Virulence traits of avian pathogenic (APEC) and fecal (AFEC) <Emphasis Type="Italic">E. coli</Emphasis> isolated from broiler chickens in Algeria

Avian pathogenic E. coli (APEC) is the etiologic agent of avian colibacillosis, the most common disease responsible for chicken morbidity in the world. Although multiple virulence-associated factors were identified, their prevalence in Algeria is still poorly known. In the present research, 92 avian pathogenic E. coli (APEC) isolates were recovered from broilers with clinical signs and lesions of colibacillosis. In addition, 32 E. coli isolates collected from feces of healthy birds (AFEC) were included for comparison. All isolates were investigated by PCR for the presence of a total of 11 virulence-associated genes described for avian pathogenic (iroN, ompT, hlyF, iss, iutA, and fimC) and diarrheagenic E. coli (eae, stx, elt/est, ipaH, and aggR). The sensitivity of 39 APEC isolates to 16 antibiotics was also determined using antimicrobial pretreated microplates. Here, we report that 98% of the examined isolates host at least one of the tested virulence factors. The most prevalent genes in APEC were iutA (90.6%), ompT (86.9%), and iss (85.8%); whereas, iutA (78.1%), fimC (78.1%), and iroN (68.7%) were the highest prevalent genes in AFEC. Our data showed that none of the AFEC isolates harbor any of the tested diarrheagenic genes. Moreover, only elt/est (5.4%), stx (2.1%), and ipaH (2.1%) genes were carried by APEC isolates. We further established that ceftazodime, ceftiofur, mequindox, amoxicillin/clavulanic acid, and meropenem were the most efficient antibiotics against the analyzed APEC isolates. Overall, our findings provide more insights about APEC and AFEC virulence potential in Algeria which could participate in the fight against colibacillosis.     

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.     

Bovine blood neutrophil acyloxyacyl hydrolase (AOAH) activity during endotoxin and coliform mastitis

The dynamics of blood neutrophil acyloxyacyl hydrolase (AOAH) activity, the appearance of endotoxin (lipopolysaccharide, LPS) in blood and the role of blood neutrophil AOAH in the severity of Escherichia coli and endotoxin mastitis were investigated in early postpartum dairy cows experimentally challenged with either endotoxin (n = 6) or E. coli (n = 6). The AOAH activity of blood neutrophils started to decrease significantly at post challenge hours (PCH) 6-24 and 12-24 in the endotoxin and E. coli-challenged groups, respectively; it returned to pre-challenged values at PCH 48 in both endotoxin- and E. coli-challenged groups. The cows were classified as moderate and severe responders according to milk production loss in the non-challenged quarters at PCH 48. There were no severe responders in the endotoxin-challenged group. In the E. coli-challenged group, only 1 severe responder was identified. The pre-challenge neutrophil AOAH activity of the severe responder was approximately 30% lower than that of moderate responders. No LPS was detected in the plasma of endotoxin-challenged cows; neither was it found in the plasma of moderate responders in the E. coli-challenged group at any PCH. However, at PCH 6, a remarkable amount of LPS was detected in the plasma of the severe responder from the E. coli-challenged group. Furthermore, neutrophil AOAH activity was increased by approximately 70% in the severe responder at PCH 6, but it increased by only approximately 15% in moderate responders. This was followed by a decreased neutrophil AOAH activity at PCH 12-24 and 24-72 in moderate and severe responders, respectively; the decreased AOAH activity at those PCH was more pronounced in the severe responder. The pronounced decreased neutrophil AOAH activity during mastitis often coincided with extreme leukopenia, neutropenia and a maximal number of immature neutrophils in the blood. Our results demonstrate that a decrease in neutrophil AOAH activity results in the appearance of LPS in the blood, and low blood neutrophil deacylation activity could be considered as a risk factor for severe clinical coliform mastitis.     

Unravelling the impacts of disturbance type and regeneration on movement of threatened species

Landscape Ecology - Anthropogenic disturbance alters animal movements. Large mammals require vast areas to meet their needs, and they encounter anthropogenic disturbances frequently during daily...     

