice infected with B. melitensis strain 0331 (group 1) from both of group 2 and group 3, and recorded 100% sensitivity and 100% specificity. Conclusions The results of this study suggested that rOMPs of B. melitensis has potential diagnostic ability to differentiate the FPSR in serological diagnosis of brucellosis.

Hemorrhagic septicemia (HS) is an acute septicemic disease principally affecting cattle and buffaloes caused by specific serotypes B:2 and E:2 of Pasteurella multocida in Asia and Africa, respectively. Despite continuing researches on pathogenesis of P. multocida for several decades, the mechanisms by which these bacteria develop the diseases are poorly understood. Although the involvement of the nervous system in the disease progress of HS is rare under natural conditions, few reports indicated the involvement of the nervous system in outbreaks of HS in cattle and buffaloes. Additionally, recent pathogenesis studies in both mouse and buffalo experimental models reported the involvement of nervous system due to P. multocida B:2, with bacteriological and histopathological evidences. In this review, we summarized and discussed the updates on the involvement of the nervous system in pathogenesis of HS focusing on clinical signs, pathological and pathophysiological changes. [J Adv Vet Anim Res 2015; 2(3.000): 252-262]