Patrizia Casagrande Proietti

Detection of Helicobacter spp. in gastric, fecal and saliva samples from swine affected by gastric ulceration.

The aim of this study was to evaluate the presence of Helicobacter (H.) spp. in swine affected by gastric ulceration. Stomachs from 400 regularly slaughtered swine were subjected to gross pathological examination to evaluate the presence of gastric ulcers. Sixty-five samples collected from ulcerated pars esophagea and 15 samples from non-ulcerated pyloric portions were submitted to histopathological and molecular analyses, to detect Helicobacter spp., H. suis and H. pylori by PCR. Feces and saliva swabs were also collected from 25 animals in order to detect in vivo the presence of Helicobacter spp.. Gastric ulcers were detected in 373 cases (93%). The presence of ulcers in association with inflammatory processes was further confirmed by histological examination. Forty-nine percent (32/65) of the ulcerated esophageal portions as well as 53% (8/15) of the non-ulcerated pyloric portions were positive for Helicobacter spp. by PCR. The Helicobacter spp. positive samples were also positive for H. suis, while H. pylori was not detected. These results were confirmed by restriction enzyme analysis. With regard to feces and saliva samples, 15/25 (60%) and 16/25 (64%) were positive for Helicobacter spp. PCR, respectively but all were negative in H. suis and H. pylori specific PCR.

A case of Candida guilliermondii abortion in an Arab mare

Ascending infections of equine uterus frequently result in placentitis and abortions; most of these infections are bacterial and are less commonly due to fungi. This report describes an abortion case in an Arab mare due to Candida guilliermondii that was diagnosed via cytological, histological, cultural and biomolecular assays. The histological lesions found were severe necrotizing placentitis associated with fetal pneumonia. To our knowledge this is the first case of C. guilliermondii abortion reported in equine species.

Cloning and expression of pigeon IFN-γ gene

This is the first paper describing the cloning of pigeon IFN-γ gene (PiIFN-γ) and the analysis of the in vitro expressed recombinant protein. The PiIFN-γ gene was identified by RT-PCR as a 498bp, fragment coding for a precursor protein of 165 amino acids instead of 164 amino acids, as observed in the other avian species. The recombinant protein was expressed in vitro by an eukaryotic system and the biological properties of the cytokine were tested using a chicken macrophage cell line. The high degree of amino acid and nucleotide identity, shared with the ChIFN-γ, and the fact that the pigeon protein was functional on chicken cells, indicates a cross-reactivity between pigeon and chicken IFN-γ. The detection of the PiIFN-γ could represent an useful instrument in understanding the role played by this cytokine in immune response related to vaccinations and infectious diseases in the pigeon.

Evaluation of intestinal bacterial flora of conventional and organic broilers using culture-based methods

Abstract The major bacteria colonizing the intestinal tract (ileum and caecum) of organic (O) and conventional (C) chickens were counted, isolated and identified by conventional methods. Chickens were obtained from 7 conventional and 7 organic chicken farms (n=203). Intestinal sampling was performed at different ages, every 10 days, starting at 20 days until 40 and from 20 days to 80 days of age, respectively, for conventional and organic birds. Statistical analysis was performed on two separate data sets (40 days of age and all ages).The comparison of C vs O systems was analyzed with univariate and multivariate procedures. There were large differences in bacterial counts in relation to the portion of intestine, the rearing system and the farms. In the ileum of conventional birds Enterobacteria were higher than in organic birds (7.03 vs 6.09 CFUxlog/g; P<0.05), whereas the contrary was observed for Lactobacilli (6.75 vs 7.07 CFUxlog/g; P<0.05). With respect to the other microflora, the effect of farm probably masked possible differences. The effect of rearing system was more visible in the caecum than in the ileum: Enterobacteria levels were higher in C than in O chickens (7.42 vs 7.05 CFUxlog/g; P<0.01), whereas Enterococci (7.65 vs 6.55 CFUxlog/g; P<0.05), Lactobacilli (7.85 vs 7.31 CFUxlog/g; P<0.05) and total aerobia (8.12 vs 7.66 CFUxlog/g; P<0.01) counts were higher in organic chickens. Multivariate analysis of caecum microflora showed the possibility of discriminating the rearing system. In the ileum of conventional birds Enterobacteria and total aerobia increased with age, while Lactobacilli decreased. In the O system, Enterobacteria, Lactobacilli and total anaerobia showed a similar trend, whereas total aerobia and Enterococci showed the opposite trend. A similar situation was observed in the caecum. Further investigations are necessary to better assess the role and effect of the enteric flora on the productive performance and on the health status of reared chickens.

Effects of oregano (Origanum vulgareL.) and rosemary (Rosmarinus officinalis L.) aqueous extracts on broiler performance, immune function and intestinal microbial population

ABSTRACT A 57-day study was performed to determine the effects of two aqueous extracts (AEs) on broiler performance, immune function and intestinal microflora. Four groups of 75 one-day-old female broilers (Ross308) received one of the following treatments: (1) a standard commercial feed (C); (2) C supplemented with 2 g/kg rosemary AE (R); (3) C supplemented with 2 g/kg oregano AE (O); (4) C supplemented with 1 g/kg oregano AE + 1 g/kg rosemary AE (OR). Individual body weight, average daily gain, average daily feed intake and feed conversion efficiency were determined at 1, 11, 22, 36 and 57 days. Sample collections for IgG titration and intestinal microflora examination were performed at 22 and 57 days. The addition of oregano AE alone or in combination with rosemary AEs improved body weight up to 36 days of age (P < .01). A time effect was recorded for total serum IgG in all groups (P < .001) and the percentage increase of the value was positively (P < .05) influenced by the AE supplementation. Lactobacilli raised (P < .001) in ileum and cecum of all groups supplemented with AEs. Staphylococcus spp. population was constantly lower in both intestinal tracts of the AE supplemented groups. On the basis of our results, AEs could improve broiler performance and immune function and contribute to a balanced gut microflora, essential for the digestion process and protection against enteropathogenic organisms.

