Camel - calves diarrhoea instigated by Escherichia coli and detection of some virulence - associated markers

Bacteriological examination of internal organ specimens including intestine, liver, spleen, lungs and kidneys [45] and faecal samples [77] from necropsied and diarrhoeic camel-calves respectively that had severe episodes of diarrhoea, revealed the isolation of Escherichia coli from 33/ 45 [73.3%] of the necropsied specimens and 58/ 77 [75.3%] of the faecal samples. Eleven sero-groups were detected, with high prevalence of O3 [19.8%], OM [17.6%] O80 [16.5%]. Two enterohaemorrhagic, enterotoxigenic O157 isolates [2.2%] were also detected one from each group of specimens. E. coli isolation from faecal specimens revealed wider serogroups range as O91, O125 and O29 were additionally detected from faecal samples and not from the organ specimens. Investigating of some virulence markers of the isolates revealed that the haemolytic activity was expressed significantly higher [p < 0.01] than the rest of investigated markers as 43/91 [47.3%] were haemolysin producers. The detection of the VT2 gene was revealed in 34/91, [37.4%], Shiga-like toxin production in 26/91, [28.6%] and Congo red binding affinity was expressed in 31/91 [34.1%] of the isolates. The highest percentage of hydrophobic activity was [81.2 +/- 5.7] by the O163 isolates, whereas the lowest activity [40.2 +/- 3.2] was recorded by the O125 isolates. It was observed that the highest expression of virulence markers was among the isolates of O157 [2 isolates], O26 [3 isolates] and O163 [2 isolates], as these isolates expressed full range of the investigated virulence markers. The rest of E. coli isolates from other serogroups revealed abridged virulence criteria. Antibiotic sensitivity testing of E coli revealed highest sensitivity against gentamicin [94.4%], trimethoprim sulphamethazole [91.2%], streptomycin [87.9%] and cephalosporine [84.6%], It was concluded that highly virulent E. coli strains induced infection in camel-calves, and their treatment with the recommended antibiotics could curb the episode of the diarrhoea in few days

Bacteriological and immunological studies on escherichia Coli Isolates recovered from diarrhoeic and contact apparently healthy sheep with histopathological changes encountered

Bacteriolological examination of organ .specimens collected from 18 necropsied local breed sheep, that had history of diarrhoea revealed the isolation of Escherichia coli in 13/18 [72.2%] of the animals, whereas in the faecal samples from diarrhoeic and contact apparently healthy sheep the incidence of E. coli was 42/59 [71.2%] and 17/35 [48.6%] respectively. The identified 72 E. coli isolates were affiliated to 9 .serogroups, O76 [13], O16 [10], O5 [10], O28 [8], O117 [7]. O145 [5], O91 [3], O6 [3], O157 [3] and untypable [10]. All serogroups adhered better on enterocytes cell-model than oil HEp-2 cells, as 53/72 [73.6%] of the isolates adhered with an average of 63.5 +/- 6.2 E. coli enterocyte, whereas on HEp-2 cells the adhesion was .significantly lower [P < 0.05] as 45/72 [62.5%] of the isolates were adherent with an average of 53.9 +/- 6.7 E, coli/ cell. In verotoxin-production assay, 41/72 [56%] of the isolates were verotoxin producer. The OMPs extract of O76, O16 and O5 [which were the most dominant serogroups] were similar and had dense peptide bands between the range of 35k Da to 42 kDa. Lighter peptide bands were also demonstrated in the higher and lower gel ends. Immunoblotting analysis, also showed strong immunogenic protein bands at 35-37kDa and 40 - 42k Da against sera from infected sheep as well us against sera from rabbits immunized with a pool from the extracted OMPs. Using sera from immunized rabbits as adhesion inhibitors in the previously mentioned cell models revealed significant inhibition [P < 0.01] of the adhesion pattern on both previously mentioned cell models. The inhibition averaged 63.7% reduction in the adhesion capacity of enterocyte cell model [23.0 +/- 5.7 E coli/cell], whereas the inhibition of adhesion averaged 52.3% in HEp-2 cell-model [20.2 +/- 3.4 E. coli/cell]. Histopathological findings of necropsied sheep revealed intestine mucosa had coagulative necrosis associated with inflammatory cells infiltrations. Diffuse desquamations and erosions in the mucosa of the villi. Aggrigation of epithelioid and lymphoid cells was also demonstrated in the lamina propria. Lung, interstitial stroma was thickened; bronchioles and alveoli had focal epithelioid stratification, desquamations and ulcerations. Liver was infiltrated by neutrophils, macrophages, lymphocytes and Kupffer cell proliferation. Kidney showed coagulative necrosis of renal tubules. Heart and spleen sections showed also pathological changes which also confirmed the systemic infection with attaching and toxigenic E. coli. It was concluded that OMPs can be used as an effective, highly immunogenic vaccine for controlling infection with E. coli in sheep as well as other animals

