Klebsiella pneumoniae infection in canaries (Serinus canaria Domestica): a case report.

Background: Klebsiella pneumoniae can cause high mortality in birds. In recent years, small farming of canaries has been developed in Iran and infectious diseases are the main obstacle in the progress of these occupations which increases the importance of identification of pathogenic bacterial agents. Case description: A flock with 250 one-year-old canaries presented a history of anorexia, lethargy, mild diarrhea, and approximately 30% mortality. Physical examination revealed that the birds were severely debilitated, cachectic, dehydrated with ruffled feathers, and wet discolored stool around the cloaca. Necropsy findings revealed enlarged liver with multiple pale white, irregular foci on the surface of parenchyma, serosal petechial hemorrhages, and enlargement of lungs, liver, and kidneys. The entire intestine was intensely reddened with fibrinonecrotic exudate content. Histopathological findings of the liver elicited multifocal hepatocellular necrosis, hemorrhage, and also basophilic bacterial colonies. The results of biochemical and molecular tests confirmed K. pneumonia as the causative agent. Findings/treatment and outcome: Based on antimicrobial susceptibility test, K. pneumonia isolates were susceptible to gentamycin and ciprofloxacin which were administrated for the considered treatment protocol. Conclusion: K. pneumoniae is one of the most important causes of mortality in canaries with multiple antibiotic resistance, therefore assessments of health conditions can supply suitable information to help decision-making about the sanitary processes, control, prevention, and treatment.

Antimicrobial Susceptibility Profile of Enterococcus Species Isolated from Companion Birds and Poultry in the Northeast of Iran.

Enterococci are Gram-positive facultative anaerobic bacteria commonly found in the gastrointestinal tract of the mammals and birds. These cocci are isolated from urinary tract infections, bacteremia, endocarditis, and burn wounds in humans. The evolution of antibiotic-resistant enterococci raised a problem due to the possibility of the transmission of these organisms between poultry and human. Regarding this, the present study was conducted to evaluate the prevalence of Enterococcus species among companion birds and poultry in the Northeastern of Iran and determine the antibiotic susceptibility profile of enterococci. To this end, oral and cloacal swabs were collected from 150 caged birds. Antibiotic susceptibility profile was determined using the standard disk diffusion method. The results revealed that out of 150 samples, 56 (37.33%) cases contained enterococci. Most of the specimens (25.33%) were Enterococcus faecalis isolated from 6.66% of the samples. Additionally, 2.66% and 1.33% of the samples were contaminated with Enterococcus mundtii and Enterococcus gallinarum, respectively. Furthermore, Enterococcus malodoratus and Enterococcus raffinosus were isolated from 0.66% of the samples. The results revealed that all of the isolates of E. faecalis and E. faecium were resistant to more than five antimicrobial agents. Most of E. faecalis and E. faecium isolates showed resistance to Cefazolin, Tiamulin, Flumequine, and Cephalexin. Accordingly, the majority of the isolates had multidrug resistance to the tested antibiotics. In conclusion, the presence of multidrug-resistant enterococci in the birds living close to humans requires thorough observations due to the transmission of these organisms to humans.

Toxigenic Clostridium difficile in retail packed chicken meat and broiler flocks in northeastern Iran.

This study was designed to evaluate the occurrence of Clostridium difficile in both broiler chicken farms and packed chicken parts sold at market places in Mashhad, the second most popular Islamic pilgrimage city after Mecca in northeastern Iran. The fresh faecal samples were obtained from broiler farms, while the chicken packs were purchased from retail outlets across the city at market places and samples were obtained from the necks, thighs, and wings. The selective culture was used for isolation of C. difficile. Out of 40 pooled fresh faecal and 65 packed chicken samples, 14 (35%) and 10 (15.3%) samples were positive, respectively. Some of the C. difficile isolates from chickens packs (7 out of 10, 70%) and faecal samples (5 out of 14, 36%) were detected as toxigenic (A, B and binary toxins) using molecular identification. From 14 isolates of the faecal samples, five isolates were tcdA and tcdB positive, and none was binary toxin positive. The results of the present study suggest that broiler chickens are a potential source of C. difficile, which may infect humans through contact or consumption of chicken meat, although the significance of food contamination is entirely unclear, the role of poultry products as a potential source of the infection should be investigated.

An outbreak of yolk sac infection and dead-in-shell mortality in common canary (Serinus canaria) caused by Klebsiella pneumoniae.

Yolk sac infection (YSI) and dead-in-shell mortality caused by Enterobacteriaceae in birds are not a rare phenomenon, however there are only a few reports indicating the association between these conditions and Klebsiella spp. among canary chicks (Serinus canaria). There have been reports of high mortality among 1-3 day old canary chicks in an indoor flock of canaries. In order to study the causative agent, yolk sac samples from dead-in-shell and day-old canary chicks were cultured. Klebsiella pneumonia was isolated and identified based on biochemical tests and using genus and species-specific multiplex PCR and later tested for their susceptibility to 13 antimicrobial agents. The isolates showed susceptibility to Gentamycin, Chloramphenicol, Florfenicol and Streptomycin.

Genomic diversity of Clostridium perfringens strains isolated from food and human sources.

