Antibiotic resistance patterns of faecal indicator organisms and occurrence of Salmonella spp. in wild boar (Sus scrofa scrofa) in Italy.

In order to monitor antibiotic resistance in faecal indicator organisms and evaluate the occurrence of Salmonella spp., faeces from 110 wild boars (Sus scrota scrota), killed during a demographic control program in two different regional parks in Bologna province, were collected from September 2002 to June 2003. A single isolate of Escherichia coli, Enterococcus faecalis and Enterococcus faecium from each sample was tested for antibiotic susceptibility using the agar diffusion method recommended by CLSI (formerly, NCCLS). A total of 110 E. coli, 48 E. faecium and 5 E. faecalis strains were isolated and submitted to antibiotic susceptibility tests. Antibiotic resistance patterns were similar in wild boar populations from both parks. Multiple antibiotic-resistance to ciprofloxacin, rifampin and erythromycin was found with h1gh frequency in Enterococcus spp strains. E. coli isolates showed a low antibiotic resistance level. Two Salmonella arizonae and one Salmonella spp. strains, isolated from wild boars of one park, didn\u27t show any resistance concerning the antibiotics tested

Prevalence of hepatitis E virus in Italian pig herds. Preliminary results

Hepatitis E virus (HEV) is the causative agent of hepatitis E, and is an unenveloped positive sense single-stranded RNA wus. Swine HEV strains are genetically closely related to human strains from the same area, suggesting the occurrence of zoonotic transmission. Recently, human cases of hepatitis E have been linked to the consumption of raw or undercooked meat or organs from deer, wild boars or p1gs. The disease is now considered an emerging food-borne transmitted zoonosis. During 2006, a pilot investigation was performed to determine the prevalence of HEV in pig farms located in Northern Italy. 274 faecal samples were collected from healthy fattening animals (3-4 and 8-9 months of age) and from healthy breeding animals (gilts and sows) from 6 different farms, and analyzed using a Nested-RT-PCR targeting the open reading frame 2 (ORF2) region. Stool samples were suspended in water, and viral RNA extraction was performed using a commercial kit. Extracted viral RNA was subjected to RT-PCR amplification using degenerate primers conA 1-conS1 for the first amplification, and degenerate primers conA2-conS2 for the nested PCR, yielding a final fragment of 145 bp. HEV RNA was detected in sixty-nine of the 274 (25.2%) examined samples. None of the six farms resulted negative and the prevalence within the farms ranged between 2% and 60.5% For the characterization of the strains, randomly selected positive samples were subjected to nucleotide sequencing, and aligned with those present in the NCBI Data Bank Sequence analysis showed that all stra1ns were Swine Hepatitis E belonging to Genotype 3. These preliminary results confirm that swine HEV is widespread in Italian swine farms

Detection of serum antibodies to hepatitis E virus in domestic pigs in Italy using a recombinant swine HEV capsid protein

Background: The hepatitis E virus (HEV) has been detected in both humans and animals, particularly pigs, worldwide. Several evidences, including human infection following consumption of raw contaminated meat, suggest a zoonotic transmission of HEV. In Italy, large circulation of genotype 3 HEV has been reported in swine, and recent studies have confirmed the involvement of this genotype in autochthonous human cases. Result: In this study 111 sera collected from healthy pigs in two Italian regions were tested for anti-HEV IgG antibodies. For specific HEV antibody detection in swine, we developed ELISA and Western blotting methods, using a truncated capsid (ORF2) protein lacking the first 111 amino acids of a swine HEV genotype 3 strain. The ORF2-based ELISA revealed anti-HEV antibodies in 104 out of 111 pigs compared with 102 detected with a commercial ELISA kit. A lower number of sera reacted with the recombinant ORF2 protein in a Western blotting format (81/111). Using a Latent class analysis (LCA), the estimated sensitivities for ELISA-ORF2 and ELISA-kit tests were 0.961 and 0.936, respectively, whereas specificities were 0.599 and 0.475. The estimated sensitivity of Western blotting was 0.775, and the specificity was 0.944. Conclusions: The overall results confirm the high prevalence of HEV seropositive healthy pigs in Italy. Through comparisons with a commercial ELISA test, the swine genotype 3 HEV antigen produced in this study was proven suitable to detect anti-HEV antibodies in pig sera by both ELISA and Western Blotting

Impiego di un programma integrato di bioattivazione per il controllo della polmonite micoplasmica del suino