Oedematous skin disease: a recurring infection in buffaloes

Eighteen Corynebacterium pseudotuberculosis were isolated from 30 specimens [9 biopsies from closed skin lesions and 21 exudate specimens from open lesions] collected from 27 buffalos with symptoms of skin infection. C. pseudotuberculosis was isolated in pure cultures from 6 out of 9 biopsies and in mixed cultures from 12 out of 21 exudate specimens. The majority of isolates were of biotype II [15 isolates], whereas only 3 isolates were of biotype I. The pattern of isolation of both biotypes I and II from biopsies and exudate specimens was similar. Both biotypes I and II differed in their course of pathogenicity in G. pigs. C. pseudotuberculosis was not isolated from blood suckling and biting flies [236 files] existed on the infected animals and in their premises. This might rule out the role of vectors in transmission of infection. In-vitro antibiotic sensitivity testing revealed that C. pseudotuberculosis isolates were most sensitive to penicillin 89%, amoxycillin 89%, erythromycin 83%, chloramphenicol 78% and ampicillin 78%. The histological examination of skin biopsies from the diseased buffalos showed inflammatory reaction due to the bacterial infection

Hyaluronic acid mediated adhesion of pasteurella multocid a to different host cells

The in vitro adhesion of 12 Pasteurella multocida [isolated from different sources] was studied on turbinate and tracheal epithelial cells from porcine and bovine as well as on HeLa-S3 cells. All serotype A isolates [8 from rabbits and one from human] adhered to all cell models with distinctively higher affinity to HeLa cells than turbinate or tracheal cells. Serotype B isolate from bovine [one isolate] failed to adhere on any cell model, whereas serotype D from rabbit [one isolate] adhered weakly to all cell models. The same serotype [D] from buffalo [one isolate] adhered weakly to HeLa cells and bovine cells and failed to adhere to the porcine cells. Altering the P. multocida outer surface characteristics using specific antiserum or protein denaturing agents as heat and acidity [95C 1 hour and pH 5], trypsin [250 units/ml], pronase-E [100 ug/ml] did not affect the bacterial ability to adhere to host cells. Only hyaluronidase treatment [50 units/ml] reduced the adhesion drastically. On the other h and, blocking the host cell receptors with N-acetyl glucosamine or D-glucuronic acid [hyaluronic acid monomeres] did not inhibit the adhesion. Only locking the HeLa cells with hyaluronic acid [10 mg/ml] reduced or stopped the ability of P. multocida to adhere to the HeLa cells

Study on virulence factors of pasteurella multocida isolated from different sources

In this study 15 Pasteurella multocida isolates [collected from different sources] were investigated for virulence factors, namely, capsular and outer membrane protein [OMP] components, binding properties to fibrinogen and fibronectin and the bacterial surface hydrophobicity. All serotype A [7 isolates] demonstrated hyaluronic acid capsule when tested by the hyaluronidase depolymerization test. The electrophoretic analysis of the OMP preparations revealed 37 kDa protein-b and s in all serotype A and E isolates, which was analogous to the major P. multocida H protein or porin H [channel-forming protein]. OMP preparations from serotype B isolates revealed 32 kDa protein b and s, similar to the characteristic protein b and s found in all P. multocida serotype B isolated from hemorrhagic septicemia-positive cases. Other protein b and s, 21, 24, 30, 31, 34, 36, 53 and 64 kDa were demonstrated in different serotypes, which could be related to the bacterial adhesion characteristics