Evidence for a primate origin of zoonotic Helicobacter suis colonizing domesticated pigs

Helicobacter suis is the second most prevalent Helicobacter species in the stomach of humans suffering from gastric disease. This bacterium mainly inhabits the stomach of domesticated pigs, in which it causes gastric disease, but it appears to be absent in wild boars. Interestingly, it also colonizes the stomach of asymptomatic rhesus and cynomolgus monkeys. The origin of modern human-, pig- or non-human primate-associated H. suis strains in these respective host populations was hitherto unknown. Here we show that H. suis in pigs possibly originates from non-human primates. Our data suggest that a host jump from macaques to pigs happened between 100 000 and 15 000 years ago and that pig domestication has had a significant impact on the spread of H. suis in the pig population, from where this pathogen occasionally infects humans. Thus, in contrast to our expectations, H. suis appears to have evolved in its main host in a completely different way than its close relative Helicobacter pylori in humans.

Is the horse a reservoir or an indicator of Coxiella burnetii infection? Systematic review and biomolecular investigation

The role of the horse in Coxiella burnetii infection has not been defined. Accordingly, a twofold approach was taken to further our knowledge on this topic: (1) conduct a systematic review of the literature to establish available evidence of C. burnetii infection in the horse; (2) undertake a biomolecular investigation of 122 cases of equine abortion, stillbirth and neonatal foal death, for the presence of C. burnetii using a PCR test targeting the IS1111 gene of C. burnetii. A review of the literature turned up seven studies that identified C. burnetii DNA in equine specimens, especially aborted fetuses, while an additional 34 studies sought to determine seroprevalence of the infection in the horse. A meta-analytical approach was taken to calculate a pooled mean seroprevalence in equines based on published studies. A seroprevalence of 15.8% (95% confidence interval: 9.6-23.0%) was obtained. This figure is comparable to those previously reported in other species, especially ruminants. None of the 122 cases of equine abortion, stillbirth or neonatal foal death were positive for C. burnetii DNA. C. burnetii has rarely been looked for in equine specimens in previous studies. Cases of equine abortion should be comprehensively investigated to assess the risk of abortion in a pregnant mare infected with C. burnetii. Consideration should also be given to the possible role of the horse as a source of the organism for other animal species including humans.

Investigation on intestinal bacterial flora and Salmonella spp. presence in organic and conventional chickens

Abstract The aim of this study was to evaluate the possible differences in the intestinal microflora composition among the different rearing systems (conventional vs organic) and the Salmonella diffusion using bacteriological techniques. The results showed that the differences between the two groups at the same age, expressed by the bacterial count, are not conclusive in showing an influence of the rearing systems. Salmonella Hadar was isolated once in caeca of conventional and once in caeca of organic ones. Though the results are preliminary and referred to a well defined geographic area in Central Italy, Salmonella detection does not seem to be common in conventional and organic chicken farms.

Ascaridia galli: a report of erratic migration

Abstract This paper describes a case of an unusual recovery of adult Ascaridia galli in hen’s egg. Several data are available on this occurrence but it appears to be the first case described in Italy. The worm was identified as an adult female, 6.8 cm in length, with three trilobed lips, cervical narrow alae, oesophagus club-shaped without posterior bulb, vulva near the middle of body, with gravid uteri containing a large number of eggs. The presence of Ascaridia galli in hen’s eggs cannot be considered as hazard for public health but may be cause of a potential consumer complaint. Moreover it is a sign of presence of ascaridiosis, parasitosis that still produces economic losses in modern poultry production system.

Phenotypic and genotypic characterization of canine pyoderma isolates of Staphylococcus pseudintermedius for biofilm formation

Biofilm-forming ability is increasingly being recognized as an important virulence factor in several Staphylococcus species. This study evaluated the biofilm-forming ability of sixty canine derived clinical isolates of S. pseudintermedius, using three phenotypic methods, microtiter plate test (MtP), Congo red agar method (CRA) and tube adherence test, and the presence and impact of biofilm-associated genes (icaA and icaD). The results showed that icaA and icaD genes were detected concomitantly in 55 (91.7%) of 60 isolates. A majority (88.3%) of the strains screened had matching results by the tube adherence test, MtP and PCR analysis. Better agreement (95%) was found between the PCR-based analysis and the CRA. Results of the icaA and icaD gene PCRs showed good agreement with CRA results, with a kappa of 0.7. Comparing the phenotypic methods, the statistical analysis showed that the agreement among the phenotypical tests using categorical data was generally good. Considering two classes (biofilm producer and biofilm non-producer), the percentage of matching results between the CRA method and the tube adherence test and between the CRA method and the MtP was 93.3%. A concordance of 100% was revealed between the MtP and the tube adherence test. The results indicate a high prevalence of the ica genes within S. pseudintermedius isolates, and their presence is associated with in vitro formation of a biofilm. A combination of phenotypic and genotypic tests is recommended for investigating biofilm formation in S. pseudintermedius.