Concentrations of metals (zinc, copper, cadmium, and mercury) in three domestic ducks in France: Pekin, Muscovy, and Mule ducks

The role of different factors such as biological material (tissues, organs) and trophic condition (overfeeding or not) in the metal accumulation was studied in three genotypes of ducks (Pekin, Muscovy, and Mule) under breeding conditions. Results showed that overfeeding decreased the concentration in Cd, Cu, and Zn through the dilution process. In contrast, mercury concentration increased with this method. A relation between lipidic metabolism of genotypes and the distribution of this metal in biological material was found. Domestic ducks were little contaminated, but a low chronic contamination in Cd was observed, probably coming from the food. Due to the low levels of contamination observed in these breeding ducks, they can be considered as a good control for further contamination studies and comparison with accumulation levels recorded in the field. The impact of feeding condition on accumulation showed the importance of taking into account the life cycle of birds before studying their contamination and the impact of pollutants.     

Comparison of genome segments 2, 7 and 10 of bluetongue viruses serotype 2 for differentiation between field isolates and the vaccine strain

Bluetongue (BT) virus serotype 2 (BTV 2) was first confirmed in Tunisia in February 2000 and has since spread northward and westward, infecting several other countries and islands, including Corsica, where clinical disease was reported in October 2000. BT was again reported on the Island in July 2001, some six months after a vaccination campaign against BTV 2. The molecular relationship between isolates of the BTV 2 Corsican wild-type viruses from 2000 and 2001, and the attenuated BTV 2 vaccine were determined by comparing corresponding sequences of genome segments 2, 7 and 10 with each other and with already published sequences available in the genome database. Complete genetic stability was observed between the isolates of the Corsican BTV 2. There was some divergence between the nucleotide sequences of segment 10 obtained from the wild-type and vaccine virus strains. Based on these differences, primers were selected that could be used in RT-PCR to differentiate between the wild-type and the vaccine viruses.     

Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

F4 receptor-independent priming of the systemic immune system of pigs by low oral doses of F4 fimbriae

Oral administration of F4 fimbriae of Escherichia coli induces intestinal mucosal immune responses in F4 receptor-positive (F4R(+)) pigs, but not in F4R(-) pigs. We examined whether F4 fimbriae in F4R(-) animals behave like a food antigen and can induce oral tolerance. Therefore, F4R(+) and F4R(-) pigs were fed 2mg of F4 and challenged i.m. to evaluate the effect of oral F4 on the systemic immune system. As control antigen, two different oral doses (2 and 600 mg) of OVA were used. Thirty days after the i.m. OVA challenge, the OVA-specific serum IgG titre in 600 mg-fed pigs was lower than that in non-fed animals, indicating that tolerance was induced. Conversely, in the 2mg-fed pigs a rapid increase of OVA-specific IgG with higher titres than those in non-fed pigs was seen following challenge, indicating a priming of the systemic immune system. A similar priming was seen in both F4-fed F4R(-) and F4R(+) pigs. Upon challenge, non-fed pigs displayed a primary immune response with a slow increase of F4-specific serum IgG, whereas F4-fed F4R(-) and F4R(+) pigs showed secondary responses with a rapid increase of serum IgG. This was expected in F4R(+) pigs, as in these animals oral F4 induces F4-specific antibody-secreting cells in the spleen, suggesting a priming of the systemic immune system. However, also the F4-fed F4R(-) pigs displayed a secondary response, despite the failure to detect a response upon oral F4 administration. These findings suggest that the F4 antigen, at a dose of 2 mg, behaves like a common food antigen in F4R(-) pigs, namely it induces a systemic priming.     

Density and Organic Matter Influence on Egg‐laying and Hatching of Branchiura sowerbyi (Oligochaeta)

This study aimed to assess the combined effects of density of adults of Branchiura sowerbyi and the source of organic matter on the production of eggs and young species. The experimental units used were 250‐mL beakers with 100 mL of fine sand (0.074–0.210 mm) and 150 mL of chlorine‐free water with constant aeration at the temperature 25 C. The experiment was conducted based on a two‐factor model with six treatments and three repetitions, applied in a completely random design. The densities 5 individuals/38.5cm² (D5; 1298/m²) and 8 individuals/38.5cm² (D8; 2078/m²) were assessed with commercial fish feed (FF), commercial poultry feed (PF), and turkey manure (TM) as sources of organic matter. A total of 1223 cocoons and 2138 eggs were produced, with average hatching of 40.0 ± 15.1%. The highest number of cocoons and eggs was found with the use of PF, D8 having produced the highest number of cocoons and eggs. The hatching rate was not influenced by the source of organic matter or density and no influence was registered from the density of adults on the survival. The PF provided the highest growth of adults, and generally the use of TM provided the worst performance.