Introduzione. La qualit\ue0 dell\u2019aria nelle strutture di allevamento dei suini dipende soprattutto dalla concentrazione dei biogas, derivati dall\u2019azione della flora microbica fecale sull\u2019urea contenuta nelle urine; il peggioramento della qualit\ue0 ambientale \ue8 alla base di molti problemi sanitari, basati in primo luogo sull\u2019impossibilit\ue0 degli animali a mantenere un bilancio organico corretto. Una soluzione possibile e innovativa \ue8 rappresentata dalla bioattivazione con miscele batterico-enzimatiche; per questa via, si pu\uf2 ottenere una riduzione delle emanazioni, controllando anche la formazione di sostanze dannose per l\u2019ambiente. Materiali e metodi. \uc8 stato programmato un intervento in due aziende suinicole (il reparto di magronaggio-ingrasso di un ciclo chiuso completo e un allevamento di solo ingrasso), per controllare il danno indotto dall\u2019infezione micoplasmica nelle fasi di produzione a flusso continuo; sono stati messi a confronto gruppi trattati e non trattati, diversi tipi di pavimentazione (grigliato completo, concreto completo e misto grigliato-concreto) e pi\uf9 schemi d\u2019intervento, utilizzando, da soli o in combinazione, bioattivatori in polvere e liquidi. Il sistema di bioattivazione impiegato comprende due bioattivatori (in polvere e soluzione) e un prodotto naturale di origine vegetale da aggiungere alla razione alimentare. La polvere \ue8 stata distribuita sulle pavimentazioni, in ragione di 0,5 Kg per 100 mq ogni 15 giorni mediante spargimento manuale, mentre la soluzione \ue8 stato aerosolizzata nell\u2019ambiente di stabulazione, mediante impianto computerizzato, in ragione di 1 litro ogni 100 mq al mese; il prodotto vegetale, per la sua composizione in grado di riequilibrare la flora intestinale, \ue8 stato invece aggiunto quotidianamente alla razione, in ragione di 500 g/t di mangime. I rilievi e le osservazioni hanno riguardato la concentrazione di azoto ammoniacale (rilevata con metodo chimico), la presenza di anticorpi specifici per M. hyopneumoniae prima e dopo il periodo di trattamento (valutata con metodo ELISA), i punteggi polmonari al macello (valutati con metodo quantitativo) e il peso medio di partita. Risultati. In tutti i gruppi trattati, la sieroprevalenza per M. hyopneumoniae \ue8 stata elevata, a dimostrazione della circolazione dell\u2019infezione nel periodo di osservazione, ma la diminuzione della concentrazione ambientale dell\u2019ammoniaca, ottenuta grazie all\u2019impiego dei bioattivatori) ha permesso di ottenere benefici evidenti sul punteggio polmonare medio, confermati dalla significativit\ue0 statistica delle differenze, e sul peso medio di partita. In rapporto ai tipi di pavimentazione, la bioattivazione ha fornito i risultati migliori combinando polvere e liquido sul grigliato completo. Conclusioni. I risultati ottenuti dimostrano che nelle situazioni di allevamento, la stabilizzazione del bilancio organico degli animali attraverso una gestione ambientale che ne garantisca anche il benessere \ue8 una soluzione attuabile, in una logica di depotenziamento delle infezioni; essi sono la conseguenza dell\u2019accresciuta reattivit\ue0 degli animali, correlata ad un miglior bilancio organico, piuttosto che l\u2019effetto di un\u2019azione diretta sui patogeni; \ue8 in corso la valutazione della reattivit\ue0 immunitaria aspecifica dei suini sottoposti a trattamento, per correlare stato sanitario e produttivit\ue0 al miglioramento della qualit\ue0 ambientale

Canine placenta histological findings and microvascular density: The histological basis of a negative neonatal outcome?

Placenta is essential for the development of the fetus, and its impaired function can lead to a negative outcome (i.e., neonatal mortality). In dogs, investigations on placenta histology and neonatal outcome in healthy bitches are lacking, and a contribution is provided in this study to emphasize the use of placenta histology in practice. Fifty-one placentas from 11 litters were collected during cesarean section, classified according to the litter size (large (L) or small (S)) and the outcome, this latter as healthy (Group 1) or dead within 7 days (Group 2). The placenta/puppy weight ratio (PPR) was calculated, and specimens were formalin-fixed and paraffin-wax embedded, and on the resulting histological slides, capillary density (CD) was quantified. Among necrosis, calcification, and intravascular leucocytes, only the presence of multifocal-confluent necrosis (significantly more frequent in Group 2) was associated with a higher risk of death within 7 days (odds ratio = 30.7). Mixed logistic regression ruled out the effect on death both of a bitch and cesarean type (programmed vs. emergency). PPR and CD values were associated with litter size; large litters had lower PPR (p < 0.01) and higher CD (p < 0.05) than small litters. The relationship between PPR and CD was negative and significant (p < 0.01). Necrosis was a frequent finding in canine placentas, but only when multifocal-confluent was it associated with a poor outcome. The litter size influenced PPR (lower in L) and CD (higher in L), and this is likely due to the plasticity of placenta adaptation

