Marisa Macagnan

DETECTION OF SALMONELLA SP. FROM PORCINE ORIGIN: A COMPARISON BETWEEN A PCR METHOD AND STANDARD MICROBIOLOGICAL TECHNIQUES

The aim of this study was to compare a polymerase chain reaction (PCR) method combined with selective enrichment in Rappaport-Vassiliadis broth (PCR-RVB) with standard microbiological techniques (SMT) for the generic detection of Salmonella in samples of porcine origin. Two hundred sixty eight field samples consisting of 42 sets of pooled porcine mandibular lymph nodes and tonsils, 44 samples of intestinal content, 38 pork sausage meat samples and 144 samples of feed collected from swine farms were submitted to the PCR-RVB and SMT protocols. Salmonella was detected in 54 samples using the PCR-RVB assay and in 42 samples by SMT, three of the SMT Salmonella-positive samples (one each of S. Derby, S. Panama and S. Typhimurium) being Salmonella-negative by PCR-RVB. For the PCR-RVB method 15 Salmonella-positive samples were negative by SMT, a significant difference according to the Mac Nemar’s chi-squared test (p=0.0153). Subsequent serological typing of the SMT isolates showed the following Salmonella serovars, the number of positive samples being given in parentheses: Typhimurium (12); Bredeney (10); Panama (5); Saint-paul (5); Minnesota (3); Mbandaka (2); Derby (1); Enteritidis (1); Orion (1) and Salmonella sp. (2). We concluded that, although the use of both PCR-RVB and SMT increased the number of positive samples, the PCR-RVB, due to its higher sensitivity and greater speed in giving results, can be implemented to detect Salmonella in samples of porcine origin.

Natural infection of turkeys by infectious laryngotracheitis virus

The infectious laryngotracheitis virus (ILTV) is an important respiratory pathogen of chickens that also infects pheasants and peafowl. Epidemiologically non-related commercial turkey flocks with clinical signs such as tracheitis, swollen sinuses, conjunctivitis and expectoration of bloody mucus were examined for the presence of the virus. Laboratory ILTV detection was performed by virus isolation in embryonated eggs and cell cultures, PCR and sequencing of amplification products, histopathology, indirect immunofluorescence and electron microscopy. One ILTV turkey isolate was also experimentally inoculated into susceptible chickens and turkeys, reproducing a mild respiratory disease. This is the first description of natural infections with ILTV in turkeys.

Prevalence of antibodies against Ornithobacterium rhinotracheale in broilers and breeders in Southern Brazil.

Abstract In this investigation, we determined the prevalence of the Ornithobacterium rhinotracheale (ORT) infection in broilers and broiler breeders in southern Brazil. We also correlated the presence of antibodies in broilers with performance. Sera from 1550 broilers from 50 flocks were collected during the slaughter time in nine companies with federal veterinary inspection of the state of Rio Grande do Sul. Sera from 480 meat-type breeders of 40 flocks from 14 companies in southern Brazil were also analyzed by enzyme-linked immunosorbent assay, and the prevalence of antibodies was determined. The prevalence of ORT antibodies in broiler flocks was 63.83%, but in each individual flock only 6.52% of the birds were positive. The prevalence in broiler breeder flocks was 100.00%, and in each individual flock 94.62% of the birds were positive. There was a positive correlation between the presence of antibodies to ORT and decreased body weight in broilers. There was no significant correlation between presence of antibodies to ORT and age, lineage, efficiency index, feed conversion, and mortality. There was a positive correlation between the presence of respiratory signs and antibodies to ORT, although the reverse correlation was not significant. These results confirm that ORT is present and widespread in broilers and broiler breeders in southern Brazil.

Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil

Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

Laryngotracheitis: reproducibility of the disease and comparison of diagnostic methods

Infectious laryngotracheitis virus (ILTV) cause mild to severe respiratory disease in chickens, the purpose of our study being to use Brazilian isolate of ILTV to reproduce ILTV disease in chickens by experimental infection and to compare three diagnostic methods (nested polymerase chain reaction (PCR), virus isolation, histopathology) for detection of ILTV. Forty-eight chickens intratracheally inoculated with ILTV and a further 48 with PBS, showing mild respiratory signs 48 hours post infection (PI) but no signs of infection after day 10 PI. Every 2 days PI, six birds were arbitrarily selected from the control and infected groups, sacrificed and the trachea collected. Both the nested PCR and virus isolation detected the virus from day 2 until day 12 PI. However, at day 12 PI, PCR detected ILTV DNA in 100% of the samples while the virus isolation method detected ILTV in only 33% of the samples. Tracheal histopathology showed intranuclear inclusion bodies on days 8 and 10 PI. The results indicate that the field-isolate of ILTV studied by us is of low pathogenicity and that our nested PCR protocol was able to detect positive samples over a longer infection period than many ILTV diagnostic test already described.

Detecção sorológica e microbiológica de Salmonella spp. em emas (Rhea americana)

The fast agglutination screening test, using S. Typhimurium as the antigen, was compared with the standard bacterial method to identify rheas (Rhea americana) contaminated with Salmonella spp.at slaughter.Seventy birds were serologically tested for Salmonella enterica Pullorum using a commercial antigen.Of these, 66 were also submitted to a macroagglutination test, using a strain of Salmonella Typhimurium isolated from rheas.All birds did not react with the commercial S. Pullorum antigen, but 37 were positive for the FAST-ST.The isolation of Salmonella spp. was verified in 66 (94.2%) birds. 85.7% were found in liver samples, 60% in feces and 42.3% in cloacal swabs.A total of 16.6% were identified as being S. enterica enterica rugosa, 35.9% as S. Typhimurium, 46.5% as S.Newport and 0.9% as S. Anatum. An insignificant concordance between the results of bacterial isolation and the serological response was observed (k=0.016).The detection of Salmonella spp. by bacteriological and serological methods in samples from rheas must be deemed important, because birds without a clinical manifestation can be significant sources of salmonellas in food infections.