Clostridium perfringens is a serious pathogen which causes enteric diseases in domestic animals and food poisoning in humans. Spores can survive cooking processes and play an important role in the possible onset of disease. In this study, RAPD-PCR and REP-PCR were used to examine the genetic diversity of 49 isolates of C. perfringens type A from three different sources. The results of RAPD-PCR revealed the most genetic diversity among poultry isolates, while human isolates showed the least genetic diversity. Cluster analysis obtained from RAPD-PCR and based on the genetic distances split the 49 strains into five distinct major clusters (A, B, C, D, and E). Cluster A and C were composed of isolates from poultry meat, cluster B was composed of isolates from human stool, cluster D was composed of isolates from minced meat, poultry meat and human stool and cluster E was composed of isolates from minced meat. Further characterization of these strains by using (GTG) 5 fingerprint repetitive sequence-based PCR analysis did not show further differentiation between various types of strains. In conclusion, RAPD-PCR method seems to be very promising for contamination source tracking in the field of food hygiene.

Isolation of Clostridium difficile and molecular detection of binary and A/B toxins in faeces of dogs.

The aim of this study was to isolate Clostridium difficile from dogs' faeces, and to study the frequency of its virulence genes. A total of 151 samples of dogs' faeces were collected. The isolation of C. difficile was performed by using the bacterial culture methods followed by DNA extraction using boiling method. Multiplex PCR method was performed for identification of tcdA, tcdB, cdtA and cdtB genes and single method was carried out for detection of tcdC. Twelve samples (7.9%) were positive in bacteriological assay and based on molecular assay, 66.7% of the isolates (8 of 12 C. difficile isolated) had shown tcdA+, tcdB+ profile. This is the first investigation on molecular assay of C. difficile in Iran's dog population.

Molecular identification and successful treatment of Chlamydophila psittaci (genotype B) in a clinically affected Congo African grey parrot (Psittacus erithacus erithacus).

Avian chlamydiosis is caused by Chlamydiophila psittaci with the highest infection rate in parrots (Psittacidae) and pigeons (Columbiformes). A two-year-old Congo African grey parrot was examined since the bird had shown clinical signs of anorexia, depression, diarrhea, and mild dyspnea and based on biochemical and hemathological analysis the bird was diagnosed as having anemia, leukocytosis, heterophilia, lymphopenia and monocytosis. With regards to clinical and paraclinical findings, the case was diagnosed to be carrying Chlamydiophila spp. In addition, choanal cleft and cloaca swabs were positive for Chlamydiophila spp. in a diagnostic polymerase chain reaction (PCR) (600 bp amplicon). Polymerase chain reaction products were typed by ompA gene-based PCR, using CTU/CTL primers (1050 bp amplicon). The PCR product sequence was compared with the sequences obtained from GenBank. The phylogenetic tree has revealed 100% identity with genotype B obtained from previous studies. The bird was hospitalized and treated with doxycycline regimen for 45 days, with a weekly sampling process to trace the presence of C. psittaci DNA in faecal and choanal swabs, this process continued to the point where the specimens turned negative after two weeks. Laboratory and radiology results were within normal limits after the treatment. Genotype B is predominantly isolated from Columbidae and there have not been any reports regarding the clinically affected African gray parrot with this genotype. Subsequently, to the best of our knowledge, this is the first report of chlamydiosis by genotype B on Congo African grey parrot.

Isolation and characterization of a very virulent Infectious bursal disease virus from turkey.

Infectious bursal disease (IBD) is a highly contagious disease of chickens caused by Infectious bursal disease virus (IBDV). In turkeys, however, infection with classical virulent IBDV strains lead only to subclinical forms of the disease. We attempted to isolate IBDV from the bursa of turkey and characterize it. Amplification of a 743-bp fragment of VP2 gene by RT-PCR and restriction analysis of the product showed a pattern compatible with very virulent IBDV (vvIBDV). Comparison of the sequence of this isolate with those of other IBDVs and phylogenetic analysis confirmed very virulent nature of the isolate. This is the first report on the isolation of vvIBDV from turkey in Iran.

Molecular characterization of Iranian infectious bursal disease viruses.

This study was conducted to characterize nine infectious bursal disease virus (IBDV) isolates from Iran. A reverse transcriptase-polymerase chain reaction (RT-PCR) procedure was used to amplify a 743-bp fragment of the VP2 gene hypervariable region from IBDV field isolates. Amplified VP2 fragments of the nine IBDV isolates were sequenced and compared with published sequences of IBDV strains from Iran and around the world, and their phylogenetic relationships were analyzed. Three isolates demonstrated close relation to classical attenuated strains of IBDVs, and six isolates showed sequences common in European and Asian strains of very virulent IBDVs (vvIBDVs). Four nucleotide changes--802A, 934A, 940A, and 1366A--were common in all Iranian vvIBDVs except in one isolate. Amino acid sequences of three Iranian vvIBDVs were 100% identical and resembled vvIBDV strains from European (UK661), Asian (HK46, GZ96), and Iranian origins (IR01, SDH1). Some unique amino acid substitutions after major hydrophilic peak A in Iranian vvIBDV field isolates were observed: 231S-L, 231S-P, and 233N-K. Phylogenetic analysis showed that the Iranian vvIBDVs were closely related to European and Asian vvIBDVs. Further comprehensive investigations will provide more information on the distribution, variability, and phylogenetic relationships of different IBDVs isolated in Iran and other parts of the world.

Explanatum explanatum (Creplin, 1847) Fukui, 1929, in buffaloes in the Ahwaz area, southwest Iran.

Explanatum explanatum (Creplin, 1847) Fukui 1929, was found in three indigenous buffaloes in the west Ahwaz area, southwest Iran for the first time. The presence of this amphistome in bile ducts caused remarkable fibrosis of submucosa ad hyperplasia of the endothelial layer, such that villus-like structures and protuberances were formed in large bile ducts of the infected livers.