Porcine Lawsonia intracellularis Ileitis in Italy and Its Association with Porcine Circovirus Type 2 (PCV2) Infection

The objective of this study was to employ a diagnostic algorithm, which involves detecting positive farms by stool PCR followed by PCR and histology/immunohistochemistry on ileum samples, for diagnosing Lawsonia intracellularis proliferative enteritis in Northern Italy. The primary aim was to examine the relationship between the gold standard of L. intracellularis diagnostics, namely histology and immunohistochemistry, and PCR in acute and chronic cases of L. intracellularis enteritides. An additional goal was to investigate the coinfection of L. intracellularis with porcine circovirus type 2 (PCV2). Twenty-eight ileum samples, including four from acute cases and 24 from chronic cases, were collected. PCR yielded positive results in 19 cases (four acute and 15 chronic cases). In comparison, immunohistochemistry was positive in 16 cases (four acute and 12 chronic cases), with an observed agreement of 89%. The findings suggest that performing the two tests in series can increase the specificity of the causal diagnosis. PCR may be used as a screening tool to identify the presence of the microorganism, and only positive cases will be examined by histology and immunohistochemistry to confirm the causative role of L. intracellularis. Co-infection with PCV2 was demonstrate in two out of four acute cases and in two out of 24 chronic cases, providing further evidence to support the hypothesis that when the infection starts with ubiquitous pathogens such as L. intracellularis, it may boost the possibility of PCV2 replication, especially in acute cases. As a result, this may trigger a transition from subclinical to clinical forms of PCV2 disease

Immunohistochemical detection of aetiological agents of proliferative and necrotizing pneumonia in Italian pigs

Proliferative and necrotizing pneumonia (PNP) is a form of interstitial pneumonia that occurs in weaning and post-weaning pigs. PNP is characterized by hypertrophy and hyperplasia of type II pneumocytes and coagulative necrosis and granular debris within alveolar spaces. Canadian and European studies suggest that the porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are the main causes of the disease, but Aujezsky's disease virus (ADV) and swine influenza virus (SIV) have also been considered as potential aetiological agents. An immunohistochemical study was carried out on the lungs of 28 Italian pigs with PNP in order to evaluate the role of PRRSV, PCV2 and ADV in PNP lesions. PRRSV infection was identified in the lungs of 11 pigs, PCV2 in the lungs of four pigs and coinfection with both viruses in the lungs of eight pigs. Neither virus was detected in the lungs of the remaining five pigs. ADV antigen was not detected in any sample. The principle aetiological agent of PNP in Italy therefore appears to be PRRSV. Coinfection with PRRSV and PCV2 is characterized by more severe microscopical changes in affected lungs

PCV2-DNA in formalin-fixed and paraffin embedded lymph nodes of wild boar (Sus scrofa ssp. scrofa): one sampling approach for two laboratory techniques

Superficial inguinal lymph nodes from 72 wild boars examined in a previous immunohistochemical (IHC) study on porcine circovirus type 2 (PCV2) were selected for a PCV2 polymerase chain reaction (PCR) analysis. Four of these lymph nodes were PCV2-IHC strongly positive with PMWS histological lesions (outcome 1), 6 weak to mild PCV2-IHC positive without PMWS histological lesions (outcome 2) and 62 PCV2-IHC negative. Considering IHC the gold standard for diagnosis, the aims of the study were to evaluate the suitability of the PCV2-DNA extraction from formalin-fixed and paraffin-embedded (FFPE) tissue and the sensitivity and specificity of PCR under two IHC interpretations criteria: (A) the sample was considered positive if the result was outcome 1; (B) the sample was considered positive if the result was outcome 1 or 2. Under (A) criteria, sensitivity and specificity of PCR were 100% and 89.7%, respectively; the Cohen's Kappa coefficient was 0.49. Under (B) criteria, sensitivity and specificity of PCR were 80.0% and 95.2%, respectively; the Cohen's Kappa coefficient was 0.72. The high Cohen's Kappa coefficient under the (B) interpretative criteria indicates good agreement between the two methods. In conclusion, 1) DNA extracted from FFPE specimens of wild boar is suitable for PCR and further represents a screening test for PCV2/PCVD (PCV2 Diseases) investigations in wild boar as well; 2) routine histological sampling can also be useful for PCV2 virological studies in wild boar